Environmental Studies
VOL.l
Water and Wastewater Analysis
ABD Publishers
Handbook of Methods in
Environmental Studies
Vol. 1: Water and Wastewater Analysis
S.K.Maiti
B.Sc (Hons), M.Sc (Calcutta University)
M.Tech. (Env. Sc. & Engg.), liT, Mumbai
Ph.D (Indian School of Mines, Dhanbad)
I: :
-
o.
AB
ABD Publishers
Jaipur (India)
By : S.K. Maiti
ISBN : 978-81-8577-34-07
ISBN 81-8577-34-0
'!~
Author
Published by :
ABD Publishers
B-46, Natraj Nagar, Imliwala Phatak
Jaipur-302 015 (Rajasthan) India
Ph. : (0) 594705 (R) 762072
Fax: 0141-597527
E-mail: abdpublishers@rediffmail.com
Distribution by :
Dedicated To
My father, Late Shri Mura/idhar Maiti
and
My mother, Smt. Mondakini Maiti .
Contents
SECTION-A: AIR POLLUTION
Air quality monitoring: Micrometeorology; Objectives, Siting criteria for AQMS, Ambi-
ent and source emission monitoring, NAAQS, EPA-USA AQS, Monitoring frequency, Reporting, AQ monitoring for existing industry and for new industry, Self test
Monitoring of particulate pollutants: Dustfall, Calibration ofHVS, monitoring ofSPM,
preparation
Analysis of physical parameters: Introduction, Coarse fractions, Texture, Bulk density
and pore space, Field moisture, WHC, Wilting point, Infiltration rate, Self test
Analysis of Chemical parameters: pH, Lime requirement, EC, Organic carbon, Organic
matter, Total nitrogen, Av. N, Av. P (Bray's and Olsen's), Total P, P-fixing characteristics
of soil, Exch. K, Exch. Na and SAR, CEC, Av. S, Exch. Ca and Mg, Chloride, Monitoring
ofPb, Fe, Cu, Mn, Zn, Ni and Cr, Cd, DTPA fractions, Hg
Soil microbiology: Collection/processing of sample, Enumeration of bacteria, actinomycetes,
Filamentous fungi, VAM infection in root, Estimation ofVAMF spores, Soil respiration- in
situ and laboratory measurement, Soil enzymes- dehydrogenase, invertase, amylase and
cellulase, Chemicals, apparatus and instruments for soil analysis laboratory
SECTION-E: ECOLOGY
Objectives, Assessment of productivity; Diversity, Plant community studies, Litter-fall and
litter decomposition, Study offaunal community; Self test.
Preface
provided at the Centre. Prof. Gurdeep Singh and Prof. N.C. Saxena deserve special
mention. Special thanks are due to Dr. I.N. Sinha and Dr. Dinesh Mohan for their
meticulous correction and mental support. I am also thankful to our laboratory staff
for their interest in the work and to Shri Ajoy Bhattacharya for careful drafting of
the figures. Thanks are also due to Dr. P.K. Goel, Reader in Deptt. of Pollution
Studies, Y.C. College of Science, Karad, for his critical suggestions for improve
ment ofthe contents and getup of the book. It is my pleasure to acknowledge the
help and assistance received from Dr. v.P. Upadhayay (MOEF, Bhubaneswar),
Dr. N.C. Karmakar (ISM, Dhanbad), Dr. Amal Kr. Pal (CU), Dr. Ram Bhagat and
Dr. B. Pati (Vidyasagar University, Midnapore), Dr. P.C. Mishra (Sambalpur
University) Dr. Arvind Kumar (S.K.University, Durnka), Shri P.C. Jha, Rajiv Kr.
and Vinita Arora (CMPDIL, Ranchi), and Shri Suresh Jain (MECON, Ranchi).
I wish to express my sincere appreciation to my wife (Sumita) and children
(Polly and Tukai) for their patience and sacrifice but for which the book would not
have seen the light of the day.
I am also grateful to my publishers for timely publication of the book.
Place: Dhanbad
Date: 162001
Contents
CHAPTER 1: WATER SAMPLING AND PRESERVATION
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
1.10
1.11
1.12
1.13
Sampling
2
2
3
7
8
9
9
10
13
'Questions
2.1
2.2
2.3
2.4
2.5
2.6
2.7
2.8
2.9
2.10
2.11
Standards
Drinking water Standards (India)
International Drinking Water Standards
Coliform Standards
Effiuent Standards
Stream Standards
Wastewater Analysis for Proposed Project
Primary Water Quality Criteria for Bathing Waters
Effiuent Standards for Coal Mines
Standards for Effiuents from Textile Industry
Questions
3.1
3.2
3.3
3.4
Acidity
Alkalinity
Biochemical Oxygen Demand (BOD)
Boron
13
15
15
15
15
20
22
24
26
27
27
28
29
30
30
33
37
48
3.5
3.6
3.7
3.8
3.9
3.10
3.11
3.11.1
3.11.2
3.11.3
3.11.4
3.12
3.13
3.14
3.15
3.16
3.17
3.18
3.19
3.19.1
3.19.2
3.20.2
3.20.3
3.21
3.22
3.23
3.24
3.25
3.26
3.27
3.27.1
3.27.2
3.27.3
3.27.4
50
52
57
60
66
70
74
74
76
77
78
79
85
91
94
96
100
104
108
109
III
113
114
116
117
120
123
123
125
128
133
138
139
141
142
142
3.28 Solids
3.28.1
3.28.2
3.28.3
3.28.4
3.28.5
142
143
144
146
148
149
150
3.29.1
3.29.2
151
154
Estimation of sodium
Estimation of Potassium
3.30 Sulphate
155
3.30.1
3.30.2
156
158
Gravimetric Method
Turbidimetric Method
3.31 Sulphide
161
3.32 Thiocyanate
164
167
169
Introduction
174
174
4.2
174
4.3
176
4.4
4.5
177
177
4.6
178
4.7
Aluminium
181
4.8
Arsenic
182
4.9
Cadmium
183
4.10 Chromium
4.11 Copper
184
185
4.12 Iron
185
4.13 Lead
186
4.14 Manganese
187
4.15 Mercury
188
4.16 Selenium
4.17 Silver
192
193
4.18 Vanadium
193
4.19 Zinc
194
195
197
5 .1
197
5.2
203
5.3
204
Estimation ofMLSS
Estimation ofMLVSS
205
205
5.4
206
5.5
209
5.3.1
5.3.2
212
6.1
Introduction
213
6.2
213
6.3
214
6.4
215
6.5
223
6.6
225
6.7
226
6.8
229
6.9
233
237
240
242
243
244
247
248
250
252
253
7.3
253
7.4
254
7.4.1
7.4.2
7.4.3
7.4.4
7.4.5
7.4.6
255
255
256
256
256
257
257
257
259
7.4.7
7.4.8
7.4.9
7.4.10
7.4.11
7.5
7.5.1
7.5.2
257
258
258
259
259
7.6
Phytoplankton Productivity
259
7.7
Estimation of Chlorophyll a
261
7.8
264
265
267
7.9
8.1
8.1.1
8.1.2
Toxicity Test
Significance of Toxicity Tests
Classification of Toxicity Tests
267
267
8.2
269
8.3
Toxic Unit
271
8.4
273
274
9.1
Radioactivity
274
9.2
275
9.3
277
280
282
286
287
287
288
288
296
304
water quality,
composition and state of the biological life present in the water bodies,
nature of the particulate matter present, and
physical description of the water body (hydrology, dimension, morphometry,
nature oflake bottom or river bed).
Location
Objectives
Baseline site
Headwater lakes
or undisturbed
upstream river
stretches
Trend site
Global river
flux site
Mouth of a major
river.
To determine fluxes of critical pollutants from river basin to ocean or regional seas.
Some trend stations on rivers also
serve as global flux stations.
1.6 Sampling
The objective of sampling is to collect a portion of water which is small enough in
volume to be transported conveniently and handled in the laboratory while still
representative of the characteristics ofthe water available in that water body. Water
is a dynamic system. Its constituents vary with time.
b. Composite samples
The term composite refers to a mixture of grab samples collected at the same sampling point at different times. A composite sample of 24 h period is considered
standard for most of the determination. It provides more meaningful data than the
grab samples.
Sometimes a composite sample representing one shift or a shorter time period or
a complete cycle of a period operation may be preferable.
Take at least 120-150mL of sample in each h, in some cases even at intervals of
30 min (if composition varies within an hour) and mix at the end of sampling
period or combine in a single bottle as collected.
A final volume of 4-5 L is sufficient.
c. Integrated samples
For certain purpose, the information needed is provided best by analyzing mixture
of grab samples collected from different points simultaneously.
Such samples are useful for rivers or streams that vary in composition across
the width and depth. For collection of integrated samples, special sampling
device is needed.
Sample is collected at a known depth without disturbing the surface water.
1.6.3 SAMPLING FREQUENCY
Water sample should be collected at intervals so that no change in water quality
could pass unnoticed. It depends on the type of data required, purpose of monitoring, availability of funds and personnel.
Number of Samples: Number of samples and how often should samples be col-
t.
Sj2
--
Where,
N = Number of samples
t = Student 't' -statistics for a given confidence level
S = overall standard deviation, and
U = acceptable level of uncertainty
For, example, if standard deviation is 0.5 mg/L, U is 0.2 mg/L, and 95% confidence level is desired, then 25 to 30 samples must be taken. (t-test is used for less
than 30 samples).
The frequency is determined by the objectives of monitoring. The following
frequencies may be adopted provisionally:
Once in a year: Generally applicable for long-term ecological evaluation ofbiological water quality. The time and place should be the same. Sometimes also used to
know the year-to-year trends, usually as a result of increased human activities in
the watershed.
Four times in a year for studying seasonal variations in water quality. Sometimes
three samples are taken in a year like, pre-monsoon, monsoon and post-monsoon
period.
Weekly samples for one year.
Round the hour sampling for 24 h (i.e. industrial effiuents)
The bottles should be soaked with 10% HCI for 24 h and then thoroughly
cleaned and rinsed with distilled water.
If metals are to be analyzed, rinse the container with solution of 1 part concentrated HN0 3 to 4 parts water followed by distilled water.
Cleaning solution; acid dichromate: Prepare a saturated water solution of potassium dichromate (K2Crp7). Add 32 mL of this K 2Crp7 solution in 1L of
concentrated H2 S04 (sp. gr. 1.84).
1.6.5 WATER SAMPLE~S
Several different type ofwater samplers are available and many of them are
designed for specific purposes. Two most commonly used water samplers are
described here.
a. Dissolved oxygen (DO) sampler
(ross sKtM)n
--'''''
WliIghted
1I-4---4-....l/.__ b'''---.:Y--''---+--J!
6
b. Depth sampler
The depth sampler, sometimes called a grab sampler, is designed in such way that it
can retrieve a sample from any predetermined depth. A simple and relatively inexpensive depth sampler suitable for moderate depth 30m) is illustrated in Fig.l.2.
1.6.6 SAMPLE COLLECTION
1. Wherever possible, the container should be rinsed 2 to 3 times with the sample
to be examined.
2. Sample where water is well mixed.
3. Avoid large non-homogeneous matter such as leaves, rags, twigs and other
floating material in the sample.
4. Provide complete information about the source and the conditions under which
the sample was coi!('cted
5. Sample preferably at 20cm depth in a shallow channel (Fig.I.3). Depths> 60cm,
collect 2 samples at 20% and 80% below the surface
6. Sampling from dug wells and similar sources: Prepare the sampling device bottle
with the help of a string and attach a weight at the bottom (Fig. 1.4).
1.6.7 LABELLING OF CONTAINER
Each sampling bottle must be provided with an identification label on which the
following information is legibly and indelibly written (Table 1.2).
1.6.8 TRANSPORTATION OF SAMPLE
Sample containing bottles should be placed in a box for transportation to the laboratory. Sturdy, insulated wooden or plastic boxes will protect samples from sunlight, prevent the breakage of bottles and should allow a temperature of 4C to be
attained and maintained during transport. Fig.I.5 shows a suitable sample transport
box.
1.2:
Name of study
Sample number/ sample station
identification no.
Source/location of sampling point
Date and time of collection
Volume of sample
Nature of collection
Given any preservative
Purpose of analysis:
Field measured parameters
:
:
:
:
Grab/composite sample
Yes/No.
Drinking water/effluent discharge.
Temperature, pH, DO, or any
other parameters.
~. '.:.'
:.
~~~~
,
H
::.
S.~ f--Irowtation
10
I!l
4oR-~'::-tt.----~ bottles
~.e..~.~
.......1III!j.~.~~""'E.~
. . . .~
:
Table 1.3:
Preservation of samples.
Parameters
Preservation Method
Maximum
holding period
Acidity
Alkalinity
Refrigerate at 4C
Refrigerate at 4C
Refrigerate at 4C
2 mL H2 S0 4 /L
Analyze immediately
Refrigerate at 4C
Analyse immediately or fix on site
Not required
Acidify with HN0 3 to pH < 2.
Analyse as soon as possible, add 2 mL
40% H2 S0 4 to pH < 2, refrigerate.
Analyse as soon as possible.
Refrigerate, add H2 S0 4 to pH < 2,
40 mg HgCI/L, refrigerates at 4C.
Refrigerate, add 2 mL 2N zinc
acetate/100 mL; add NaOH to pH> 9.
Refrigerate at 4C
Refrigerate at 4C
Sterilised bottle, no specific preservative,
refrigerates at 4C
24 h
24 h
6h
7 day
BOD
COD
Residual CI 2
Colour
DO
Fluoride
Metals
NH3-N
Nitrate
N-Kjeldahl
Phosphorus
Sulphides
Sulphate
Turbidity
Colifom bacteria
24 h
6h
7 day
6 months
24 h
7 days
24 h
7 days
7 days
36 h
Lcations - Lanions
% Difference =
Lcations + Lanions
100
10
0-3
3 - 10
10 - 800
0.2
2
5
An accurate ion balance does not necessarily mean that the analysis is correct.
There may be more than one. error and these may cancel each other. As a result
additional checks are needed. With careful work, the difference will not generally
exceed 2% of the total cations or anions in waters of moderate concentrations (250
to 1000 mg/L). The calculations to see the correctness of results of the actual data
are given in example boxes 1 and 2 later.
Calculate TDS to EC ratio: The numerical value of TDS (mg/L) should not
exceed that of electrical conductivity (~S). The relationship between the two variables is often described by a constant (commonly between 1.2 and 1.8 for freshwater) that varies according to chemical composition. For freshwater the normal range
can be calculated from the following relationship:
Conductivity = TDS x F, where F is in the range of 1.2 to 1.8.
Typically, the constant is high for chloride-rich waters and low for sulfate-rich
waters (UNEPIWHO, 1996).
TDS/conductivity = 0.55 to 0.70.
If the ratio ofTDS to EC is outside these limits, measured TDS or measured EC
is suspect; reanalyze the sample (APHA, 1998).
Introduction
Study area
Methods
Results
Analysis of
results
Significance of
results
Recommendations
Information
sources
11
Any major restrictions (personnel, access, finance, facilities, etc.) which limit the described scope of the programme
should be identified.
A summary of the geography, hydrology, and other salient features (which, for example, may include land use,
industry and population distribution) of the area under
study should be provided.
All locations, structures and features mentioned in the
text should be identified on maps.
In certain cases, profiles of river or lake systems may provide additional useful information.
A separate section should provide details of the methods
and procedures used for all aspects of the study.
A summary of the procedures to be followed for quality
assurance should be presented.
Results should be presented in graphical form whenever
possible and graphs should clearly demonstrate the relationship of the data to the monitoring objectives.
Graph should summarise entire data rather than individual
observations.
International units of measurement should be used wherever possible but if local units are used precise description and conversions should be provided.
The statistical analysis of results should be described.
The reliability ofthe statistics and its implication, should
also be given.
Analytical procedures appropriate to the problems should
be used and should focus on the goals of the study or the
programme.
Results should be presented in graphical, rather than in
tabular form.
The report should include a section devoted to interpretation of the results in terms of the monitoring objectives.
This will help ensure that the objectives are being met
and that questions are being answered.
This section should emphasise the need to provide information.
Recommendations for proposed future activities should
normally be listed in the following two categories;
a. One concerned with scientific matters, the other with management issues.
b. Ideally, these recommendations should be presented in
order of priority.
All sources of information and all literature referred to
should be correctly and fully cited so that a permanent
record is available.
12
Example - 1
Problem: An analysis of water from a surface stream yields the following
results. Whether the analysis is acceptable?
valef]cy'i,,!, m~'Ol.b.i:::
'%"'~:::_::
>,"'v
',',if
.' Concentration
'Total anions - ~
I "",,,,,,,,,,,,.,,,'
difference is 17.87%, which is more than ~5% ; :Therefore,
;" >,d hing .wrong ir{tl1is analysis
". ,,;:1,::-";'- .' ",
".J.:"'"
. ~~,::';~;< .".
~:l/<~ $.,<"
Example - 2
Problem: An analysis of water from a surface stream yields the following results.
If an error of 10% is acceptable, should the analysis is acceptable?
" Concentration
+'2'_ 60
. IL ,',,;
.C
.a mg,i"":0i'
, Mg+2= 10. mg/L .. :
;,Na+ =7mg/L
./<+ =20 mg/!.:
13
Example - 3
If standard derivation is 0.5 mg/L, test values for 95% confidence level is
1..9 6 and acceptance leve.1 of uncertainty is 0 .2 mg/L (i .e acceptance error), <d
then number of sample needed to meet 95% confidence level is,
. '.
N~
96
X0'J2
0.2
Also calculate the number of sample needed for 99% confidence level
(t 2 .575) for S
0.5 mg/L and acceptable level of uncertainty (UI
0.2
mg/L
N .~
1.13 Questions
I.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
14
12. Describe the salient points to be considered for the preparation of a standard
monitoring report.
l3. How to prepare 6N, IN, O.lN and O.02NHCI, H 2S04 andHNO] solution. (Hints:
See annexures).
14. How to prepare 15N, 6N, IN and O.lN NaOH solutions. (Hints: Seeannexures).
15. How to prepare phenolphthalein and methyl orange indicator solution. (Hints:
See annexures).
16. An analysis of water from a surface stream yields the following results. If an
error of 10% is acceptable, should the analysis be considered complete. Ca+2 =
32 mgIL; Mg+2 = 42.59 mgIL; Na += 173.57mgIL; K+ = 24.20 mgIL; HC~= 186.46
mg/L; CI- = 275.64 mgIL; SO~2 = 84.25 mg/L; NO; = 9.80 mg/L; F- = 1.24.
(Ans: 3.66%).
17. An analysis of water from a surface stream yields the following results. If an
error of 10% is acceptable, should the analysis be considered complete. Ca+2 =
55 mg/L; Mg+2 =18 mgIL; Na+ = 98 mglL; HCO; = 250 mgIL; CI- = 89 mgIL;
SO~2= 60mgIL. (Ans: 8%).
Water Quality
Standards
2.1 Standards
Standards are the concentration to be maintained to achieve the stated objectives. The term standard applies to any define rule, principle or measurement
established by the authority or that has official backing. As per Californian
Federal Act (1967), standard has been defined as " pollution level that can not
legally be exceeded during a specific time period in a specific geographical
area." The primary standards are related to human health and secondary standards are related to protect human welfare. Three types of standards are of
interest in water quality monitoring study.
Effluent standards
Stream standards
16
Characteristics
in mg/L
Indian Council of
Medical Research
(1963)
Desirable
concen.
25
10
25
Physical
1. Colour
(hazen units)
2. Odour(TON)
3. Turbidity(NTU)
4. TS
5. DS
Nothing disagreeable
5
25
1500
500
Chemicals
6. pH
7. Total hardness
(as CaCO a)
8. Calcium
9. Magnesium
10. Iron (as Fe)
11. Copper(as CuI
12. Manganese
13. Sulphate(SO.)
14. Chlorides
1 5. Nitrates
16. Fluoride(as F)
1 7. Phenolics
18.Zinc
7-8.5
6.5 - 9.2
6.5 - 8.5
7 - 8.5
6.5 - 9.2
300
75
50
0.3
1.0
0.1
200
200
20
1.0
0.001
-
600
200
150
1.0
3.0
0.5
400
1000
50
1.5
0.002
300
75
30
0.3 (max)
0.05
0.1 (max)
150
250
45
0.06-1.2
0.001
5
200
75
30
0.1
0.05
0.05
200
200
45
1.0
0.001
5
600
200
150
1.0
1.5
0.5
400
1000
45
1.5
0.002
15
0.05
0.05
0.01
0.05
0.05
0.01
0.05
0.1
0.001
0.01
0.05
0.05
0.01
0.05
0.1
0.001
0.01
0.05
0.05
0.01
0.05
0.1
0.001
0.01
0.2
0.2
0.01
0.2
0.01
0.2
0.3
0.2
3
30
3
30
Toxic
19.Arsenic
20. Cyanides
21. Cadmium
22.Chromium(6+) 23.Lead
24.Mercury
25.Selenium
0.1
0.0001
0.01
Others
.26. Anionic detergents
(as MBAS)
27.Residual CI
28.Mineral oils
29.PABs
30. Pesticides
31. Radioactive
materials
a. Alpha emitters
b. Beta emitters
3 pCi/L
30 pCi/L
17
i.
n.
Table 2.2a:
WHO guidelines for drinking water quality: Microbiological and
biological quality.
Unit
Organisms
Guideline
values
Remarks
1. MICROBIOLOGICAL QUALITY
A. Piped water supplies
No./100 mL
No'/100 mL
No./100 mL
Coliform
organisms
No./100 mL
0
0
No./100 mL
No./100 mL
0
0
Coliform
organisms
No./100 mL
No./100 mL
No./100 mL
o
10
18
No./100 mL
Coliform
organisms
No./100 mL
No./l00 mL
Coliform
organisms
No./l00mL
ii)
iii)
Table 2.2b:
WHO guidelines for drinking water quality: Inorganic constituents
of health significance.
Constituents
Units
Guideline value
1. Arsenic
mg/L
0.05
2.
Asbestos
3.
Barium
4.
Beryllium
5.
Cadmium
mg/L
0.005
6.
Chromium
mg/L
0.05
7.
Cyanide
mg/L
0.1
8. Fluoride*
9. Hardness
mg/L
1.5
10. Lead
mg/L
11. Mercury
mg/L
0.001
12. Nickel
1 3. N itrate-N
mg/L
10
14. Nitrite
1 5. Selenium
mg/L
16. Silver
17. Sodium
19
Unit
Guideline Remarks
values
Bq/L
0.1
Bq/L
Table 2.2d:
WHO guidelines for drinking water quality: Aesthetic quality.
Constituents or
characteristics
Unit
Guideline
values
Remarks
1. Aluminum
mg/L
0.2
2. Chloride
mg/L
250
No guideline
value set
3. Chlorobenzenes and
chlorophenols
4. Colour
TCU
5. Copper
mg/L
6. Detergents
7. Hardness
8. Hydrogen sulfide
mg/L
15
No guideline
value set
500
Not detectable
by consumers
9. Iron
mg/L
0.3
10. Manganese
mg/L
0.1
No guideline
value set
6.5-8.5
13. Sodium
mg/L
200
14. TDS
mg/L
1000
15. Sulfate
mg/L
400
12. pH
Inoffensive to
most consumers
17. Temperature
No guideline
value set
1 8. Turbidity
NTU
19. Zinc
mg/L
Secondary Drinking
Water Standards
Constituents
MCl, mg/l
Constituents
SMCl, (mg/l)
Inorganics
1. Arsenic
2. Barium
3. Cadmium
4. Chromium
5. Fluoride
6. Lead
7. Mercury
8. Nitrate
9. Selenium
10.Silver
0.05
1
0.01
0.05
4
0.05
0.002
10
0.01
0.05
1. Chloride
2. Colour
3. Copper
4. Corrosivity
5. Fluoride
6. MBAS
7. Iron
8. Manganese
9. Odor, TON
10. pH
11. Sulfate
12.TDS
13. Zinc
14. Sodium
250
1 5 colour unit
1
Noncorrosive
2
0.5
0.3
0.05
3
6.5-8.5
250
500
5.0
20
Microbiological
Coliforms
1/100 ml
Physical Charl!cteristics
Turbidity, NTU
1-5
21
Treated water: Throughout any month in 90% of the samples examined, colifonn
bacteria should be absent in 1OOmL of sample. No sample should contain E. coli
in 100 mL. No sample should contain more than 10 colifonn organisms per 100
mL. An MPN of colifonn bacteria of 10 per 100 mL should not occur in consecutive samples.
Treatment requirement
I.
Nil
No treatment is required
11.
Less than 50
Simple chlorination.
Ill.
N.
22
Annex.-1
100
4.
* Dissolved solids,
mg/L. Max.
5. pH value
6. Temperature, C,
Max
Land for
irrigation
Marine
coastal
areas
600
Annex.-1
200
Shall pass
850 micron
IS Sieve
Annex.-1
*a.
*b.
Floatable
solids, max
3 mm
Settleable
solids,
max 850
microns
2100
2100
2100
5.5-9.0
5.5 to 9.0
Shall not
exceed 5C
above the
receiving
water temp.
5.5-9.0
20
10
5.5-9.0
Shall not
exceed
5C
above the
receiving
water
temp.
20
8.
1.0
50
50
100
9.
10.
11.
10
mg/L, Max
Total residual
1.0
C1 2, mg/L, Max
Ammonical N,
50
mg/L, Max.
TKN (as N),
100
mg/L, Max.
Free ammonia
5
(as NH3 , mg/L, Max.
78ble Cont...
23
30
350
100
100
250
0.2
0.2
0.2
250
0.2
0.01
0.01
0.01
0.1
0.1
2.0
2.0
1.0
2.0
0.1
2.0
1.0
2.0
2.0
2.0
3.0
3.0
3.0
5.0
15
15
0.05
0.05
0.05
3.0
3.0
5.0
2
60
5
0.2
2.0
0.2
1000
1000
600
2.0
15
0.2
15
5.0
1000
1000
1000
2.0
5.0
1.0
5.0
10- 7
10-7
10.8
10-7
10-6
10-8
10-7
10-6
90% survival
of fish after
96h in 100%
effluent
90% survival
of fish after
96h in 100%
effluent
90%
90%
survival of survival of
fish after
fish after
96h in
96h in
100%
100%
effluent
effluent
Table Cant ...
5.0
24
0.2
0.2
20
10
10
10
10
450
10
10
10
10
10
10
9600
50
1000
30
2300
7300
780
10
10
10
10
450
10
10
10
10
10
10
9600
50
1000
30
2300
7300
780
Class
Remarks (parameters)
2
3
B
C
25
Table 2.6:
Quality of water for different uses.
Characteristics
1.
2.
3.
4.
pH
DO,mg/L,Min
BOD,mg/L,Max.
Total Coliform,
MPN/l00 mL,Max
5. Colour, hazen units,Max.
6. Odour
7. Taste
8. TDS, mg/L, Max.
9. Total hardness
(as CaC0 3 ),mg/L
10. Ca Hardness
(as CaC0 3).mg/L
11. Mg hardness
(as CaC0 3 ).mg/L
12. Copper (as CuI,
mg/L,Max.
13. Iron (as Fe).mg/L, Max.
14. Manganese (an Mn).
mg/L, Max.
15. Chlorides (as CIl,
mg/L, Max.
1 6. Sulphates (as SO.).
mg/L,Max.
17. Nitrates (as N03 ',
mg/L,Max.
18. Fluorides (as F),
mg/L, Max.
19. Phenolic compounds
(as CeHsOH).mg/L,Max.
20. Mercury (as Hg).
mg/L, Max.
21. Cadmium (as Cd),
mg/L, Max.
22. Selenium (as Se),
mg/L, Max.
23. Arsenic (as As),
mg/L, Max.
24. Cyanide (as CN),
mg/L, Max.
25. Lead (as Pb),mg/L,Max.
26. Zinc (as Zn).mg/L,Max.
27. Chromium (as Cr),
mg/L, Max.
28. Anionic detergents
as MABS, (mg/L,Max.)
29. PAH, mg/L, Max.
30. Mineral oil, mg/L, Max.
31. Barium (as Ba).
mg/L, Max.
6.5-8.5
6
2
50
6.5-8.5
5
3
500
6.5-8.5
4
3
5000
10
Unobjec
tionable
Tasteless
500
300
300
-
300
-
6.5-8.5 6.0-8.0
4
1500
-
200
100
1.5
1.5
0.3
0.5
50
250
600
600
400
400
1000
20
50
1.5
1.5
1.5
0.002
0.005
0.001
0.01
0.01
0.05
0.2
0.2
0.05
0.05
0.1
15
0.05
0.1
15
0.05
0.2
1.0
0.2
0.2
0.01
2100
-
26
0.05
Absent
10.9
26
2
1.2
1.0
2.25
6.
7.
8.
9.
10.
11.
1 2.
1 3.
14.
15.
16 .
17.
1 8.
19.
20.
21 .
22.
23.
24.
25.
Temperature
Colour (Hazen Units)
Odour (TON)
Turbidity
Total suspeneded solids (TSS)' mg/L
Total Dissolved solids (TDS), mg/L
pH
Total alkalinity (as CaC0 3 ), mg/L
P value
M value
Total hardness as CaC0 3 mg/L
Chloride as CI, mg/L
Sulfate as SO 4' mg/L
Fluorides as F, mg/L
Percent sodium (% Na)
Phosphate as PO 4' mg/L
Nitrate as N0 3 , mg/L
Total Kjeldahl nitrogen (TKN), mg/L
Total ammonical nitrogen (as N), mg/L
BOD, 20C, 5 days, mg/L
COD, mg/L
Disolved oxygen demand (DO), mg/L
Total residual chlorine as C1 2 , mg/L
Oil and Grease, mg/L
Chromium (Cr 6 +), mg/L
27
Rationale
28
Table 2.9:
Standards for effluents from coalmines to be discharged into sewer,
stream or land (MOEF, 2000).
Parameter
Standard
pH
Chemical Oxygen Demand (COD)
Total Suspended Solids (TSS)
5.5 to 9.0
250 mg/L
100 mg/L
200 mg/L (Land for irrigation)
10 mg/L
OPTIONAL PARAMETERS
All other parameters indicated in the general standards for discharge of environmental pollutants under Schedule VI, shall be in addition to the effluent standards
specified under clause 3. Monitoring frequency shall be once in a year for the
optional parameters.
pH
Total suspended solids
Bio-chemical oxygen demand (BOD)
Chemical oxygen demand (COD)
Total residual chlorine
Oil and grease
Total chromium as Cr
Sulphide as S
5.5-9.0
100
30
250
1
10
2
2
1
Note:
1. Where the treated effluent is discharged into municipal sewer leading to terminal
treatment plant, the BOD may be relaxed to 100 mg/L and COD to 400 mg/L.
2. The quantity of effluent (litre per kilogram of product) shall not exceed 100, 250
and 80 in composite cotton textile industry, composite woollen textile industry
and textile processing industry, respectively.
29.
2.11 Questions
1. Defme the standards ?
2. In WHO {I 984) guidelines drinking water standards are divided into 5 categories ? Name those categories.
3. What is MCL and SMCL ?
4. List the different drinking water standards available in India.
5. Name the different effluent standards proposed in India.
6. How effluent standards are different from stream standards.
7. How may classes of water has been listed under stream based standard?
8. Name the organisms considered under biological quality of drinking water.
9. How does percent sodium is calculated?
Introduction
Acidity of water is its quantitative capacity to react with a strong base to a
designated pH. All waters having a pH lower than 8.5 contain acidity.
There are two types acidity:
Methyl orange acidity also known as mineral acidity (PH < 4.0). (Bromocresol
blue is now recommended as it has a sharper colour change at pH 3.7).
Phenolphthalein acidity or CO 2 acidity, which is due to dissolution of CO 2 in
water and algal photosynthesis.
Both CO 2 and mineral acidity can be measured by means of standard solution of
alkaline reagents. Mineral acids are measured by titration to a pH about 3.7 i.e.
methyl orange acidity. Titration ofa sample to the phenolphthalein end point of pH
8.3 , measured both mineral acidity plus due to week acids, also known as phenolphthalein acidity. Two types of acidity are shown in Fig. 3.1.
10
31
Measurement of Acidity
PRINCIPLE
The concentration of mineral acids present and contributing the mineral acidity
can be calculated by titrating or neutralising samples to pH 4.3 . The CO 2 and
bicarbonates (carbonic acid) present in the sample can be neutralised completely
by continuing the titration to pH 8.3. Since CaCO] has an equivalent weight of 50,
N/50 NaOH is used as titrating agent so that I mL N/50 NaOH = I mg of acidity.
CHEMICALS AND REAGENTS
Chemicals
I . Standard NaOH titrant
4. Potassium biphtalate
2. Phenolpthalin indicatorr
3. Methyl orange indicator
5. Ethyl alcohol
6. Methyl alcohol
Reagents
1. Carbon dioxide free water: Prepare all stock and standard solutions and
AxB
NormalityofNaOH = -- - 204.2 x C
Where,
A
32
1. Measure suitable volume of sample (50 or 100 mL) in a 250 mL conical flask or
beaker depending upon the method to be followed.
2. Add 2 drops of methyl orange and see the colour. If colour turns yellow, methyl
orange acidity is absent. If the colour turns pink titrate with standard 0.02N
NaOH till colour changes to yellow, characteristic of pH 4.4 - 4.3 . Note down
the volume ofNaOH required (A).
3. Add 2-3 drops phenolphthalein indicator and continue titration with NaOH till
faint pink colour appears indicatiing pH 8.3. Note down the additional volume
ofNaOH required (8).
CALCULATION
mLofSample
Where,
A = mL NaOH required for sample to raise pH up to 4.4 -4.3 by methyl orange.
B = mL NaOH required for sample to raise pH from 4.4 to 8.3 by phenolphthalein.
N = Normality ofNaOH used .
3. For measuring total acidity use A + B instead of only A or B in the calculation.
O'()2 N
StlndMd
NaOH
f1
.
33
Methyl orange acidity (pH <4) also known as ............... ..... .. .acidity.
Phenolphthalein acidity (pH < 8.5) is due to (a) and (b) .
How to stored NaOH titrant in the laboratory?
A 100mL sample of water has an initial pH of8.4. 20 mL ofO.02N NaOH is
required to titrate the sample to pH 4.3 . What is the total acidity in mglL as
CaC0J"
3.2 Alkalinity
Drinking water
IS: 105000 (1991) = 200 mgIL (maximum), beyond this limit taste becomes
unpleasant. 600 mgIL (permissible in the absence of alternate source).
Effluent discharge
MOEF (1993)= No Standard
Introduction
The alkalinity of a water is a measure of its capacity to neutralise aCIds. Alkalinity is
a measure of the water ability to absorbed W without significant pH change. That
is, alkalinity is a measure of buffering capacity of water. It is the sum of all the
titrable bases.
The major portion of alkalinity in natural waters is caused by hydroxide, carbon
ate, and bicarbonate. For practical purposes, alkalinity due to other materials in
natural water is insignificant and may be ignored. Where bicarbonate concentra
tion is high Ca and Mg are also high.
In natural water, most of the alkalinity is caused due to CO 2 The free CO2
dissolves in water to form carbonic acid (H2COJ), which fuI1her dissociates into H+
and HCOJ. The HCO J thus formed further dissociates into H+ and COJ ~ .
CO 2 + Hp ~ H 2CO J (dissolved CO 2 and carbonic acid)
H2CO J ~ W+ HCO; (bicarbonate)
HCO J ~ W + CO;- (carbonate)
Thus, alkalinity meqlL = [HCO;] + [CO;-] + [OH-] - [W]
Where the quantities in parenthesis are concentration in meqlL or mgIL as
CacoJ
34
Environmental Significance
Alkalinity in itself is not harmful to human beings, still the water supplies with less
than 100 mglL are desirable for domestic use. In large quantities, alkalinity imparts
bitter taste of water. The other applications of alkalinity data are:
Estimation of Alkalinity
PRINCIPLE
Alkalinity of a sample can be estimated by titrating with standard sulphuric acid.
Titration to pH 8.3 or de-colourization of phenolphthalein indicator will indicate
complete neutralisation ofOH and Y2 of COl while to pH 4.5 or sharp change from
yellow to pink of methyl orange indicator, that will indicate total alkalinity (complete
neutralisation of OH, COl' HCOJ The alkalinity titration curve is shown in Fig.
3.3. Wherever possible, the titration should be carried out on filtered water at the
point of sampling.
14
__________ L ____________ _
"
:. t co;-. Heoi
OH- + C0 2 - + HCo;
o~--------
__________
___________
Milliliter. of titront
INTERFERENCE
Colour, turbidity, iron, aluminium and residual chlorine are prime sources of
interference. Colour and turbidity can be avoided using potentiometric titration .
Residual chlorine can be removed by adding ofa small amount (usually one drop)
of 0.1 mollL sodium thiosulfate solution.
35
Chemicals
I. Sulphuric acid, H 2 S0 4 (sp. gr. 1.84)
2. Phenoplthalein indicator
3. Sodium carbonate, Na2CO]
4. Sodium hydroxide, NaOH
5. Methyl orange indicator
6. Phenolpthalin indicator
Reagents
l. COz-free distilled water must be used for preparation of all stock and standard
solution. If the distilled water has a pH lower than 6.0, it should be freshly boiled
for 15 min and cooled to room temperature. Deionised water may be substituted
for distilled water provided it has a conductance ofless than 0.2 mS/m and a pH
greater than 6.0.
2. Sodium carbonate solution (0.05 N): Dry 5 to 7 g primary standard Na2 CO] at
250C for 4 h and cool in a desiccator. Weigh 5.300g, transfer to a 1000-mL
volumetric flask, and fill to the mark with distilled water. Do not keep longer than
I week.
3. Standard H zS04 solution (0.02N): Prepare O.IN H 2S04 by diluting 3.0 mL conc.
H 2S0 4 (sp. gr. = 1.84) to 1000mL. Standardise it against standard Na2C03 (0.05
N). Dilute appropriate volume of H2S0 4 to 1000 mL to obtain standard 0.02 N
H2S04,
4. Phenolphthalein indicator (pH 8.3 indicator): Dissolve 0.5 g in 500 mL 95%
ethyl alcohol. Add 500 mL distilled water. Add drop wise 0.02 N NaOH till pink
colour appears.
5. Methyl orange indicator: Dissolve 0.5 g of methyl orange in 100mL of water
and dilute to 1ObOmL with CO 2 free distilled water.
36
il
0.02 N
Standard
H.SO.
37
Table 3.1:
Relationship between hydroxide (OHI. carbonate (CO]I
an~ bicarbonate (HCO]I alkalinities.
Results of
titration
OH alkalinity as
CaCO]
P =0
P<Yz T
P=Yz T
P>Yz T
P=T
CO~- alkalinity as
CaCO]
HCOi alkalinity as
CaCO]
0
0
0
2P
T
T-2P
0
2P-T
T
2P
2 (T-PI
0
0
0
3.
4.
5.
6.
7.
Drinking water
No BOD standard has been prescribed by WHO (1984), IS: 10500 (1983,1991),
MHW (1975) orUSEPA (1974) orICMR(l963)
Emuent discharge
30 mgIL, (MOEF, 1993); IS: 2490 (1981) = In inland surface waters; 350 mgIL
(Public Sewers); 160 mgIL (land for irrigation and marine costal areas).
In India, Central Pollution Control Board (CPCB) recommends BOD measurement at 27C for 3 days.
For bathing water (MOEF, 2000) = 3 mgIL or less
38
Introduction
Biochemical oxygen demand (BOD) is defined as the amount of oxygen required by
bacteria in decomposing organic material in a sample under "aerobic condition at
20 "C over a period of 5 days.
C6H.P6 + 602
Microbes
Type of sample
BOD (mg/L)
Remarks
Water
1-3
reasonable
River
5-20
tolerable
Sewage
50-100
very bad
Industrial waste
100-10,000
Extremely poor
39
-e
...J
C7I
0'
o
co
Carbonateous oxygen
(C BOD)
10
11
12
de~
13
14
15
Time, days
3.
4.
5.
6.
APPARATUS
1. Incubation BOD bottles, 300 mL capacity: Clean bottles with a detergent
(or chromic acid), rinse thoroughly, and drain before use so that bottles are free
40
from organic matter. As a precaution, against drawing air into the dilution bottle
during incubation, use a water seal. Place a paper or plastic cup over flared
mouth of bottle to reduce evaporation of the water seal during incubation.
2. Incubator: Thermostatically controlled at 20"C. Exclude all light to prevent
possibility of photosynthetic production of DO.
Reagents
All reagents prepared for DO determination also required for BOD test. In addition,
prepare following stock solution.
41
"[M.easure
'----...,VlFlnal DO
Incubate
20C .S da"
IHea$Ure mital
00
Fig. 3.6: BOD-test (Direct method) (300 mL sample is taken in BOD bottle and DO in
the beginning and after incubation for 5 days at 20C is measured).
Dilution method (sample having high BOD) (Figs. 3.7 and 3.8)
For polluted water, domestic sewage, industrial effiuents, where BOD values are
generally high, dilution of the sample is needed to provide sufficient dissolved
oxygen. The important pre-requisite steps to be followed are:
I. Preparation of dilution
wat~r
1. Aeration: Aerate the required volume of distilled water in a container by bubbling compressed air for 1-2 days to attain DO saturation. Try to maintain the
temperature near 20 OC.
2 pH: Neutral pH (7.2) should be maintained. It is customary to buffer the dilution
water by means of phosphate buffer at about pH 7.2. The buffer is essential to
maintained favorable condition at all times
3. Addition of nutrients: Add -I mL each of phosphate buffer, magnesium sulphate, calcium chloride and ferric chloride solution for each litre of dilution
water and mix well. These are added to provide proper osmotic conditions and
essential micronutrients.
4. Seeding: Some samples may not contain a sufficient microbial population,
such as some untreated industrial wastes, disinfected wastes, high temperature
wastes or wastes with extreme pH. In such cases seed the dilution water by
42
adding population of microorganisms. In the case of the wastes that are not
expected to have sufficient bacterial populations, add seed to dilution water.
Generally, 2 mL settled sewage or any other given below is considered sufficient for 1L of dilution water.
Seed source
It is necessary to have present populations of microbes capable of oxidising the
biodegradable organic matter present in the sample. Common sources of seeds are:
Domestic wastewater
Unchlorinated or undisinfected effluents from biological wastewater treatment.
plant
Surface water receiving wastewater discharge
Use supernatant from domestic wastewater after settling at room temperature
for at least 1 h but not later than 36 h
When effluent from biological treatment process is used, inhibition ofnitrification is recommended.
II. Dilution of sample
1. Neutralize the sample to pH around 7.2 if it is highly alkaline or acidic.
2 The sample should be free from residual chlorine, ifit is present, remove it by
43
Dilution method (with seeded) i.e when dilution water seeded with bacteria. An
outline of the test procedure is given in Fig. 3.8.
Table 3.3:
Recommended dilution for the BOD& test.
Range of BOD values
to be determined
(mg/LI
0-6
4 - 12
10 - 30
20 - 60
40 - 120
100 - 300
200 - 600
400 - 1200
1000 - 3000
2000 - 6000
R
C
E
S
I
Sample
volume
(mLI
Dilution water
volume (mLI
Undiluted
0
500
800
900
950
980
990
995
998
999
500
200
100
50
20
10
5
2
1
_ Source of
Dilution
factor Md M
sample
1
2
5
10
20
50
100
200
500
1000
R
R,E
R,E
E,S
S
S,C
S,C
I,C
I
I
= River water
Crude (raw sewage)
Biologically purified sewage effluent
Settled sewage or weak industrial wastewater
Strong industrial wastewater.
=
=
=
=
1. In preparing dilutions for BOD test, siphon-off or pour careful~ the standard
Dilution water
Air
nutrients
(Jl()mL-VS
Glassbottle
(20 L)
Air --H:lt-L.o
stone
Measure iniAl DO
Dilution water
lunseeded)
800 bottle
44
5. Fix the bottles kept for DO determination and blank by adding 2 mL MnSO.
followed by 2 mL NaOH + KI + NaN3 as described in the estimation of DO.
6. Determine DO in the sample and in the blank on initial (0 day) and after 5 days
(end of test day).
Oilution water
( sHdeda
"'UUl"t
SEEDED TnT
$ANPLE
tiM' DO I
CALCULATION
1. When BOD is determined in an undiluted sample (Direct method):
BODs (mgIL) = DO before incubation (mgIL) - DO after incubation (mgIL)
0.-02
p
Where,
DO of the sample bottle on O-day (before incubation).
D.
O2
DO of the sample bottle after 5th day.
B.
DO of the blank bottle on O-day.
B2
DO of the blank bottle after 5th day.
f
ratio of seed in diluted sample to seed in seed control
45
46
47
equal to 2 mg/L, while that containing just seeded dilution water has DO equal
to 8 mg/L. Find the BODs of the waste.
14. A mixture consisting of30 mL of waste and 270 mL of seeded dilution water has
an initial DO of8.5 mg/L. After 5 days, it has a final DO of2.4 mg/L. Another
bottle containing just the seeded dilution water has an initial DO of8.75.mg/L
and a fmal DO of8.53 mg/L. Find the BODs of the waste.
15. Determining BODs of an unknown sample: The BODs ofa wastewater is s~s
peeted to range from 50 to 200 mg/L. Three dilutions are prepared to cover the
range, i.e. 5 mL, 10 mL and 20 mL waste diluted to 300 mL with dilution water. The
initial 00 and final 00 as measured in each bottle are given below. Calculate the
BODs of the wastewater.
SolutionofQ.15:
DO after 5 2consumed
Wastewater Dilution . Initial
water, mL DO, mg/L daysmg/L
mglL
mL
BODs
mg/L
295
92
6.9
23
0.0167 l38*
10
290
4.4
4.7
0.033
142
280
9.1
8.9
1.5
7.4
0.067
llO
wastewate~mL
8.9
1.5
10
.5
9.1
92
2.5
5.8
92
7.5
48
3.4 Boron
(Curcumin colorimetric-extraction method: range 0.1 to J mg/L)
Drinking water
IS: 10, 500 (1991) = I mgIL (5 mgIL is pennissible in the absence of alternate
source)
WHO (1993) = 0.3 mgIL (Guide line value).
EC (1980) = I mgIL (Guide level)
Emuent discharge
MOEF, (1993) = 2 mgIL (omitted by GSR, 80 (E), dated 31st December, 1993
(w.e.f. 31.12.93)
Stream standards (CPCB, 1979) = 2 mgIL (E-c1ass water Irrigation purposes).
Introduction
In most natural waters boron is rarely found in concentrations greater than 1 mgIL,
but even this low concentration can have deleterious effects on agricUlture crops.
Boron (B) is essential to plant growth, but at high concentration it becomes deleterious. B concentration in Indian soil ranged from 2 to 8 mgIL.
At high concentration, it effects the central nervous system, while extended
consumption may lead to a condition known as borism. Different methods for
boron estimation are:
I. Colorimetric-curcumin method: O. 10 to I mgIL.
2. Colorimetric-carmine method: 1 to 10 mgIL.
Estimation of Boron
PRINCIPLE
Water sample containing boron is acidified with Hel and evaporated in the presence ofcurcumin (a dye extracted from tunneric), a red coloured complex rosocyanine
is fonned. The colour product is dissolved in isopropyl alcohol and absorption is
measured at 540 nm in spectrophotometer.
APPARATUS
1. Hot-water bath, with temperature control at 55C.
2 Spectrophotometer.
3. Evaporating dish, 100 to 150 mL capacity.
INTERFERENCES
Nitrate concentration above 20 mgIL begins to interfere. Hardness level about 100
mglL as CaC03 give high results because of the turbidity caused by the insolubility
49
Reagents
1. Boron stock solution (ImL = 1 mg B): Dry about 109 of boric acid (H]BO])
2.
3.
4.
S.
powder in a desiccator containing a silica gel for 24h. Dissolve 5. 719g of the dry
boric acid in water and dilute to IL. Store the solution in a plastic bottle.
Boron standard solution (ImL = 0.01 mg B): Dilute IOmL of stock boron solution to IL with water. Store in a plastic bottle.
Curcumin solution: Dissolve 40 mg offmely ground curcumin powder and 5 g
of oxalic acid in 80mL of isopropyl alcohol. Add 4 mL of cone. HCI and make up
to I OOmL with isopropyl alcohol.
Hydrochloric acid (1+ 19): Add I volume ofHCI (cone.) to 19 volumes of water.
Sodium hydroxide solution (20gIL): Dissolve 2 g fNaOH in water and dilute to
lOOmL.
SAMPLING
Samples should be collected and stored in polythene bottles. Filter the sample
through a 0.45-~m membrane filter as soon as possible after sampling. No preservation is required.
PROCEDURE
1. Calibration of standards: Prepare a series of standard boron solution cover-
2
3.
4.
5.
6.
50
CALCULATION
Calculations are not required, as the B concentration can be read directly from the
calibration curve provided that no dilution of the original sample was made. In case
of dilution, mUltiply the reading from the graph by the dilution factor.
Emuents discharge
MOEF (1993) = No Standards.
Introduction
Free carbon dioxide in the water accumulates due to microbial activity and respiration of organisms. Surface waters normally contain less than 10 mg free CO2 per litre
while some groundwater may easily exceed that concentration up to 30 to 50 mglL.
This imparts the acidity to the water because of the formation of carbonic acid (C02
+ Hp = H2COJ The CO2 content of a water may contribute significantly to corrosion (e.g concentration of CO 2 >20 mgIL is bad). Free CO2 is determined by titrating
the sample using a strong alkali to pH 8.3 .
Estimation of Free CO 2
PRINCIPLE
Free CO2 reacts with sodium carbonate to form sodium bicarbonate, or with so~ium
hydroxide to form sodium carbonate. Completion of reaction is indicated by the
development of the pink colour characteristics of phenolphthalein indicator at the
equivalence pH of8.3.
51
CO2 + N~C03 +
CO2 + 2 NaOH
~O =
2NaHC03
N~C03 + ~O
REAGENTS
indicator.
2 The colour change to pink indicates the absence offree COl"
0.05 N
Standard NaOH
PIMI(
( f nO Point I
51
CALCULATION
AxNx44x 1000
Free C02mg/L
= --------
mLofsample
3. A sample of water collected in the field had a pH of6.8. By the time the water
sample reached the laboratory for analysis, the pH had increased to 7.5. Give a
possible explanation for this change.
Introduction
Pure water is a poor conductor of electricity. Acids, bases and salts in water make it
relatively good conductor of electricity and such substances are called electrolytes.
The electrolyte in solution dissociates with positive (cations) and negative (anions)
ions and imparts electrical conductivity. Thus, higher the concentration of electrolyte
in water, the moreis its electrical conductance (i.e'. less the resistance to the flow of
electricity). This gives a rapid method to get an idea about dissolved solids in water.
The important ions that impart conductivity in water are:
i. anions: CI- , S~, CO;, HCO; and NO;
ii. cations: Ca 2+, Mg 2+, Na+ and K+
Conductivity is customarily reported in micromhos per centimeter (/lmhos/cm). In
the International System of units (SI) the reciprocal of ohm is the Siemens (S) and
53
Estimation of Conductivity
PRINCIPLE
A
G=k L
k is called "conductivity" (preferred to "specific conductance"). The unit of k is
1I0hm-cm or mho per centimetre. It can be defmed as the conductance of a conductor
Icm in length and 1 cm 2 in cross-sectional area. The specific conductance depends
on the nature of the conductor (the solution between the electrodes), the ion
concentration, and pressure.
The conductance of a solution measured by conductivity cell depends on cell
parameters, A and L. Although theoretically one could calculate the specific conductance from measurements ofG (or R) and values of A and L, it is easier, and more
accurate to use a solution of known specific conductance, measure G, and calculate
54
meter conductance
0
0.0001
0.001
0.01
0.1
1
Equivalent conductance,
mho-cm 2 /equivalent
149.9
148.8
146.9
141.1
128.9
111.9
E,
Specific conductance,
k,pmho/cm
14.9
146.9
1412
12890
111900
55
vary from 0.19 to 0.55 depending on the soluble components and on the temperature
of measurement.
Conductivity measurement gives rapid and practical estimation of the variations in the dissolved mineral contents of a water supply. The most commonly
applied relationship is a linear proportionality, as given in the following equationDissolved solids = A x Specific conductance;
A water quality survey conducted by US Dept of Interior (1997), found that the
value of A varies from 0.60 to 0.74. A higher value of A is associated with high
concentration of sulphate ions. The relationship between dissolved solids, specific
conductance is summarised in Table 3.5.
Table3.5:
Relationship between dissolved solids and specific conductance.
Source
Surface
Ground
Ground
Ground
Dissolved solids
(DS)(mg/L)
water
water
water
water
107
236
80
173
Specific conductance
(SC) IpS/cm)
171
392
99
270
Ratio (A)
(DS/SC)
0.64
0.61
0.74
0.65
APPARATUS
Conductivity meter.
REAGENTS
Standard potassium chloride, (O.OlM): Dissolve 745.60 mganhydrous KCl in double
distilled water or deionised water and make up to 1000 mL at 25C. This is standard
reference solution, which at 25C has a specific conductance of 1412 ~mhos/cm. It
is satisfactory for most samples when cell has a constant between 1 and 2 cm'.
INTERFERENCE
Conductivity measurement is affected by:
nature ofthe various ions, their relative concentration and the ionic strength of
water;
dissolved CO 2; turbidity, and
temperature (the conductivity must be determined at 25C).
PROCEDURE
The measurement of conductivity should be made in-situ or in the field immediately
after water sample has been obtained, because conductivity changes with storage
time. Conductivity is also temperature dependant, thus, if the conductivity meter is
not equipped with automatic temperature correction, the temperature of the sample
should be measured and recorded.
56
1. Rinse the conductivity cell at least three portions of standard KCI solutions.
2. Adjust the temperature of a fourth portion to 25C 1C
3. Immerse the conductivity cell in the standard KCI solution. (Note: the cell should
not be closer than 2 cm to the sides and bottom of the container).
4. Observe and record the temperature of the KCL solution to the nearest O.toC.
Some meters have builtin thennometerl or automatic temperature compensation.
5. Turn the meter on and follow manufacturer's interactions and record the meter
reading.
6. Calculate the cell c.:;n.,tant.
Kc = (1412/C K )
[1 +0.019(t-25)]
Where,
Measured conductivity, flmhos/cm.
CKti
t = Observed temperature, C
K c = Cell constant (em-I).
Measurement of sample conductivity
1. Rinse the conductivity cell with at least three portion of the sample.
2. Adjust the temperature of the portion of the sample to 25C 1C.
3. Immerse the conductivity cell in the solution. There should not be any air
bubble clinging to the electrodes and cell should not be closer than 2 cm to the
sides and bottom of the container. A typical conductivity measurement
procedure is shown in Fig. 3.10.
4. Observe and record the temperature of the KCL solution to the nearest O.IC.
Some meters have built in thennometerl or automatic temperature compensation.
5. Tum the meter on and follow manufacturer's instructions and record the meter
reading.
ConductiVity
Ctll
~."d. to
Canductwa
IridgO
57
6. When conductivity of the sample has been measured, the conductivity can be
calculated as follows:
C xK
Conductivity =1 +0.0191 (tt_25)
Where, C m = measured conductivity in units of ~mho/cm at tOe.
REPORTING
Report the reading and the temperature of the sample at the time of reading. Report
the conductivity at 25e.
3.7 Colour
Standards for Colour
Drinking water
IS: 10500 (1983) = Max. I 0 Hz units;
IS: 10500 (1991) = Max. 5 Hz (above 5 Hz consumer acceptance decreases. (25 Hz
max. permissible limit in the absence of alternate source).
MWH (1975); ICMR (1963) = 5 Hz units is permissible and maximum permissible
is 25 units.
WHO (1984) = Maximum 10Hz units;
WHO (1993)= 15 TCU (time colour unit)
USEPA(1974)= 15 Hz units;
EC (1980) = 1Hz (guide level) and 20 Hz (MAC, maximum admissible concentration)
Note: The standard of 15 Teu is based on the fact that at 15 TeU no colour is detectable
visually. Therefore, consumers' acceptance decreases and colour standards are not based on
health effect (John De Zhane, 1977).
Effluents Discharge
MOEF, 1993 = All efforts should be made to remove colour as far as possible.
58
Introduction
Colour in water may result from the presence of natural metallic ions (iron and
manganese), humus and peat materials, plankton, weeds. For example, Iron oxides
cause reddish water and Manganese oxides cause brown or brackish water. Industrial
wastes from textile and dying operation, pulp and paper production, food processing,
mining and coal processing operations, refinery, slaughter house operation may
add substantial colouration to water in receiving stream.
Colour is removed to make water suitable for general and industrial applications. Coloured industrial wastewater may require removal of colour discharge into
watercourses.
Colour can be described relative to type and density; the type being related to
whether it is true colour (dissolved) caused by colloidal particles or apparent colour
(filterable), caused by suspended matter. It is express in Hazen units.
Environmental Significance
Coloured water is not aesthetically acceptable to the general public.
True colour caused by organic compounds may exert chlorine demand during
disinfection process and thereby seriously reduce the effectiveness of chlorine
as a disinfectant.
Many industrial wastes are highly coloured, and some contain coloured
substances that are quite resistant to biological destruction.
Estimation of Colour
PRINCIPLE
Waters containing natural colour are yellow-brownish in appearance. It has been
found that potassium chloroplatinate (~PtCI6) tinted with small amounts of cobalt
chloride (CoCI) yield colours that are very much like the natural colours of water.
ImgIL ofPt (KltCI6) gives standard 1 unit of colour. Colour intensity increases
with increase in pH. For this reason recording pH along with colour is advisable.
1mgIL Pt (~PtCI6) = 1 true colour unit (TCU) = 1 Hz
REAGENTS
1. Stock standard colour solution: Dissolve 1.246g KltCl 6(equivalent to 500 mg
metallic platinum) and Ig crystallized CoCI 2 6Hp (equivalentto about 250 mg
59
metallic cobalt) in distilled water. Filter it to remove any slight turbidity. Add
100mL conc. HCI and dilute with distilled water to 1000mL. This stock standard
solution, has a colour of 500 Hazen unit.
2 Working standard colour solution: Prepare standards having colour of following
units in 50mL ofNessler's tube. Add 0.5, 1.0, 1.5,2.0,2.5,3.0,3.5,4.0,4.5,5.0,6.0
and 7mL of stock coloured solution in 50mL Nessler's tube and filled with
distilled water up to the mark which will give 5, 10, 15,20,25, 30, 35, 40, 45,50, 60
and 70 Hazen units. These standards can serve for several months, provided
that these are protected from dust and evaporation.
PROCEDURE
1. Filter the sample through a glass fibre filter paper to remove the turbidity.
2 Measure the optical density (OD) with a spectrophotometer at a wavelength
between 385 and 470nm. Put filtered water in the reference cell.
3. Prepare calibration graph (prepared in the range of 10 to 70 units).
4. Record as colour to the nearest whole number.
ESTIMATION OF COLOUR IN INTACT SAMPLE
The colour of the sample is observed by filling a matched Nessler's tube to the 50mL mark with sample and comparing it with standards. Look vertically downwards
through tubes towards a white or specyular surface placed at such an angle that
light is reflected upward through the columns ofliquid. If turbidity is present and
has not been removed, reported as "apparent colour". If the colour exceeds 70
units, dilute sample with distilled water in known proportions until the colour is
within the range of the standards. Measure the pH of the each sample.
CALCULATION
3. What is the difference between true colour and apparent colour? How to estimate
colour?
4. "1 mg fL ofPt. gives 1 unit of colour" (true/false).
Effluent discharge:
250 mg/L, into inland surface waters and marine coastal areas (MOEF, 1993;
IS: 2490, 1981)
Environmental Significance
Chemical Oxygen Demand (COD) test determines the oxygen required for chemical
oxidation of most organic matter and oxidizable inorganic substances with the help
of strong chemical oxidant. The test can be employed for the same purpose as the
BOD test taking into account its limitations.
The intrinsic limitation of the test lies in its inability to differentiate between the
biologically oxidisable and biologically insert material.
COD determination has an advantage over BOD determination in that the result
can be obtained in about 3 h, as compared to 5 days required for BOD test. Further
the test is relatively easy, gives reproducible results and is not affected by
interference's as the BOD test.
Measurement of COD
NOTES ON COD TEST BY REFLUX DIGESTION METHOD
1. COD determines the amount of oxygen required under specific test conditions
for the oxidation ofwaterbome organic and inorganic matter.
2. The reflux methods are limited by the reagents employed to a maximum COD of
800 mg/L. Simple with higher COD may be processed by appropriate dilution of
the sample.
3. If the sample is diluted, it must consume at least 5 mL of dichromate. Dilute the
sample if more than 20 mL of the potassium dichromate solution is consumed by
the reaction.
61
4. COD results are not accurate if the sample contains more than 1000 mg/L chloride. Consequently, the reflux method should not be applied to samples such as
seawater and brines.
PRINCIPLE
The organic matter and oxidisable inorganic substances present in water or wastewater get oxidised completely by standard potassium dichromate (K zCrp7) in the
presence ofHzS04 to produce COz + Hp. The excess K ZCrp7 remaining after the
reaction is titrated with ferrous ammonium sulphate [Fe(NH4MS04)Z]. The dichromate consumed gives the 02 required for oxidation of the organic matter. The
contents are refluxed for 2 h.
2~Crp7 + 8HzS04~
6Fe3++2Cr3++7HP
Comments: It is based on the principle that all organic compounds can be oxidized
by strong oxidizing agents in acidic media. Hence:
COD values are always greater than the BOD values. COD values are too large,
if greater amounts of biologically resistant organic matter is present, i.e. lignin.
COD value do not provide any information on the rate at which actual
bio-degradation will take place.
Reflux condensers are used to prevent loss of volatile organic compounds.
INTERFERENCES
1. Fe (II) and hydrogen sulfide exert COD of 0.14 mglmg Fe2+and 0.47 mglmg ~S
respectively.
2. The chlorides interference is eliminated by addition of mercury salt (mercury
sulfate), which precipitated chlorides as HgCI 2: Hgz+ + 2Cl- = HgCI 2. Although
1g HgS04 is specified for 50 mL sample, a lesser amount may be used where
sample chloride concentration is known to be less than 2000 mg/L or as long as
a 10: I ratio ofHgS04: Cl- is maintained.
3. Addition of A~SO4to conc. H ZS04acts as a catalyst to stimulates the oxidation
of straight chain aliphatic and aromatic compounds.
4. Nitrite (N0z-) exerts a COD of 1.1 mglmg NOz-N. Sulphamic acid is added at 10
mg sulfamic acid for every Img ofNOz-N in the refluxing flask. Add sulfamic
acid to the standard K ZCrp7' since it must be included in the distilled water
blank.
62
SAMPLE HANDLING
Natural, not very heavily polluted water should be analyzed on the same day or at
least within 24h. If there is to be a delay before analysis, the sample may be preserved by adding sulfuric acid, about 2mL H2S04 (d = 1.84) diluted to 1 + 2 to each
100 mL of sample. If samples are to be stored for longer than 24h, deep-freezing is
recommended.
Safety precautions:
1. Exercise extreme care when handling conc. H2S04 , especially at the start of
refluxing step.
2. Silver sulfate (Ag2S04 ) is poisonous; avoid contact with the chemical and its
solution.
3. Mercuric sulfate (HgS04 ) is very toxic; avoid contact with chemical and its
solution.
APPARATUS
1. COD Reflux apparatus consisting of a flat bottom 500-mL capacity flask with
condenser.
2. A heating mantle
1.
2.
3.
5.
6.
7.
Reagents
1. Standard potassium dichromate, 0.25N or 0.0417 M: Dissolve 12.259 g of
K2Crp7 (dried at 103C for 24 h) in distilled water and dilute to lL. Add about
120 mg ofsulphamic acid to take care of6 mg/L ofN02-N.
2. Standard ~Cr207solution, 0.025N or 0.00417M: Dilute 100 mL of0.25N
K2Crp7solution to lL. (This solution is required only if COD is determined in
the range of 10 to 50 mg/L).
3. Sulphuric acid - silver sulphate reagent: Add 109 A~SO4 to 1000 mL of conc.
H 2S0 4 and keep overnight for dissolution.
4. Standard ferrous ammonium sulphate (FAS), approx. O.25N: Dissolve 98 g of
[Fe(NH4)iS04)2' 6HPl in about 400 mL distilled water and add 20 mL ofconc.
H2S04, cool and dilute to lL. Standardise the solution daily against the standard K2Crp7'
5. Standard FAS, O.025N: Dilute 100 mL of standard FAS solution to IL. (or dissolve 9.8 g ofFAS in lL distilled water with 20 mL ofconc. sulphuric acid).
63
N=(A x B)/C
Where,
N = normality of the FAS solution
A = volume ofK2Crp7solution, mL
B = normality ofK2Crp7solution
C = volume ofFAS, mL
6. Ferroin indicator: Dissolve 1.485 g of 1,1 O-phenonthroline monohydrate,
together with 695 mg ofFeS04.7Hp in water and dilute to 100mL. This indicator
solution is also commercially available.
7. Mercuric sulphate (HgS04): Analytical grade crystals
Estimation of COD
SAMPLES WITH HIGH CHLORIDE CONCENTRATION
1. If the sample contains more than 100 mgIL chloride after dilution, proceed as
follows:
2. To 20 mL of sample or aliquot in the flask add 0.5 g mercuric sulfate and shake
thoroughly. This addition is sufficient to complex 40 mg of chloride or 2000
mgIL when 20 mL of sample is used. If more chloride is present, add more HgS04
to maintain a HgS04 : CI- ratio of 10 : 1. It is not important if a slight precipitate
appears as it hardly affects the COD determination.
Table 3.6:
Reagent quantities for different sample sizes.
Sample size
(mL)
Standard
K2 Cr2 0 7
(mL)
H2SO. with
A9 2SO.
(mL)
10
20
30
40
5.0
10.0
15.0
20.0
15
30
45
60
50
25.0
75
Cone. of
FAS(moI/L)
Final vol.
Before titration
(mL)
0.2
0.4
0.6
0.8
0.05
0.10
0.15
0.20
70
140
210
280
1.0
0.25
350
HgSO.
(g)
64
0.2SN
!>t~rd
FA!.
6
:~
0.4g HgS04
+201llL !>amp"
+10 filL 0.25M 1(2(r20,
Afttr 2 h R.flux
BLUE GREEN -
~
HfAT
Wine
Rid
For samples having low COD of less than 50 mgIL the potassium dichromate is
taken with a diluted normality of 0.025 N and the contents are titrated with the
corresponding 0.025 N FAS. On the other hand, for samples containing more than
50 mgIL COD, the higher strength of 0.25 N of these reagents is used.
Steps in COD determination
1. Take a 500 mL capacity flat-bottom conical flask and add 0.4 g ofHgSO4'
2. Add 20 mL of sample or an aliquot of sample diluted to 20 mL with distilled
water and mix well.
3. Add few glass beads followed by 10 mL of 0.025 Nor 0.25 N potassium dichromate depending upon the expected COD.
4. Add slowly 30 mL conc. H2S04 + A~SO 4 reagent and mix thoroughly. The slow
addition along with swirling prevents fatty acids to escape out due to high
temperature.
5. Mix well. If the colour turns green, take either fresh sample with lesser aliquot or
add more dichromate and H2 S04 , Connect the flask to condenser. Mix the contents before heating. Improper mixing will result in bumping and sample may be
blown out.
6. Reflux for a minimum of 2 h. Cool and then wash down the condenser with
distilled water.
65
7. Dilute for a minimum of 150 mL (about 300 mL), cool to room temperature and
titrate excess K 2Crp7 remaining after retluxing with corresponding standard
ferrous ammonium sulfate using ferroin as an indicator (8-10 drops). Sharp
colour change from blue green to wine red indicates end point or completion of
the titration.
8. Perform blank in the same manner using distilled water instead or sample.
CALCULATION
(A-B) x N x 8 x 1000
COD (mgIL) = - - - - - - mL of the sample
Where,
A = mL offerrous ammonium sulfate used for blank
B = mL offerrous ammonium sulfate used for sample
N = Normality offerrous ammonium sulfate
8 = Milliequivalent weight of oxygen.
Stock solution of500 mg/L COD: Dissolve 0.4251 g of potassium acid pthalate in
distilled water and dilute to 1L for a 500 mgIL COD solution. A recovery of 98-100%
of the theoretical oxygen demand can be expected.
Recovered COD,
mg/L
12 .3
40 . 2
92.0
270
12 .34
37.9
88.6
257
Bias,mg/L
+0.04
- 2 .3
- 3.4
- 13
Statistically significance
no
yes
yes
yes
66
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
BOD test
COD test
Drinking water
IS: 10500.(1991) = 0.2 mgIL (residual free chlorine, minimum). To be applicable
only when water is chlorinated. Tested at consumer end. When protection
against viral infection is required, it should be minimum 0.5 mgIL.
Effiuent discharge
MOEF (1993) = 1.0 mgIL (total residual C1 2, maximum), for discharge in inland
surface water and marine coastal areas.
67
Introduction
Chlorine is primarily added to the water (i.e., chlorination) for destroying the
harmful microorganisms in water and wastewater. It is become common practice to
refer to chlorine, hypochlorous acid, and hypochlorite ions are "free chlorine residuals", and the chloramines are called "combined chlorine residuals". An uncontrolled excess of chlorine in water, whether free, available or combined, can adversely affect the subsequent use of water. The smallest amount of residual chlorine considered significant is 0.1 mg/L.
Disinfectant residuals are used universally in disinfection practice to control
addition of disinfectant as to ensure effective disinfection without waste of chemicals.
12 (liberated) + 2 KCl
(colourless solutIOn)
INTERFERENCE
Oxidized form of manganese and other oxidizing agents interfere.
Reducing agents such as organic sulphides also interfere.
Use only acetic acid for acid titration; H 2S04 will only increase interference.
Never use hydrochloric acid.
68
Orthotolidin test: The Orthotolidin test is quick and easy method. In 100rnL sample,
ImL of orthotolidin is added. If yellow colour develops, it shows the presence of
chlorine. By comparing the intensity of colour with the standard sample, the residual
chlorine can be find out.
Reagents
1. Standard sodium thiosulphate, O.lN: Dissolved 25g Na2Sp3.5 ~O in 1000 rnL
of freshly boiled and cooled distilled water and standardise against potassium
bi-iodate or potassium dichromate after at least 2 weeks storage. This initial
storage is necessary to allow oxidation of lUly bisulphite ion present. Add
about 5 rnL of chloroform as a preservative to minimize bacterial decomposition.
4. Starch indicator (lOg/L): Prepare slurry by adding small quantity of water to
1.0 g starch powder. Add 100 mL boiling water to it and continue boiling for a
few minutes until solution becomes clear. The solution is cooled and preserved
by adding 1.25 g of salicylic acid or a few drops of toluene or chloroform.
Standard sodium thiosulphate, O.OlN: Improve the stability ofO.01N Na2 Sp3
solution by diluting aged O.1N solution. ImL ofO.OlN Na2Sp3 = 354.5 Ilg CI as
CljrnL.
(f)
0.01 N
5tMd.rd N.2 51 0,
[J
100 mL Sallpl'
+Pnch of KI
1'5 ilL aClllc ecid
+StaI'th indicator
SlUE COLDR -
Color Loss
lEnd Point)
(A - B) x N x 35.45 x 1000
---------mLofsample
Example-1
For treating 3,000 m3 of water, 12.25 kg of CI-gas used. The chlorine demand
of the water is measured to be 2.6 mg/L. What is the residual CI concentration in the treated water?
Solution:
Total Dosage = 12.25 kg/3,OOO m 3 =4.084 mg/L
Residual chlorine = 4.084 mg/L - 2.6 mg/L = 1.484 mg/L (Ans.).
70
Introduction
Chlorine demand is defined as the difference between the amount of chlorine
applied and the amount of free chlorine remaining at the end of the contact period.
The chlorine demand is different for different waters. For effective disinfection,
doses of chlorine, optimum contact time and residual chlorine are required to be
determined. The smallest amount of residual chlorine considered significant is 0.1
mg/L. Temperature, pH and initial chlorine dosages are important variables in
estimating optimum chlorination practice.
Chlorine combines with water to form hypochlorous acid (HOC}) and hydrochloric acid (HCI). Hypochlorous acid (HOC I) dissociates to give the OC}- and W
ions.
CI 2 +HpB HOCI+W+CIHOCI B W + OCI-
71
INTERFERENCE
The major interfering substances are oxidisable organic and inorganic matter. In
addition, chlorine and hypochlorous (HOCI) acid also react with wide variety of
substances, including ammonia and naturally occurring humic substances. The
reactions of chlorine with ammonia are as follows:
NH3 + HOCI -+- NH2CI + Hp (monochloramine)
NH2CI + HOCI-+- NHCI 2 + Hp (dichloramine)
NHCI 2 + HOCI -+- NCI3 + ~O (trichloromine)
Reagents
I. Standard sodium thiosulphate titrant (O.IN): Prepared as given in determination
of residual chlorine,
2. Starch indicator: Same as in residual chlorine
72
Where,
A = Volume of standard Na2Sp3' mL
B = Normality of standard Na2Sp3
V = Chlorine solution titrated, mL.
PROCEDURE
1. Take 1000 mL of sample in 10 stoppered bottles.
2 Add standard chlorine solution in ascending order. If chlorine demand of treated
water to be examined, use dose of 0.0 to'300 f,1g CljL. In case ofpoUuted water
sample and treated effluents, use dose of 0.1 mg to 3 mg C~ IL.
3. Allow a contact period of30 minutes for drinking water and suitable higher time
for polluted water, or secondary effluents.
4. Estimate residual chlorine as described earlier.
4. Plot the residual chlorine versus chlorine added. In case of organically polluted
samples, a distinct breakpoint can be obtained. But in case of treated water
sample, it is possible that only a straight line is obtained in absence of any
ammonium compound. A typical break point curve is shown in Fig. 3.13.
5. A residual of 0.2 mg Cl/L after the breakpoint is recommended.
CALCULATION
Calculate chlorine dosage in mg/L for each treated sample solution as follows:
Chlorine dosage, mg/L = 2AB
Where,
A = chlorinating solution added to each sample aliquot, mL
B = available chlorine/mL of chlorinating solution, mg
73
OS
1S
11
~ 10
:=
.....
.,e
C>I
.E
~
7
6
~
... 5
;g 10
:J
'0
'ta:
3
2
12310561811
(hlorlnt dosagt mg/littr
Example-1
What is the daily amount of chlorine needed to treat .30,280 m3 (8MGD) of
water to satisfy 2.8 mg/L residual chlorine demand and provide 0 .5 mg/L
residual chlorine.
74
3.11 Cyanide
Drinking water
IS: 10500 (1983, 1991) = 0.05 mgIL (no relaxation)
ICMR (1963) = 0.05 mgIL
MWH (1975) = 0.05 mgIL
WHO (1984) = 0.1 mgIL
WHO (1993) = 0.07 mgIL
EC (1980) = 0.05 mgIL
Effluent discharge
MOEF (1993) = 0.2 mgIL (Inland surface water, land for irrigation and marine
costal areas); 2 mgIL (Public sewers)
APPARATUS
Distillation Apparatus (Fig. 3.14)
Boiling flask, IL with inlet and provision for water-cooled condenser. The inlet
tube shall be a funnel with 8-mm diameter stem that extended to within 6mm of the
bottom of the flask. The condenser shall be connected to a vacuum-type absorber
which shall be in turn connected to a vacuum line which has provision for fme
control. The flask shall be heated with an electric heater.
75
Chemicals
1. Sodium hydroxide, NaOH
2. Sulphuric acid, H2S04
3. Sulfamic acid, NH2S03H
4. Magnesium chloride, MgCl2
5. Lead carbonate, PbC03
1. Sodium hydroxide solution: Dissolve 40g ofNaOH in water and dilute to IL.
2. Magnesium chloride reagent: Dissolve 510g MgClr 6Hp in water and dilute to IL.
3. Sulfuric acid (1+1): Slowly add Ivol of H2S04 (sp.gr 1.84) to Ivol of water,
stirring and cooling the solution during the addition.
4. Lead carbonate (PbCO): Powder
5. Sulfamic acid (NHzSOJH)
Allihn
wat~r-cool~
Water in
Need!e
valve
38-mmx2~m
tnt tube
PROCEDURE
76
PROCEDURE
1. Place 100 mL of the absorption solution or an accurately measured aliquot
dilute to 100 mL with NaOH solution (1 .6g/L) in a beaker of conical flask. For
sample with low cyanide concentration 5 mg/L) do not dilute_
2. Add 0.5 mL indicator solution.
3. Titrate with standard AgN0 3 solution to the first change from yellow to salmon
pink.
4. Titrate a blank of 100 mL NaOH solution.
5. Record the results of titration and calculate the cyanide concentration in the
original sample.
CALCULATION
(A-B) x 1000
Cyanide, rnglL =
250
x -------
mL of original sample
A = AgN0 3 solution to titrate sample, mL
B = AgN03 solution to titrate blank, mL
mL portion used
77
REAGENTS
1. Chloramine-T solution: Dissolve Ig of white-coloured, water soluble grade
Chloramine-T powder in 100 mL of water. Prepare fresh weekly and store in
refrigerator.
2. Cyanide stock solution (ImL = 1 mg CN-): Dissolve approximately 1.6g
NaOH and 2.51g KCN in IL distilled water (Caution: KCN is highly toxic,
avoid contact or inhalation). Mix thoroughly. Standardise against standard
AgNO) titrant, using 25 mL KCN solution. Check the titer weekly because the
solution gradually loses str-ength.
3. Cyanide standard solution (lmL = 10 Ilg CN-): Based on the concentration
for the KCN stock solution, calculate the volume required (approx. 10 mL) to
prepare IL ofa 10 I1g cyanide/mL. Dilute with NaOH solution.
4. Cyanide solution (ImL = 1 IlgCN): Dilute 10 mL of 10 I1g CN-/mL solution
to 100 mL with NaOH (1.6g1L) solution. Prepare fresh daily and keep in a
glass-stoppered bottle.
5. Pyridine-barbituric acid reagent: Place 15g of barbituric acid in a 250 mL
volumetric flask and just add water to wash the side of the flask and wet the
barbituric acid. Add 75 mL of pyridine and mix. Add 15 mL of conc. HCr and
cool to room temperature. Dilute to the mark with water and mix until all of the
barbituric acid is dissolved. This solution is stable for approximately 6 months
if stored in an amber bottle under refrigeration. Discard if precipitate develops.
6. Acetate buffer: Dissolve 41 Og of sodium acetate trihydrate (NaC2~02.3HP)
in 500 mL of water. Add glacial acetic acid to yield a solution of pH 4.5,
approximately 500 mL.
PROCEDURE
Preparation of Standard Curve
Pipette a series of standards containing I to 10 Ilg of cyanide into 50-mL volumetric flasks (0.02 to 0.2 Ilg cyanide/mL). Dilute to 40 mL with NaOH solution (1.6
gIL). Use 40 mL NaOH dilution solution as blank.
Colour development: Add I mL acetate buffer and 2 mL chloramine-T solution,
stopper, and mix by inversion twice. Let stand exactly for 2 min. Add 5 rnL pyridine-barbituric acid reagent, dilute to the mark with distilled water, mix thoroughly
78
and let stand exactly for 8 min. Measure absorbance against distilled water at 578
run. Measure absorbance of blank (0.0 mg CNIL) using 40 mL NaOH dilution
solution and follow the same procedure of colour development.
Estimation of Cyanide In Water Sample
Pipet a portion of absorption solution into- a 50-mL volumetric flask (such that
concentration falls within the standardisation range) and dilute to 40 mL with NaOH
solution. Follow the colour development procedure.,
CALCULATION
Slope and intercept ofstandard curve: Calculate the slope on the standard curve,
m, and the intercept b, using following equations.
n:I:ca - :I:c:I:a
m=-------
n:I:a2- (:I:a)2
:I:a2:I:c - :I:c:I:ac
b=------
Where,
a = absorbance of standard solution;
c = concentration of cyanide in standard, mgIL
n = number of standard solutions.
m = slope of standard curve, and
b = intercept on "e" axis
The blank concentration, 0.0 mg/L, and the absorbance of the blank must be
included in the calculation of slope and intercept.
Calculate the concentration of cyanide as follows:
50
250
x -- x - -
Where,
a = Absorbance of sample solution
X = Aliquot of absorbance solution, mL, and
Y = Original sample, mL
79
stirring bar in the solution, stir at a predetermined constant rate, and maintain
constant temperature.
3. Insert the cyanide specific electrode and the reference electrode in the solution
and measure potential or the cyanide concentration following the manufacturer's
instructions.
4 . Pipet 10 and SO-mL aliquots of standard solution into 2S0-mL beakers and
dilute to 100 mL with NaOH solution (l .6g1L). Follow step-2 and step-3 starting with lowest concentration.
S. Plot concentration values of the standardising solutions on the logarithmic axis
of semi-logarithmic graph paper versus the potentials developed in the
standardising solutions on the linear axis.
PROCEDURE
1. Place 100 mL of the absorption solution (or an accurately measured aliquot
diluted to 100 mL with NaOH solution (1.6g1L) in a 2S0 mL beaker.
2. Follow step 2 and step3.
3. Use values found from the graph or direct reading ion meter to calculate the
concentration in the original sample.
CALCULATION
Calculate the concentration of cyanide in mgIL as follows:
Cyanide,mgIL = Cyanide mglL from graph or meter x (I OO/aliquot) x
(2S0/mL of original sample)
Effluent discharge
MOEF (1993) = No Standards
80
Introduction
1. One of the most important water quality parameters is the amount of dissolved
oxygen (DO) present. The DO levels in. natural and wastewater depend on the
physical, chemical, and biological activities in the water body.
2. Oxygen is considered as poorly soluble in water. Its solubility is related to
pressure and temperature.
3. In fresh water, DO reaches 14.6 mgIL at 0"C and approximately 9.1,8.3 and 7
mgIL at 20"C, 25"C and 35C, and 1 atmospheric pressure respectively. At temperature of20-30C, the level of saturated DO is 9 to 7 mgIL.
4. Minimum amount required for health fish population may be as high as 5 to 8
mgIL.
5. The oxygen depleting substances reduces the available DO. During summer
months, the rate of biological oxidation is highly increased, while, unfortunately, the DO concentration is at its minimum due to higher temperature.
Environmental Significance
1. To evaluate the pollution strength of domestic and industrial effluents.
2. For corrosion control: Oxygen is a significant factor in corrosion of iron and
steel pipes in water distribution system and steam boilers. Removal of DO from
boiler-feed water by physical and chemicals means is a common practice in
power industry. So DO test serves as the means of control.
3. DO measures are vital for maintaining aerobic conditions in natural waters that
receive polluted matter.
4. For controlling the rate of aeration during biological treatment of effluents.
5. Rate of biochemical oxidation can be measured by determining residual dissolved oxygen in a system at various time interval.
6. For maintenance of desire aquatic life. As DO drops, fish and other aquatic life
are threatened; in the extreme case, killed.
7. As Do levels falls; undesirable odours, tastes and colours reduce the acceptability of water.
81
or
INTERFERENCE
Nitrite (NO~) interference is overcome by the use of sodium azide (NaN). When
HzSO. is added, the following reaction occurs and the NO~ is destroyed.
NaN) + H+ -+ HN) + Na+
HN) + NOz + H+ -+ N z + Np + Hp
APPLICABILITY
Use the azide modification for most wastewater, effluent, and stream samples, especially if samples contain more than 50 J.lg NOz-NIL and not more than lmg
ferrous ironiL.
Other reducing or oxidizing materials should be absent. If ImL KF solution is
added before the sample is acidified and there is no delay in titration, the method is
applicable in the presence of 100 to 200 mg ferric ironIL.
GLASSWARE
1.
2.
3.
4.
5.
82
Reagents
1. Manganous sulfate solution (364g/L): Dissolve 364 g of monohydrate manganous sulfate (MnS04.HP) in distilled water (filter it, if necessary) and dilute
to lL. This solution should not give colour with starch solution when added to
an acidified solution of potassium iodide (KI).
2. Alkaline-iodide-azide solution: Dissolve 500 g ofNaOH and 150 g ofKI (or
135 g NaI) and dilute to 950 mL. Add 109 of sodium azide (NaN 3) dissolved in
40mL of distilled water. Cool the solution and make up the volume to 1000 mL.
This solution should not give colour with starch solution when diluted and acidified. Store the solution in a dark, rubber stoppered bottle.
3. Concentrated H1SO.: ImL neutralise about 3 mL of alkaline-iodide reagent.
4. Starch indicator: To prepare an aqueous solution take 5g of arrowroot or
soluble starch to approximately 800 mL of boiling water, with stirring. Dilute
to 1L, boil a few minutes, and leave overnight. Use clear supernatant. Preserve
by adding a few drops oftoluene or formalin. Store in a glass-stoppered bottle.
5. Standard sodium thiosulfate solution (O.025N): Dissolve directly 6.025 g
Na2Sp3.5Hp in a previously boiled 1L cooled distilled water, which will give
0.025N. Add 1.5 mL 6NNaOH or 0.4 g solid NaOH per lL. Store in brown
bottle.
This solution will have to be standardised against standard potassium dichromate ~Crp7) solution for each set of titration.
6. Standard potassium dichromate, O.25N: Dry ~Crp7 at 103C for 2 h and
take accurately 12.259 g, and dilute to lL.
7. Potassium fluoride solution (400g/L): Dissolve 40 g ofKF.2HP in distilled
water and dilute to 100 mL. This solution is used in the procedure for determining ferric ion interferences. Store this solution in a plastic bottle.
PROCEDURE (Fig. 3.15)
1. Collect the sample in a BOD bottle (300 mL capacity) taking care to avoid any
bubbling. Fill the sample to the neck of the bottle. Be sure that air bubbles have
not been trapped under the stopper and maintain a water seal around the stopper until ready for the next step of analysis.
2. Add I mL of manganous sulphate followed by I mL of alkali-iodide-azide
83
300mL S . .plt
to. SOlllinuh
Iter
~~nat lilt
Allow to prtcip. t.
Add
starth
indiCator
20111!l
84
solution. The tip of the pipette should be below the liquid level while adding
these reagents.
3. Place the stopper carefully to exclude air bubbles and mix by inverting the
bottle repeatedly for at least 15 minutes. An equivalent amount of2 mL of the
contents will come out of the bottle after placing the stopper. Allow the precipitate to settle leaving about 150 mL of clear supernatant.
4. Carefully remove the stopper and immediately add 1 mL of conc. H2S04, close
the bottle and mix with gentle inversion until the precipitate completely
dissolves.
5. Titrate 100 mL or 250 mL contents of the bottle with sodium thiosulphate solution using starch as an indicator. At the end point the blue colour turns to
colourless. The starch indicator is usually added towards the end of the titration
when a straw pale colour is obtained.
Standardisation of sodium thiosulfate
It can be done either by standard bi-iodate solution or standard potassium dichromate solution.
1.
2.
4.
5.
CALCULATION
When only a p~rt of the sample of the content is titrated, i.e. 100 or 250 mL
85
REAGENTS
All the reagents required are as discussed in DO estimation. In addition, the
following two are also required:
PROCEDURE
1. Collect sample in a glass-stoppered bottle of 500 to 1000 mL capacity, using
the same precaution as for regular DO samples.
2. Add 10 mL alum solution and 1 to 2 mL conc. NHpH.
3. Stopper the bottle and invert gently for about 1 min. Let sample settle for about
10 min and siphon clear supernatant into 300-mL DO bottle.
3.13 Fluoride
(SPANDS Method)
Drinking water
IS: 10500: (1991); ICMR, (1963) = 1.0 mglL (maximum permissible). Fluoride
may be kept as low as possible. High fluorides may cause fluorosis. 1.5 mgIL
(maximum permissible limit in the absence of alternate source).
WHO (1984) = The guideline value of 1.5 mgIL in drinking water has been
proposed as excessive amounts of fluorides (more than 1.5 mgIL) causes
disfigurement of teeth known as mottled enamel (dental fluorosis).
Effluent discharge
MOEF (1993) = Guideline value of2 mgIL set for discharge into inland surface
water and 15 mgIL for discharge into public sewers and marine coastal areas.
86
Introduction
Fluorides is more common in ground water than in surface water. The main sources
of fluoride in ground water are different fluoride bearing rocks. In rare instances
the fluoride concentration of naturally occurring water may approach 10 mgIL.
Such water should be defluoridated.
A fluoride concentration of approximately 1 mgIL in drinking water effectively
reduces dental caries or tooth decay without any harmful effect on health.
The actual concentration of fluoride in drinking water depends on the air temperature, because ambient air temperature influences the amount of water that people
drink. The USEPA (1975), published the fluoride primary standards related with
ambient temperature (Table 3.8).
Table 3.8:
USEPA primary drinking water Mel for fluoride (1975) .
Annual average of maximum daily
air temperature of community in
which water system is situated (OC)
12.0 or
12. 1 to
14.7 to
17.7 to
21.5 to
26.3 to
below
14.6
17.6
21.4
26.2
32.5
2.4
2.2
2.0
1.8
1.6
1.4
Average F
conc., mg/L
State (name of
district)
Tamilnadu
Salem
4.5
Madurai
4.5
Coimbatore
4.5
Gujrat
Surat
4.0
Mehsana
4.0
Karnataka
Belgaum
0.8-1.2
Rajasthan (Banswara)
1.6-2.0
Anandpuri
Garhi
1.6-3.3
Ghatola
3.4-4.7
Talwara
1.8-3.1
Choubisa (1997):IJEH,39(4):281-28B.
Andhra Pradesh
Nalangonda
Veliore
Anantpur
Punjab
Bhatinda
Ferozpur
Haryana
Kamal
Average F
conc., mg/l
0.8-1.2
0.~-1.2
.0. 8- 1.2
3.0
3.0
3.6
87
Estimation of Fluoride
APHA (1995) recommends the following three methods for fluoride estimation:
a. Ion-selective electrode method: 0.1 to more than 10 mg FIL.
b. SPANDS method (colorimetric): 0 to 1.4 mg FIL.
c. Automated complexone method (colorimetric): 0 to 2.0 mg FIL.
PRINCIPLE (SPANOS method)
Under acidic condition fluorides (HF) react with zirconium-SPANDS solution and
the colour (colour of SPANDS reagent) gets bleached due to formation of a
colourless complex anion ZrF;-. Since bleaching is a function of fluoride ions, it is
directly proportional to the concentration offluoride (i.e. as the amount of fluoride
increases, the colour produced becomes progressively lighter).
INTERFERENCES
Chlorine, colour and turbidity interfere and must be removed by distillation.
Interference caused by alkalinity, chloride, iron, phosphate and sulphate is not linear
and hence cannot be accounted mathematically.
Samples and standards should be at the same temperature or at most have a 2C
temperature difference. Temperature is maintained nearly constant throughout colour
development. Different calibration curves are prepared for different temperature
ranges.
APPARATUS
1. Spectrophotometer for use at 570 nm, providing a light path of at least 1 cm or
longer.
2. Nessler's tubes with capacity of 100 mL.
CHEMICALS AND REAGENTS
Chemicals
I. Sodium fluoride
2.
3.
4.
5.
Sodium arsenite
ConcHCI
SPANOS
Zirconyl chloride octahydrate
Reagents
88
4.
S.
6.
7.
PROCEDURE
1. Preparation ofstandard curve: Prepare fluoride standards in the range of 0 to
lAO mg/L by diluting appropriate quantities of standard fluoride solution to 50
mL with distilled water (Table 3.10).
2. In each 50 mL of Nessler tube, add 10 mL mix solution (or 5 mL of SPANOS
and 5 mL zirconyl acid reagent). Mix well. Take absorbance immediately of the
bleached colour at 570 nm using reference solution for setting zero absorbance.
3. Plot standard graph between concentration and absorbance (optical density).
Table 3.10:
Preparation of Fluoride standard solutions (Fluoride stock solution
1mL = 10 I'g F (Final volume: 50 mL)
F stock solution, mL
Fin I'g
1
2.
4
6
8
10
20
40
60
80
0.2
0.4
0.8
1.2
1.6
89
CALCULATION
Fluoride, mgIL =
mg F determined photometrically
x 1000 x - -
mL sample used
The ratio B/C applies only when a sample is diluted to a volume B, and a
portion C taken from it for colour development.
REAGENTS
I. Sulphuric acid, ~SO4 ' concentrated.
2. Silver sulphate, A~S04' crystals.
APPARATUS
Distillation assembly: Glassware consisting of a IL round bottom, borosilicate
boiling flask, an adaptor with a thermometer opening, a connecting tube, a condenser
and a thermometer reading up to 200 "C, assemble as shown in Fig. 3.16.
90
PROCEDURE
1. Preparation of acid mixture:
a. Place 400 mL distilled water in the boiling flask and carefully add 200 mL
conc H2S04
b. Swirl until the flask contents are homogeneous.
c. Add 25-35 glass beads. Make sure that all joints are tight.
d. Start heating slowly at first, then as rapidlyre efficiency of the condenser
permits (the distillate must be cool) until the temperature of the flask contents
reach exactly 180C.
e. Discard the distillate. The process removes fluoride contamination and adjusts
the acid: water ratio for subsequent distillation.
2. After cooling the acid mixture remaining after step 1 (to 120C or below), add
300 mL of sample, mix thoroughly, and distil as before until the temperature
reaches 180C. To prevent sulphate carry-over, do not heat above 180C. When
high-chloride samples are distilled, add Ag2S04 to the boiling flask at the rate
of 5 mg/mg of chloride.
3. Use H2 S04 solution in the flask repeatedly until the contaminants from samples
accumulate to such as extent that recovery is affected or interferences appear in
the distillate. Check suitability of the acid periodically by distilling standard
fluoride samples, flush the distillation apparatus with 300 mL distilled water.
4. Use the distillate to conduct fluoride analysis. Correct the volumereiationship
if the distillate volume differs from that of the original sample.
91
Introduction
Hardness is defined as the concentration of multivalent metallic cations in solution.
Hard water are generally considered to be those waters that requires considerable
amounts of soap to produce a foam and that also produce scale in hot-water pipes,
heaters, boilers, and other units in which the temperature of the water is increased.
The hardness of waters varies from place to place. In general, surface waters are
softer than groundwater's. The hardness of water reflects the nature of geological
fonnation with which it has been in contact. Water are commonly classified in
tenns of degree of hardness as in the Table 3.11.
Table 3.11 :
Classification of hard water
Hardness range (mg/L as CaC0 3 )
0-75
75-150
150-300
above 300
Degree of hardness
Soft
moderately hard
Hard
Very hard
Causes and sources of hardness: Hardness is caused by the presence of multi valent metallic cations in water. The principle hardness causing cations are divalent
Ca2+, Mg2+, Sr2+ Fe2+ and Mn2+ and anions are HCOl ' S04' CI, NO l and SiOl .
Types of hardness: Carbonate hardness was fonnerly called temporary hardness,
because it can be removed by boiling and is caused by dissolved Ca and Mg bicarbonate. Removal process of Ca and Mg bicarbonate is as follows:
92
Carbonate hardness is especially important since it leads to scaling. Noncarbonate hardness was formerly called permanent hardness, because it cannot be
removed by boiling. Non-carbonate hardness cations are associated with sulphate,
chloride and nitrate ions of Ca and Mg.
Environmental Significance
Estimation of Hardness
PRINCIPLE
In alkaline condition ethylene-diamine-tetra-acetic (EDTA) acid or its sodium salt
(Na2EDTA) reacts with Ca and Mg to form a soluble chelated complex. Ca and Mg
ions develop wine red colour when small amount of dye such as Eriochrome Black
T is added under alkaline condition. When EDTA is added as titrant, the Ca and
Mg will be complexed with EDTA resulting in sharp change from wine red to blue,
which indicates end-point of titration. Hardness is normally expressed as CaCOJ
M2+ + Eriochrome Black T -+ (M. Eriochrome Black.T)complcx
Wine red
M2+ + EDTA -+ [M.EDTA] compcx
I.
Blue
The sharpness of the end point increases with increasing pH. However, pH can
not be increased beyond 10 because of the danger of precipitation ofCaCO J or Mg
(OH)2. The pH for this titration has been recommended at 10.0 0.1. A limit of 5
minutes is set for the duration of the titration to minimise the tendency of CaCO)
precipitation.
CHEMICALS AND REAGENTS
Chemicals
1. Ammonium chloride, NH4Cl
3. EDTA Mg salt
93
6. HCl (1+1)
8. Hydroxylamine hydrochloride
10. Methyl red indicator
Reagents
1. Buffer solution (pH 10): Dissolve 16.90 gNH4Cl in 143 rnL ofconc NHpH.
Add 1.25 g EDTA Mg salt to obtain sharp change in indicator and dilute to 250
rnL. Alternatively, instead of EDTA Mg salt, 1.179g Na2EDTA plus 0.780g
MgS04 .7Hp can also be used. This has to be titrated with standard calcium
solution to avoid interference produced by EDTA to the buffer. Store in a
plastic or borosilicate glass container for no longer than I month. Stopper tightly
to prevent loss of ammonia (NH3) or pick up of carbon dioxide (C0 2) from air.
2. Eriochrome black T indicator: Mix 0.5 g dye with 100 g NaCI to prepare dry
powder. or dissolve 0.5 g of indicator in 100 rnL of ethyl alcohol.
3. Standard calcium solution (O.OlN) (1 gIL): Weigh accurately 1.0 gAR grade
CaC03 (dried at 180 OC for Ih before weighing) and transfer to 250 mL conical
flask. Place a funnel in the neck of a flask and add 1+1 HCl till CaC0 3dissolves
completely. Add 200 mL distilled water and boil for 20-30 min to remove COl"
Cool and add methyl red indicator. Add NHpH (3N) till intermediate orange
colour develops. Dilute to 1000 rnL to obtain I rnL = I mg CaC03, (IrnL =
0.40 I mg Ca or 0.243 mg Mg).
4. Standard EDTA solution (O.OIM): Dissolve 3.723 g EDTA-disodium salt
and dilute to IL. Standarised against standard calcium solution. I rnL = I mg
CaC03 The reagent is stable for several weeks.
s. Inhibitor: Dissolve 1.5 g hydroxylamine hydrochloride in 100rnL 95% ethyl
alcohol or isopropyl alcohol.
PROCEDURE (Fig. 3.17)
I. Select a sample volume that requires less than 15 mL EDTA titrant and complete
titration within 5 minutes, measured from time of buffer addition.
2. Take 50 mL well mixed sample in a conical flask.
3. Add 1-2mL buffer solution followed by I rnL inhibitor. Usually 1-2 mL buffer
solution is sufficient to give a pH of 10
4. Add a pinch of Eriochrome black T and titrate with standard EDTA (O.OIM)
till wine red colour changes to blue. Note down the volume of EDTA consumed
(A).
5. Run a reagent blank with distilled water. Note the volume of EDTA consumed
(B).
6. Calculate the volume of EDTA required by sample; C = (A - B).
Low hardness sample: For softened water or natural water oflow hardness (less
than 5 mgIL), take a large sample, say 100 mL to 1000 rnL for titration. Add
proportionately larger amount of buffer, inhibitor and indicator.
Observation-I: Standardisation of EDTA solution.
Observation-II: Estimation of Total Hardness against standard EDTA
solution.
94
0.01 N
Sbndrd
EOTA
!J
50 -L
.'-1
_L
.'.l
$MIpie
lilt.... lOIutioll
illllillitor
+Pinch of 8T "indicator
"
CALCULATION
CxDxlOOO
Total hardness (as CaC01 mgIL) = - - - - mL of sample
Where,
C = mL of EDTA required by sample for titnltion
D = mg CaC01 equivalent to ImL EDTA titrant (lmg for O.OIM EDTA used
here).
.
95
... cont.
absence of alternate source)
EC (1980) = 100 mgIL (guide level).
Effluent discharge
MOEF (1993) = No guideline value set.
Introduction
Calcium and magnesium are common constituents of natural water and important
contributor to the hardness of water. The source of Ca and Mg is the rocks from
which it is leached. Being a important contributors of hardness, it reduces the utility
of water for domestic use. Some CaC03 is desirable for domestic water because it
provides a coating in the pipes which protects them against corrosion.
REAGENTS
1. Sodium hydroxide solution, 8% (80 gIL): Dissolve 8 g ofNaOH in distilled
water to prepare 100 mL of solution.
2. Murexide (ammonium purpurate) indicator: The indicator changes from
pink to purple at the end point. Mix 0.2 g of ammonium purpurate and 100 g of
sodium chloride and grind thoroughly to fine powder (i.e. one that passes through
a 40-mesh sieve).
3. EDTA solution (0.01 M): Dissolve 3.723 g ofN a2EDTA in 1L of distilled water.
1 mL = 0.401 mg Ca; ImL = 0.243 mg Mg.
PROCEDURE
1. Fill the burette with O.OIM EDTA standard solution. (Fig. 3.18)
2. Take 50 mL or a smaller portion diluted to 50 mL so that the Ca content is about
5 to 10 mg. in an Erlenmeyer flask.
3. Add 1 mL ofNaOH to raise pH to 12.0 and a pinch of murexide indicator (0.1
to 0.2 mg) or 1-2 drops of indicator solution. (Note: Titration should be
completed within 5 min of addition ofNaOH solution.)
4. Titrate immediately against EDTA solution till pink colour changes to purple.
Note the volume of EDTA used (A). Confirm the end points by adding 1 to 2
drops oftitrants in excess to make certain that no further colour changes occurs.
96
(Note: If 15mL or more titrant is consumed, take a s'maller sample aliquot and
repeat the test).
5. Run a reagent blank with distilled water. Note the mL of EDTA required and
keep it aside to compare endpoints of the sample titration.
0.01 N
St~d.,.d
EOTA
[j
SO .. l 5. ....
p'.
+l .. l 11I0Il
-+ Pinch at l1uruid. indicator
CALCULATION
T
Calcium (mglL)
400.5
1.05
T x 1000 x 1.05
V
Where,
T = volume of titrant, mL
V = volume of sample. mL.
Cont.
97
...cont.
WHO (1984, 1993) = No specific guideline set for Mg.
IS: 10500 (1983, 1991) = 30 mgIL (maximum, beyond this limit, encrustation
to water supply structure and adverse effects on domestic use) - 100 mgIL
(permissible limit in the absence of alternate source).
EC (1980) = 30 mg/L (guide level) and 50 mg/L (MAC).
Taste threshold for magnesium as sulphate has been reported as 100 mgIL and
500 mgIL for the average individuals.
Effluents discharge
MOEF (1993) = No guideline value set for Mg.
Introduction
Magnesium has been considered as non-toxic to humans at the concentration
expected in water. Magnesium salts have a laxative and diuretic effect particularly
for individuals not accustoms to high dosage.
Example
~1
50 mg/L MgC03
b.
c.
50 mg/L CaSO.
d.
55 mg/L CaCl z
,,,';;'0'
.""'>h;:;::;;
',,,e,'.,.
Solution:
y;1':~j,
,.
The concentratiopl>"gittle above. express~(rin ,mo a
c,
,d. ,.
98
Example-2
A sample of groundwater has 100 mg/L of Ca 2+ and 10 mg/L of Mg2 +. Express
its hardness in units of meq/L and mg/L as CaC0 3
Removal of Hardness
In surface water, hardness values generally are 150-200 mg/L and softening is not
usually needed. However, in case of groundwaters, where the hardness level is
more than 1000 mg/L, softening is needed. The softening can be performed by
either lime soda process or ion-exchange process.
99
Example - 3
Water has the following composition: Calcium = 82mg/L, magnesium = 33mgl
L, sodium = 14 mg/L, bicarbonate = 280 mg/L, sulfate = 36mg/L and chloride
= 82 mg/L Determine carbonate hardness, noncarbonate hardness and total hardness all in terms of mg/L of CaC0 3
.. ,'
, ,
SolUtion:
'
Step~ 1 : Convert anc()l)c~:mtration (rng/L) to meq/L and mglL as CaC0 3. Construct
'.'~ a 'table asfollows: . f ,,>;!'
Jo'n ,species Mol. Wt. ;';Equivalent
'
Ca 2 + .,
, MgH
";" Na"
)'Total cations
HCOJ61
CI, 35.5
96:1 ,"
SO"
Total anions
~t.
Concentration
meq/L . mg/L as CaC0 3
mglL
20.0
12.2
23
82
32
14
200
1,35
, ":'i;' 30
, 365
230
50
85'
, ",.
, 280
36
82
0.
365
200
Ca 2
' HC03-
335
I, SO..
230
Mg2+
365
Na+
Y,:
CI-
"I
280
10. A sample of water has the following concentration of ions with the pH near
neutral.
Cation
Ca 2+
Mg2+
mg/L
95
Na+
15
26
Anion
HC03'
S04CI-
mglL
160
135
73
100
= 131.14 (Aos).
Emuents discharge
MOEF (1993) = No guideline value set
Environmental Significance
I. The determination of methylene blue active substances gives an idea of the
presence of detergents in water.
2. The widespread use and discharge of detergents into surface waters can result
in lowering of its aesthetic quality by foam formation and by causing toxicity to
aquatic wildlife.
.
3. Biodegradable linear alkyl benzene sulfonates (LAS) have replaced the branched
chain alkyl benzene sulfonates (ABS) in detergent formulations, which were
more resistant to biodegradation. Differentiation between linear and branched
chain alkyl benzene sulfonates, as well as differentiation of the various positional isomers of either type, is not possible by this test method. While the
methylene blue method may be employed to monitor studies designed to mf?asure biodegradability, it cannot be used to predict this quality.
Estimation of MBAS
This test method covers determination of compounds that react with methylene
blue under the conditions specified in the test procedure. They are referred to as
methylene blue active substances (MBAS), and are calculated and reported in terms
of the reference material, linear alkyl benzene sulfonate (LAS).
101
Chemicals
1.
2.
3.
4.
5.
6.
7.
Chloroform
Methylene blue
Phenolphthalein Indicator
Sodium dihydrogen phosphate, NaHlO4.Hp
Sodium dioctyl sulfo succinate
H2S04 solution - 14%, v/v
NaOH solution
Reagents
1. Linear Alkyl Benzene Sulfonate Solution Stock (1 mL= 1 mg LAS): Weigh
the amount of reference material (Le., sodium dioctyl sulfo succinate, Ig in 1L)
necessary to provide the equivalent of 1 g of LAS on a 10% active basis.
102
2.
3.
4.
5.
6.
7.
8.
Dissolve in water and dilute to lL, mixing gently to prevent foam formation.
Record the molecular weight of the LAS reference material as supplied. The
stock solution may be stored at 4C in the dark for 12 months in a well-stoppered flask without deterioration.
Linear Alkyl Benzene Sulfonate Standard Solution, (lmL= O.Olmg LAS):
Dilute 10.0 mL of the foam-free stock solution to lL with water that has been
previously adjusted to pH 2 with sulfuric acid and mix. The standard solution
may be stored at 4C in the dark for at least 12 months in a well-stoppered flask
without deterioration.
Methylene Blue Solution (30 mglL): Dissolve O.lg of methylene blue chloride in 100 mL of water. Transfer 30 mL of this solution to a l-L volumetric
flasks and add 500 mL of water. Add carefully 50 mL of 14% sulfuric acid
stock solution and 50 g of sodium dihydrogen phosphate monohydrate
(NaHl04.HP). Shake until solution is complete and then dilute to lL with
water and mix.
Sulfuric Acid Stock Solution (14 % volume per volume): Add carefully 140
mL of cone H 2 S04 (sp gr 1.84) to 700 mL of cold (0 to SoC) water with good
stirring, dilute to I L with water and mix.
Sulfuric Acid Solution, Dilute (0.7% volume! volume): Dilute carefully 50mL
of 14% H 2S04 stock solution to IL with water and mix.
Phenolphthalein Indicator Solution (5g1L): Dissolve 0.5 g ofphenolphthalein in SO mL of 9S% ethyl alcohol and dilute to 100 mL with water and mix.
Sodium Hydroxide Solution (10glL): Dissolve 10 g ofNaOH in water, dilute
to 1L and mix.
Phosphate Wash Solution: Dissolve SO g sodium dibydrogen phosphate monohydrate (NaHl04.HP) in 500 mL of water in a I-L volumetric flask. Add
carefully 50 mL cone H 2 S04 stock solution and dilute to volume with water and
mix. The solution has a pH of approximately 1.8.
PROCEDURE
Preparation of calibration curve
Prepare a series of standards by adding the standard solution (2) from 25-mL
burette to a series of 250-mL separatory funnels and dilute the standards to 100
mL volume with water, yielding solutions as follows:
Standard, mL (lmL=O.OI mg LAS)
0.00 (blank)
0.00
1.00
0.01
3.00
0.03
S.OO
O.OS
7.00
0.07
9.00
0.09
12.00
0.12
103
1. Add 3 drops of phenolphthalein indicator solution (6) and just enough NaOH
(7) to produce a pink colour. Add dilute H2 S04 solution, in small increments
until the pink colour is barely discharged.
2. Add 25 mL of methylene blue solution (3) and mix. Add 25 mL of chloroform
and mix thoroughly for 30s with shaking. Vent carefully, permit the phases to
separate and then drain the chloroform layer into a second 250-mL separatory
funnel. Leave ~y emulsion layer in the first separatory funnel. Repeat the extraction, serially, with two additional 25-mL portions of chloroform.
3. Add 50 mL of phosphate wash solution (8) to the combin-:d chloroform extract
in the second separatory funnel and shake vigorously for 30 seconds. Hold the
separatory funnel in a vertical position and swirl the contents. Allow to settle
for I minute. Filter the chloroform layer through a glass wool plug into a conditioned 100-mL volumetric flask.
4. Add 20 mL of chloroform to the second separatory funnel and repeat the shaking, swirling, and settling step. Combine the chloroform layer through the glass
wool into the volumetric flask. Add additional chloroform as needed to bring
the flask to 100-mL volume and mix thoroughly.
5. Using a 50-mm light-path cell, at 650-nm wavelength, set the photometer to
zero absorbance with the extract of the calibration blank.
6. Measure the absorbance of each of the extracts. Because of a tendency to fade
slowly, the absorbance of the extracted methylene blue complex should be measured within 30 min. Prepare a calibration curve by plotting photometer reading in absorbance against concentration of LAS in mgl100 mL of extract on
rectilinear graph paper, and record the molecular weight of the LAS reference
material, as supplied, on the graph.
CALCULATION
Calculate and express as MBAS, the apparent concentration of linear alkyl benzene sulfonate (ABS) as follows:
MBAS, mgIL = W
1000/S
Where,
S = sample volume selected.
W = LAS in the sample from the calibration graph, mg.
REPORTING OF RESULTS
Include in the report the molecular weight (mw) of the LAS used to prepare the
calibration curve. Report results as:
MBAS (calculated as LAS, mw.......... ) = ..........mgIL.
104
Drinking water
MOEF (1993) = 100 mgIL (maximum) for discharge into inland surface water
and marine coastal areas .
Introduction
. Four forms of nitrogen namely; nitrate nitrogen (N03-N), nitrite nitrogen (N0z-N),
ammonical nitrogen'(NH3-N) and organic nitrogen may be present in waters and
wastewaters. Analytically, organic nitrogen and ammonical nitrogen can be determined together and have been referred to as total nitrogen or more correctly total
Kjeldahl nitrogen (TKN).
The presence of total nitrogen is accepted as a chemical evidence of recent
organic pollution particularly of animal origin. The nitrogen forms cause:
Algal blooms
Eutrophication
Toxicity to fish and other organisms
Health disorders by nitrate and nitrites such as methemoglobinemia.
Total Kjeldahl nitrogen (TKN) is the sum of nitrogen contained in the free
ammonia and other nitrogen compounds which are converted to ammonium sulfate
under the specific digestion conditions (ASTM, 1995). The most reliable results
are obtained with fresh samples. Commonly occurring concentration of TKN in
domestic effluents, according to Metcalf and Eddy (1995) is:
1. For strong sewage: TKN = 85 mgIL (organic, 35 mgIL + NH,-N, SO mgIL)
2. Medium strength: TKN = 40 mgIL (organic, 15 mgIL + NH,-N, 25 mgIL)
3. Low strength: TKN = 20 mgIL (organic, 8 mgIL + NH,-N, 12 mgIL)
Used for measuring organic nitrogen and ammonical nitrogen which are essential nutrients for growth.
105
Estimation of TKN
SAMPLE HANDLING
Conversion of organic nitrogen to ammonia may occur in samples between collection and analysis. This effect can be decreased by addition of2 mL H2SO. (sp. gr.
= 1.84) per litre and storage at 4"C. However, it is prudent to analyse all samples as
soon as possible after collection.
PRINCIPLE
so mL
+so mL
Samplp
di9.~ton
rp~gent
Heat
106
Digestion of sample
Distillation of sample
107
Sample size, mL
500
250
100
50
25
Distillation
1. Take 50 mL of boric acid cum indicator solution (pink colour solution) in 500
mL of flask and place it below the condenser of the distillation assembly so that
the lower open end of the condenser is dipped in solution.
2. Collect at least 200 mL of distillate in flask (Pink solution turns to green due to
absorption of ammonia). It is good, to put a mark at the level of250 mL in the
flask, so that one could easily know when 200 mL distillate will be collected.
Remove the flask with distillate.
3. Titrate the distillate with O.2N H 2S04 to its original colour (i.e.,change of green
to pink colour indicates the end point of titration)
4. Run distilled water blank in the same manner.
CALCULATION
(A - B) x 280
Total-N (TKN) ,mg/L= - - - - mL of Sample
Where,
A= volume of H2S04 required for sample, mL
B= volume of H 2S04 required for blank, mL
108
Drinking water
IS: 10500 (1983, 1991) = No standards
Canadian MACL (1987) = 0.5 mgIL.
Effluents discharge
MOEF (1993) = 50 mgIL (max.) for inland surface water, public sewers and
marine coastal areas; Free ammonia (as NH) ) = 5 mgIL for inland surface
water and marine coastal areas.
Ammonia is present naturally in surface waters and wastewater. Its concentration is generally low in groundwater because it is absorbs by soil particles and
clays and not leached readily from soils. Ammonia concentration encountered in
water varies from less than 10 Ilg ammonia- NIL in some natural surface and groundwater to more than 30 mgIL in some wastewater.
When nitrogenous organic matter is destroyed by microbial activity; ammonia
is produced and is therefore found many surface and groundwater. The properties
of the two forms of ammonia nitrogen i.e. free ammonia and ammonium ions,
depends on the pH as below.
pH
10
11
%NH)
25
78
96
% NH;
100
99
96
75
22
109
If it is not possible to carry out the detennination soon after sampling, the sample
should be refrigerated at 4C. Chemical preservation can be achieved by adding
either 20-40 mg HgCI2 or I mL HzSO4 to I L of sample.
Four different methods for detennining ammonia-N are included in APHA
(1993). The selection of methods depends on the concentration of ammonia and
presence of interferences. These methods are:
I. Two colorimetric methods: The Nesslerisation and phenate method.
The Nesslerization method has been dropped as a standard method (APHA,
1995), although it has been considered as a classic water quality measurement
for more than a century. The use of mercury in this test and its problems of
disposal were the reasons of deletion.
The phenate method is used to analyse both freshwater and seawater for low
NH)-N concentration.
2. One volumetric method: The distillation followed by titration is used for NH)N concentration greater than 5 mgIL.
3. The instrumental method using an ammonia-selective electrode is applicable
over range from 0.03 to 1400 mg N~-NIL.
PRINCIPLE
Ammonia produces a yellowish-brown coloured compound when reacted with
alkaline Nessler reagent (KzHgI4 or 2KI + HgI z)' provided the sample is clarified
properly. Pretreatment with ZnSO4 and NaOH precipitates Ca, Fe, Mg, and sulfide,
which fonn turbidity and apparent colour. Addition of EDTA (before Nessler's
reagent) or Rochelle salt solution prevents precipitation of residual Ca and Mg in
presence of the alkaline Nessler's reagent.
2KzHgI4 + NH) + 3KOH -+ I-Hg-O-Hg-NHz + 7KI + 2H zO
(yellow-brown colour)
INTERFERENCE
Colour, turbidity, Ca, Mg, salts and Fe in the sample constitute the prime sources
of interference.
110
Chemicals
1.
2.
3.
4.
5.
6.
7.
Zinc sulfate
Sodium hydroxide
Potassium iodide, KI
Rochelle salt (potassium sodium tartrate tetrahydrate)
EDTA
Mercuric iodide
Ammonium chloride
Reagents
1. Zinc sulphate solution: Dissolve 100 g ofZnS04 .7Hp in distilled water and
dilute to 100 mL.
2. Sodium hydroxide, 6N: Dissolve 24 g ofNaOH and dilute to 100 mL.
3. EDTA reagent: Dissolve 50 g of EDTA in 60 mL of water containing 109
NaOH. If necessary, apply gentle heat to complete dissolution. Cool to room
temperature and dilute to 100 mL.
4. Rochelle salt solution: Dissolve 50 g of potassium sodium tartrate tetrahydrate
(KNaC4Hp6.4HP) in 100 mL of water. Remove NH3 by boiling off 30 mL
solution. After cooling dilute to 100 mL.
5. Nessler reagent: Dissolve 1OOg HgCl2 and 70 g KI in a small quantity ofwater.
Add this mixture to a cooled solution of 160 g NaOH in 500 mL of water and
dilute to 1L. Keep overnight. Filter through a glass fibre filter before using.
Store supernatant in a coloured bottle.
6. Stock ammonia solution: Dissolve 3.819 g anhydrous NH4Cl, dried at 100C,
dilute to IL. 1 mL = 1 mg N= 1.22 mg NH 3
7. Standard ammonia solution: Dilute 10 mL of stock ammonia solution to IL
with distilled water. ImL=lO ~g N = 12.2 ~g NH3
PROCEDURE
1. Take 100 mL of sample in a flask
2. Add 1 mL of ZnS04 solution and or 0.50 mL NaOH to obtain a pH of 10.5.
Allow to settle and filter the supernatant through Whatman filter paper No. 42.
3. Take a suitable aliquot of sample and dilute to 50 mL.
4. Add 3 drop of Rochelle salt solution or 1 drop of EDTA. Mix well.
5. Add 3 mL Nessler's reagent, ifEDTA is used or add 1 mL Nessler's reagent if
Rochells salt solution is used. Make up to 100 mL.
6. Mix well and read absorbance (optical density) after 10 minutes at 410 nm
against a blank prepared in the same way using distilled water instead of sample.
7. Prepare a calibration curve using suitable aliquots of standard solution in the
range of 5 to 120 ~g/I 00 mL (0.5 to 1.2 mgIL). For reference follow the same
procedure as per steps I to 5 above, but use the standard solution in place of
sample.
8. The value of ammonia is obtained from the standard graph and multiplied by
the dilution factor.
111
When sample contains more than 2 mgIL of ammonia-N, as in the case with
domestic sewage and many industrial wastes, the concentration can be determined
by titration with a standard solution of H2S04 after distillation and absorption of
ammonia in boric acid solution.
SELECTION OF SAMPLE SIZE
The Table 3.12 is useful in selecting sample volume for the distillation followed
by titration method.
Table 3.12:
Selection of sample volume for ammonia estimation.
Ammonia-N in sample. mg/L
5-10
10-20
20-50
50-100
Sample volume. mL
250
100
50
25
APPARATUS
1. Sodium tetraborate
2. Sodium thiosulfate, Na2Sp3.5Hp
3. Ethyl alcohol
4. Boric acid, H3B03
5. Sodium carbonate, Na2C03
6. Sodium hydroxide, NaOH
7. Methyl red indicator
8. Methyl.blue
9. H2 S04
Reagents
1. Use ammonia-free water in making all reagents and dilution: This should be
prepared for each batch of samples.
112
3.
4.
S.
6.
7.
a.
b.
Distillation of NH3
1. Take 100 mL of sample in a 500 mL ofKjeldahl flask and add 20 mL of borate
2.
3.
4.
5.
6.
buffer and adjust pH to 9.0 with 6N NaOH solution. Add a few glass-beads of
boiling chips.
The contents is heated on low heat for first 10-30 minutes and raise the heat
thereafter for about an 1 h to released all the residual ammonia.
Take 50 mL of boric acid cum indicator solution (pink colour solution) in 500
mL of flask and place it below the condenser of the distillation assembly so that
the lower open end of the condenser is dipped in solution.
Collect at least 200 mL of distillate in a flask (Pink solution turns to green due
to absorption of ammonia). It is good, to put a mark at the level of 250 mL in
the flask, so that one could easily know when 200 mL distillate is collected.
Remove the flask with distillate. Titrate the distillate with 0.02N Hl S04, to its
original colour (i.e. green to pink colour indicates the end point of titration).
Run a distilled water blank in the same manner.
CALCULATION
(T J - T l ) x 280
NH)-N,mgIL= - - - - mLofSample
Ifnormality of HlS04 is other than 0.02N, use following formula:
NH)-N, mgIL =
(T
Tl )
N x 14
1000
J
--=---=-------
mL of Sample
113
Where,
T. = Volume of H2S04 titrant used against sample (mL)
T2 = Volume of H S0 titrant used against distilled water Blank (mL)
2
4
N = Nonnality of H2S04 titrant.
100llll ~ ..
20 fill borat.
buff"
Gr..n _ Pink
(tnd
point)
(1995).
4.
5.
6.
7.
114
... cont.
Effluents discharge:
MOEF (1993) = 10 mgIL (inland surface waters) and 20 mgIL (marine coastal
areas).
Stream standards:
CPCB (1979) = 20 mgIL (Class A); and 50 mgIL (Class C)
Environmental Significance
1. Nitrate nitrogen (NO]-N) is the highest oxidisable form of nitrogen and occurs
in trace quantities in surface waters but may attain high levels in some groundwater.
2. Nitrate is an important plant nutrient and causes eutrophicatiori in receiving
water bodies.
3. High concentration in drinking water (>40 mgIL) may cause blue-baby disease.
Certain bacteria commonly found in the intestinal tract of infants can convert
nitrates (NO;) to highly toxic nitrites (NO;). Nitrites have a greater affmity for
haemoglobin in the blood stream than does oxygen, they replace that needed
oxygen, a condition is known as methemoglobinemia. The resulting oxygen
starvation causes a bluish discoloration of the infant and known as blue-baby
syndrome. In extreme. cases, the victims may die from suffocation. Usually
after the age of about 6 months, the digestive system of a child is sufficiently
developed that this syndrome does not occur.
Estimation of Nitrate
Following methods are available for determination of nitrate:
a. Ultraviolet spectrophotometric method
,
b. Ion chromatographic method
c. Nitrate electrode method
d. Cadmium reduction method (NO; is reduced to NO;).
e. Devarda's alloy method (reduction to ammonia) (UNEP/WHO, 1996).
f. Nitrate can also be determined colorimetrically by phenol disulphonic acid
method and Brucine method.
STORAGE OF SAMPLE
Start nitrate determination promptly after sampling. If storage is necessary, store
for up to 24 hr at 4C; for longer storage preserve with 2mL conc. H2SO/L and
store at 4OC. When sample is preseryed with acid, NO; and NO; cannot be determined individually.
115
Spectrophotometer, for use at 220 nm and 275 nm with matched silica cell of Icm
or longer path.
REAGENTS
1. Stock nitrate solution (ImL= 100 Ilg N03-N): Dry potassium nitrate (KNO])
in an oven at 105C for 24 h. Dissolve 721.8 mg in water and dilute to 1000 mL.
Preserve with 2 mL chloroform per L. This solution is stable for at least
6 months.
1. Treatment of sample
Take 50 mL of clear sample and (filter it, if necessary) add I mL HCl solution
and mix thoroughly.
I. For samples and standards, subtract two times the absorbance reading at 275
nm from the reading at 220 nm to obtain absorbance due to NOJ-N only.
2. Construct a standard curve by plotting absorbance due to nitrate against
NOJ-N concentration.
3. Obtain sample concentration directly from the standard c' .rve.
Note: If correction value is more than 10% of the reading at 220 nm, do not use this
method.
116
This method is suitable for nitrate concentration exceeding I mgIL especially for
wastewater and polluted surface waters. This analysis may be carried out on the
original sample or on the residue from the detennination of ammonia.
Nitrate is reduced to ammonia by nascent hydrogen by the use of Devarda's
alloy (59% AI, 39% Cu, 2% Zn). The resulting ammonia is distilled and its concentration detennined by titration.
INTERFERENCES
Ammonia must be removed from the sample before the main procedure is started.
This is achieved either by pre-treatment or by using the distillation residue from
the detennination of ammonia The air in the laboratory and the distilled water
used for solutions and during the procedure should be free of ammonia.
APPARATUS
The same glassware and distilling apparatus as for the ammonia nitrogen detennination.
CHEMICALS AND REAGENTS
The following reagents are required in addition to those used for the ammonia
nitrogen detennination.
1. Mixed indicator solution
2. Boric acid, 2% (20 gIL)
3. Devarda's alloy, powdered: If fme powder is not available, the material
should be ground to pass through a 0.07-0.1 mm sieve (200-140 mesh). It should
be as free as possible from nitrogen.
4. Hel, 0.00714 N: Dilute 6 mL conc. HCI (12N) to 1000 mL and standardise
against sodium carbonate solution.
5. Sodium hydroxide, ION: 400 gIL.
PROCEDURE
117
CALCULATION
(A - B)
14
1000
-n
Where,
A = volume of 0.0714 N acid used for titration of the distillate of the sample,
mL
B = volume of 0.0714 N acid used for titration of the distillate of the blank, mL
V = volume of the undiluted sample, mL.
n = concentration ofnitrite-N, in mgIL determined separately.
Note: The same calculation without subtracting nitrite gives the result for total
oxidised nitrogen, i.e. nitrate + nitrite.
The result is reported as nitrate nitrogen (as N) mgIL and should be rounded to
two significant figures.
118
The yellow colour follows Beer's law and is proportional to the concentration of
nitrate present in the sample.
INTERFERENCE
Chloride and nitrite are the two main courses of interferences. Pretreatment of
sample is necessary when the interfering radicals are present.
APPARATUS
Chemicals
1.
2.
3.
4.
5.
6.
7.
8.
Ag2S04
Conc. H2S04 (sp. gr., 1.84)
KOH
EDTA
Phenol
NHpH, Conc.
KN0 3
Potash alum, Aluminium hydroxide
Reagents
1. Standard silver sulfate: Dissolve 4.40 g A~S04 in distilled water and dilute
to 1000 mL; 1 mL = 1 mg Cl.
2. Phenol disulfonic acid (PDA): Dissolve 25 g white phenol in 150 mL conc.
H2 S04 , Add 75 mL fuming H2 S04(15 % free S03)' stir well and heat for 2 h on
water-bath. If fuming sulfuric acid is not available, add additional 85 mL conc.
H2 S04 to the 150 mL H2 S04 , stir well and heat for 2 h.
3. Ammonium hydroxide: Concentrated
4. Potassium hydroxide (12 N): Dissolve 673 g KOH in distilled water and
make up to 1000 mL with distilled water.
5. Stock nitrate solution: Dissolve 721.8 mg anhydrous potassium nitrate and
dilute to 1000 mL with distilled water; 1mL = 100 Ilg N03-N
6. Standard nitrate solution: Evaporate 50 mL stock nitrate solution to dryness
on water-bath. Dissolve residue in 2 mL PDA reagent and dilute to 500 mL.
1 mL = 10 Ilg N.
7. EDTA: Weigh 50 g EDTA and make a paste with 20 mL distilled water. Add 60
mL NHpH and mix well.
8. Aluminium hydroxide: Dissolve 125 g potash alum in 1000 mL. Heat to 60C
and add 55-60 mL NHpH and allow to stand for 1 h. Decant the supernatant
and wash the precipitate a number of times till it is free from CI or nitrite.
119
PROCEDURE
I. Pre-treatment of sample
1. Colour removal: If the sample has a colour in excess of 10units, add 3 mL
aluminium hydroxide to 150 mL of sample. Stir well and allow to settle for a
few minutes. Filter and use the filtrate discarding the first portion of the
filtrate.
2. Nitrite removal: (a) Generally nitrite occurs along with ammonia and gets eliPlinated in the routine test due to decomposition of nitrite and NH; to N2 (b)
Oxidise NO~ to NO; under acidic condition using KMn04 (c) Add sulfamic
acid to the sample to suppress nitrite interference.
3. Chloride removal: Determine the chloride content of the sample and precipitate out it as AgCI by using silver sulphate solution at the rate of ImL to remove 1mg of chloride. One should be very careful while adding A~SO 4 because excess Ag will precipitates as silver oxide when alkali is added to the
sample.
CALCULATION
Construct a standard curve by plotting the nitrate-nitrogen in mglL against
absorbance.
2. Obtain sample concentration directly from the standard curve.
I.
120
INTRODUCTION
Nitrite (NO~) is fonned in waters by oxidation of ammonia compounds (by aerobic
nitrifying bacteria, e.g. l'!itrosomonas) or by reduction of nitrate (by facultative
anaerobic denitrifyiub bacteria, e.g. Pseudomonas). Such oxidation and reduction
may occur in wastewater treatment plants, water distribution system, and natural
waters. As an intennediate stage in nitrogen cycle, it is unstable. Very high nitrite
levels are usually associated with waters having unsatisfactory microbiological
activity.
APHA (1995) described two methods of nitrite-nitrogen estimation:
1. Colorimetric method: Suitable for concentration of 5 to 1000 Ilg NOz-NIL (applicable in the range of 0.05 - Img/L of N0 2-N). Sample containing higher
concentration must be diluted. (UNEPIWHO, 1996 also recommended this
method)
2. Ion Chromatography method: Not discussed here.
INTERFERENCE
Chlorine in the samples or nitrogen trichloride which nonnally coexist with nitrite
produce a false colour. This can be minimised by addition of naphthyl amine hydrochloride. Nitrite detennination should be carried out in filtered and turbidity free
samples. Presences of strong oxidants or reductants in the sample readily affect the
nitrite concentration. High alkalinity (> 600 mglL as CaC03) gives low results
owing to a shift in pH.
121
STORAGE OF SAMPLE
Never use acid preservation for samples to be analysed for nitrite. Make determination promptly on fresh samples to prevent bacterial conversion of nitrite to nitrate or ammonia. For short-term preservation of 1 to 2 days, store at 4C.
APPARATUS
1. Spectrophotometer for use at 543 nm, providing a light path of l-cm or longer.
2. Nessler's tubes or 100 mL capacity volumetric flasks.
CHEMICALS AND REAGENTS
Chemicals
1.
2.
3.
4.
5.
6.
Sulfanilamide
Sodium nitrite, KN02
Chloroform, CHCl3
N-( I-naphthyl)-ethylenediamine dihydrochloride (NED dihydrochloride)
Phosphoric acid, Hl0 4, 85%
Conc H2S04
Reagents
1. Colour reagent: To 800 mL water add 100 mL of 85% phosphoric acid and 10
g sulfanilamide. After dissolving sulfanilamide completely, add 1 g NEDdihydrochloride. Mix to dissolve and dilute to 1000 mL with water. Solution is
stable for about a month when stored in a dark bottle in refrigerator.
2. Preparation o/standard nitrite solution: The standard nitrite solution could be
prepared by anyone of the following two methods .
122
0.0
0.02
0.04
0.08
0.10
0.20
0.30
PROCEDURE
1. Take 50 mL of sample in a 100 mL volumetric flask.
2. Removal of suspended solids: If sample contains suspended solids, filter it
through Whatman no. 42 filter paper.
3. Check the pH of the solution. If pH is not between 5 to 9, neutralise pH by
adding 1 N HCI or NHpH as required.
4. Add 2 mL of colour reagent, mix well and makeup the volume to 100 mL. At
this stage pH should be 2-2.5.
5. Measure the colour after 10 min at 543 nm. Prepare calibration curve in the
range of 0 to 1 mg N02-NIL at the interval ot(f.1 mg N02-NIL.
6. Prepare a blank in the same way by using distilled water instead of the sample.
7. Calculate and record as N0 2-N in mgIL.
CALCULATION
Read the concentration ofN02-N in samples directly from the calibration curve.
If less than 50 mL of sample is taken, calculate the concentration as follows: .
mgIL from standard curve x 50
Nitrite (as N), mgIL = - - - - - - - - - - mLsample
123
Effluents
All efforts should be made to remove unpleasant odour as far as possible
(Ministry of Environment and Forests, 1993).
Introduction
SOURCES OF ODOUR IN DRINKING WATER
Many substances with which water comes into contact in nature of human use may
impart perceptible taste and odour. These includes minerals, metals and salts from
soil, inorganic substances are more likely to produce taste unaccompanied by odour.
Organic substance on the other hand, is likely to produce both taste and odour.
Principles among are the reduced product of sulphur as H 2S, which imparts rotten
egg odours. Certain species of algae secret substances may results both taste and
odour problems. The combination of two more substances, neither of which would
produce taste or odour by itself, but may produce odour ifmixes. This synergistic
effect may noted either in case of organic and chlorine.
124
Environmental Significance
...
APPARATUS
1.
2.
3.
4.
5.
PROCEDURE
TON =
(A+B)
A
125
Table 3.13 :
Threshold odour number (TON) corresponding to various dilution.
Sample volume diluted to
200mL
200
100
50
25
8 .3
2 .8
1.4
TON
1
2
4
8
24
70
140
140
70
35
17
4
2
1
TON
1.4
3
6
12
50
100
200
Example-1
A sample of water requires 150 mL of odour free water to render the odour
barely detectable. What is the volume of sample and what is the TON .
. Volume of sampl~ (Art Odour free water (8) = 200 mL
,' A+i50 F 200 "'::;..v
' Tbertlfor~, A-=
TQN= (At B)!
(50+ 1
=4
. =
Drinking water
IS: 10500 (I983, 1991) = 0.01 mgIL (beyond this limit, undesirable taste and
cont...
126
... cont.
odour after chlorination is produced). 0.03 mgIL (permissible in absence of
alternate source).
MHW (1975) = 0.01 mgIL (acceptable); 0.03 mgIL (cause of rejection)
WHO (1984) = No guideline value set.
Effluent discharge
MOEF (1993) = 10 mgIL (inland surface discharge and land for irrigation) and
20 mgIL (public sewer and marine coastal areas).
Effluent discharge from coal mine (MOEF 2000) = 10 mgIL
Introduction
In the determination of oil and grease, an absolute quantity of a specific substance
is not measured. Rather a group of substances with similar physical characteristics
are determined quantitatively on the basis of their solubility in an organic extracting
solvent. Thus, oil and grease are a class distinguished on the basis of its solubility
characteristics, not is chemical composition. Oil and grease is defmed as any material
recovered as a substance soluble in the solvent. This test is applicable to natural
water and domestic wastewater. This method is also suitable for most industrial
wastewater.
The 12th edition of APHA (1971) standard methods prescribes the use of
petroleum ether as the solvent for natural and treated waters and n-hexane for polluted water.
In the 14th, 17th, and 19th editions of APHA (1981, 1989, 1995) standard
methods only trichlorofluroethane has been recommended.
Two test methods are referred in APHA (1995):
1. Liquid-liquid extraction (partition gravimetric method) : for 4 to 100 mgIL oil
and grease concentration in water and wastewater.
2. Soxhlet extraction (20 to 200 mgIL in water and wastewater).
Environmental Significance
Oil and grease determination on raw and settled wastewater gives a measure of
the effectiveness of primary settling tanks.
When discharged in wastewater, they may cause surface films and shoreline
deposits leading to environmental degradation.
In this method an acidified water sample is extracted with petroleum ether solvent
in a separatory funnel.
In the gravimetric portion of the procedure, the petroleum ether solvent
127
PROCEDURE
1. Take 250 mL of sample in a 500-mL beaker.
2. Add 5 mL of concentrated (1+ I) HCI to make pH 2 (acidification is needed to
maintain the integrity of the sample) ..
3. Transfer the sample to separating funnel. Add 30 mL of petroleum ether and
shake vigorously for 5 to 10 minutes. Invert the separatory funnel and vent with
stopcock to relive pressure building during extraction. Allow the layer to
separate. If a clear solvent layer cannot be obtained due to emulsion,with water,
add up to 100g ofNaCI to separatory funnel. Shake to dissolve the salt frequently.
This will break the emulsion.
4. The upper one petroleum ether and lower one is of water become distinct. Discard
the lower water portions from separating funnel. The systematic procedure is
shown in Fig. 3.21.
AQUEOUS
LAYER
128
Along with the results, mention the solvent used for extraction.
Effluent discharge
IS: 2490 (1981) = 5.5 to 9.0
MOEF, Schedule - VI, (1993) = 5.5 to 9.0
Introduction
pH of most natural water falls within the range of 4 to 9. The majority of waters are
slightly basic (Le. generally over 7.0) because of the presence of carbonate and
bicarbonates. The pH increases (acidic) during daytime due to photosynthetic activity
because of consumption of CO2, whereas it declines (alkaline) at night due to
respiratory activity. By defmition, pH is the negative logarithm of hydrogen ion
concentration, more precisely hydrogen ion-activity.
129
or,
1
pH = log - 10 [W]
Alkaline range
Acidic range
7
pH scale
14
at 25C
pH
Bases
at 25C
pH
HCI
H2 SO.
Acetic acid
Boric acid
O.lN
O.lN
O.lN
O.lN
1.1
1.2
2.9
5.2
NaOH
Ammonia
Sodium biocarbonate
O.lN
O.lN
O.lN
13.0
11.1
8.4
Environmental Significance
It is important in almost every phase of environmental engineering practice, such
as of water supplies, water softening, disinfection and corrosion control. Low pH
causes corrosion, high pH causes taste, soapy feel; pH < 8.0 is preferable for effective
disinfection with chlorine.
Example - 1
Calculate the pH values for two solutions (a) in which [OH-) is 0.001 moles; (b)
[OH-) is 2 .5 x 10-' moles.
sdi:rtIon; ,
=1.0 x 10-1
1 .O)()O~J, ~ ,>
--
1.0 x ;, 10-14
10
0.001
(AI"l~)
= 4 .0x 10-5 M
,,2.5 x 10- 10
pH,;= -Jog (4.0x JO-5) = -(log4+ log 10-5 )
' (Ans)
"
-(13.602-5.00)
= 4.40
(acidic)
130
Estimation of pH
Detennination of pH of water should be made in situ. If this is not possible, for
example with well water or when access to a lake or river is very difficult, the
measurement should be made immediately after the sample has been obtained.
Two common methods used for pH measurement are:
Use ofpH indicator paper: Simple, inexpensive, inaccurate.
Use ofelectronic pH meter: Accurate and free from interferences, gives reading
with an accuracy of 0.05 pH.
1. ELECTRONIC METHOD
Principle
.,. (1)
Mn+
Where,
E = Half-cell potential
T = Absolute temperature
F = Faraday constant
M = Activity of ions to be measured
R
n
K
Gas constant
Valence
Constant
The E (half-cell potential) can not be measured alone. If the glass electrode is
placed against a reference electrode (usually the calomel electrode), the potential
difference between two electrodes is measurable. (E - Ecal).
Reference electrode consists of a half cell that provides a constant electrode
potential. Commonly used reference electrodes are Calomel and silver: silver-chloride electrode. Calomel electrodes are of 3 types, nonnal, tenth nonnal and saturated depending on the concentration ofKCI solution used in preparing them. The
potential develops on each electrode is a function ofKCI concentration. In general,
saturated type of calomel electrode is employed.
Before any pH measurement, two electrodes have to be placed first in a solution of known pH (e.g H+ concentration = 19!L). This is called standardization of
electrode and pH meter. The overall potential of the total cell Eo equals to E-Eca1'
and is given as:
RT
Eo =/ ~
K
log -1- - Eca1
... (2)
131
... (3)
E-E =
o
RT
F
RT
F
(.Og
~ -logJ
(W)
log-W
RTpH
F
For H- ions, RTIF = 0.0591 at 25C; therefore
E -E
pH = _0_ _0
0.0591
Apparatus
1. pH meter: It consists of a potentiometer, a glass electrode, a reference electrode,
and a temperature compensating device (Fig. 3.22)
2. Beakers
3. Stirrer: Use either magnetic, TFF- coated stirring bar or a mechanical stirrer
with inert plastic-coated impeller.
Reagents
Buffer solutions of pH of 4.0, 7.0 and 9.2. Standard buffer tablets are commercially
available. Buffer of different pH can be prepared by dissolving standard pH tablets
in distilled water. The above buffer solution can be prepared as follows :
Procedure
Calibration of pH meter
I. Read carefully the operational manual of pH meter to be used.
2. Calibrate the pH meter against the standard buffer solution of pH 4.0, 7.0, and
9.2 according to the requirement as follows .
l32
yjth Agel
Wash the electrode with distilled water and dip the electrode in buffer of pH
7.0 and move the temperature knob to the required or room temperature and
adjust the meter accordingly.
Wash the electrode with distilled water and dip the electrode in buffer of pH
4.0 and move the temperature knob to the required or room temperature and
adjust the meter accordingly.
Wash the electrode with distilled water and dip the electrode in buffer of pH
9.2 and move the temperature knob to the required or room temperature and
adjust the meter accordingly
Sample analysis
1. After calibration, rinse the probe with distilled water, blot dry with a soft tissue
paper, place in sample solution.
2. Establish equilibrium between electrodes and sample by stirring sample to ensure
homogeneity; stirring gently to minimize CO2 entrapment.
3. Keep the electrode dipped in storage solution when not in use.
133
Reporting
Report the pH with precision of O.Oland the temperature of the sample at the
time the measurement.
pH Electrode Maintenance
G lass electrodes (sensing electrode) fail because of scratches, deterioration or accumulation of debris on the glass surface. The electrode in rejuvenated by alternately immersing it 3 times each in O.IN HCI and O.IN NaOH. If this fails, immerse tip in KF solution for 30 sec. After rejuvenation, soak in pH 7.0 buffer
overnight. Rinse and store in pH 7.0 buffer. Rinse again with distilled water before
use. The pH electrode actually requires little maintenance. The electrode should be
stored in water. If allowed to dry out, the electrode needs to be hydrated for a
couple of hours prior to use. The level of concentrated KCI must be monitored and
replenished by adding saturated KCI through the stoppered orifice in the side of the
electrode. The correct liquid level is approximately 5 mm below the bottom edge
of the hole. The electrodes used for measurement generally need replacing periodically (e.g. yearly). Old or poor quality electrode often shows a slow drift in the
reading.
Electrode should not be left out for long periods, and be always kept immersed
in distilled water or pH 7.0 buffer.
134
Introduction
Phenols are the most important groups of aromatic compounds. It is commonly
know as carbolic acid. It has been used widely as a germicide and disinfectants.
Phenol can be treated biologically up to 700 mgIL aerobically and up to 200 mgIL
anaerobically. Phenolic compounds are sometimes found in surface waters from
natural and industrial sources.
Generally, the level of phenol in domestic wastewater is low. However, it has
been reported in the range of 10 to I 00 ~gIL in Mumbai sewage (Igole and Kadam,
1999). But many industries produces high concentration of phenol in their effluents, like Petrochemical (10-30 mglL phenol); Petroleum refineries (10-200
mglL); Coke oven effluent (30-1000 mgIL) etc. Phenol removal processes in water
treatment include superchlorination, chlorine dioxide or chloramine treatment,
ozonation and activated carbon absorption.
Environmental Significance
Phenols are highly toxic to aquatic organisms. For human, phenol is toxic by ingestion and inhalation. Above 1 ppb concentration of phenol are highly objectionable
with respect to taste and odour problems in water. They react with chlorine which
may produce chlorophenol that are odouriferous and are of noticeable medicinal
taste in the drinking water. Their presence in streams and other waterways will
cause off flavour in fish tissues and other aquatic food.
1. Both test methods are photometric procedures based on the reaction of steam
distillable phenolic compounds with 4-arnminopyrine.
2. Test method (A) differs from (B) mainly in that the sample is extracted with
chloroform, thereby providing 20-fold greater sensitivity.
3. Both the procedures involve separation of phenolic compounds from the background matrix by distillation.
PRESERVATION AND STORAGE OF SAMPLE
13S
Apparatus
1. Distillation apparatus: All glass, consisting of l-L borosilicate glass distilling apparatus with Graham condenser.
2. pH meter.
Reagents
Prepare all reagents with distilled water free of phenols and chlorine.
1. Phosphoric acid, H 3PO. (1+9): Dilute 10 mL 8S% Hl0 4 to 100 mL with
water.
.
2. Methyl orange indicator
3. Special reagents for turbid distillates:
a. Sulphuric acid, H 2S04, IN
b. Sodium chloride, NaCI
c. Chloroform, CHCl 3
d. Sodium hydroxide. NaOH, 2.5N: Dissolve 10 g NaOH pellets in 100 mL
water.
Procedure
Preparation of distillation mixture
1. Measure SOO mL of the sample into a beaker.
2. Adjust pH of the sample to 4.0 with Hl0 4 solution using methyl orange or a
pH meter.
3. Transfer the mixture to the distillation apparatus.
4. Use a SOO-mL graduated cylinder as a receiver.
S. Omit adding Hl0 4 and adjust pH to 4 with 2.S N NaOH, if sample was preserved as described earlier.
Distillation
6. Collect about 4S0 mL distillate. When distillation is finished, and, boiling is
ceased, add SO mL warm water to the distilling flask. Next, continue the distillation until a total of SOO mL ha!: been collected.
7. Ifthe distillate is turbid, a second distillation may be helpful. Acidify the turbid
distillate with H2 S04 solution (1 +9) and repeat the previously described distillation. The second distillate usually eliminates the turbidity.
136
stock solution to 100 mL with freshly boiled and cooled water. Prepare this
solution fresh on the day it is used.
3. Phenol solution, standard (1 mL = 1Jig of phenol): Dilute 50 mL of the intermediate solution to 500 mL with freshly boiled and cooled water. Prepare this
solution fresh within 2 h of use.
ESTIMATION OF PHENOL BY DIRECT PHOTOMETRIC METHOD
This test method is applicable to water that contains more than 0.1 mg/L of phenolic compounds.
Principle
This is a photometric test method, based on the reaction of steam-distillate phenolic compounds which react with 4- aminoantipyrine at pH 10 2 in the presence
of potassium ferricyanide [~Fe(CN)6] to form a coloured antipyrine dye. The antipyrine colour formed in an aqueous solution is measured at 510 om. The concentration of phenolic compounds in the sample is expressed in terms of mg/L of
phenol (C6HPH).
Apparatus
Photometric equipment: Spectrophotometer equipped with absorption cell
providing light paths 1 to 5 cm for use at 500 om.
Chemicals
1.
2.
3.
4.
Reagents
1. Ammonium chloride (NH.CI) solution: Dissolve 20 g ofNH4CI in water and
dilute to I L.
2. Ammonium hydroxide (NHPH): Concentrated (sp. gr. 0.90)
3. 4-aminoantipyrine solution: Dissolve 2 g of 4-aminoantipyrine in water and
dilute to 100mL. Prepare the reagent fresh as used.
4. Potassium ferricyanide: Dissolve 8g of [~Fe(CN)6] in water and dilute to
100mL. Filter if necessary. Prepare fresh daily.
Procedure
1. Calibration standards: Prepare a 100mL distilled water blank and series of
100-mL phenol standards containing 0.1, 0.2, 0.3, 0.4 and 0.5 mg phenollL.
2. Sample: Transfer 100-mL distillate, or a portion not containing more than 0.5
mg phenol diluted to 100mL, in a 250-mL beaker. Also prepare a blank consisting 100 mL of water.
137
3. Add 5 L NHpH solution immediately. Adjust the pH between 9.8 and 10.2
withNHpH.
4. Add 2mL of 4-aminoantipyrine solution, mix well; add 2mL ~Fe(CN)6 solution and mix well.
5. After 15 min, transfer the solution to absorption cells and read absorbance of
sample and standards against the reagent blank at 51 Onm.
Calculation
Use of calibration curve: Estimate sample phenol content from photometric reading by using a calibration curve.
A
mgIL phenol = - - x 1000
Where,
A = mg phenol in sample, from calibration curve
B = mL original sample.
Use of single phenol standard:
mgIL phenol =
ex D x 1000
-------
ExB
Where,
C = mg standard phenol solution,
D = absorbance of sample
E = absorbance of standard phenol solution
Test data of photometric method (ASTM, 1995) as follows:
Amounts added
mgIL
Amount found
mgIL
Bias,
mgIL
Bias,
Statistically
significant
7.154
35.768
71.535
6.930
34.430
68.777
- 0.224
- 1.38
- 2.758
-3.1
- 3.7
- 3.9
yes
yes
yes
138
3.27 Phosphorus
.of phosphorus
Drinking water
IS: 10500 (1983, 1991) = No standards.
WHO (1984) = Not issued any regulation or guidelines for phosphorus.
U.S.EPA (1974) = Phosphorus has not been regulated not even in secondary
drinking water standards
Effluent discharge
MOEF (Schedule-VI, \993) = Dissolved phosphate (as P), 5 mglL (max.) for
discharge in inland surface water bodies.
Introduction
Phosphorus occurs in waters and wastewater almost solely as phosphates. These
are classified as (a) orthophosphate; (b) condensed phosphates and (c) organically
bound phosphates. They occur in solution, in particulates or detritus or in the body
of aquatic organisms.
Water supplies may contain phosphate derived from natural contact with minerals or through pollution from application offertiliser, sewage and industrial waste.
Groundwater, therefore, is more likely to have higher phosphate concentration.
In sewage, concentration o.f phosphorus (total as P) has been reported in the
range of4 mglL (organic- P, 3 mglL and inorganic- P, I mg/ L) to 15 mglL (organicP, 5 mglL and inorganic-P, 10 mglL).
The daily requirement of P is equal to that of Ca. The recommended dietary
allowance (RDA) is 800 mg for adults . Dietary deficiency is not expected because
of abundance of phosphorus in the diet.
Environmental Significance
The crucial level of phosphorus that causes eutrophication problems is somewhere
near 0.005 mglL in summer growing conditions. Studies suggest that 0.0 I mglL of
phosphorus is acceptable while 0.02 mglL is excessive.
The presence of phosphorus in large quantities in freshwater indicates pollution through sewage and industrial wastes.
Though phosphorus poses problems in surface waters, its presence is necessary
for biological degradation of wastewater.
Phosphorus can be classified as total-P, inorganic-P (orthophosphate,
HlO~, HP01-, PO; ) and organic-P
Only inorganic compounds of phosphorus are of significant environmental
importance.
l39
Estimation of Phosphorus
METHODS OF PHOSPHATE ESTIMATION
APHA (1995) lists three preliminary steps and the four methods for examination of
phosphorus.
Preliminary steps
a. Preliminary filtration step.
b. Preliminary acid hydrolysis step for acid-hydrolyzable phosphorus.
c. Preliminary digestion step for total phosphorus.
Methods
a. Vanadomolybdo phosphoric acid colorimetric method
b. Stannous chloride method
c. Ascorbic acid method
d. Automated ascorbic acid reduction method.
ANALYTICAL SCHEME FOR DIFFERENTIATION OF PHOSPHORUS
FORMS
A scheme for the differentiation of various phosphorus forms which can be determined in water is given in Fig. 3.23.
ISample I
Unfiltered
-4
,l.
Filtrate
No sample digestion -4 Dissolved orthophosphate
[
Acid digestion -4 Dissolved orthophosphate + hydrolysable phosphate
Acid-persulfate digestion -4 Dissolved total recoverable phosphate
140
141
CALCULATION
Calculate phosphate concentration as follows:
mg phosphate/L =
mL of sample
REAGENTS
1. Conc. sulphuric acid (H 2S04)
2. Phenolphthalein indicator
3. Conc. nitric acid (HNO))
4. Sodium hydroxide (NaOH), 1N: Dissolve 40 g NaOH in 1L of distilled water.
PROCEDURE
Digestion
142
the [mal solution. Each 1C increase in temperature producing about 1% increase in colour. Hence, hold samples, standards, and reagent within 2C of
one another and in the temperature range between 20 and 30C.
Colour measurement
1. After 10 min, but before 12 min, measure the absorbance of the colour
spectrometric ally at 690 nm and compare with a calibration curve, using distilled water blank.
CALCULATION
mg ofP (in approx 104.5 mL [mal volume)
Phosphate (mgIL) = - - - - - - - - - - - - - - - - x 1000
mLsample
3.28 Solids
Solids refer to matter suspended or dissolved in water or wastewater. Solids may
affect water or effluent quality adversely in a number of ways. Waters with high
dissolved solids generally are of inferior palatability. For this reason, a maximum
limit of 500 mg dissolved solidslL is permissible for drinking water. Three types of
solids are analyzed for water and wastewater.
143
Total Solids (TS) is the measure of all kinds of solids i.e. suspended, dissolved and
volatile solids. Total solids can be determined as the residue left after evaporation
at 103 to 105C of the unfiltered sample. It consists of two parts: Total Suspended
Solids (TSS) and Total Dissolved Solids (TDS).
Each fraction is again divided into volatile suspended solids (VSS) and fixed
solids after heating at 550C. VSS is the organic fraction that lost as CO 2 and fixed
solids are the ash portion that remains.
Total solids
by filtration (1 micron pore size filter paper)
Total Dissolved solids (TDS)
Volatile solids
(VSS)
Organic
Fixed solids
(FS)
Inorgainc
Volatile solids
(VSS)
Organic
Fixed solids
(FS)
Inorgainc
APPARATUS
1. Take an evaporating dish or clean beaker (400mL capacity) of suitable size and
dry at 103 to 105C for Ih. Store and cool the dish in desiccator until needed.
Weigh immediately before use. Note the initial weight (Wi) in mg.
2. Put 250-300 mL unfiltered well-mixed sample in it.
3. Put in hot air oven at j 03 C to 105C for 2 h up to dryness.
4. Cool in desiccator and take the fmal weight (Wt) in mg.
5. Repeat cycle of drying, cooling, desiccating and weighing until a constant weight
is obtained, or weight change is less than 4% of previous weight or 0.5 mg,
which ever is less. When weighing dried sample, be alert to change in weight
due to air exposure and lor sample degradation. Duplicate determination should
be within 5% of the average.
144
CALCULATION
(Wf - Wi) x 1000
Total Solids mglL = - - - -- - - Volume of sample, mL
145
PRECAUTIONS
1. Results may be questionable if the TSS include oils, grease or other volatile
matter.
2. Exclude large floating particles or submerged agglomerates of non-homogeneous
material from the sample.
3. In case of highly polluted water, sewage or wastewater, excessive residue on
the filter paper may form a water-entrapping crust, in such case, limit the sample
size so that yielding no more than 200 mg residue.
4. Prolonged filtration times resulting from filter clogging may produce high results
owing to increased colloidal materials captured on the clogged filter.
APPARATUS
In addition to the ones listed in section 3.28.1 for total solids (page 143) the following
apparatus are required:
1.
2.
3.
4.
PROCEDURE
Sample size
146
CALCULATION
(Wf - Wi) x 1000
Total Suspended Solids mgIL = - - - - - - - Volume of sample, mL
Report the results as: Total SS dried at .......... .."C, .. .......... mglL.
Total suspended solids can also be obtained as a difference between TS and
TDS
TSS = TS-TDS
Drinking water
ICMR (1963) = 500 mgIL (minimum) to 1500 mgIL (maximum)
WHO (1984) = 1000 mgIL (maximum)
WHO (1993) = 1000 mgIL (consumer complaint is taste)
IS: 10500 (1983,1991) = 500 mgIL (maximum) (beyond 500 mgIL
palatability decreases and may cause gastrointestinal irritation). 2000 mgIL
(permissible limit in the absence ofaltemate source)
MHW (1975) = 500 mgIL (acceptable) and 1500 mglL (cause for rejection)
Effluent discharge
IS: 2490 (1981) = 2100 mgIL (maximum).
MOEF, Schedule - VI (1993): 2100 mgIL - inland surface water. Omitted by
GSR 80 (E) dated 31st Dec, 1993 (w.e.f 31.12.93)
Introduction
A large number of solids are found dissolved in natural waters, the common ones
are carbonates, bicarbonates, chlorides, sulphates, phosphates, and nitrates of
calcium, magnesium, sodium, potassium, iron, magnesium etc. In other words, TDS
is simply the sum of the cations and anions concentration expressed in mgIL.
The results of a determination of TDS can be used to check the accuracy of
analyses. This is accomplished by comparing the value of calculated TDS with the
measured value. TDS can be calculated by adding the ionic concentrations and
express the results as mgIL as follows:
Calculated TDS = 0.6 (alkalinity) + Na + K + Ca + Mg + Cl + S04 + SiO J
+NO J + F
The measured TDS should be higher than the calculated value, because a significant
contributor may not be included in the calculation. If the measured value is less than
the calculated value, all values are under suspicion. The acceptable ratio is:
147
1.0 <
measured TDS
calculated TDS
<1.2
ENVIRONMENTAL SIGNIFICANCE
A high content of dissolved solids elevates the density of water, influences osmoregulation of freshwater organisms, reduces solubility of gases (like oxygen) and
reduces utility of water for drinking, irrigation and industrial purposes. TDS
concentration beyond 500 mg/L, decreases palatability and may cause
gastrointestinal irritation.
PRINCIPLE
A well mixed, measured portion of sample is filtered through a standard glass-fibre
filter and the filtrate portion is evaporated to dryness at 180 2 C and that gives
the amount of total dissolved solids. The reason for higher temperature used is to
remove all mechanically occluded water. Where organic matter is generally very
low in concentration, the losses due to higher drying temperature will be negligible.
APPARATUS
it clean, dry at 103-105 C, cool in a desiccator and note the initial weight (W I)'
2. Filter about 100mL to 250mL of sample through filter paper (Whatman No.
41) and take the filtrate in the evaporating dish. (Note: Choose the sample
volume to yield 10 to 200 mg of dried residue. If more than 10 minutes are
required for complete filtration, decrease the sample volume.)
3. Evaporate the sample in a hot air oven at 180 2 C and after the whole water
is evaporated, cool the evaporating dish in a desiccator and take the fmal
weight(W).
CALCULATION
W-W
2
Where,
WI
W2
I X 1000 x 1000
148
Example-1
Problem: Find out TDS with the following results.
Example-2
An analysis of a sample of water with pH 7.5 has produced the following
concentrations (mg/L). Find the total dissolved solids (TDS) in mg/L.
ENVIRONMENTAL SIGNIFICANCE
To determine the need for and design of primary settling tank.
To determine the efficiency of sedimentation units.
To study the physical behaviour of waste streams entering natural water bodies.
149
APPARATUS
The volumetric test requires only an Imhoff cone.
PROCEDURE
I. Fill an Imhoff cone to the l-L mark with a well-mixed sample (Fig. 3.24).
2. Settle for 4S-min, gently agitate sample near the sides of the cone with a rod or
by spinning, settle 15 min longer, and record volume of settleable solids in the
cone as mUL.
3. If the settled matter contains pockets of liquid between large settled particles,
estimate volume of these and subtract from volume of settled solids.
4. The practical lower limit of measurement depends on sample composition and
generally is in the range of 0.1 to 1.0 mLlL. where a separation of settleable and
floating materials occurs, do not estimate the floating material as settleable
matter.
5. Replicates usually are not required.
RESULTS
Report the results as mL of settleable solids IL (volumetric method).
r'l\l
I
~
Fig. 3.24: Imhoff cones used to measure settleable solids.
ISO
CALCULATION
Settleable Solids, mgIL = TSS, mgIL - Non-settleable Solids, mgIL
Drinking water
IS: 10500 (I991) =No standards for sodium
USEPA (1974) = No MeL
USEPA (I980) = 20 mgIL recommended.
WHO (1984, aesthetic quality) = No maximum contaminant level has been set
for Na but 200 mgIL was fixed based on taste
Ee (1980) = 20 mgIL guide level
Emuent discharge
MOEF (1993) = No standards for Na
Introduction
Sodium (Na) ranks 6th among the elements in order of abundance and is present in
most natural waters. Sodium is generally found in lower concentration than cal-
151
cium and magnesium in freshwater. The salts ofNa are highly soluble in water and
impart softness (in contrast to hardness). In food, USEPA recommends daily intake
levels of 1600-9600 mg for adults and 69-92 mg/kg/day for infants based on measurement of sodium excretion from urine (sodium is mostly excreted in urine).
Food intake of sodium averages 90% or more of the total sodium, estimated at 28g/day.
Na levels may vary from < I mg/L to more than 500 mg/L.
In ground water, it varies widely but normally range between 6 mg/L and 130
mg/L.
In surface waters, sodium concentration may be less than 1 mg/L or may exceed 300 mg/L depending upon the geographical area.
The average level ofNa is > 100 mg/L.
Environmental Significance
1. High Na content, in the form of chloride and sulphate, make the water salty in
taste and unfit for human consumption (i. e. unpalatable).
2. High amount ofNa wiII render the boiler operations difficult.
3. High Na concentration also leads to hypertension.
4. A water with high Na content is not suitable for agricultural use as it tends to
deteriorate the soil.
5. As such there is no standards ofNa in drinking water. But for the effluent to be
discharged on land for irrigation purpose, maximum allowable percent ofNa is
60. (However, the standard of sodium has been omitted by GSR, 80 (E), 31.12.93
MOEF - schedule - VI).
152
Slit
Phototube
LPG Gis
I. Burner unit: The main unit, consists of an atomizer (for aspiration of solution)
mixing chamber, burner, optical lens, light filter, photo-detector and control
valves.
2. Compressor unit: Compressed air from the compressor unit is applied to the
atomizer.
3. Electronic metering unit to read out readings.
4. LPG (liquid petroleum gas) or laboratory gas.
II. Working principle of flame photometer
1.
2.
3.
4.
153
REAGENTS
1. Deionised distilled water: Use deionised distilled water to prepare all reagents
and calibration standards, and as dilution water.
2. Stock sodium solution: Dissolve 2.5418 g NaCI dried at 140 "C and dilute to
1000 mL with water; 1 mL = I mg Na.
3. Intermediate sodium solution: Dilute 10 mL of stock solution with water to
100 mL; 1 mL= 100 Ilg Na. Use this intennediate solution to prepare calibration
curve in sodium range of 1 to 10 mgIL.
4. Standard sodium solution: Dilute 10 mL intennediate sodium solution with
water to 100 mL; ImL = 10 Ilg Na. Use this solution to prepare calibration
curve in sodium range of 0.1 to 1 mgIL
PROCEDURE
Pretreatment of Sample
I. For non-polluted samples, filter the sample to remove any suspended particles
which otherwise may clog the capillary of the instrument.
II. For highly polluted samples and wastewater samples, pretreatment is required.
154
7. Now feed different standard sodium solutions within the range (i.e. 0-1, 0-10
or 0-100 mg NaiL) as desired.
8. Plot a standard curve between concentration and emission of standard sodium
concentration.
9. Put off the fuel supply first followed by air and then main switch.
CALCULATION
For direct reference to the calibration curve:
mg NaIL = (mg NaIL in portion)
D = dilution ratio =
ENVIRONMENTAL SIGNIFICANCE
1. It is not present in high concentration and ranks 7th among the elements in
order of abundance and constitutes 2.4% by weight of the earth's crust. It is
never found free in nature. Most potassium minerals are insoluble.
2. Its concentration in drinking waters seldom reaches 20 mgIL. As a rule ratio of
Na to K is 10:1 or 20: 1.
3. The method of analysis is flame photometry at 770 nm.
4. It is a naturally occurring element, however concentration remain quite lower
compared to Na, Ca and Mg.
5. It has got more or less similar chemistry like Na and remains mostly in solution
without undergoing any precipitation.
SELECTION OF METHODS
APHA (1995) recommend four methods for the determination of potassium. All
are rapid, sensitive and accurate, and selection depends on instrument availability
and analyst choice.
1. Atomic absorption spectrometric method
2. Inductively coupled plasma spectroscopy method
3. Flame photometric method
4. Potassium-selective electrode method.
ISS
INTERFERENCE
Interference may occur at sodium to potassium ratios of 5: 1 or greater. Calcium
may interfere if calcium-to-potassium ratio is 10: 1 or more. Magnesium begins to
interfere when the magnesium-to-potassium ratio exceeds 100: 1.
APPARATUS
Flame photometer
REAGENTS
1. Deionised distilled water: Use this water for preparing all reagents and calibration standards, and as dilution water.
2. Stock potassium solution (1.907g1L): Dissolve 1.907 g of dry (11 OOC) KCI in
1000 mL of distilled water; 1 mL = 1 mg K.
3. Intermediate potassium solution: Dilute IOmL of stock K solution with water
to 100 mL. ImL = IOOllg K. Use this intennediate solution to prepare calibration curve in K range of 1 to 10 mgIL.
4. Standard potassium solution: Dilute 10 mL intennediate potassium solution
with water to 100 mL; I mL = 10 Ilg K. Use this solution to prepare calibration
curve in K of 0.1 to 1 mgIL.
To minimize potassium pickup, store all solutions in plastic bottles.
S.
3.30 Sulphate
Drinking water
USEPA, SMCL (1974) = 2S0 mgIL
Canadian MAL (1987) = 500 mgIL
WHO (1984) = 400 mgIL (aesthetic quality)
coni.
156
...cont.
ICMR (1963) = 200 mWL - 400 mWL (max. pennissible)
IS: 10500 (1991) = 200 mWL (max) - 400 mWL (max pennissible in the
absence of alternate source)
MWH (1975) = 200 mWL (acceptable - 400 mWL (cause of rejection)
Effluent standards
MOEF (1993) = 1000 mWL (maximum limit) is for discharge in inland
surface water, p~blic sewer and water for irrigation.
Introduction
1. The sulphate ion is one of the major anions occurring in natural waters.
2. It is of importance in public water supplies because of its cathartic effect upon
humans when present in excessive amounts.
3. Sulphate is one of the least toxic anions and WHO (1984) does recommend
guideline value of 400 mWL in drinking water. However, catharsis, dehydration
. and gastrointestinal irritation have been observed at high concentration in
drinking water and WHO therefore suggests the health authorities should be
notified when concentration of sulphate in drinking water exceeds 500 mWL.
4. Waters with about 300-400 mWL sulphate have a bitter taste and those with
1000 mWL or more of sulphate may cause intestinal disorders (laxative).
Environmental Significance
Estimation of Sulphate
Sulphate can be estimated by either gravimetric or turbidimetric methods (APHA,
1995)
105C
Principle
157
I. Drying oven, equipped with thermostat control Muffle furnace with heat
indicator.
2. Desiccator, Crucible
3. Analytical balance, capable of weighing upto 0.1 mg.
4. Filter paper: Acid -washed, ash-less hard-frnish filter paper sufficiently retentive
for frne precipitates.
Chemicals and Reagents
Chemicals
1. Methyl red indicator
2. Hydrochloric acid, HCI
3. Conc. HN03
4. Barium chloride, BaClz
5. Silver nitrate, AgN03
Reagents
1. Methyl red indicator solution: Dissolve 100 mg methyl red-sodium salt in
distilled water and dilute to 100 mL.
2. Hydrochloric acid, HCI 1+1: Dilute conc. HCI with distilled water in 1: 1
ratio.
3. Barium chloride solution: Dissolve 100 g BaCI2' 2Hp in 1000 mL of distilled
water. Filter through a membrane filter before use. 1 mL ofthis reagent is capable
of precipitating approximately 40 mg S04'
4. Silver nitrate-nitric acid reagent: Dissolve 8.5 g AgN03 and 0.5 mL conc
HN03 in 500 mL distilled water.
158
Procedure
I. Removal of silica
1. If the sample contains more than 25 mgIL silica, it has to be removed prior to
the sulphate analysis. Forremoval ofsilica, evaporate suitable volume of sample
near to dryness in a platinum dish on a water bath. Add 1 mL HCI , tilt, and
rotate dish until the acid comes in complete contact with the residue. Continue
evaporation to dryness. If organic matter is present dry in hot air oven at
180C. Moistened the residue with 2 mL of distilled water and 2 mL of HCl.
Transfer the residue in hot water and filter it. Wash insoluble silica with several
small portions of hot distilled water. Combine filtrate and washings. Discard
residue.
II. Precipitation of barium sulphate (BaSOJ
1. Adjust the volume of clarified sample (treat if necessary to remove interfering
substances) which contains approximately 50 mg sulphate ion in a 250-mL
volume.
2. Acidity with HCI to pH 4.5 to 5.0, using a pH meter or orange colour of methyl
red indicator.
3. Then add an additional 1 to 2 mL HCl.
4. Heat the solution to boiling and with stirring gently, and warm BaC~ solution
slowly precipitation appears to be complete; then add 2 rot in excess.
5. If the amount of precipitate is small, add a total of SmL BaC~ solution.
6. Digest the precipitate at 80 to 900c, preferably overnight but not less than 2 h.
IlL Preparation of filter
Place a filter on a watch glass and dry in a conventional oven of 103 to 105C. Cool
in a Desiccator and weigh the filter paper.
Iv. Filtration and weighing
1. Mix a small amount of ash-less filter paper pulp with the BaSO. and filter at
room temperature. The pulp aids filtration and reduces the tendency of the
precipitate to creep.
2. Wash the precipitate with small portions of warm distilled water until the
washings are free of chlorides, as indicated by testing with silver nitric acid
reagent.
3. Dry the filter with precipitate by the same procedure used in preparation of
filter. Cool in a desiccator and weigh.
Calculation
mg BaSO. x 411.5
mgIL SO.
mLsample
159
Principle
Sulphate ion (SO;-) is precipitated in an acetic acid medium with barium chloride
(BaCI 2) so as to form barium sulphate (BaSO4) crystals of uniform size.
BaCI2 (excess) + SO:- --+ BaS04 (precipitate)
The crystal formation is enhanced in the presence of an acetic acid buffer solution containing magnesium chloride, potassium nitrate, sodium acetate and acetic
acid.
Light absorbance of the BaSO4 suspension is measured by a photometer and
sulphate concentration is determined by comparison of the reading with a standard
curve. Minimum detectable concentration is approximately 1 mg sulphate IL.
Interference
Apparatus
1. Magnetic stirrer: Use a constant stirring speed. Use magnets ofidentica1 shape
and size. The exact speed of stirring is not critical, but keep it constant for each
run of samples and standards and adjust it to prevent splashing.
2. Nephelometer or Spectrophotometer (use at 420 nm, providing a light path of
2.5 to 10 cm).
3. Stopwatch.
4. Measuring spoon, capacity 0.2 to 0.3 mL.
Reagents
160
Procedure
1. Formation of barium sulphate turbidity
Calculation
mg SO~- x 1000
mg SO~-IL = - - - - - mLsample
161
3.31 Sulphide
(Iodometric Method)
Drinking water
WHO, (1984) = No guideline value set for drinking water, however, it should
not be detectable by consumers
IS: 10500 (1991) = No guideline value set for drinking water.
Effluent discharge
MOEF (1993) = Max 2mgIL and 5mgIL has been set for discharge to inland
surface water and marine coastal areas respectively
Introduction
1. Sulphide is often present in groundwaters, especially in hot springs.
2. Its common presence in wastewater comes partially from the decomposition of
organic matter, sometimes form industrial wastes, but mostly from the bacterial
reduction of sulphate.
3. Hydrogen sulphide escaping into the air from sulphide-containing wastewater
causes odor nuisance. The threshold odour concentration ofH2 S in clean water
is 0.025 and 0.25 mgIL.
162
Environmental Significance
1. Gaseous H2S is very toxic in sewers.
2. H2S attacks metals directly and indirectly and causes serious corrosion of
concrete sewers.
3. Dissolved H2S is toxic to fish and other aquatic organisms.
Estimation of Sulphide
PRINCIPLE
In acidic medium excess iodine is added which oxidizes H2S. The remaining iodine
is titrated against standard thiosulphate solution using starch as an indicator.
H2S + 12 ~ 2W + 21- + S
12 + Sp:' ~ 21- + S.o:
(lmL 0.025N 12 = 0.4 mg H2S)
Hydrochloric acid
Iodine
Starch
Potassium iodide, KI
Sodium thiosulphate, Na2SPr5HP
Formaldehyde as preservative
Reagents
1. Hydrochloric acid; Hel, 6N: Dilute conc. HCl (I2N) two times
2. Standard iodine solution, O.025N: Dissolve 20 to 25 g KI in a little water and
add 3.2 g iodine. After iodine has dissolved, dilute to 1000 mL and standardize
against 0.025N Na2Sp) using starch solution as indicator.
3. Standard sodium thiosulphate solution (O.02SN): Dissolve directly 6.025 g
Na2Sp).5Hp in a previously boiled 1L cooled distilled water, which will give
0.025N. Add 1.5 mL 6N NaOH or 0.4 g solid NaOH per litre . . Store in
brown bottle. This solution will have to be standardised against standard
potassium dichromate (K2Crp,) solution for each set of titration.
163
O02SN
Stlnd ..d
HI,S, OJ
/1
~O.02S
CALCULATION
1 mL 0.025 N iodine solution reacts with 0.4 mg S2-
[(A
B)-(C
D)]
16
1000
164
3.32 Thiocyanate
Drinking water: No guideline value set (IS: 10500, 1983, 1991; WHO, 1984,
1993).
Emuent discharge: No guideline value set (MOEF, 1993).
Environmental Significance
Many natural waters contain thiocyanate from organic decomposition products and
wastewater discharges. Some industrial waste, such as those from the steel industry, petroleum refining, and coal gasification, may contain significant concentration of thiocyanate. Thiocyanate is not recognised as a toxic chemical compound.
However, when chlorinated, thiocyanate is converted to the highly toxic and volatile cyanogen chloride.
Estimation of Thiocyanate
This test method covers the detennination of dissolved thiocyanate in water, wastewater, and saline water in the range from 0.1 to 2.0 mgIL. For higher concentrations, use an aliquot from the diluted sample.
PRINCIPLE
1. Thiocyanate reacts with ferric ions at a pH of <2. to fonn a coloured complex
which is detennined colorimetrically at 460 om and adheres to Beer's Law.
2. Industrial waste may be highly coloured and contain various organic compounds
which must be removed by absorption on macroreticular resin prior to analysis.
PRECAUTIONS
Many samples contain cyanide. Because of the toxicity of cyanide, great care must
be exercised in its handling. Acidification of cyanide solutions produces toxic
hydocyanic acid (HCN). All manipulations must be done in the hood so that any
HCN gas that might escape is safely vented. Residual sample remains could be
toxic hence these should be disposed off properly.
SAMPLING
I. Thiocyanate is stable in both the acid and alkaline pH range.
2. If the sample is to be preserve for cyanide, remove the sulfide before stabilisation
at a high pH. Cyanide can be converted into thiocyanate in the presence of
sulfide at a high pH.
3. Thiocyanate is biodegradable. Samples that may contain bacteria should be
preserved at pH < 2 by the addition of mineral acid and refrigerated.
165
APPARATUS
I. Acetone
2. Hexane
3. Methyl alcohol
4. KSCN
5. Macroreticular resin, 18 to 50-mesh or equivalent
6. Ferric nitrate solution
7. HPz solution
8. Conc HN03 , sp. gr. 1.42
9. NaOH
Reagents
166
d. Pour off methanol and add 3 times the resin volume ofO.IN NaOH. Stir for 15
min.
e. Pour offNaOH solution and add 3 times the resign volume ofO.IN HN03 Stir
for 15 min.
t: Pour off HN03 solution and add 3 times volume of distilled water. Stir for 15
min. Drain excess water and use purified resin to fill the column. Store excess
purified resin after converting it with distilled water. Keep in a closed jar.
PROCEDURE
Preparation of calibration curve
l. Prepare a series of thiocyanate standards containing 0.0 to 2 mg SCN-IL by
pipetting 0 (blank) to 40 mL aliquots of standard thiocyanate solution into
200 mL volumetric flask. Dilute to volume with water and mix thoroughly.
167
APPARATUS
Secchi disk attached to a string
The disc is made of rigid plastic or metal, but the details
of its design are variable. It may be 20 to 30 cm or
even larger in diameter and is usually painted white.
Alternatively, it may be painted with black and white
quadrants. The disc is suspended with a light rope or
chain so that it remains horizontal when it is lowered
Fig. 3.28: The Secchi disc. into the water. The suspension rope is graduated at
intervals of 0.1 and 1 meter from the level of disc
itself and usually does not need to be more than 30 m length. A weight fastened
below the disc helps to keep the suspension rope vertical while a measurement is
being made. Fig. 3.28 shows a typical secchi disc. The same size and pattern of
disc should be used at any given sampling station so that a series of measurements
made over a number of years are free as possible from distortions arising from
differences in apparatus.
INTERFERENCES
Suspended algae, microscopic animals, suspended matter (silt, clay and organic
matter) and water-colour are factors that interfere with light penetration into the
water column and reuce Sec chi disc transparency.
APPLICATION
Combined with other field observations, Secchi disc reading may furnish
informations on:
1. Suitable habitat for fish and other aquatic life;
2. The lake's water quality and esthetics;
3. The state of the lake's nutrient enrichment; and
4. Problems with and potential solutions for the lake's water quality and recreational
use impairment.
168
PROCEDURE
1. The observation should be made early in the morning or late in the afternoon.
2. Lower the Secchi disc, where possible, through a shaded area of water surface.
As the disc is lowered, note the depth at which it just disappears from view.
Note the point on string at upper surface of water (L.).
3. Lower the disc a little further, then lift up slowly till it just reappears. Note the
point on the string at upper surface of water (L2). Calculate as follows:
L. +L2
Secchi disc Transparency = - - 2
REPORTING
The report must also state the diameter of the disc and the pattern, if any, on the
upper surface of the disc.
Sec:chi disc (SO) transparency value is used to state the lake tropic status index
(TSJ) as below.
Lou tropic state
Oligotropic
Mesotropic
Eutropic
Hypereutropic
SD transperancy, m
<4
2-4
0.5-2
<0.5
Chl-a (pg/L)
<2.6
2.6-7.2
7.2-55.5
> 55.5
Total P (pg/L)
< 12
12-24
24-96
>96
1'81
<40
40-50
50-70
>70
The TSI can be calculated for SO transparency, Cha a (ChI) in Ilg/L and total P
(TP) in Ilg/L as follows:
On the basis of SO tranaparency; TSI = 60-14.4 In (SO)
On the basis of Chi: TSI = 9.81 In (Chi)) + 30.6
. On the basis ofTP, TSI = 14.42 In (TP) + 4.15
Example - 1
Lake monitoring shows that the Secchi disc transparency is 1.96m, total P 33 mg/L
and chi-a 3.4 mg/L. Calculate TSI using the above equation.
169
Drinking water:
WHO (1984) = 5 NTU (preferably <1 for disinfection efficiency).
WHO (1993) = 5 NTU (median turbidity <1 NTU, single sample <5 NTU)
USEPA (1974) = 1 to 5 NTU
Canada (MAC, max allowable conc, 1987) = 5 NTU
IS: 10500 (1983) = 10 NTU (max);
IS: 10500 (1991) = 5 NTU (above 5 NTU consumers acceptance decreases),
10 NTU (maximum permissible in the absence of alternate source.
MHW (1975) = 5 NTU (acceptable) - 25 NTU (cause of rejection).
Emuent sample
MOEF (1993) = No guideline value set.
Sources of Turbidity
Turbidity is caused by wide variety of suspended and colloidal materials. Run-off
from barren areas during rain is the most natural contributor of turbidity, particularly
silt and clay. The discharge of untreated industrial and domestic effluents also adds
great quantities of turbidity. Organic material reaching water bodies serves as food
for bacteria, resulting the enhancement of bacteria and other microorganisms feed
upon bacteria. Effluents from sewage treatment plants, rich in nitrogen and
phosphorus, as well as agricultural run-off stimulate growth of algae and also
contribute to turbidity.
Environmental Significance
Turbidity is an important consideration in public water supplies for three major
reasons Aesthetics - turbidity in drinking water is automatically associated with possible
wastewater pollution.
Filterability - Filtration of water is rendered more difficult and costly when
turbidity increases.
Disinfection - many ofthe pathogenic organisms may be encased in the particles
and protected from the disinfectant.
Estimation of Turbidity
PRINCIPLE
Turbidity may be defmed as interference to the passage of light by suspended particles in water. It can be measured by its effect on the transmission of light, which is
termed as Turbidimetry or by its effect on the scattering oflight, which is termed as
170
Nephelometry (Fig.3.29). Turbidimeter can be used for sample with moderate turbidity and Nephelometer for samples with low Turbidity. Higher the intensity of
scattered light higher the turbidity.
The original standard chosen for turbidity was silicon dioxide (Si02).
1 mg Si02 1L = 1 unit of turbidity.
The Si02 was originally used to calibrate the Jackson candle turbidimeter. But
Jackson candle turbidimetric unit (JTV) has been removed as standard method of
turbidity measurement in 17th edition of Standard methods (1989). The formazine
standard of 40 NTV is equivalent to 40 JTU. The lowest turbidity measured by
Jackson candle turbidimeter is 25 JTV.
Li ....
"'' .7
Sht
c:::::I
c:::o
......'.~l
Pl'Iototube
\.:.J
Tumidim.tt'r
N~ph~lorn~
.r
INTERFERENCES
Dissolved material that imparts a color to the water may cause serious errors in
nephelometric readings. Floating or suspended large particles and entrained air
bubbles will give false or unstable readings. Certain turbulent motions also create
unstable reading conditions of nephelometer.
PRECAUTIONS
Scratches, fmger marks, or dirt on the wall of the sample cell may give erroneous
readings. Cells should be kept scrupulously clean both inside and outside and discard
when their is a scratch.
STORAGE OF SAMPLE
Determine the turbidity on the day the sample is taken. If this is not feasible, store
the sample in the dark for up to 24 h and refrigerate at 4"C if possible, but do not
freeze. Prolonged storage is not recommended because of irreversible changes.
PREPARATION OF SAMPLE
Bring the sample to room temperature and shake sample vigorously for at least one
.min. Let the sample stand 2 to 3 min to allow air bubbles to disappear, then gently
invert the sample several times or swirl, mix before examination.
171
PROCEDURE
Distilled water, mL
97.5
100
2.5
95
100
5
100
10
90
100
12.5
87.5
100
15
85
80
100
20
100
75
25
2. Calibration o/instruments (Fig. 3.30):
NTU instrument reading
NTU standard value
10.010.0
8.1
8.0
6.0
4.0
2.0
-Instrument is adjusted to read this value.
6.3
4.7
2.8
1
2
4
5
6
8
10
172
10r-----------------~
o~~~~~~~~~~
2
4
6
3. Turbidity measurement ofsample: Gently agitate the sample and wait till all air
bubbles disappear.
4. The Nephelometer reading is noted and NTU value is determined from the
standard curve (Fig. 3.30).
5. If turbidity is exceeded 40 NTU, dilute the sample. In such case, apply correction
factor. (for example, if the sample is diluted to 10 times and the Nephelometer
reading is 9.5 NTU, the turbidity of unknown solution is 9.5 x 10 = 95 NTU).
Turbidity Summary
Caused by: Suspended and colloidal matter such as clay, silt, organic and
ganic particulate, plankton and other micro-organisms.
inor~
Concentration expected: From almost zero (0.05 NTU) in distilled water to over
1000 NTU in highly turbid water.
Indicator of: Necessity of treatment, potential contamination by pathogen; poor
treatment plant efficiency - Coagulant dosage, filter run timing, contamination in
distribution system.
Test: A physical and microbiological parameter, simple, inexpensive, mandated
by public health regulation; expressed in the Nephelometric turbidity unit (NTU).
NTU: Turbidity unit measurd by Nephelometric standard method.
Regulation: 0.1 NTU as a goal; less than INTU as a Standard; 5 NTU as an
exception for potable water.
173
Analysis of Metals in
Water and Effluents
4.1 Introduction
HEAVY METALS
The kidney contains million of excretory units called nephrons. Chemicals that are
toxic to the kidney are called nephrotoxins. Metals like Cd, Pb and Hg are
nephrotoxins.
1. Heavy metals: Cd, Cr, Cu, Mn, Hg, Pb, Ag, Zn
2. Non-metallic: As, Se
3. Others: Na, K, Ca, Mg, Fe
IS 10.500
(1991)
WHO
(1984)
1. Arsenic
0.05 (no
relaxation)
0.05
0.05
0.05 (may be
relaxed upto
1.5)
1.0
(permissible)1.5
(excessive)
1.00"
(As)
2. Copper
(Cu)
USEPA
(1974)
Remarks
Cont...
175
0.005
0.01
Kidney effect.
0.05
0.05
Liver/kidney effect.
0.3
0.30
Taste/appearance are
affected.
0.05
0.05
0.1
0.05
Hardness,taste,
gastrointestinal irritation
in presence of sulfate.
0.001
0.002
NVS
NVS
0.05
0.01 (no
relaxation)
5 (may be
extended upto
15)
0.01
0.01
Skin
discoloration
(Argyria).
Gastrointestinal effect.
5 (permissible)
Desirable.
(Ag)
10. Selenium
(Se)
11 . Zinc (Zn)
Inland
surface
water
Public
sewers
Discharge on
Land for
irrigation
Marine
coastal
areas
1.
2.
3.
4.
5.
6.
0.2
2
3
2
0.1
2
0.2
2
3
1
2
2
0.2
2
0.2
3
2
1
2
Cont...
176
... Cont.
7. Iron (as Fe)
S. Lead (as Pb), Max.
9.Manganese (an Mn)
10.Mercury (as Hg), Max.
11. Nickel (as Nil, max.
12. % sodium (% Nal, max
13.Selenium (as Se), Max.
14. Zinc (as Znl, Max.
15. Vanadium (as V)
3
0.1
2
0.Q1
3
3
1
2
0.01
3
0.05
5
0.2
0.05
15
0.2
3
2
2
0.01
5
60
0.05
15
0.2
Boron,Nickel and % Sodium have been omitted by G.S.R. SO(EI, dated 31st
December,1993 (w.e.f. 31.12.1993).
% Na =
Ca
Na x 100
+ Mg + Na + K
~m
SUSPENDED METALS
177
ACID-EXTRACTABLE METALS
1. Hot plate
2. Conical (erlenmeyer) flasks, 125-mL, or Griffin beakers, 150-mL, acid-washed
and rinsed with water.
3. Volumetric flasks, 100-mL.
REAGENTS
1. Mix samples and transfers a suitable volume (50 to 100 mL) to a beaker. Add 5
mL conc. HN03 , cover with glass.
2. Bring to a slow boil and evaporate on a hot plate to the lowest volume possible
(about 10 to 20 mL) before precipitation occurs.
3. Continue heating and adding conc. HN03 as necessary until qigestion is complete as shown by a light-coloured, clear solution.
4. Do not let the sample dry during digestion. Wash down the walls of the beaker
and watch -glass with distilled water and then filter.
5. Transfer the filtrate to a 100 mL volumetric flask, cool and makeup the volume.
Use portions of this solution for required metal analysis.
1. Hot plate
2. Conical (erlenmeyer) flasks, 125-mL, or Griffm beakers, 150 mL, acid-washed
and rinsed with water.
3. Volumetric flasks, 100-mL.
REAGENTS
Samples are acidified with HN03 and HCI and heated on a hotplate to reduce the
volume to a defmed level. After filtration, the samples are ready for analysis by
atomic absorption spectrophotometer (AAS).
178
1. Measure 100 mL ofa well-mixed sample into a ISO mL beaker or flask. Add
0.5 mL of conc. HN03 (sp. gr. 1.42). (Note: If the sample has been preserved at
the recommended level of 5 mL of conc. HN03 per liter of sample, the addition
51it-s
Chopper
R.adout
Monothromator
---L:J---
. . "'"" . .-'-.- D-
o.t.ttor
Oxidiz.r Gas
Samplf Cup
Fig. 4.1 : A basic Atomic Absorption Spectrometer (AAS)
179
Element
Wavelength (nm)
Instrument detection
limit (mg/L)
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
Silver (Ag)
Aluminum (AI)
Calcium (Ca)
Cadmium (Cd)
Cobalt (Co)
Chromium (Cr)
Copper (Cu)
Iron (Fe)
Magnesium
Manganese (Mn)
Molybdenum (Mo)
Sodium (Na)
Nickel (Ni)
Lead (Pb)
Tin (Sn)
Zinc (Zn)
328.1
309.3
422.7
228.8
240.7
357.9
324.7
248.3
285.2
279.5
313.3
589.0
232.0
283.3
224.6
213.9
0.01 (A-Ac)
0.1 (N-Ac)
0.003 (A-Ac)
0.002 (A-Ac)
0.03 (A-Ac)
0.02 (A-Ac)
0.01 (A-Ac)
0.02 (A-Ac)
0.0005 (A-Ac)
0.01 (A-Ac)
0.1 (N-Ac)
0.002 (A-Ac)
0.02 (A-Ac)
0.05 (A-Ac)
0.8 (A-Ac)
0.005 (A-Ac)
A-Ac
180
4. Chromium (Cr) (I mL = 100 "g Cr): Dissolve 0.1923 g crOJ in water, dissolved completely, acidify with 10 mL conc. HNOJ and dilute to 1 L.
Chromium stock solution (I mL = 1 mg Cr): Dissolve 2.828 g of primary
standard potassium dichromate (K2Crp7) in 200 mL of water and dilute to 1 L.
Chromium solution, standard (I mL = 0.1 mg Cr): Dilute 100 mL of Cr
stock solution and 1 mL conc. HNOJ to 1 L with water.
5. Copper (Cu) (ImL = 100 "g Cu): Dissolve 0.1 g copper metal in 2 mL conc.
HNOJ , add 10 mL conc. HNOJ and dilute to 1 L with water.
6. Iron (Fe) (ImL = 100 mg Fe): Dissolve 0.1 giron wire in a mixlure ofl0 mL
(1+1) HCI and 3 mL conc. HN0 3. Add 5 mL conc. HN03 and dilute to 1 L.
7. Lead (Pb) (I mL = 100 "g Pb): Dissolve 0.1598 g lead nitrate, Pb(N03)2' in a
minimum amount of (1 + 1) HN0 3, add 10 mL conc HN03, and dilute to 1 L
with water.
8. Manganese (I mL = 100 "gMn.): Dissolve O.1gMn metal in 10 mLconc HCI
mixed with 1 mL conc HNOJ Dilute to 1 L with water.
Manganese solution, stock (I mL = ImgMn): Dissolve 3.076 g of Manganous sulfate monohydrate (MnS04 HP) in a mixture of 10mL of conc. HNOJ
(sp.gr. 1.42) and 100 mL of water. Dilute to 1 L with water.
Manganese solution, standard (I mL = O.lmg = 100 "g Mn): Dilute 100
mL ofMn stock solution to 1 L with water.
9. Silver (Ag) solution, stock (I mL = 100"gAg): Crush approximately 2 g of
silver nitrate (AgN03) crystals and dry to constant mass at 40C. Dissolve
0.1575 g of AgN0 3 in water containing 5 mL of conc HNOJ dilute to 1 L. Store
in a amber glass bottle.
Silver (Ag) solution, intermediate (I mL = I"g Ag): Dilute 10 mL of stock
silver solution and 5 mL of conc HNOJ to 1 L with water. Store in an amber
glass bottle.
Silver solution, Standard (1 mL = 0.1 "g Ag): Dilute 100 mL of silver intermediate solution and 5 mL of conc. HN03 to 1 L with water. Prepare fresh
before use.
10. Zinc (Zn) ( 1 mL = 100 "g Zn): Dissolve 0.1 g zinc metal in 20 mL 1+ 1 HCI
and dilute to 1000 mL with water.
Zinc stock solution (1 mL = 1 mg Zn): Dissolve 1.245 g of Zinc oxide (ZnO)
in a mixture of 10 mL conc. HNOJ and 10 mL of water. Dilute to 1 L of water.
Zinc solution, standard (1 mL = 100"g Zn): Dilute 100 mL ofZn stock solution and 1 mL ofconc. HN0 3 to 1 L of water.
The Arsenic (As) and Selenium (Se) are analyzed by hydride generation atomic
absorption spectrometric method.
B
x --
181
Where,
A = Concentration of metal in digested volume, mgIL
B = Final volume of digested solution, mL and
C = Sample size, mL.
Report the results as follows: (Solid sample, i.e soil,sludge, etc)
AxB
Metal concentration, mg/kg (wet - weight basis) = - - g sample
Where,
A = Concentration of metal in digested volume, mgIL
B = Final volume of digested solution, mL
Drinking water
IS: 10500 (1991) = 0.03 mgIL (0.2 mgIL is permissible in the absence of
alternate source).
WHO (1993) = 0.2 mgIL
EC (1980) = 0.05 mgIL (guide level) and 0.2 mgIL (MAC, max admissible
concentration).
Emuent discharge
MOEF, (Schedule VI, 1993) = No Standards.
There is little information available regarding the toxicological effects of aluminum of human health. Al is ljI1onitored in boiler make-up water, where alum has
been used, to determine whether aluminum is present after pretreatment. Residual
Al may consume ion exchange capacity. Al is monitored in cooling water make-up,
since its presence may result in deactivation of anionic substances in scale or corrosion inhibitor treatment chemicals or both.
Estimation of Aluminium
1. AAS, direct - 0.3 mgIL to 5.0 mgIL
2. AAS, Chelation-extraction - 0.01 mgIL (10 flgIL) to 0.3 mgIL (300 flgIL)
3. Colorimetric: Eriochrome Cyanine R method.
182
4.8 Arsenic
Drinking water
WHO (1984); USEPA (1974); EC (1980) = 0.05 mgIL
WHO (1993) = 0.01 mgIL.
IS:I0500 (1991); ICMR (1963); MHW, (1975) = Indian standard: 0.05 mgIL
Effluent discharge
MOEF, Schedule VI (1993) = 0:2 mgIL max. (for inland surface water, public
sewers, land of irrigation and marine/costal areas).
Herbicides, insecticides, and many industrial effluents contain arsenic and are potential sources of water pollution. Arsenic is significant because ofits adverse physiological effects on human.
Estimation of Arsenic
Preserve the sample with HNO) (sp. gr. 1.42) to a pH of2 or less immediately at the
time of collection, normally about 2 mLIL is required. If only dissolved arsenic is
to be determined, filter the sample through a 0.45 J,lm membrane filter before acidification . Arsenic in most waters and wastewater is determined by three test methods as given in Standard Methods (1995):
1. Colorimetric silver diethyldithio carbamate: 5 to 250 Ilg As/L
2. AAS, Hydride generator (I to 20 Ilg As/L)
3. AAS, Graphite furnace (5 to 100 Ilg As/L).
HYDRIDE GENERATION METHOD
Principle
Organic arsenic containing compounds are decomposed by adding sulfuric and
nitric acid and repeatedly evaporating the sample to fumes of sulfur trioxide .
The arsenic (V) so produced, together with inorganic arsenic originally present,
is subsequently reduced to arsenic (III) by potassium iodide and stannous chloride,
and finally to gaseous arsine by zinc in HCI solution.
Alternatively, the arsenic is converted to arsine by sodium borohydride (NaBH 4 )
in HCI solution. The arsine is removed from solution by aeration and swept by a
flow of nitrogen into a hydrogen flame where it is determined by AAS at 193.7 nm.
Determination of Arsenic with sodium borohydrate
I. Clean all glassware before use by rinsing first with hot HNO) (1 +.1) and then
with water.
2. Pipette a volume of well-mixed acidified sample containing less than 1.0 Ilg of
3.
4.
5.
6.
7.
8.
183
arsenic (50 mL maximum) into a 200-mL Berzelius beaker (or similar apparatus) and dilute to approximately 50 mL.
To each beaker, add 7 mL~SO,,(1 + 1) and 5 mLofconc. HN03 Add a small
boiling chip and carefully evaporate to fumes ofS03, maintaining an excess of
HN03 until all the organic matter is destroyed. This prevents darkening of the
solution and possible reduction and loss of arsenic. Cool, add 25 mL of water,
.
and again evaporate to fumes ofS0 3, to expel oxides of nitrogen.
Cool, and adjust the volume in each beaker to approximately 50 mL with water.
Add 8 mL conc. HCI and mix.
Attach one beaker at a time to the rubber stopper containing the gas dispersion
tube.
Fill the dropper or syringe with 0.5 mL of sodium borohydrate solution (Dissolve 4 g of NaBH4 in 100 mL of water. Prepare fresh before each use) and
insert into the hole in the rubber stopper.
Add the sodium borohydrate solution to the sample solution. After the absorbance has reached a maximum and has returned to the baseline remove the
beaker. Rinse the gas dispersion tube with water before proceeding to the next
sample. Treat each succeeding sample, blank, and standard in a like manner.
Removal of As
There are following treatment options under evaluation for treatment of arsenic
(De Zuane, 1977). These are:
Lime softening
Activated alumina
Reverse osmosis
Electrodialysis
Drinking water
WHO (1971) = 0.01 mgIL
WHO (1984); EC (1980); Canadian MAL; (i987) = 0.005 mgIL
WHO (1993) = revised to 0.003 mgIL
IS: 10500 (1991); ICMR (1963); MHW (1975) = 0.01 mgIL
Emuent discharge
MOEF (Schedule VI, 1993) = 2 mgIL (Inland surface and marine/coastal
areas) and 1 mgIL public sewer.
184
Estimation of Cadmium
1. AAS, Direct - 0.05 to 2 mgIL
2. AAS, Chelation-extcaction: 5 to 200 J.lg/L
3. AAS, Graphite furnace: 0.5 to 10 J.lgIL
4. Colorimetric: Dithizone method.
Drinking water
IS: 10500 (1991) = 0.05 mgIL (may be carcinogenic above this limit)
ICMR (1963); MHW (1975) = 0.05 mgIL.
WHO (1984); WHO (1993); EC (1980); USEPA (1974) = 0.05 mgIL
Emuent discharge
MOEF (l993) = 2 mgIL Total Cr (max) (as Cr) - public sewers; inland surface
water; and marine/costal areas.
Hexavalent chromium (VI) salts are used extensively in metal finishing and plating
application, in anodizing aluminum, and in the manufacture of paints, dyes, explosives and ceramics. Cr (VI) is toxic to human, animals and aquatic life. It can
produce lung tumor when inhaled and readily induces skin sensitization. There is
no evidence to indicate that Cr (III) form is detrimental to human health.
Estimation of Chromium
1. AAS, Direct - 0.1 to 10 mgIL
2. AAS, Graphite-furnace: 5 to 100 J.lglL
3. Colorimetric Diphenylcarbohydrazide method - 0.01 to 0.5 mgIL
185
Drinking water
IS: 10500 (1991) = 0.05 mgIL (may be relaxed up to 1.5 mgIL in the absence
of alternate source).
ICMR (1963) = 1 mgIL (desirable) - 3 mgIL (max. permissible).
WHO (1984, 1993) = 1 mgIL (health -based provisional guideline value is
2 mgIL).
USEPA (1974) = I mglL.
EC (1980) = 0.1 mgIL (at plant) and 3 mgIL at supply point. Above 3 mgIL,
astringent taste, discolouration, and corrosion may occur.
Emuent discharge
MOEF (1993) = 3 mgIL (Inland surface water, public sewer and marine coastal
areas).
Copper salts are used in water supply systems to control biological growth in reservoirs and distribution pipes and to catalyze oxidation of manganese. Corrosion of
copper-containing alloys in pipe fitting may introduce measurable amounts of copper into the water in a pipe system.
In small amount, it is not detrimental to health, but it will impart an undesirable
taste to the water. It causes Wilson's disease. Copper is essential to human, the
adult daily requirement has been estimated at 2 mg.
Estimation of Copper
1.
2.
3.
4.
186
... cont.
USEPA (1974) = 0.3 mgIL
Effluent discharge
MOEF (l993) = 3 mgIL (for inland surface water, public sewers and marine
costal areas).
Surface water generally contain < 1 mgll of Fe. Some groundwaters contain much
higher level of Fe. Water containing Fe >2 mgIL, causes staining of clothes (while
washing) and sanitary wares and imparts bitter astringent taste. Taste and odour
problems may be caused by filamentous organisms that prey on iron compounds
(Frenothrix, Galionella and Laptothrix are called "iron bacteria") originating another consumers objections (red water).
It is a known fact that iron in trace amounts is essential for nutrition. In case of
Fe deficiency anaemia, larger doses are taken for therapeutic reason. The daily
requirement is 1-2 mg, with dietary ranges of 7-35 mglday, with an average of 16
mglday. Fe from ambient air constitutes a negligible exposure. With a daily food
contribution of 16 mg and water contribution of 0.14 mg, it is clear that water
contributes to the diet between less than 1-15%.
Estimation of Iron
1.
2.
3.
4.
~gIL
IRON REMOVAL
Fe removal can be accomplished using aeration, sedimentation/filtration with pH
adjustment, ozone or chlorine treatment, Chemical precipitation/filtration, ion exchange (softening), potassium permanganate oxidation with precipitation/filtration,
well water pretreatment with sequestering agents like polyphosphate and sodium
silicates and lime softening.
4.13 Lead
Drinking water
IS: 10500 (l991) = 0.05 mgIL (maximum); no relaxation.
WHO (1971) = 0.1 mgIL
WHO (1984) = 0.05 mgIL
WHO (1993) = 0.01 mgIL
cont..
187
... cont.
WHO (European, 1984) = 0.10 mgIL - 0.05 mgIL (max. for running water);
Netherlands and Germany (in pipes) = 0.3 mgIL (water still for 24h).
USEPA (1974) = 0.05 mgIL.
Emuent discharge
MOEF (1993) = 0.1 mgIL (Inland surface water), 0.1 mgIL (public sewers) and
2 mgIL (marine/coastal areas)
Lead concentration in surface and ground raw water range from traces to 0.04
mgIL, averaging about 0.01 mgIL. Industrial and mining sources may contribute to
some localized Pb pollution. Use ofPb pipes in water supplies may provide high
Pb at consumers' points. In most cases, higher Pb concentration is expected at the
consumers tap than that of water treatment plant.
The health effects of Pb are neurotoxic and three human systems are most
affected: blood-forming system, nervous system (which include irreversible brain
damage) and renal system. Such a toxic level is reached when the blood level exceeds 100-120 JlgIL.
Estimation of Lead
1. AAS-Direct: Range 1 to 10 mg PblL. This test method is applicable for the
determination of dissolved and total recoverable Pb in most waters and wastewaters.
2. AAS, Chelation -extraction: 100 to 1000 JlgIL
3. AAS, Graphite furnace: 5 to 100 JlgIL.
4. Colorimetric: Dithizone method - 1.0 Jlg Pb/l 0 mL dithizone solution.
-
188
cont...
Emuent discbarge
MOEF (1993) = 2.0 mgIL (for Inland surface water, public sewers, land of
irrigation and marine/coastal areas.
The concentration ofMn in water is less than that of Fe. A mean expected value is
around 0.06 mgIL. Mn is an essential trace nutrient for plants and animals. Nutritional deficiency in man has not been evaluated as a health hazard. Also minimum
requirements for nutrition have not yet been established. WHO estimates an average daily requirement for normal physiological function of 3-5 mg. Undesirable
effects may be taste, staining of laundry and discoloration of water at 0.15 mgIL.
Estimation of Manganese
1. AAS, Direct: 0.02 to 5 mgIL
189
HEALTH EFFECTS
The principal target organ of inorganic Hg is kidney with neurological and renal
disturbance. Methyl mercury compounds are very toxic to the central nervous system; they are also the major source of environmental contamination. Fatal dose for
man varies between 3-30 g. Four to 12 mg of mercury per day may be safe, and
fatal doses of mercury would be 75-300 mgIL.
Estimation of Mercury
Standard Methods (1995) advises to preserve samples by treatment with HNO) to
reduce the pH to less than 2. For all samples the Cold vapour Atomic Absorption
Spectroscopy is the method of choice, with the "Dithizone method" as the method
to be used in potable waters for high levels of mercury.
FLAME LESS ATOMIC ABSORPTION SPECTROMETRY OR COLD
VAPOUR ATOMIC ABSORPTION SPECTROSCOPY
Principle
The specific technique is based on room temperature reduction of Hg to Hgo (element) by SnCll in solution, followed by sweeping ofHgo by air into an absorption
cell. In absorption cell, liberated Hg is irradiated by a low pressure mercury lamp,
which emit 253.7 nm wavelength. Now the Hg vapour, in atomic form, absorbs the
253.7 nm radiation and causes a change in transmittance, which can be correlated
to the total mercury content in the sample solution.
Chemicals and Reagents
Chemicals
1. Potassium permanganate (KMnOJ
2. Conc. HNO)
3. Stannous chloride (SnCll)
4. Potassium dichromate (KzCrl0 7)
5. Conc. HlSO.
6. NaOH pellets
7. HgCll
Reagents
I. Potassium permanganate solution (KMnOJ, 1% (w/v) ): Dissolve 5 g of
KMnO. in water and carefully add to it 50 mL of conc. HlSO. make up a volume of500 mL using distilled water. (Fill this solution in RI and R5 vessels).
2. Sodium hydroxide, (NaOH), 200/. solution: Dissolve 50 g ofNaOH pellets
in distilled water and make up a volume of 250 mL. (Fill this solution in R3
vessels and function is to absorbed acid vapour).
3. Cone. H1SO.: Filled in R4 vessels. Function is to absorb moisture.
4. SnC11 20% solution (w/v): Take 20 g ofSnCll in a clean beaker. Add 10mL
190
5.
6.
7.
8.
conc. HCl and dissolve while it over a burner. Boil for one min, cool and dilute
with distilled water to make 100 mL.
RN03 , 10% solution: Add 20 mL ofHNO) to water and make up 200 mL.
Stock mercury solution (1 mL = 1000 pg): Dissolve 0.1354 g mercuric chloride (HgCI2) in about 70 mL water, add 1 mL conc. HNO), and dilute to 100 mL
with water; ImL = Img Hg or 1000 ~g.
Intermediate stock solution: Dilute 1mL of stock mercury solution to 100 mL
with distilled water. 1mL = 10 ~g.
Standard mercury solution: Prepare a series C?f standard mercury solutions
containing 0 to 500 nanogram ImL by appropriate dilution of stock mercury
solution with water containing 10 mL conc. HNO/L (dilution water). Prepare
standards daily.
Dilution water,
mL
99.8
99.5
99
98
97
96
95
Final volume,
mL
100
100
100
100
100
100
100
~g).
Concentration of
Hg, nglmL
20
50
100
200
300
400
500
Procedure
Analytical method used with the FAAS method involves three important steps:
a. Sample preparation,
b. Vapour generation, and
c. Absorption measurement.
A typical mercury analyzer is shown in Fig. 4.2.
1. Fill the RI and R5 vessels with 1% KMn04 solution (16 to 20 mL),just sufficient enough to dip the inlet. R3 contain 4 mL of20% NaOH. R4 contain 4 mL
of conc. H2 S04 ,
2. Standards or sample solution is placed in the reaction vessel R2.
3. Add the required amount ofHNO) to maintain a volume of 10 mL + 2 mL
Snel2 solution and replace the stopper immediately (Max volume 12 mL).
4. Switch on the magnetic stirrer immediately and stir vigorously for about 5 min.
(purpose is to generate mercury vapour).
5. Pump the resulting Hg vapour through 20% NaOH solution (R3) (act as acid
trap i.e. absorb acid) and 50% H2 S04 solution (R4) (act as moisture trap) and
then absorption cell.
191
D~t~ttor
Amplifi~r
Syst~m
FUNCTION A DIAGRAM
MA 5800 DIE
to Purify Air
to Generate to Merwry Vapour from Sample Solution
R3 to Absorb Acid Vapour
Rio to Absorb Moisture
RS to Absorb (Gtnerated)twrcury Vapour
R,
R2
Signal
~ui ...l~nt
to 100 ng 01 Hg
1()
20
30
(Tim~ (5~t)
6. Measure the absorbance at 253.7 run using Hg-hollow cathode lamp as the light
source.
7. At the end of measurement absorb Hg vapour in 1% KMnO4 in 10% H2 S04 in
the outlet.
Repeat the measurement for standard 20, 50, 100, 200, 300, 400 and 500 nanogram ofHglmL. Prepare calibration graph by plotting the absorbance Vs concentration ofHg in nanogram/mL.
Note: Never fill the traps with more liquid than specified above, otherwise the
liquid may be carried over the next trap or to the absorption cell.
Start the pump and allow the air to flow through for a minute.
The RI and R5 traps should be replaced at least once in 3 days.
Clear R3 (alkali) and R4 (acid) traps and refill then with respective reagents
daily.
192
Drinking water
MOEF, Schedule VI (1993) = 0.05 mgIL (Inland surface water, public sewers
and marine/costal areas).
Selenium is a priority pollutant and all public water agencies are required to monitot its concentration. In animals Se prevent bone formation and causes "alkali disease" and "blind staggers". The problem is critical with animals because they are
dependent upon the local plants for food.
Estimation of Selenium
1. AAS, Gaseous hydride (applicable: 1 to 20 JigIL).
2. AAS, Graphite furnace (applicable: 2 to 100 JigIL).
3. Colorimetric: Minimum detectable quantity: 10 Jig SelL.
SAMPLING
When determining only dissolved selenium, filter the sample through a 0.45 J1M
membrane filter as soon as possible after sampling. Add HNOl to the filtrate to
bring the pH to < 2.
.
When determining total recoverable selenium, add HNOl to the unfiltered
sample to a pH of < 2.
PROCEDURE
:
193
Estimation of Silver
1.
2.
3.
4.
Vanadium has an atomic number of23, an atomic weight of50.94, and valances of
2, 3,4, and 5. The dominant form of vanadium in natural waters is V$+. It is consid-
194
ered as nonessential for most higher plants and animals. Vanadium complexes have
been noted in coal and petroleum deposits. Vanadium is used as a catalyst in the
production of sulfuric acid and synthetic rubber. Vanadium can be found in waste
that result from chemical cleaning of components in which the metals is alloyed.
Vanadium pentaoxide dust causes gastrointestinal and respiratory disturbances. The
United Nations Food and Agriculture Organization (UNFAO) recommended maximum level for irrigation waters is 0.1 mgIL.
Estimation of Vanadium
1. Graphite furnace: ASS: 10-200 IlgIL of V based on a 20-IlL sample size. (Minimum detection limit: 4 IlgIL).
2. Colorimetric: Gallic acid method - 21lg VIL.
3. ASS, ICP and Gallic acid methods are suitable for potable water sample, where
as ASS and ICP preferred for polluted water sample.
Zinc (Zn) is an essential and beneficial element for human bodies. However, above
S mgIL, cause bitter taste and opalescence in alkaline waters. The Zn concentration
of U.S. drinking water ranges from 0.06 to 7 mgIL with a mean of 1.3 mgIL. Zn
enters the domestic supply from deterioration of galvanized Fe and de-zincification
of brass, besides industrial waste.
Estimation of Zinc
1. AAS, Direct: O.oI to 2 mgIL
195
Colorimetric method
Aluminium (AI)
Boron (B)
Neocuproin method
Chromium (Cr)
Copper (Cu)
Curcumin method
Iron (Fe)
Dithione method
Lead (Pb)
Phenoanthroaline method
Manganese (Mn)
Silver (Ag)
Dithione method
Zinc (Zn)
Persulfate method
Vanadium (V)
Diethyldithio-carbamate method
196
19. The heavy metals are known to be nephrotoxin elements and in addition cause
diseases in nervous system in human. Match the following diseases with the
respective metal.
Metals
Types ofDiseases/symptoms
1.
2.
3.
4.
5.
6.
7.
a. Itai-Itai disease
b. Wilsons's disease
c. Argyria (darkening ofthe skin and eyes)
d. Minamata disease
e. "Alkali disease" and "blind staggers"
f. Anemia
g. Taste and staining of clothes and
discoloration of water
h. Nervous system and kidney
i. Skin disorder and lungs tumor
j. Gastrointestinal and respiratory
disturbance
k. Bitter stringent test and an opalescence
Chromium (Cr)
Cadmium (Cd)
Copper (Cu)
Iron (Fe)
Lead (Pb)
Mercury (Hg)
Manganese (Mn)
8. Selenium (Se)
9. Silver (Ag)
10. Zinc (Zn)
11. Vanadium (V)
in alkaline water.
Hints: l(i); 2(a); 3(b); 4(f); 5(h); 6(d); 7(g); 8(e); 9(c); lQ(k); 110>.
Treatabili 4y Studies of
Wastewater
1. Alum [Aluminium sulphate, AI1(S04)3.18HP1: When alwn is added in water containing Ca and Mg bicarbonate alkalinity, the reaction is:
Al2(S04)3.l8H20 + 3Ca(HCO)2 ~ 2Al(OH)3,j, + 3CaS04+ 6Hp + 18Hp
... (1)
The insoluble aluminiwn hydroxide is a gelatinous floc that settles slowly through
the water, swiping out the suspended materials. Most effective in the pH range of
5.5 to 8.0
... (3)
... (4)
4. Ferric chloride (FeCI3.6HP): Ferric chloride is used primarily in the coagulation of wastewater and industrial waste.
2FeC13 + 3Ca(OH)2 ~ 3CaCl 2+ 2Fe(OH)3,j,
... (5)
198
...(6)
Example - 1
What amount of alkalinity is required to react with 10 mg/L alum?
' Solution;
}O mg/l x (3 x 100 g.mol- 1 /666 .7 g.mol-1 ) = 4 .5 mg/L CaC0 3
.If alkalinity is le~s than 4 .5 mglL CaC0 3, then additional lime is to be added.
,.
,.. '
Example - 2
What is the amount of natural alkalinity required for coagulation of raw water
with dosage of 15.0 mg/L of ferric chloride?
Solution:
Step-l: Write the reaction equation and calculate molecular weight (MW) .
.2FeCl z+ 3Ca(HC0 3)z ~ 2Fe(OH)3 + 3CaCI z + 6CO z
2[55 .85 + (2 x 35 .45)] 3[40.08 + 2(1 + 12 +48))
253.5
486.2
The above equation suggests that 2 moles of ferric chloride react with 3 moles of
calcium bicarbonate.
Step-2: Determine the alkalinity needed for X
mg/L Ca(HC0 3 )z
~----
mg/l FeCl z
486.2
= --=
1.92
253 .5
Therefore, X
icc
199
Example - 3
Given that liquid alum is used as a coagulant. Sp . gravity of alum is 1.33. One
gallon (3 .785L) of alum weights 5.03 kg and contains 2 .44 kg of dry alum .
Determine:
(a) mL of liquid alum required to prepare a 100 mL solution of 20,000 mg/L alum
concentration.
(b) The dose concentration of 1 mL of stock solution in a 2000 mL jar sample.
Solution:
Step-l: Determine alum concentration in mg/L
Alum concentration = 2.44 kg/3.785 L = 0 .644 kg/L = 645 mg/L.
Step-2: Prepare 100 mL stock solution having 20,000 mg/L alum concentration.
Let X = mg of alum required to prepare 100mL stock solution.
X
20,OOOmg
= 2000 mg
1000mL
100mL
2000
- - - Y = 3 .10 mL (Ans. a) .
lmL
645
Step-4: Find lmL of alum concentration (Z) in 2000 mL sample (Jar sample).
(Z mg/L) (2000 mL) = (20000) (1 mL)
20000
Z = --10 mg/L. (Ans).
2000
Note: Actual final volume in the Jar after addition of alum is 2001 mL, using
2000 mL sample, which is reasonable.
Coagulant aids
Coagulant aids are used sometimes to produce tougher and better settling floes.
Some common coagulant aids are: polyelectrolytes, activated silica and bentonitic
clays.
Polyelectrolytes: These are classified according to their type of charge on the
polymer chain. There are three forms.
Cationic (Positive): Magnaflocs LTlI ; Chitosan, Zetag 63
Anionic (Negative): Setlyte AP50, EA 1533, Sufloc, Magnafloc ISS
Non-ionic (no-charge): Setyle.
Activated silica and clays: These offer advantages of increased rate of chemical
reaction, reduced coagulant dose, extended optimum pH range and production of a
faster settling and tougher floes. Activated silica is normally used with aluminium
coagulants at a dosage between 7 and II percent of alum dose, expressed as mgIL
ofSi0 2
Bentonitic clays: These have been used as coagulant aids in conjunction with Fe
and Al primary coagulants in treating waters containing high colour, low turbidity
and mineral content.
200
This practice covers a general procedure for the evaluation of a treatment to reduce
dissolved, suspended, colloidal, and non-settleable mater from water/effluent by
chemical coagulation- flocculation, followed by gravity settling. The procedure
may be used to evaluate colour, turbidity, and hardness reduction.
This practice permits the evaluation of various coagulants and coagulant aids
used in the treatment of water and wastewater.
The effects of concentration of the coagulants and coagulant aids and their
order of addition can also be evaluated by this practice.
PRINCIPLE
The coagulation-flocculation test is carried out to determine the chemicals, dosages, and conditions required achieve optimum results. The primary variables to
be investigated using the recommended practice include, but-are not limited to:
1. Chemical additives
2. pH
3. Temperature
4. Order of addition and mixing conditions
APPARATUS (Fig. 5.1)
REAGENTS
Purity ofReagents: Reagent grade chemicals shall be used in all tests. The chemicals and additives typical of those used for test solution and suspensions are given
in Table 5.1.
201
Table 5.1:
Prime chemicals use in coagulation process
Name
Chemical formula
Dosage required,
ppm
Remarks,/most
effective pH
1.Alum
AI2(SO.'31SH20
40-90
treatment pH:6-S.5
35-40 with lime
2. Ferric
Fe 2(SO.'3. x Hp
sulphate
25-30
3. Ferric
FeC13~6H20
chloride
4. Ferrous
FeSO. 7H 2O
sulphate
5. Lime,
Ca(OH'2
hydrated
Coagulants aids - Polyelectrolytes (anionic, cationic, non-ionic'
Sewage treatment
pH:S-S.5
Sewage treatment
pH:5.5-7
PROCEDURE
1. Nonnally 6 beakers are used, one Jar is used as control and rest 5 beakers are
filled with sample.
2. Measure equal volwne (500 mL) of sample into each of the beakers. Place the
beakers such that the paddles are in centre.
3. Record the sample temperature at the start of the test. In addition, record the
initial pH, turbidity, COD, and colour (in case of coloured eflluents).
4. Start the multiple stirrer operation at the ''flash mix" speed of approximately
120-rpm.
5. Add different chemical dosages, e.g. 200 mg/L, 300 mg/L, 400 mg/L, 700 mg/
L etc. Flash mix for approximately 1 min after the addition of chemical. Records
the flash mix time and speed.
6. The flash mixing is followed by slow mixing for about 20 minutes with a speed
of30rpm.
Record the time for the first visible floc fonnation.
Every 5 minutes (during the slow mix period), record relative floc size and
mixer speed.
If coagulant aids are used, mixing speed is critical because excessive stirring
tends to break up early floc fonnation and may redisperse.
7. After the slow mix period, withdraw the paddles and observe settling of floc
particles. Record the time required for the bulk of the particles to settle. In most
cases this time will be that required for the particles to settle to the bottom of
the beaker; however, in some cases there may be interfering convection currents. If so, the recorded settling time should be that at which the unsettled or
residual particles appear to be moving equally upward and downward. .
8. After 15 minutes of settling, record the appearance of floc on the beaker bottom. Record the sample temperate. By means of a pipette or siphon, withdraw
an adequate sample volwne of supernatant liquor from the jar at a point one
half of the depth of the sample.
202
9. The supernatant liquid is tested for colour, turbidity, pH, COD and other required analysis.
REPRODUCIBILITY
It has been well recognised that reproducibility of results is important. To demonstrate reproducibility, the so-called 3 and 3 procedure is suggested. In this procedure, duplicate sets of 3 jars are treated simultaneously with the same chemical
dosages.
Jar test data-sheet (ASTM, 1995)
1.
2.
3.
4.
5.
6.
7.
8.
9.
Date.................... ..
Sample No. and name:
Location: ............ ..
Sample size: .............. mL
Initial pH: ............ ..
Initial Turbidity (NTU): ............ ..
Initial Colour (Hz): ............ ..
Initial Temperature ("C): ............ ..
Initial COD (mgIL): ............ ..
Parameters
Chemicals, mgIL
Jar Number
a.
b.
c.
d.
Flash Mix Speed, rpm
Flash Mix Time, min
Slow Mix Speed, rpm
Slow Mix Time, min
Temperature, C
Time First Floc, min
Size Floc
Settling rate
q'urbidity, NTU
Colour
pH
COD,mgIL
203
pH
COD (mg/L)
Initial
Final
Turbidity, NTU
Initial
Final
Colour, Hz
Initial
Final
1.
2
3
4
5.
F/M value
Conventional ASP
Oxidation ditch
Sequential batch reactor (SBR)
204
FIM ratio =
So.Q
So
V.X
S.X
Where,
So = Influent BODs or COD concentration, mgIL
e = (V/Q) = hydraulic retention time (HRT), days
X =MLVSS, mgIL
V = Volume of aeration tank, m3
Q = Influent wastewater flow rate, m3/day
FIM = Food-to-microorganisms ratio, dayl.
Measure the influent BODs concentration by standard method. Also calculate
MLVSS in the aeration tank. Calculate the HRT of the system. The FIM ratio can
be calculated by using the above formula and express in per day (i.e., l/day).
20S
APPARATUS
Filtration apparatus, Vacuum pump, Hot air oven 103C, Analytical balance, Glassfibre filter disc, Desiccator.
PROCEDURE
1. Place a glass fibre filter disk, rough side up, in a filter container and place in
filtration apparatus.
2. Wet the filter with a small volume of reagent grade water.
3. Depending upon the MLSS content, filter 2S, SO or 100 mL portion ofa wellmixed sample. Filter the measure volume by applying suction for about 3 min
after filtration is complete.
4. Remove the filter paper from the filtration apparatus and transfer to an aluminium-weighting disk as a support.
S. Dry for at least lh in the range 103 to 10SoC in Hot air oven, cool in a desiccator to balance temperature and weigh. Repeat the cycle of drying, cooling, desiccating and weighing until a constant weight is obtained.
CALCULATION
(A-B)
1000
MLSS, mgIL=
sample volume, mL
Where,
A = Weight of filter paper + dried residue, mg
B = Weight of filter paper, mg.
206
APPARATUS
Muffie furnace, 550C in addition to the apparatus used for MLSS detennination.
PROCEDURE
1. Put the filter paper containing MLSS residues in a muffie furnace at a temperature of 550C. Allow furnace to raise the temperature before inserting the sample.
Usually, 15 to 20 min ignition are required for 200 mg residues.
2. Let dish or filter disk cool partially in air until most of the heat has been dissipated.
3. Transfer to a desiccator for final cooling in a dry atmosphere.
4. Weight disks or dish as soon as it has cooled to room temperature.
5. Repeat cycle of igniting, cooling, desiccating and weighing until a constant
weight is obtained or until weight change is less than 4% or 0.5 mg, which ever
is less. Duplicate detennination should agree within 5% of their average.
CALCULATION
MLVSS, mgIL =
(A- B) x 1000
sample volume, mL
Where,
A = weight of residue + dish before ignition, mg
B = weight of residue + dish after ignition, mg
207
Example-1
To maintain 3000 mg/l (0.3%) MlSS concentration in aeration tank with SVI value
of 100; what should be percent sludge that be returned?
100
------- =
100/(0.3 x 100) - 1
43%
208
Example-2
To maintain 2000 mg/L of MLSS in aeration tank with SVI of 100 mL/g, calculate the
recirculation ratio.
Estimation of SVI
APPARATUS
1000
Defme SVI.
How SVI is measured in the laboratory?
The SVI value is closely related with settling characteristics of sludge. (TIF).
What should be the ideal value of SVI?
What is sludge density index (SDI)? How SDI is determined? What should be
the acceptable value of SDI?
6. "Rate of return activated sludge (RAS) calculation is based on SVI value"justify your answer.
209
7. The MLSS concentration in the aeration tank is 2800 mgIL; sludge volume
(SV) is 285 mgIL. Calculate SVI and estimate MLSS concentration in RAS
and required returned sludge ratio. (Ans. SVI = 120 mUg, MLSS in RAS =9804
mg/L; return sludge ratio = 0.4)
8. Compute the return activated sludge (RAS) flow rate in m3/day as a percentage
of influent flow of 10 MGD (37,850 m3/day). The laboratory result shows that
SVI = 110 mUg and MLSS = 2500 mgIL (Ans. 37.9%).
9. Determine the RAS flow as a % of influent flow of 10 MGD. The sludge set
tling volume in 30 min is 255 mL. (Ans: 34% 12,900 mJ/day).
simple substrate
VFAs
Acetic acid
CH4 +
CO 2
210
APPARATUS
1. Centrifuge, with head to carry 50-rnL tubes or 250-rnL tubes.
2. Distillation assembly: Distillation flask, 500-rnL capacity; condenser; adapter
(Fig. 5.2).
100 mL Sample
+ 100 ml water
+ I) mL H2S04
(ollect
150 mL
di!> till ate
r-ol
Heat
Fig. 5.2: Distillation assembly for VFA estimation.
REAGENTS
1. Sulphuric acid (1+1)
2. Standard NaOH titrant, O.IN
3. Phenolphthalein indicator
PROCEDURE
1. Take 200 rnL sample and centrifuge for 5 minutes (or filter it).
2. Collect supernatant, and place exactly 100 mL supernatant liquor in a 500 rnL
distillation flask.
3. Add 100 rnL distilled water, few glass beads or similar materials to prevent
bumping, and 5 rnL H 2S04
4. Mix well so that acid does not remain in the bottom of the flask.
5. Connect distillation flask to a condenser and adapter and distill at the rate of
about 5 rnL/min.
6. Discard first 15 ml distillate (Note: H 2S and CO 2, liberated during distillation
and titration, will give a positive error. Discarding first 15 rnL of distillate eliminates this error).
211
Where,
N = Normality ofNaOH
Note: The distillation method is empirical and gives incomplete and somewhat
variable results. Factors such as heating rate and proportion of sample recovered as
distillate affect the results. Therefore, recovery factor is 4:alculated for each distilling apparatus and set of operating condition. The recovery factor is calculated by
diluting approximate volume of acetic acid stock solution to 250 mL' in a volumetric flask to approximate the expected sample concentration and distil as for a sample.
Recovery factor can be calculated as follows:
a
f=b
Where,
a = volatile acid concentration recovered in distillate, mgIL
b = volatile acid concentration in standard solution used, mgIL
If recovery factor is consider, the VFA is calculated as:
mL NaOH x N x 60,000
VFA as acetic acid, mgIL = - - - - - - - - mL of sample x f
Where,
f = recovery factor.
2.
3.
4.
5.
Microbiological
Analysis of Water
Organisms
Guideline value
1OO-mL sample.
Treated water entering the distribution
system:
In the case oflarge supplies, where sufficient samples are examined, must not
be present in 95% of samples taken throughout any 12-month period
MOEF (2000) notification for bathing water
213
6.1 Introduction
Pathogenic bacteria, pathogenic protozoan cysts, and viruses have been isolated
from wastewater and natural waters. The sources of these pathogens are the faces
of human and of wild and domestic animals. Identification and enumeration of these
disease causing organisms in water and wastewater are not recommended because
no single technique is currently available to isolate and identify all the pathogens.
In fact, concentrations of these pathogens are generally low in water and wastewater. In addition, the method for identification and enumeration of pathogens are
labour intensive and expensive. Instead of direct isolation and enumeration of
pathogens, total coliform (TC) has long been used as indicator of pathogens contamination of a water that poses a public health risk.
Fecal coliform (FC), which is more fecal specific, has been adopted as a standard indicator of contaminations in natural waters. Both TC and FC tests are used
in standards for drinking water and natural water. Fecal streptococci (FS) is used as
a pollution indicator in Europe. The FCIFS ratios have been employed for identifying pollution sources. The FS are present in the intestines of warm-blooded animals
and of insects, and they are present in the environment (water, soil and vegetation)
for long period of time. E. coli bacteria have also been used as an indicator.
The concentration of pathogenic microorganisms in untreated municipal wastewater are presented in Table 6.1
Table 6.1:
Microorganism concentration in untreated municipal waste water.
Organisms
1. Total coliforms
Fecal coliforms
Fecal streptococci
Protozoan cysts
Shigella
Salmonella
Helminth ova
Enteric virus
Giardia lamsdia cysts
10. Entamoeba histolytica cysts
11. Cryptosporidium oocysts
2.
3.
4.
5.
6.
7.
8.
9.
214
4.
5.
6.
7.
1.
2.
3.
4.
SOURCES OF COLIFORMS
The discharge of~dstewater from municipal sewers is one of the most important
sources of coliform in drinking water. Municipal sewage contains human feces and
water contaminated with these effluents may contain pathogenic (disease-causing)
organisms and consequently, may be hazardous to human health ifused as drinking water. In rural areas, open-defecation in the field, bathing and washing of clothes
215
etc. are some of the common sources of coliform contamination. Fecal contamination of water is routinely detected by microbiological analysis. Pathogenic microorganism are transmitted by water include bacteria, viruses and protozoa.
The intestinal tract of man contains countless rod-shaped bacteria known as
coli/arms. Each person discharges from 100 to 400 billion coliform organisms per
day, in addition to other kind of bacteria (Metcaff and Eddy, 1985). The coliform
bacteria include the genera Escherichia and Aerobacter.
216
Sample bottle: Use sterilized bottles of glass or plastic of any suitable size and
shape (preferably use 250 mL capacity). Plastic bottles are sterilized in autoclave at
121C for 15 min. The non-plastic bottles (glass) sterilized in hot-air oven at 170C
fOF Ih.
Dechlorination of Sample
Chlorinated effluent discharge: Add sufficient sodium thiosulphate solution to a
sterilized sample-bottle to give a concentration of 100 mg/L of sample. For example,
in a 120-mLcapacity sample-bottle, add 0.1 mL oflO% Na2 Sp3' thatwiIl neutralized
a sample containing 15 mg/L residual chlorine.
217
Presumptive test
Confirmatory test
Completed test
218
J,
J;
'l-
J;
'l-
Gas produced,
Positive test
J,
Gas produced."" Coliform
group confirmed + ve test.
J,
No gas produced. Negative test.
Coliform group absent.
(2)
J,
Negative colonies
watery colonies
Coliform group absent.
219
2. Lactose
3. K2 HPO.
4. KH 2 PO.
5. NaCI
6. Sodium lauryl sulfate
7. Distilled water
Total ingredients
Final pH after sterilisation
20.00
5.00
2.75
2.75
5.00
9
9
9
9
9
0.1 9
1.0 L
35.06 g
6.8
/\ I
10"'l
I mL
lmL
lml
I ml
UU U U U
IO"'L
lmL
I mL
Olml
001 ml
Fig. 6.1: Procedure for decimal dilution techniques used for MPN
count in wastewater or domestic sewage.
4. Care should be taken that no air bubbles should remain inside the inverted
Durham's tubes. The air bubbles trapped in the Durham's tubes are removed by
gently inverting the test tubes after closing their mouth by thumb or a fmger.
220
When 10 replicates are used: Pour 10 mL medium (71.2 gIL) in each test tubes
and add 10 mL of sample.
When 100 mL portion is used: Pour 50 mL medium (1 06.8g1L) and add 100 mL of
sample in one bottle.
Table 6.3:
Preparation of Lauryl tryptose (L TB) broth.
Inoculum,
mL
Amount of medium
in tube, mL
10
10
20
100
100
100
10 or
10
20
10
50
35
20
mo~e
Volume of medium
mL
+ inoculum,
11 or more
20
30
30
150
135
120
Dehydrated lauryl
tryptose broth
required, gIL
35.6
71.2
53.4
106.B
106.B
137.1
213.6
For drinking water, when 10-mL portion sample is used in each of 10 test tube or
fermentation tube (10 mL water sample + 10 mL medium) , prepared LT broth as
per Table -4.1 but dissolve 35.6g ingredients in 500 mL of distilled water instead
of 1000 mL of distilled water .
For drinking water, when 20-mL portion sample is used in each of 5 test tube or
fermentation tube (20 mL water sample + 10 mL medium), in such case prepare
LT broth as per Table -4.1, but dissolve 35.6g ingredients in 334 mL of distilled
water instead of 1000 mL of distilled water.
Incubation
Incubate the inoculated fennentation tube at 35 O.soC. At the end of 24 2 h
shake each tube gently and examine it. If no gas is fonned repeat this for another 48
3 h. Record the gas fonnation.
Interpretation
The appearance of gas bubble must not be confused with actual gas production. If
the gas is fonned as a result of fennentation, the broth medium becomes cloudy.
The absence of gas fonnation at the end of 48 h of incubation constitute negative
test.
Since this reaction may also be produced by the organisms other than the
colifonns, the positive tubes from the presumptive test are subjected to confinnatory test.
Results
Record the number of positive tubes and refer Table 6.10 or 6 .11 for drinking water
or Table 6.12 for effiuent.
Confirmatory test (Stage-II)
Transfer one loopful of medium from positive (+) tubes of presumptive test to
Brilliant Green Lactose Bile (BGLB) broth medium (Table 6.4). The BGLB broth, in
221
addition to containing lactose, also contains two components inhibitory to grampositive bacteria. Brilliant green is a dye related to crystal violet and belongs to the
triphenylmethane dye series. Oxgall is a surface active agent which also inhibits the
growth of gram-positive bacteria. Gas formation in 24 or 48 h confirms the results of
the presumptive step.
Table 6.4:
Media composition of BGLB broth
10.0 9
1.Peptone
10.0 9
2.Lactose
3.0xgall
20.0 9
4.Brilliant Green
13.3 mg
5.Distilled water
1.0 L
Final pH after sterilization
7.2
Note: 5 g sodium taurocholate can also be used in place of 20 g oxgall
tubes. The numbers of tubes to be prepared are equal to all positive tests in the
presumptive test.
4. Shake gently, the fermentation tube with positive results and transfer one loopful
of medium to BGLB broth
5. Incubate the tubes at 35-37C for 482 h. and record the tubes with gas formation
as + ve number. The wrong concentration ofBGLB or the exposure of the media
to excessive heat or light may give false positive tests.
6. To confirm presence of coliform bacteria, the completed test is carried out. For
this test, inoculum from each positive tube of the conflrmatory test is streaked
on a plate ofEMB (Eosin Methylene Blue) or Endo agar plate.
Results
Record the number of positive tubes and refer Table 6.10 or 6.11 for drinking water
or Table 6.12 for effluent.
Completed test (Stage-III)
222
No9rowttl
or grow til
without
No9rowll\
g..
Growth tr_,.rrotl
by wlr. loop
~
'.GrOWlh
~
. or growttl
Without
&1 3S'C
.9&S
Intubation
8rill0lll1
gr n
LAclo..
~I
3SoC
bi .. brolh
N~t;v.,~t
:" .a'
FOs.tl'W' s.t
Fig. 6.2: Total coliform by MPN technique (Presumptive test and confirmed test).
Prepare Endo agar or Eosin Methylene Blue (EMB) agar petri-plates (Table 6.5). The
number of petri-plates to be prepared is the same as of tubes showing gas production in BGLB medium. Label the plates corresponding to the tubes of confirmatory
test.
Table 6.5:
Media composition of EMB and Endo agar
EMB AGAR
EN DO AGAR
1. Peptone
10.0 g
2. Lactose
10.0 9
3. K2HPO.
2.0 9
4. Agar
15.0 9
5. Eosin Y
0.4 9
6. Methylene blue
0.065 9
7. Distilled water
1 L
pH after sterilisation 7.1-7.2
10.0 g
1. Peptone
2. Lactose
10.0 g
3. K2HPO.
3.5 9
4. Agar
15.0 9
5. Sodium sulphate
2.5 9
6. Basic fuchsin
0.5 9
1 L
7. Distilled water
pH after sterilisation 7.4
223
'each plate into the tubes of MacConkey broth and record the gas production (a
repetition of presumptive test but with the colonies) within 48 h. at 37C.
4. Also examine the colonies by gram-staining. For this transfer the colonies to a
nutrient agar slant. Subject the colonies from agar slants to the gram staining.
S. If organisms appear rod (bacilli) shaped, red stained, occurring singly, in pairs
or in short chains, the test is confirmed. Since the coliform organisms are gramnegative.
P-A broth medium is commercially available in dehydrated and in sterile concentrated form. The culture medium may be prepared by mixing ingredients as per
Table 6.6.
1. Make this formulation triple strength (3X) when 100-mL sample is examined.
(e.g. dissolve triple the quantities of ingredients in 1000 mL water or the
quantities as per Table 6.6 in 334 mL water).
2 Dissolve the P-A broth medium in water without heating, using a stirring device.
3. Pour SO mL prepared medium into a screw-cap 2S0-mL dilution bottle. A fermentation tube insertion is not necessary.
4. Autoclave for 12 min at 121C with the total time in the autoclave limited to 30
minor less.
S. The pH should be 6.8 0.2 after sterilisation.
Table 6.6:
Beef extract
Peptone
Lactose
Tryptose
5. K2 HPO.
6. KH 2 PO.
7.
8.
9.
10.
Sodium chloride
Sodium lauryl sulfate
Bromocresol purple
Reagent grade distilled water
3.0 9
5.0 9
7.46 9
9.83 9
1.35 9
1.35 9
2.46 9
0.05 9
0.0085 9
1.0 L
224
~~.,-.
Dlutian bottle 250 ml
Incubate at 35 'f SC \
for 1 to 2 days
~.
Contents turn to
di.tinct yellow colour
POSITIVE PRfSUHTlVf
TEST
gas production
Po ..m-. Cont,mo.li..
t t
PROCEDURE
1. Shake sample vigorously for 5 sec (approximately 25 times) and inoculate 100
mL sample into a P-A culture bottle.
2 Mix thoroughly by inverting bottle once or twice to achieve even distribution
of the triple-strength medium though out the sample. Incubate at 35 5C and
inspect after 24 and 48 hour for acid reactions.
Interpretation
1. A distinct yellow colour forms in the medium when acid conditions exist following lactose fermentation.
2 If gas is also produced, a gentle shake of the bottle will result in a foaming
reaction.
3. Any amount of gas and/or acid constitutes a positive presumptive test requiring confirmation.
CONFIRMATORY TEST
Culture medium: Use brilliant green lactose bile (BGLB) broth fermentation tube.
Transfer all cultures that show acid reaction or acid and gas reaction to BGLB broth
for incubation at 35 O.soC.
225
Gas production in the BGLB broth culture within 48 h confirms the presence of
coliform bacteria. Report result as presence-absence test positive or negative for
total coliforms in 100 mL of sample.
COMPLETED TEST
The completed test is same as for total coliforms as given in earlier section.
1.
2.
3.
4.
Tryptose
Lactose
Bile salts mixture or bile salt No.3
K2 HPO.
5.
KH 2 PO.
6.
NaCI
20.0
5.0
1.5
4.0
1.5
5.0
9
9
9
9
9
9
226
Interpretation
RESULTS
Record the number of positive tubes and refer Tables 6.10 or 6.11 for drinking water
and Table 6.12 for efiluent.
STEPSINFECALCOLJFORMTEST
Step-I:
Select all positive tubes of presumptive test
from total coliform MPN test
Step-2:
Transfer one loopful of inoculum to
EC medium. Incubate at 45.5 O.2C for 24 2 h
~~
'I27
and S. gallinarum and all have been isolated from the feces of warm-blooded
animals.
4. The normal habitat of fecal streptococci is the gastrointestinal tract of warmblooded animals.
5. Fecal Streptococci - two stains of fecal streptococci, namely - S. faecalis and
S. faceium are the most human specific members of the FS group. The fecal
streptococci are frequently used in identification of fecal pollution in water.
They are unable to multiply significantly in open water and do not survive long,
thus their presence in high numbers indicates recent pollution.
6. The fecal streptococci test have been used with fecal coliforms test to differentiate human fecal contamination from that of other warm-blooded animals. The
ratio offecal coliforms (FC) to fecal streptococci (FS) could provide information
about the sources of contamination.
FC(10 6 /g feces)
FS(10 6 /g feces)
Ratio (FC/FS)
Chicken
Cow
Human
Sheep
1.3
0.23
13.0
16.0
3.4
1.3
3.0
38.0
1.3/3.4 = 0.4
0.23/1.3 =0.2
13/3= 4.4
16/38 = 0.4
If the ratios are obtained in the range of 1 to 2, interpretations are uncertain. Use
ofFCIFS ratio can be very helpful in establishing the source of pollution in rainfallrun-off studies and in pollution studies conducted in rural areas, especially where
septic tanks are used. In many situation where human pollution is suspected on the
basis of coliform test results, the actual pollution may, in fact be caused by animal
discharges.
PRINCIPLE
The FS can be estimated by multiple tube MPN techniques and membrane filter
techniques. The multiple tube techniques is discussed here. The multiple-tube test
228
consists of preswnptive test and confmned test. The FS grown in a mediwn containing sodiwn azide, at a temperature of 35C. Routine methods may gives false
positive and additional confmnatory tests may be required. A flow diagram of fecal
streptococci (FS) test is given Fig. 6.5.
PROCEDURE
Presumptive FS-test by using azide dextrose broth
1. Inoculate a series of tubes of azide dextrose (AD) (Table 6.9) broth with appropriate graduated quantities of sample.
2 Sample size for drinking water: Use sample of 10 mL portion or less. Use doublestrength (2X Le. 69.4g1L) broth for 10-mL inocula.
3. Sample size for wastewater: Use only decimal multiples oft mL (1 .. 0, 0.1 and 0.01
mL) for wastewater.
4. Prepare AD broth according to the following compositions.
Table 6.9:
Media composition of Azide dextrose broth for FS test.
1.
2.
3.
4.
5.
6.
7.
Beef extract
Tryptone
Glucose
NaCI
NaN 3
Distilled water
pH a iter sterilisation
Total ingredients
4.5 9
15.0 9
7.5 9
7.5 9
0.2 9
1.0 L
INCUBATION
1. Incubate inoculated tubes at 35 O.5C.
2. Examine each tube for turbidity at the end of24 2 h.
3. If no definite turbidity is present, re-incubate, and read again at the end of
48 3h.
4. Calculate the fecal streptocotci density from the MPN index table.
CONFIRMATORY TEST
The test is used to observe the bacterial colonies.
1. All the positive tubes after 24 h may be subjected to confirmatory test. Streak a
portion of growth from each positive AD broth tube on PSE (Pfizer selective
enterococcus) agar.
229
Drinking
Water
Sample
Azide dellOse
broth 12 III
lI
for 24 to 4& h
No gas: FS absent
Gas formahon: PoSlt.V@ F S
tut
positive tublils
Coo'" d , .. ,
0.r
inocula from
PSf
.'!\.....C
9(/1
Brownl~-Black
cobni" with
brown hole
'"'l4,"i.
to
-
o
230
When 5 test tubes are used and each contains 20 mL of water sample - Table
6.10 (for drinking water).
When 10 test tubes are used and each contain 10 mL of water sample - Table
6.11 (for drinking water)
For decimal dilution (10mL, ImL and 0.1 mL) - Table 6.12 (efiluent).
When the series of decimal dilution is different from that in the Table 6.12, select
the MPN value from Table 6.12 for the combination of positive tubes and compute the MPN index using the following formula:
10
MPNIl 00 mL = MPN value from table x
If the MPN comh;nation does not appear in Table 6.12, or for other combinations of tubes or dilution or when MPN tables are not available, in that case Thomas
formula can be used.
No. of positive tubes
100
MPNIIOOmL
..J (mL sample in negative tubes) x (mL sample in all tubes)
Table 6.10:
MPN index and 95% confidence limits for various combinations of positive and
negative results when five 20-mL portions are used.
MPN index/
100 mL
95% confidence
limits (approximate)
Lower
Upper
< 1.1
1 .1
0.05
6.3
2.6
0.3
9.6
4.6
0.8
14.7
3.0
0.0
8.0
1.7
26.4
>8.0
4.0
infinite
Table 6.11:
MPN index and 95% confidence limits for various combinations of positive and
negative results when ten 10-mL portions are used.
No of tubes giving positive
reaction out of 5 of
20 mL each
MPN indexl
100 mL
95% confidence
limits (approximate)
Lower
Upper
< 1.1
0.0
3.0
1 .1
0.03
5.9
2.2
0.26
8.1
Cont...
231
.. . Cont.
3
3.6
0 .69
10.6
5.1
1.3
13.4
6.9
2.1
16.8
9.2
3.1
21.1
12.0
4.3
27.1
16. 1
5.9
36.8
23.0
8.1
10
>23.0
13 .5
59.5
infinite
Table 6 . 12:
MPN index and 95% confidence limits for various combinations of positive results
when five tubes are used per dilution (10 mL. 1.0 mL 0.1 mL) .
Combination
of positive
tubes
MPN
index!
100 mL
0-0-0
0-0-1
0-1-0
0-2-0
<2
2
2
4
1.0
1.0
1.0
10
10
13
1-0-0
1-0-1
1-1 -0
1-1-1
1-2-0
2
4
4
6
6
1.0
1.0
1.0
2.0
2.0
11
15
15
18
18
2-0-0
2-0-1
2-1 -0
2-1-1
2-2-0
2-3-0
4
7
7
9
9
12
1.0
2.0
2 .0
3.0
3.0
5 .0
17
20
21
24
25
29
3-0-0
3-0- 1
3-1-0
3-1-1
3-2-0
3-2-1
8
11
11
14
14
17
3.0
4 .0
4 .0
6 .0
6.0
7 .0
24
29
29
35
35
40
4-0-0
4-0-1
4 - 1-0
4- 1-1
4-1 -2
4-2-0
4-2- 1
4-3-0
13
17
17
21
26
22
26
27
5.0
7 .0
7 .0
9.0
12
9
12
12
38
45
46
55
63
56
65
67
77
4-3 - 1
4-4-0
33
34
15
16
80
5-0-0
5-0-1
5-0 -2
23
30
40
9
10
20
86
110
140
5-1-0
5-1-1
5-1-2
30
50
60
10
20
30
120
150
180
5-2-0
5-2- 1
5-2-2
50
70
90
20
30
40
170
210
250
5-3-0
5-3-1
5-3 -2
5-3-3
80
110
140
170
30
40
60
80
250
300
360
410
5-4-0
54-1
5-4-2
5-4-3
5-4-4
130
170
220
280
350
50
70
100
120
160
390
480
580
690
820
5-5-0
5-5- 1
5-5-2
5-5-3
5-5 -4
5-5 -5
240
300
500
900
1600
> 1600
100
100
200
300
600
940
1300
2000
2900
5300
Cont ...
232
Example - 2
The following combinations of positive and negative tubes were found for treated
sewage. Calculate tilt: MPN index.
Size of sample portion
No of positive tubes
No of negative tubes
10
1
0.1
0.01
1
1
4
2
3
5
Example-3
Estimate the MPN index for the following six samples.
Sample
A. raw
5/5
water
B x 10-3 5/5
C
5/5
0
E
5/5
4/5
0/5
Combination
positive tubes
5/5
3/5
1/5
5-3-1
5/5
3/5
3/5
3/5
1/5
3/5
2/5
1/5
1/5
0/5
1/5
0/5
1/5
0/5
0/5
5-3-1
5-3-2
5-3-2
4-3-1
0-1-0
MPN indexl
100 mL.
11,000
11,000,000
1,400
1,400
330
20
'
SalTle as sample, -
?(3.:~I\S~.'
'
233
Mt1owe~~r,
it is >poJlJ't-ed water
.
Y.r'~:'<!'<'''/.
{0:3
11,000,000 (Ans).
': Sa~ple
~nd
6:12,
MPN
140..
--d;..
.1: .~
sa~f)I~'Mf
. ::>~~0-':"9"'(f')
-(~
\33;;~ il~
, .'~~~,.'..
33 .
= 330 (Ans)
. ,h,
_.-:V
.. JO
xf '
2.
20(Ans).
The MF method gives a direct count of total coliform and fecal coliform present in
the given sample. A measured volume of water sample is filtered though a membrane filters with a pore size of about 0.4,5 l1m, which traps the bacteria on its
surface. The membrane filter is then placed on a suitable medium in a sterile container and incubated at an appropriate temperature. Ifcoliforms and/or fecal coliforms
are present in the water sample, characteristic colonies are formed and that can be
counted directly. The standard volume to be filtered for drinking water samples is
lOOmL.
The success of this method depends on using effective differential or selective
media that will enable easy identification of colonies. The method has advantage
over the traditional MPN water analysis because it is more direct and quicker (given
results in 18-24 h) and can easily test large volumes of water (hence yielding more
accurate results).
Limitations: Unsuitable for water containing very high levels of suspended
materials, sludge and sediments, all of which could block the filter.
234
Table 6.13:
Sample type
100
10
l'
0.1'2
0 .01'2 0.001' 2
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
'Small volumes should be added to the filtration apparatus together with a minimum of 9 mL of sterile diluent to ensure adequate dispersal across the surface of
the filter membrane.
21 .0, 0.1, 0.01, 0.001-mL volumes are filtered after first preparing serial dilutions of the sample. To filter:
1.0 mL of sample, use 10 mL of 1/10 dilution.
0 . 1mL of sample, use 10mL of 1/100 dilution.
0.01mL of sample, use 10 mL of 1/1,000 dilution.
0.001 mL sample, use 10mL of 1/10,000 dilution.
235
5. Forceps, blunt-edged: sterilised before use by dipping 95% ethyl alcohol and
flaming
6. Ethyl alcohol for flame sterilisation
7. Petri dishes and wax pencil for labeling them.
8. Pipette and pipette cans for sterilising
9. Measuring cylinders, capacity 100 mL and 250 mL.
10. Flask for containing culture media.
11. Digital colony cOWlter or magnifying lens.
CULTURE MEDIA
For total coliform estimation: Use M-Endo mediwn (Table 6.14) and incubate the
petri plates at 35C for 24h 2.
For Fecal coliform estimation: Use M-FC mediwn (Table 6.14) and incubate the
pertiplates at 44SC for 24h 2.
Table 6.14:
Composition of M-Endo and M-FC media .
1. Tryptose or polypeptone
Thiopeptone or thiotone
Casitone or trypticase
Yeast extract
lactose
6 . Sodium chloride,NaCI
7 . K2HPO.
8 . KH 2PO.
9. Sodium lauryl sulfate
10. Sodium desoxycholate
11 . Sodium sulfite, Na 2 S0 3
1 2 . Basic fuchsin
13 . Agar (optional)
1 4. Reagent grade water
2.
3.
4.
5.
1.
2.
3.
4.
5.
6.
7.
8.
9.
Typtose or biosate
Polypeptone
Yeast extract
Sodium chloride
lactose
Bile salts mixture
Aniline blue
Agar (optional)
Reagent grade water
10.0 9
5.0 9
5 .0 9
5 .0 9
12.5 9
1.5 9
0.1 9
15 .0 9
1.0 L
236
cessed. Sterile pads may be placed in the petridishes with sterile forceps or with
an automatic dispenser (Fig 6.6a).
2 Soaking pad with nutrients: Pipette 2.5 mL of liquid nutrient media (Use mEndo
broth for total coliform and MFC broth for Fecal coliform) on to each pad and
replace the cover (Fig. 6.6b).
Note: Absorbent pads soaked in liquid medium may be replaced by medium
solidified by agar. In this cases, petridishes should be prepared in advance and
stored in a refrigerator.
3. Assemble the filter funnel on the flask.
4. Sterilise the tips of the blunt-ended forceps in a flame and allow them to cool
(Fig.6.6c).
5. Carefully remove a sterile membrane filter(MF) from its package, holding it only
by its edge as shown in Fig. 6.6d.
6. Place the MF in the filter apparatus (Fig. 6.6e) and clamp it in place. If the
apparatus has been disinfected by boiling, ensure that it has cooled down
before inserting the MF.
7. Mix the sample by inverting its container several times. Pour or pipette the
desired volume ofsample into the filter funnel (Fig. 6.60. This volume should be
chosen in the light of previous experience, but suggested volumes are given in
Table 6.13. If the volume to be filtered less than 10 mL, it should be made up at
least 10 mL with sterile diluent so that sample will be distributed evenly across
the filter during filtration. Alternately, the sample may be diluted as suggested
in footnote of the Table 6.13.
8. Apply the vacuum to the suction flask gently and draw the sample though filter.
Just as the liquid level approaches the filter, rinse the sides with a small amount
of the sterile distilled water, and let the vacuum draw aU of the water though
filter (Fig. 6.6g).
9. Unclamp the funnel top with vacuum still applied. With a sterile forceps remove
the filter (Fig. 6.6h), taking care to touch only edge of the filter. Place the filter in
a previously prepared petridish, grid side uppermost (Fig. 6.6i).
lO. Replace the lid of the Petridish and mark it with the sample number and sample
volume. (i.e. 10 mL). Use wax pencil or waterproof pen for writing on Petridishes.
Pipette 1 mL, 10 mL and 100 mL sample respectively in filter as described in
3 replicates.
11. Incubate the M-Endo-MF plated at 35C for 24 h.
12. Insert the M-FC plates inverted (bottom up) into a watertight, resalable plastic
bags and incubates at 44.5C for 24 h immersed in a thermostatically controlled
water bath. If incubator is used, put a small container with a moist pad in the
137
base, humid atmosphere will maintain inside the incubator. This ensures that
the pad does not dry out during the incubation period.
SECOND PERIOD
Materials
1. Incubated plates
2 Simple microscope (1 0-15x) or magnifying glass.
3. Examine the M-Endo broth-MF plates using a low power microscope. Coliforms
colonies are red or pink showing a bright golden-red metallic sheen. Colonies
without the golden sheen are non-coliform. Count the coliform colonies and
record the results.
4. Examine the M-FC plates in the same manner. Fecal coliform colonies are blue
regardless of shade. All others are not coliforms. Count and record the results.
Note: that coliform results are usually reported "per 100 mL" rather than ''per
milliliter" .
CALCULATION
No. ofcoloniesll 00 mL =
No. of colonies
x 100
Volume filtered, mL
MEDIUM
M-7 h Fe agar: This medium may not be available in dehydrated form and may
require preparation from the basic ingredients (Table 6.15).
Table 6.15:
Media composition of M-7h
Polypeptone
Lactose
Sodium chloride
Sodium desoxycholate
Phenol red
Reagent grade water
5.0 9
10.0 9
7.5 9
0.1 9
0.3g
1.0 L
Fe
agar.
Yeast extract
d-Mannitol
Sodium lauryl sulfate
Bromocresol purple
Agar
3.0g
5.0g
0.2g
0 .35 9
15.0 9
1. Heat in boiling water bath. After ingredients are dissolved heat additional 5 min.
Cool to 55 to 60C and adjust the pH to 7.3 with O.IN NaOH (0.35
mLlI.. usually required).
238
Example-4
An unknown water sample is analyzed for the estimation of coliform organisms.
Three sample portions, 1 mL, 10 mL and 100 mL are used for filtration. Each of
these portions is filtered through five filter membranes using the membrane filtration technique. After incubation, in each petridish, the .colonies are counted. The
counts are as follows:
SI. No.
Sample portion
Number of colonies
1-mL
10-mL
100-mL
2
3
5, 7, 5, 8, 6
Example-5
Counts with acceptable limits: Assume that filtration of volumes 100, 50, 25,
10, 3 and 1 mL produced FC colony counts of 200, 110, 75, 35, 11, 6, and 3
respectively. What is the FC density for the sample?
Example-6
Filtration volume 40, 15, 6, 2 mL produced FC colony counts of 1, 0, and .0
respectively. Estimate the FC density for the sample.
~~
@ So_
)ad
in nutriMt
t.((~S
medium
and
coot
@)Rl'mCIIIl' m~bran<!
_".
pac.k~
lortpp,
tl)
.u(t,on
<D Add
.amp'. to
f,ltrat,on apparatus
..... CD
In f,ltratlOl"l apparatll!
~.@
U~avp to rpsu(ltatP
and then inc.ubate
CD Count
colon,.s "ft...
full in(.ubat'on
240
2 Cool to about 45C and dispense in 4- to 5-mL to perti dishes with tight-fitting
covers.
3. Store at 2 to 10C. Discard after 30 days.
PROCEDURE
1. Filter an appropriate sample volume though a membrane filter, place filter on the
surface of a plate containing M-7 h FC agar medium.
2 Incubate at 41.5C for 7 h. Fecal coliform colonies ere yellow (indicative of
lactose fermentation).
241
Sample size,
No. of positive tubes
No. of negative tubes
out of 5 tubes
out of 5 tubes
mL
0.01
5
o
4
0.001
1
0.0001
2
3
0.00001
2
3
0.000001
o
1
Detennine the MPN and range of coliform organisms per 100 mL at the 95
percent confidence level.
Biological Monitoring of
Waters
Introduction
BIOLOGICAL MONITORING
Physical and chemical characteristics of water bodies affect the abundance, species composition, stability, productivity, and physiological condition of aquatic
organisms. Biological methods used for assessing water quality include the collection, counting, and identification of aquatic organisms; biomass measurement;
measurements of metabolic activity rates; measurement of the toxicity,
bioconcentration, and bioaccumulation of pollutants; and processing and interpretation of biological data.
SIGNIFICANCE OF BIOLOGICAL MONITORING
243
4. It is usually essential to find additional sites which can act as control, i.e. the
water is uncontaminated and the organisms found there are not subjected to
the pollutional stress (such as upstream of discharges to a river).
5. Recovery sites (beyond the mixing zone of contamination) should also be selected, as well as sites which cover the full range of the suspected pollution
gradient, e.g. intervals downstream of a discharge to a river, or in a grid radiating outwards from a point source in a lake.
6. Water samples for bioassays should also be taken, whenever possible, especially when the environmental dilution of contaminants is at its lowest, such as
during low flow periods in river.
7. In rivers there is a vertical and horizontal mixing, so collect the sample at midstream 0.5 to 1m below the surface.
8. In lakes, reservoirs and estuaries where plankton population varies with depth,
collect samples from all major depth zones or water masses. In shallow area of2
to 3 m depth, sub-surface samples collected at 0.5 to 1m may be adequate.
1.
2
3.
4.
5.
6.
7.
Date of sampling:
Sampling station name and no.:
Study area (river, lake, reservoir):
Type of sample:
Depth of sampling:
Meteorological conditions:
Turbidity, water temperature, salinity or any other significant parameters:
244
Taxonomic group
Clean water
(Class-I)
Mild
pollution
(Class-II)
Moderately
polluted
(Class-III)
Prawn (Crustacea)
Beetles (Coleoptera) Riffle beetle (Stene/mis, Elmidae);
Dineutus (Gyrinidae); Hydrophilus (Hydrophilidae);
Dytiscus (Dytiscidae).
Bugs (Hemiptera)-Lethocerus (Belostomidae);
Notonecta (Notonectidae); Sigera (Corixidae);
Hydrometra (Hydrometridae); Gerris (Gerrldae).
Highly
polluted
water
(Class-IV)
Severely
Chironomus
polluted water Tubificidae (Tubifex sp.-sludge worm); Tubifera
(Class-V)
(Rat-tailed maggot).
Tolerance
rating
10
245
a. One reference station (remote station in the extreme upstream (u/s) side).
b. Locate at least one station immediately downstream (dis) of discharge point or
affected zone; The station should be ecologically similar i.e. all the stations
should have similar bottom substrate, depth, stream width, flow velocity etc.
SAMPLING METHOD
Pole
Net
f
l
-~:...
--
Fr~
Fig. 7.1: Various methods of sampling invertebrates:
a. Hand net, b. Surber sampler, c. grab (Ekman grab).
246
Surber sampler (Fig. 7.1 b): Quantitative samples may be obtained in water with a
flow rate greater than 10 cm/sec and water depth up to 60 cm (2 feet). The sampler is
made up of a strong, closely woven fabric (0.595 mm opening), approximately 69 cm
long. Smaller mesh sizes cause backwash due to clogging. This net is held open by
a square-foot metal frame (30.5 cm x 30.5 cm). Place the net opening facing upstream, using the current hold the net open. Push the horizontal frame into the
stream bottom material.
Ekman grab sampler (Fig. 7.1 c): A grab has jaws which close beneath a known
area of sediment when triggered by a messenger released down through the suspension wire from the boat. The entire content of the grab represents a quantitative
sample from a known area. Do not use grab sampler on rocky or sandy bottoms,
because small pebbles or grit may prevent closing of the jaws. The grab is made of
12 to 20 gauge brass or stainless steel and weighs approximately 3.2 kg. The grab is
made in three sizes: 15 xI::; em, 23 x 23 cm and 30 x 30 cm.
Artificial substrate sampler: This is applied where the sample can not be collected
by using traditional methods, examples include deep, fast flowing rivers or where it
is difficult to find suitable sites with similar physical characteristics. Two types of
artificial substrate sampler are shown in Fig. 7 .2a and 7.2b.
Hook for
suspersion
Stones
__itt!
rtl":~
Wlremesh basket
Woodel"l plates
Fig. 7.2: Two types of artificial substrates samplers for
sampling aquatic organisms.
'2A7
may stick on the surface. In such case, soft-bristled toot brass may be used to
remove attached macro invertebrates.
Next a series of sieve are placed in decreasing order starting from 0.5 to 0.6 mm
and finer one of 0.2 mm. Sieving yields residue-a mixture of animals and sediments.
The organisms are picked by using forceps and pipettes. Macroinvertebrates can
be preserved in 10% formalin solution. Use 70% ethyl alcohol solution, if organisms
are having calcareous shells or exoskeleton..
For qualitative analysis, rocks, leaves, or other objects are put in a white pan
partially filled with water. Organisms may be float free from these objects or use
forceps to remove the organisms and identify them under stereo zoom microscoI'e
or compound microscope.
MBI = L (niti)IN
i=!
Where,
ni = number of individuals in each taxon i;
ti = tolerance rating assigned to that taxon i;
N = total number of individuals in the sediment sample.
Example -1
The number of macroinvertebrates in a river sediment samples are 70, 30, 50,
150, and 30 organisms/m 2 for Elmidae, Gyrinidae, Hydrophilidae, Chironomidae
and Tubificidae respectively. The tolerance values for these organisms are 6,6,6,8
and 10 respectively . Compute MBI for this sample.
248
TYPES OF ALGAE
The algae divided into ::ight classes based on colour of pigments (chlorophyll).
The freshwater algae mainly belongs to Blue-green algae (Cyanophyta), Green
algae (Chlorophyta), the diatoms (Bacillariophyta), Yellow green algae (Xanthophyta)
and Flagillates (Euglenophyta).
IMPORTANCE OF ALGAE
1. Presence of algae in drinking water supply or sources causes the problems of
tastes and odours; imparts coloration of water and degrade the palatability of
water.
2 Growth of algae in filter bed causes the hindrance of filter operation and
degrades the palatability of water.
3. Growth of algae in clariflocculator, reservoir wall etc. is totally unwanted, and
which is controlled by use of copper sulfate solution (CuSOJ
4. Algae are also used as indicator oflevel of water pollution.
5. Algae also used as source of oxygen supply for the bacterial oxidation of
sewage/wastewater in oxidation pond.
Environmental problems
Type of algae
... Cont.
4. Indicator of water pollution
249
PRESERVATION OF PHYTOPLANKTON
The most suitable phytoplankton preservative is Lugol's solution. To preserve
samples with Lugol's solution add 0.3 mL Lugol's solution to 100 mL sample and
store in the dark. For long -term storage add 0.7 mL Lugol's solution per 100 mL
sample.
Preparation of Lugol 's solution: Dissolve 20g potassium iodide (KI) and 109 of
iodine crystal in 200mL distilled water containing 20mL glacial acetic acid.
OTHER PRESERVATIVES
Formalin: To preserve samples with formalin, add 40 mL buffered formalin (20g
sodium borate, N~BP7 + lL 37% formaldehyde) to lL of sample immediately after
collection.
Other commonly used preservative includes 95% alcohol, and 6-3-1 preservation (6 parts water, 3 parts 95% alcohol and 1 part formalin). Use equal volume of
preservative and sample.
To retain colour in preserve phytoplankton, store samples in the dark or add
ImL saturated copper sulfate (CuS0 4 ) solutionIL of sample.
STAINING OF ALGAE
As algae are green in colour due to the chlorophyll, staining is not required. How-
250
ever, to stain the algae, methyl blue, gentin violet or acid fuchsin (up to 1% solution) can be used.
COUNTING UNITS
Some phytoplankton are unicellular while others are multicellular (colonial) and
some are filamentous (which may be broken or complete). So the variety of configuration poses problems in counting. For example; should 4 celled Scenedesmus be
counted as one colony or four individual cell. Use this guidelines for counting and
reporting of phytoplankton (APHA, 1998). In case of filamentous algae like
Oscillatoria , a filament length of 10m can be counted as one unit.
Enumeration method
Counting unit
One cell
One organism (any unicellular
or natural colony)
400mm2
Reporting unit
CellslmL
UnitslmL
Units/mL.
(* Areal standard unit equals area of 4-small squares in Whipple grid at a magnification of200x).
251
2 Filling of cell: Shake the concentrated plankton sample and quickly transfer
ImL of sample in the cavity of S-R cell with the help of a dropper. Cover the
S-R cell with a cover slip taking care to avoid trapping of air bubbles inside. It
would be better to keep the cover slip obliquely placed on the cell as shown in
Fig. 7.3 and then pour the sample through the opening in the side comer. Do not
overfill the cavity because this would yield a depth greater than 1 mm and
produce a invalid count.
3. Wait for at least 15 minutes and allow the plankton to settle.
4. Wait at least 15 min. and allow the plankton to settle.
5. Count the plankton. For counting the microscope is first calibrated with the
help of a Whipple grid (ocular micrometer) and a stage micrometer. Calibration
has to be done separately for each different microscope. The Whipple grid is
placed in the eyepiece of the microscope and count is made in a suitable strip
across the length of the cell, i.e. 50 mm. Alternatively, at low resolution, count
can also be made in the area of the Whipple grid. For lOx objective and lOx
eyepiece, the area of Whipple grid is usually 1mm2, but it has to be calibrated
exactly with the help of stage micrometer for the particular microscope. Study a
good number of replicates and calculate the average count.
CALCULATION
When counting is made in a strip
NxCx1000
Phytoplankton (unitslmL) = - - - - - - Where,
N = Number of organisms counted in 1 mL of concentrated sample
L = Length of strip of S-R Cell in which counting has been made, (5Omm)
D = Depth ofstrip in S-RCell,mm (Imm)
W = Width of strip counted by Whipple grid, mm
S = Number of strips counted
C = Total volume of concentrated sample (mL)
V = Total volume of sample concentrated (roL)
Nx CX 1000
AxDxFxV
Where,
N = Number of organisms counted in 1 mL of concentrated sample
A = Area offield in Whipple grid in which counting has been made
D = Depth ofarea in S-RCell,mm (lmm)
F = Number of fields counted
C = Total volume of concentrated sample (mL)
V = Total volume of sample concentrated (mL)
252
DESCRIPTION OF HAEMOCYTOMETER .
The central chamber ("+") is divided into 25 subchambers. Each subchambers is
further divided into 16 chambers. Thus total number of chambers is 400 (25 x 16).
Rest of the four subchambers are further divided into 16 each. The phytoplankton
are contained in the central cubical chamber having 400 sub-chambers.
PROCEDURE
Place a drop of well-agitated sample (natural or concentrated) onto the counting
chamber. Put the special type of cover slip provided with the Haemocytometer.
There should be any overflow, in case of overflow, fill the chamber again. Wait for
a few minutes just to allow the cells to settle. Use 40x objective or greater magnification for phytoplankton count. Express the result as:
No. of organisms counted x 104
units/mL=
253
PROCEDURE
1.
2
3.
4.
5.
6.
7.
Put exactly 0.1 mL volume of the sample by using a calibrated medical doser
onto a glass slide.
Place a coverslip of known area, avoiding any air bubbles inside of the
coverslip.
Put the slide Wlder microscope.
Measure the area of the microscopic field.
COWlt the no of species in each microscopic field and record as one
microtransect. COWlt the several field by moving the slide both vertical and
horizontal direction.
Note: COWlt must be quick to avoid drying of the sample (in case of water
mOWlt); therefore, to avoid the drying use semi-permanent mOWlting
(glycerin mOWlt).
Calculate the phytoplankton as follows:
No.lmL=
Where,
C = Number of organisms cOWlted,
At = Area of coverslip, mm2
As = Area of one strip (microtransect), mm2
S = Number of fields cOWlted
V = Volume of water Wlder coverslip, mL.
254
COLLECTION OF ZOOPLANKTON
Several types of nets can be used for zooplankton collection. The size preferably is
8-mesh. In flowing or shallow water 20-30L of water is collected and filters through
the net. The volume of water filtered is noted.
PRESERVATION OF ZOOPLANKTON
Preserve zooplankton sample with 70% ethanol or 5% buffered formalin. Ethanol
preservative is preferred for materials to be stained in permanent mount or stored.
Formalin may be used for the 48h preservation with subsequent transferred to
70% ethanol. In turbid samples, differentiate animal and detrital material by adding
0.04% rose-bengal stain, which intensely stains the shell of zooplankton and is a
good general cytoplasmic stain.
ZOOPLANKTON MOUNTS
For zooplankton analysis, withdraw 5-mL sub-sample form the concentrated and
dilute or concentrated further as necessary. Transfer sample to the counting cell or
chamber or ordinary glass slide for mounting.
Counting of Zooplankton
Zooplankton can be counted by using Sedgwick-Rafter cell just like phytoplankton
(see Section 7.2) and density is represented as organismslL or organismsll OOmL.
255
Chironomus larvae and Tubifex etc. are increased their number in organically enriched
conditions. As tolerant organisms may be found either in clean or polluted situations,
therefore their presence in not definitive. Thus, a population of tolerant organisms
combined with an absence ofintolerant ones is a good indication of the presence of
pollution (APHA, 1998).
Species diversity index is a ratio between the number of species and "importance value" (no. of biomass, productivity and so on) of individuals. Some of the
numerical indices are useful in characterising and describing the aquatic community. These indices are based on structural and functional stability of the system.
R =.L
P Inp
1=1
I
Where,
S = Total number of species.
PI = n/N = proportion of individuals of the total sample belonging to the ith
species.
n , = Number of individuals (N) belonging to the ith species.
N = Total number of individuals of all the species.
This index can be used to calculate the diversity of phytoplankton, zooplankton and benthic macro invertebrates. This formula assumes an infinite sample, but
as long as sample size is large, the bias is likely to be small if n/N is used to
approximate pi. The higher the value of R, the greater is the diversity. The maximum
value of R can be more than 1. The decline in the value of R is taken as an evidence
of pollution. According to Wilham and Dorris (1968), a value of this index above
3 will indicate clean water, whereas values fewer than this would indicate pollution.
However, the use of this index should be made with great care, as wrong interpretations can be frequently drawn in the conditions of mild pollution or when the
system is under transition from one season to another (Goel et al., 1989).
J=-
InS
J ranges from 0 to 1.
256
D=I-r.p2
i=l '
Where,
= Simpson's index
S = Number of species
The Simpson's index ranges between a value of 0 (low diversity) and a maximum of 1 - 1/S.
S-1
D=-InN
Where,
D = Margalefindex
S = Number of species in sample
N = Total number of individuals in a sample
D=
r. (n)2
i=\
'
Where,
D = Mcintosh index
S = number of species
n, = number of individuals in the ith species
257
A I -A 2
Al
Where,
y+y
_1_2
258
Where,
Y, = abundance of the most abundant species.
Y2 = abundance of the second-most abundant species.
Y = total abundance of all species.
2C
S=-A+B
Where,
A = Number of species in sample A
B = Number of species in sample B
C = Number of species common to both A and B
NUMERICAL VALUES OF AI
Some AI values are reported below. The normal AI values ranges from 50 to 200.
Sample
AI
Algal culture
Marine phytoplankton
Pond water
Marine seston
Lake seston
40-96
76-200
44-221
40-146
457
259
260
MATERIALS
1. BOD bottles of 300 mL capacity (some clear colourless and some painted fully
black)
2. Float, Rope, thermometer
3. Reagents for dissolved oxygen (DO) determination
PROCEDURE
1. Take the sample from the depth in question and fix the DO.
2. Fill the same sample in clear as well as dark bottles and suspend these paired
bottles at the depth in question for at least 2 h. DO will deplete in the dark bottle
and will increase in clear bottle.
3. On completion of incubation period, take out the bottles and measure DO in
both clear and dark bottles.
CALCULATION
The increase in O 2 concentration in lighted bottle during incubation period is a
measure of net production, which is because of concurrence use of O 2 in respiration. It is somewhat less than the total (or gross) production. The loss of0 2 in dark
bottle is used as an estimate of total plankton respiration.
Net photosynthesis = Light bottle DO - initial DO
Respiration = Initial DO - dark bottle DO
Gross photosynthesis = Light bottle DO - dark bottle DO
Thus,
NPP = [(DO in light bottle - initial DO) x 0.375]IT
R = [(Initial 00- DO in dark bottle) x 0.375]/T
GPP = [(DO in light bottle - DO in dark bottle) x 0.3 75]1T
Where,
GPP = Gross Primary Productivity in gC/m) Ih or mgC/L1h
NPP = Net Primary Productivity in gC/mlfh or mgC/L1h
R = Respiration gC/mJIh or mgC/LIh
T = Time period ofincubation (h)
0.375 is a factor (i.e. 12/32 = 0.375) used to convert oxygen to carbon. Under ideal
conditions 1 mole of O 2 (32g) is released for each mole ofC (12g) fixed.
261
SAMPLING
Samples should be collected with an appropriate sampler, such as depth or grab
sampler. For nutrient-poor (high transparency) water up to 61itres will be required.
For eutrophic waters, 1-21itres are usually adequate.
PRINCIPLE
1. Three types of chlorophylls (chlorophyll a, b, and c) are found in phytoplankton and may be extracted with acetone. Each type has a characteristic light
absorption spectrum with a particular peak absorbance. The acetone extract is
analysed in a spectrophotometer at these peaks. The peak height indicates
chlorophyll concentration.
2. When samples are concentrated by filtration for the purpose of analysis, the
phytoplankton cells die. Consequently, the chlorophyll immediately starts to
degrade and its concentration is thus reduced. The degradation product of
chlorophyll a, phaeophytin, fluoresces in the spectral region, and this can lead
to errors in results. It is, therefore, essential to measure the concentration of
phaeophytin-a and to make appropriate correction to analytical results.
APPARATUS
1. Spectrophotometer with a spectral width between 0.5 and 2 mm
2. Cuvettes, 1 cm or with longer path-length
t
2.500
~o
350.0
550.0
750.8
REAGENTS
1. Magnesium carbonate suspension: Add Ig MgC0 3 in 100mL distilled
water. Shake before use.
2. Acetone solution, 90%: 90 mL acetone + 10 mL distilled water
3. Hydrochloric acid: 0.1 N
PROCEDURE
1. After recording the initial water volume, separate the cells from the water by
filtration. Filter continuously and do not allow the filter to dry during filtration
ofa single sample. As filtration ends, add 0.2 mL ofMgC0 3 suspension to the
fmal few millilitres of water in the filter cup. If extraction is delayed at this point.
filters should be placed in individual labelled bags or plastic petri dishes and
stored at - 20DC in darkness. Samples may be transported in this form.
2. Place the filter paper in the tissue-grinder, add 2-3 mL of 90% acetone, and grind
until the filter fibres are separated. Pour the acetone and put the ground filter
into a centrifuge tube; rinse out the grinding tube with another 2 mL of 90%
acetone and add this to the centrifuge tube. Make up the total volume in the
centrifuge tube to 10 mL with 90% acetone. Label, and store in darkness at 4 DC
for 10-! 2 h. Samples may also be transported in this form.
3. Centrifuge for 15 minutes at 3000 rpm to clarify the samples. Decant the clear
supernatant into a clean tube and record the volume.
4. Fill a cuvette with 900/0 acetone. Record absorbance on the spectrophotometer
at 750 nm and 663 nm. Zero on this blank ifpossible otherwise record the absorbance and subtract it from the sample reading. A typical absorbance graph of
chlorophyll is shown in Fig 7.4.
263
5. Place sample in the cuvette and record absorbance at 750 run and 663 run (750a
and 663a)
6. Add two drops of 0.1 N HCI in sample in I-cm cuvette (increased acid in propoi'tion to volume for larger cuvettes). Agitate gently for Imin and record absorbance at 750 run and 665 nm (750b and 665b).
7. Repeat the procedure for all samples. Some preliminary samples may need to be
taken to assess the best sample volume.
CALCULATION
1 Determination of Chlorophyll a in the presence of phaeophytin
i.
VsxL
26.73 [l.7(665b)-663a] x Ve
Phaeophytin a, mg/m3 = - - - - - - - - - Vs x L
Where,
Ve = Volume ofacetone extract (L) or (mL)
Vs = Volume of water ~ample (m3) or (L)
L = Light path length of cuvette or width of cuvette (cm)
264
Note: Volumes of extract and sample can also used in (mL) and (L) respectively
in the above calculation.
ChI a (mglml)
ChI a peaks (mglml)
Oligotrophic
Mesotrophic
Eutrophic
l.7 (0.3-4.5)
4.2 (1.3-10.6)
4.7(3-11)
16.1(4.9-99.5)
14.3(3-78)
42.6(9.5-275)
Source: Wetzel, R.G. (1983). Limnology, 2nd ed., Saunders College Pub.,
Philadelphia.
265
Rotifers
Rotifers are aerobic, heterotrophic and multicellular animals and metabolized solid
foods. Its name is derived from the fact that it has two sets of rotating cilia on its
head, which are used for mobility and capturing food. They are found in natural
waters, stabilisation ponds, and extended aeration basins under low organic loading. Rotifers are very effective in consuming disperse~ and flocculated bacteria and
small particles of organic matter. Their presence in an effluent indicates a highly
efficient aerobic biological purification process. Two of the most common crustaceans are of interest to the sanitary engineer. These are: Daphnia and Cyclops. The
crustaceans are strict aerobes, which feed on bacteria and algae. They are important as a source of food for fish. Their presence indicates water has low organic
load content (low BOD) and high dissolved oxygen (DO).
Monitoring.
1.
2.
3.
4.
266
l.Colpidium camphylum
2.Paramecium caudatum
3.Aspidisca costata
23. Presence of Daphnia and Cyclops indicates low BOD and high DO conditions
(TIF).
Answers:Q.19T; Q.20T;Q.2IF;Q.22 I (a),2(c),3(b);Q.23T
Effluent discharge
90% survival offish after 96 hours in 100% effluent (MOEF, Schedule VI, 1993)
268
of the lifetime of the organisms. The concentration is higher and the impact on the
organisms is severe, usually death. The test is usually completed in less than
4 days (96 hours). It is expressed as Le lo (96h), i.e. that concentration of effluent,
which causes mortality to 50% of the test population after 96 hours of exposure.
Basically it is a short term, survival determination test. It involves exposure of a
selected test organism, such as fish, to a known dilution or concentration of sample
for a specific period, typically 48 hours or 96 hours, but occasionally as short as 24
hours.
b. Chronic tests
Test organisms are exposed to lower concentration for a long period of time, preferably for the entire reproductive life cycle of the organisms. Typically 7-day period
is assumed unless specified otherwise.
c. Renewal test
The renewal test is similar to the static test except that the test solution and control
water are periodically renewed and the test organisms are transferred to the chambers with freshly prepared test mixtures or by replacing the test mixtures in the
original chambers.
269
different proportions. Flow-through tests are desirable for high BOD samples and
for those containing volatile or unstable substances.
SELECTION OF TEST ORGANISMS
The prime considerations in the selection of the test organisms for toxicity tests are:
1.
2.
3.
4.
a. Generally smaller organisms not over 5 to 8 cm long and heaving shorter life
cycle are desirable for toxicity tests.
b. The most sensitive locally available species and most sensitive life cycle stages.
c. Use test organisms that are nearly uniform in size.
d. Use organisms of the same age group or life stage.
e. Determine the past history of the test organisms including when and where
they were collected and method of collection, handling and transportation.
f Test organisms must not be collected from polluted areas where the organisms
are in poor condition or where they have usually high body burden of potential
toxicant.
g. Organisms are not to be collected from areas where disease and parasites are
prevalent or where deformed individuals are found.
h. Collect certain stage of life stages of selected organisms.
i Knowledge of environmental requirements and food habits is important in the
selection of test organisms.
The U .S.EPA (1991) recommends for a minimum of three species (for example,
a vertebrate like fish, an invertebrate and a plant) to be tested for evaluating toxicity,
but it should not include most sensitive species.
1.
2
3.
4.
5.
270
271
PROCEDURE
1.
2.
3.
4.
S.
6.
7.
8.
9.
10.
II
~u
0..
,00().()1
.1
test results. In toxic unit approach, a TU concentration is established for the protection of aquatic life.
The Toxic unit acute (TUa) is the reciprocal of the effluent concentration in the
dilution water in % tenns that causes 50% of the organisms to die by the end of the
acute exposure period.
100
TUa=-LC so
1
or, - - x 100
LC so
273
Radioactivity
Measurement
9.1 Radioactivity
. StandardSohadidaclivity .. .... ;.
Drinking water
Gross a-activity
Gross b-activity
ICMR(1963)
MWH(1975)
IS-1O,500 (1991)
WHO (1984)
WHO (1993)
3 pCi/L (max)
3 pCi/L (max)
10-8 ~Ci!mL (max)
0.1 Bq/L
0.1 BqIL
30 pCi/L (max).
30 pCi/L (max).
10-7 ~Ci!mL(max)
IBq/L
1 Bq/L
Remarks (WHO, 1993): If a screening value is exceeded, more detailed radionuclide analysis is necessary. Higher values do not necessarily imply that the
water is unsuitable for human consumption.
Effluent discharge
IS: 2490 (1981)
MOEF (1993)
1O-7~Ci!mL(max)
1O.o~Ci!mL (max)
1O.o~Ci!mL (max) for
discharge into surface water,
public sewers and marine
coastal areas; and 10-7~Ci/
mL (max) for irrigation.
9.1.1 Introduction
The radioactivity in water and wastewater originates from natural sources and
human activities. The a- activity is mainly due to rocks and minerals and p-activity
is mainly due to potassium content in water.
Radiations have dangerous effects on living beings_ Low level exposure can
also cause somatic/genetic change. Somatic effects include risk of cancer, leukemia,
sterility, cataracts and a reduction in life span. Genetic damage is caused by increasing the mutation rate in chromosomes and genes that will affect future generations.
The p-particles have more penetrating power than a-particles. The p-emitters
external to the body are more damaging than a-emitters, as p-radiation can pass
275
Radioactivity Measurement
through human skin. The a-radiation is the least penetrating type of radiations and
can be stopped by a thin layer of clothing or by a sheet of paper and barely
penetrates the human skin. The a -emitters are taken into human body by inhalation
or along with food or water and emit radiation inside the body.
Regular measurement of gross alpha and beta acti"ity in water is inexpensive,
can be completed quickly, and is useful to determine whether further analysis for
specific radionuclides is needed.
9.1.3 Sampling
Use plastic (polyethylene) or glass container for collection of sample. The sample
container may vary in size from 0.5 L to 18 L. Collect the sample and preserve in
radioactivity homogeneous state by addition of conc. HN0 3 to bring pH down
to<2.
276
using a proportional or Geiger-Mueller counter. The test sample is reduced to minimum weight of solid material having measurable B-activity by evaporation
techniques.
INTERFERENCES
Material interposed between the test sample and the instrument detector, as well as
increasing density in the sample containing beta emitters, produce significant losses
in sample counting rates. Liquid sample must be evaporated to dryness in dishes
that allow the sample to be seen directly by the detector. Most beta radiation
counters are sensitive to alpha, gamma and X-ray radiations.
APPARATUS
1. Beta-particle counter: It consists of detector, detector shield and scalar (mechanical register, power supply and amplifier are contained in a single chassis).
2. Sample mounting dishes: (3.2 mm side wall, flat bottom, noncorrosive with
uniform surface density).
3. Alpha particle absorber: Aluminum or plastic, having a uniform density.
PROCEDURE
1. Place an approximate volume of the test sample in a glass beaker, make 0.5 N
with HN0 3 and evaporate to 1-2 mL.
2. Transfer it to mounting dish and evaporate to dryness. (avoid spattering or
boiling) using a ring heater. Uniform spreading of residual salts is necessary for
reliable comparative data. Hygroscopic solids should be cooled in a dry atmosphere and stored in a desiccator until the start of counting.
3. Place the sample in the counter and count beta-activity for a length of time
sufficient to obtain desired reliabilitY.
Radioactivity Measurement
'2:77
1(6)
278
APPARATUS
REAGENTS
I. Same as described for beta-radioactivity in water. The radioactivity of the reagents may be considered as background and subtract from the test sample
counting rate.
2. Nitric acid (1 + 30): Mix 1 vol. of con. HN03 (sp. gr. 1.42) with 30 volumes of
water.
PROCEDURE
1. Place an appropriate volume of the test solution in a glass beaker, add 3 mL of
conc. HN03 (sp gr 1.42) for each 100 rnL of solution, and evaporate to 1 to 2 mL.
2. Quantitatively transfer to the mounting dish and evaporate to dryness (avoid
spattering/boiling) using a ring heater. After drying, heat the dish to dull
redness for a few seconds, using a burner.
3. Place the sample in the counter and count for a time interval sufficient till the
desired statistical reliability is obtained.
4. Record the reading.
5. Precipitation methods may be used expediently to concentrate the radioactive
material into sfall amounts of precipitate by centrifugation or filtration.
CALCULATION
Results may be expressed as counts per minute per mL (cpm) or in terms of alpha
disintegration rate, using the efficiency determined by use of the calibration standard. Calculate the results as follows:
Radioactivity Measurement
3.
4.
5.
6.
7.
8.
9.
References on VVater
Pollution
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
APHA (1995) Standard methods for the examination o/water andwastewater. American Public Health Association, American Water Works Association
and Water Pollution Con~rol Federation, 19th edition.
APHA (1998) Standard methods/or the examination 0/water and wastewater. American Public Health Association, American Water Works Association
and Water Pollution Control Federation, 20th edition.
ASTM (American Society for Testing and Materials)( 1995): Water and Environmental Technology, Vol. 11.02 Water (II). ASTM, 1916 Race Street, Philadelphia, PA, 19103-1187, USA.
ASTM (American Society for Testing and Materials)(1995) Water and Environmental Technology, Vol. 11.01 Water (I). ASTM, 1916 Race Street, Philadelphia, PA, 19103-1187, USA.
Central Pollution Control Board (CPCB) Scheme/or zoning and classification
ofIndian rivers, estuaries and coastal areas, ADSORBS/3/78-79. CPCB, New
Delhi.
Choubisa, S.L. (1997) Fluoride distribution and fluorosis in some villages of
Banswara district of Rajasthan. IJEH, (39),4: 281-288
Handbook 0/ Environmental procedures and guidelines (1994) Ministry of
environment and forest (MoEF), GOI, New Delhi.
John De Zuane (1977) Handbook o/Drinking water quality (2nd Ed). P.E.Van
Nostrand Reinhold.
Indian Council of Medical Research (lCMR) (1963) Manual ofmethods for the
examination of water, sewage and industrial waste, Special report services,
No. 47,ICMR.
lSI Specification/or Drinking Water: 10,500 (1983). lSI, New Delhi (1983).
lSI Specification/or Drinking Water: 10,500 (1993). lSI, New Delhi (1993).
Khopkar, S.M (1994). Environmental pollution analysis. Wiley Eastern Limited, New Delhi.
Lal:S Commentaries on water and air pollution laws (1996). 3rd edition. Delhi
law house, Delhi, India.
Masters Gilbert, M. (1994). Introduction to Environmental Engineering and
Science. Prentice-Hall of India Private Limited, New Delhi.
Metcalf and Eddy Inc. (1991). (3rd Ed). Wastewater Engineering - treatment,
disposal and reuse. Tata McGraw-Hill Publishing Company Limited. New Delhi.
Peavy, Howard, S. Rowe, Donald R. and Tchobanoglous, George (1985). Envi-
281
Appendices
SOOmL
SOOmL
2SOg
SOOg
103.4S
96.63
90.94
S1.16p
SOOg
2Sg
SOOg
SOOml
l00g
SOOg
SOOg
Sg
SOOg
10 tablets
2Sg
SOOg
SOOg
SOOg
l00g
SOOg
SOOmL
SOOg
l00g
l00g
SOOmL
SOOmL
73.89
454.74
140.00
64.00
280.00
260.00
15S.00
150.00
330.00
60.00
25S.00
14S.00
S6.00
7730
37S.00
96.00
170.00
181.89
227.00
218.00
170.52
S7S.00
80.00
100.00
AlK(SOJl12~O
S.
6.
7.
8.
9.
10.
11.
12.
13.
14.
IS.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
l00g
l00g
Cont...
283
Appendices
29.
30.
31.
32.
33.
34.
35.
36.
37.
38.
39.
40.
Eriochrome black T
Ferrous ammonium sulfate,
Ferric chloride, FeC~. 6Hp
Ferric nitrate, Fe(N03)3' 9Hp
Ferric sulfate, Fe2(SOA x Hp
Ferrous sulfate, FeS04. 7Hp
Ferrion indicator (solution)
Formaldehyde (31-40%; w/v)
Glycerol
Hexane
Hexamethylene tetramine
Hydrochloric acid, HCl
41.
42.
43.
44.
45.
46.
47.
48.
49.
50.
51.
52.
53.
54.
55.
56.
57.
58.
57.
58.
59.
62.
63.
64.
65.
66.
67.
68.
69.
70.
71.
25g
500g
500g
500g
500g
500g
lOOml
500mL
500mL
500mL
90.00
160.00
72.00
79.00
185.00
205.00
602.00
48.88
210.00
85.00
500mL
2.5 L
500mL
l00g
89.00
300.00
118.00
175.00
l00g
428.00
125ml
125ml
25g
500mL
500g
500g
250g
l00g
l00g
250g
500mL
l00g
125mL
500g
500g
500g
250g
500g
500g
500g
500g
500g
500g
l00g
500g
500g
40.00
40.00
125.00
70.00
120.00
74.00
95.00
245.00
312.00
420.00
145.00
154.00
50.00
278.00
278.00
135.00
560.00
120.00
221.00
233.00
433.00
90.00
830.00
250.00
130.00
180.00
284
12.
73.
74.
75.
76.
n.
78.
79.
80.
81.
82.
83.
84.
85.
86.
87.
88.
89.
90.
91.
92.
93.
94.
95.
Ig
lOOg
500g
500mL
25g
Ig
5g
25g
500g
500g
lOOg
lOOg
500g
500g
500g
500g
500g
500g
500g
500g
500g
500g
500g
500g
500g
Silver nitrate
Silver sulfate
Sodium acetate, CHJCOONa.3Hp
Sodium arsenite, NaAS0 2
Sodium azide, NaN J
Sodium bi-carbonate. NaHCOJ
Sodium carbonate, Na2CO]
Sodium chloride, NaCI
Sodium fluoride, NaF
Sodium hydroxide, pellets, purified
Sodium oxalate
Oi-sodium hydrogen phosphate, Na2HP04
Oi-hydrogen sodium phosphate, NaH 2P04
Sodium sulfate, Na 2S04
Sodium sulfite, Na2 SOJ
Sodium thiosulfate, Na 2SPJ.5HP, AR
Sodium potassium terterate
Sodium tetraborate decahydrate, (Borax)
Na2Bp7 IOH p
96. Stach powder (soluble)
500g
97. Stannous chloride, SnCI 2 , AR
lOOg
98. Sulphuric acid
500mL
2.5L
99. Sulfanilamide, C6HgNP2S
lOOgl500g
100. Zinc sulfate, ZnS04
500g
JOI. Zironyl chloride octahydrate, ZrOCI 2.8Hp lOOg
1250.00
140.00
170.00
185.00
273.00
227.36
886.70
454.12
125.00
93.22
350.00
483.00
130.00
68.00
48.00
432.00
6821
170.52
159.15
115.95
125.00
170.52
204.62
670.71
79.58
426.00
126.42
98.67
245.55
165.00
455.00
I.
Name
Capacity
50mL
lOOmL
250mL
500mL
28.00
31.50
39.00
66.00
285
Appendices
, '0
9.
10.
11.
12.
13.
Crucible
Desiccators
Distilling apparatus
Durhams tubes
Funnels
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
Petridishes
Test tube
Centrifuge tube
Separating funnel
Stands (burette, pipette
and test tube)
Filter paper
Glass slide and cover slip
Glass rods
Glass beads
Rubber tubing (ordinary and pressure)
Pestle and motor
Asbestos sheet
l000mL
300mL
159.00
276.00
l00rnr!
25mL
50mL
l00mL
medium
l00mL
250mL
500mL
lOOOmL
small
135.00
80.00
120.00
250.00
490.00
41.00
64.00
89.00
146.00
400.00
3390.00
958.00
30.00
61.00
73.00
75.00
210.00
290.00
328.00
519.00
710.00
837.00
100.00
44.00
48.00
55.00
59.00
83.00
110.00
350
200mm
10mUmin
100nos.
50mmdia
65mmdia
75mmdia
IOmL
50mL
lOOmL
250mL
500mL
l000mL
50mL
ImL
2mL
5mL
lOmL
25mL
10crn
lOmm
15mL
125mL
unit price
436.00
300.00
80.00
800.00
286
I.
2.
3.
4.
5.
6.
7.
8.
9.
10.
II.
12.
13.
14.
15.
Name
Remarks
Autoclave
BOD incubator
Bacteriological incubator
pH meter
Conductivity- meter
Centrifuge
Flame photometer
Heating mentle
Hot plate
Kjeldhal distillation
assembly
Jar-test apparatus
Laminar flow chamber
Muffle furnace
Magnetic stirrer
Mechanical stirrer
25,000
30,000
29,000
8,000
4,000
35,000
40,000
6,000
2,000
1,200
Sterilization purpose
BOD test
microbiological test
pH measurements
conductivity test
miscellaneous purpose
Na and K analysis
COD test
miscellaneous purpose
TKN and NH) estimation
15,000
60,000
10,000
3,000
2,500
Coagulation-flocculation test
Microbiology lab.
VSS estimation
miscellaneous purpose
miscellaneous purpose
12,000 - 2 lakhs
16. Microscope (counting
and measuring device;
camera lucida)
15,000
17. Nephlometer
18. Precision analytical
40,000
balance (Metler)
1,20,000
19. Spectrophotometer
500 per disk
20. Secchi disk
21. Thermometer (0-1 OO'C)
500
22. Water distillation apparatus 5,000-15,000
23. Water monitoring kit (field) 35,000
24.
25.
Water bath
Vacuum pump
5,000
8,000 - 20,000
miscellaneous purpose
turbidity estimation
miscellaneous purpose
miscellaneous purpose
transparency measurements
miscellaneous purpose
production of distilled water
Field monitoring of water
quality parameters.
miscellaneous purpose
miscellaneous purpose
Name
I.
UV -Visible spectrophotometer
4.5 lakhs
miscellaneous purpose
Cont...
287
Appendices
2.
3.
4.
5.
6.
20- 35lakhs
3.5 -5lakhs
61akhs
55,000
2.50 - 3 lakhs
metals analysis
mIscellaneous purpose
organic carbon analysis
mercury analysis
miscellaneous purpose
1.
2.
3.
4.
Characteristics
Specific gravity
1.174-1.189
11-12
11-12
Normality
Molarity
1.834-1.836
96-98
36
18
Nitric acid,
(HNO)
1.409-1.418
69-70
15-16
15-16
1.6N
2. IN
3. O.lN
Preparation 0.02N acid
solution from IN stock
solution.
Nitric acid,
(HNO)
500mlA..
83mlA..
64mlA..
8.3mUL
20mLoflN
solution in 1L
distilled water
168mlA..
28mlA..
2.8mlA..
6.4mlA..
20mLoflN
solution in lL
distilled water
20mLoflN
solution in lL
distilled water
380mlA..
15N or IS M
6Nor6M
INor 1M
O.lN orO.lM
600
240
40
4
400
07
6.7
Cont...
288
Stock sodium hydroxide solution: NaOH, 15N. (for preparing 6N, IN and O.IN
solution). Dissolve 600 g solid NaOH in 800 mL distilled water and dilute to 1000 mL.
Store NaOH in a polythene bottle with screw caps.
Ammonium hydroxide solution, NHpH: Prepare 5N, 3N, and 0.2N NHpH solution
by diluting 333 m, 200 mL, and 13 mL, respectively, of the concentrated reagent
(sp. gr. 0.90, 29%, 15N)to 1000 mL with distilled water.
Indicator solutions
Quantity
Length
Mass
Time
Electric current
Temperature
Amount of current
Luminous intensity
Base SI unils
Meter
. kilogram
second
ampere
Kelvin
mole
candela
Symbol
m
kg
s
A
K
mol
cd
7.2 Concentration
In environmental analysis whereever possible choose the units on the basis of:
1. Mass of analyte/ unit volume in case of water and air
2. Mass of analYlel unit mass in case of soils.
Typical units would be than:
Appendices
Water
289
mgIL
~gIL
Air
mg'm3
~g'm3
Soils
mg/kg
~g/kg
The alternative units sometimes found in environmental literature are based on:
Note:
I million = 106
I billion = 109
I trillion = 1012
7.3 Conversion of ppm, ppb and ppt of chemicals to concentration
expressed in SI units.
Medium
Conversion to Sf units
I ppm
I ppb
I ppt
I g'ml = I mg/L
I mg'ml
1 ~g'ml
Air
ppm
ppb
ppt
I ppm
I ppb
1 ppt
I x Ml22.4 mg'ml
I x Ml22.4 ~g'ml
1 x Ml22.4 ng'ml
Soil
ppm
ppb
ppt
I ppm
I ppb
1 ppt
1 mg/kgsoil
1 ~glkg soil
1 ng/kg soil
290
Soils
lO-6gIL
1O.9gIL
ppm(v/v) = 10-6 ml/ml
1O.9m3/m l = 10.1 cml/ml
ppb (v/v)
1O.2 m
mn = 10-1 m
mn = lO-6 m
an
S. Pressure
Pa
M = mollL= mol/dml
Nlm2
atm = 101352N/m2
bar = 10~/m2.
Torr = mmHg=133.322N/m2
lQ3mol/ml
Water
mgIL
mgIL
ngIL
10-6g1g = ppm
10.9 gig = ppb
10.12 gig = ppt = cml/ml
mglkg
mglkg
nglkg
Air
mglml
mglml
cal = 4.184J
erg
10.1 gldml = mglkg = ppm
10-6 gldml = mglkg = ppb
10.9 gldml = nglkg = ppt
100'J
eV = 1.602 x 10.19 J
Quantity
Frequency
Force
Pressure, stress
Energy or work
A quantity of heat
Power, radiant tkux
Electric charge
Electric potential
Potential difference
Electromotive force
Capacitance
Electric resistance
Conductance
SIunit
symbol
Name
Units
Hz
hertz
newton
pascal
Joule
Joule
watt
Coulomb
volt
volt
volt
farad
ohm
siemens
lis
kg.mls2
kglm.s2or N/m 2
kg.m2/s2 or N.m
kg. m2/s2 or N.m
kg. m2/ sl or J/s
A.s
N
pa
J
J
W
C
V
V
V
W
S
W/A
W/A
W/A
CN
VIA
AN
... Cont.
Cont...
291
Appendices
... Cont.
Magnatic flux
Magnetic flux density
Inductance
Luminous flux
illuminance
Activity (radionuclides)
Adsorbed dose
Wb
T
H
hl
k
Bq
Gy
weber
tesla
henry
lumen
lux
becquerel
gray
Y.s
Wb/m 2
Wb/A
cd.sr
cd.sr/m2 or lmIm2
lis
m 2/s2 or J/kg
1 inch = 2.540 cm
1 foot = 0.3048 m
1 yard=0.9144m
1 mile = 1.6093 km
1 meter = 3.2808 ft = 39.37 in
1 kilometer = 0.6214 mile
2. Area
9. Density
... Cont.
6. Linear Velocity
1 foot per second =
=
1 mile per hour
=
=
=
1 meter per second =
=
0.6818 mph
0.3048m1s
1.467 ftls
0.4470mls
1.609 kmlhr
3.280 ftls
2.237 mph
7. Mass
=
=
=
1 ton (short) =
=
=
1 ton (metric) =
=
=
1 pound
1kilogram
0.453592 kg
2.205 lb.
35.273960z
2000 lb.
907.2kg
9072 ton (metric)
1000 kg
2204.622 lb.
1.1023 ton (short).
8.Flowrate
10. Power
1 cubic foot/second = 0.028316 ml/s
I kilowatt
= 1000 J/s
=488.8 gal (U.S)/min (gpm)
= 3412 Btulhr
1 cubic foot/ minute = 4.72 x 104 ml/s
= 1.340hp
= 7.4805 gpm
I
horsepower
=746W
1 gallon (U.S) Imin = 6.31 x 10-5 ml/s
= 550 ft-Ib/s
1 million gallon/day = 0.0438 ml/s
I quadrillion Btu! year
1 million acre feet/year = 39.107 ml/s
=0.471 million barrels
1 cubicmeterlsec=35.315 ftl/s(cfs)
of oil per day
=2118.9 ftl/min (cfin)
=0.03345TW
= 22.83 x 106 gaVd
= 70.07 Ac-ftld
Appendices
293
Elevation above
sea level, m
Atmospheric
pressure, kPa
500
1000
1500
101.3
95.6
90.1
84.8
2000
79.8
2500
73.3
70.3
66.1
3000
3500
Atmospheric
pressure expressed
as a column of:
Mercury
Water
(m)
(mm)
Specific weight
of air at 200 C,
10.33
9.47
9.19
8.64
8.13
7.47
7.17
6.74
O.oII8
0.0111
0.0105
0.0099
0.0093
0.0085
0.0082
0.0077
760
717
676
636
598
550
527
496
kN/IIf.
Unit or quantity
Symbol
Application
Becquerel
Bq
SI quantity of radioactivity
Bq = 1 disintegrationls
Bq = 2.7 X 10.11 Ci
Curie
Ci
Quantity of radioactivity
1 Ci = 3.7 x 10 10 disintegrations/s
1 Ci=3.7 x IOIOBq
Electron volt
eV
Unit of energy
1 eV= 1.6 x 1O.12 erg
1 eV = 1.6 x 10.19 J
Quantity factor
Gray
Gy
Rad
rad
Rem
rem
Sievert
Sv
294
7.10 Light
a. Approximate wavelength ranges for the various region ofthe electromagnetic
spectrum.
Name of radiation
Wavelength range
y-rays
X-rays
Far ultraviolet
Ultraviolet
Visible
Near infrared
Infrared
Far infra red
Microwave
radar
Very high frequency
Ultrahigh frequency
Radio waves
0.003 -0.3 N
0.3-100N
100-20ooN
200400nm
400-800nm
0.8-2.5 11m
2.5 - 15 !lID
15-200 11m
0.2-7mm
7-100mm
1O-1000cm
10-1oom
10-1O,OOOm
b. Conversions oflight
To convert
Multiply by
einsteins
footcandles
footcandles
footcandles
lumens
lumens/ft2
lux
quanta
candella
6.024 x 1023
1.0
10.764
10.764
0.318
1.0
0.0929
1.66 x 10.24
3.l416
To obtain
quanta
lumens/ft2
lumens/m2
lux
candellas
footcandelles
footcandles
einsteins
lumens
Properties
Value
997.075
99820
958.40
1000.000
3.940
0.890 x 10.1
0.89 X 10-6
0.0000
100.00
Cont...
Appendices
295
.. .Cont.
Latent heat ofice, kllmol
Latent heat of evaporation, kJ/mol
Specific heat capacity (1 5C), Jlkg."C
Thermal conductivity (25C)m, J/cm.s.oC
Heat of vaporization, Jlkg
Surface tension (25C), N/m
Surface tension (20C), N/m
Surface tension (OC), N/m
Dielectric constant (25C)
Vapour pressure (20C), torr
6.0104
40.66
4186
0.00569
2.435 x 1()6
71.97 x 10']
72.75 X 1O.J
75.64 >~ 10']
78.54
17.535
0
5
10
15
20
21
22
23
24
25
26
Z7
28
29
30
5000
10000
15000
14.62
12.77
1129
10.08
9.09
8.99
8.83
8.68
8.53
838
822
8.07
7.CJ2
7.77
7.63
13.73
12.02
10.66
9.54
8.62
8.57
8.42
827
8.12
7.96
7.81
7.67
7.53
739
725
12.89
11.32
10.06
9.03
8.17
8.14
7.99
7.85
7.71
7.56
7.42
728
7.14
7.00
6.86
12.10
10.66
9.49
8.54
7.75
7.71
7.57
7.43
730
7.15
7.02
6.88
6.75
6.62
6.49
296
Permissible
limits in the
absence 0/
alternate
source
Above 5,
25
consumer acceptance
decreases
Remarks
Extended up to 25
only if toxic
substances are not
suspected, in absence
of alternate source
1. Colour, Hazen
units, Max.
2. Odour
Unobjectionable
3. Taste
Agreeable
4. Turbidity
10
Above 5, consumer
acceptance decreases
5. pH value
6.5 to 8.5
6. Total hardness
(as CaCO,),
mg!L, max
300
0.3
1.0
Beyond this limit
taste/ appearance are
affected, has adverse
effect on domestic
uses and water supply
structures, and
promotes iron bacteria
250
9. Residual, free
chlorine, mglL,
max
0.2
Test to be conducted
only after safety had
been established
1000
To be applicable
only when water is
chlorinated. Tested
'at consumer end.
When protection
against viral infection is required, it
should be min. 0.5
mglL.
Cont...
Appendices
10. Dissolved solids, 500
mgIL, max
12. Magnesium
(as Ca)*
mglL, max
30
13. Copper
(as Cu),
mgIL, max
0.05
14. Manganese
(as Mg),
mgIL, max
0.1
15. Sulphate
200
400
gastrointestinal irrit-
(asSOJ
45
May be extended up
to 400 provided, as
Mg does not exceed
30
100
0.002
No relaxation -
20. Cadmium
(as Cd)
(mglL)
0.01
21. Selenium
(as Se)
(mg/L)
0.05
22. Arsenic
(as As)
(mg/L)
0.05
23. Cyanide
(as CN)
(mg/L)
0.05
18. Phenolic
compounds
(as C.HPH)
0.001
298
24. Lead
(as Pb)
(mglL)
0.05
I5
To be tested when
pollution is
suspected
26. Anionic
detergents
(as MBAS),
mg/L, max
0.2
I. 0
To be tested when
pollution IS
suspected
27. Chromium
(as C~)
mgIL, max
0.05
28. Polynuclear
aromatic
hydrocarbons
(as PAH),
mg/L, Max.
May be carcinogenic -
0.01
0.03
30. Pesticides,
mg/L, Max.
Absent
Toxic
0.001
0.1
31. Radioactive
materials:
a. Alpha emiters,
Bq/L, Max.
b. Beta emiters,
pC ilL, Max.
32. Alkalinity,
mgIL, max
200
600
33 Aluminium
(as AI),
mglL, max.
0.03
Cumulative effect is
reported to cause
dementia
0.2
To be tested when
pollution is
suspected. Gas
chromatographic
mathod may be used.
Magnesium (Mg) has been added in IS: 10500: 1991 Drinking water-specification after
amendment No. I January, 1993.
Biological Examination
1. Biological examination is of value in determining the causes of objectionable
tastes and odours in water and controlling remedial treatments, in helping to
interpret the results of various chemical analysis and in explaining the causes of
clogging in distribution pipes and filters. In some instances, it may be of use in
demonstrating that water from one source has been mixed with that from another.
299
Appendices
2 The biological qualities of a water are of greater importance when the conventional flocculation and filtration processes, since increased growth of methaneutilizing bacteria on biological slimes in pipes may then be expected, and the
development of bryzoal growths such as Plumatella may cause operational
difficulties.
3. Some of the animalcules found in water mains may be free-living in the water,
but others such as Dreissena and Asellus are more or less firmly attached to the
inside of the mains. Though these animalcules are not themselves pathogenic,
they may harbour pathogenic organisms or virus in their intestines, thus protecting these pathogens from destruction by chlorination.
4. Chlorination, at the dosages normally employed in waterworks, is ineffective
against certain parasites, including amoebic cysts, they can be excluded only
by effective filtration or by higher chlorine doses than can be tolerated without
subsequent dechlorination. Amoebiasis can be conveyed by water completely
free from enteric bacteria; microscopic examination after concentration is, therefore, the only safe methods of identification.
5. Strict precautions against back-syphonage and cross-connections are required
if amoebic cysts are found in a distribution system containing tested water.
6. The cercariae ofschistosomiasis can be detected by similar microscopic examination, but there is, in any case, no evidence to suggest that this disease is
normally spread through piped water supplies.
7. The cyclops vector of the embryos of Dracunculus medinensis, which causes
dracontiasis or Guinea-worm disease, can be found in open wells in a number of
tropical areas. They are identified by microscopic examination. Such well supplies are frequently used untreated, but the parasite can be relatively easily
excluded by simple physical improvements in the form of curbs, drainage, and
apron surrounds and other measures, which prevent physical contact with the
water source.
8. The drinking water shall be free from microscopic organisms such as algae,
zooplanktons, flagillates, parasites and toxin-producing organisms. An illustrative (and not exhaustive) Jist is given in the Table below.
Classijication
microscopic
organisms
ALGAE
a) Chlorophyceae
Species of Coelastrum, Gomphospherium,
Polluted water, Impart
Micractimum, Mougeotla, Oocyst/s, Euastrum, impounded
colouration,
Scenedesmus, Actmastrum, Gonium,
sources
Eudorina,Pandorina, Pediastrum, Zygnema,
Chlamydomonas, Chlorella, Chroococcus,
Spirogyra, Chlorogonium, Stigeoclonium.
Species of Pandorina, Volvox,
Gomphosphaerium, Staurastrum,
Hydrodictyon, Nitella
Habitat
Effect ofthe of
organisms
and signijicallce
Cont...
300
Clean water
b) Cyanophyceae
Species of Anacyslis and Cylmdrospermum
Indicate clean
conditions
Species of Rivularia
Calcareous
Causes matted
waters and also growth
rocks
Clean waters
c) Diatoms (8acillariopbyceae)
Species of Fragillaria. Slephanodiscus.
Slauroneis
Indicators of
purification
Cause
discolouration
Moderately
Cause
polluted waters discolouration
Rivers and
Clog
streams impo- tilters and
unded sources cause operational difficulties
Clean water
Indicators of
purification
Coni...
301
Appendices
d) Xanthophyceae
Species of Botryococcus
zooPLANKTON a) Protozoa
Amoeba. Giardia. Lamblia. Arcolla. Difflugia. Polluted water
Actinophrys. Entamoeba histolytica
Sewage and
activated
sludge
b) Ciliates
Paramoccium. Vorticella. Carchesium.
Stentor,Colpidium. Coleps. Euplotes.
Colopoda.Bodo
c) Crustacea
Bosmina. Daphnia. Cyclops
d) Rotifen
Anurea. Rotaria. Philodina
e) Flagellates
Cerallum. Glenodinium. Peridmium.
Dinobryon.
Pollution
indicators,
Parasitic and
pathogenic
Highly
Bacteria eaters
polluted waters,
sewage and
activated sludge
Stagnant
polluted water
Step wells in
tropical
climate
Indicator of
pollution.
Carrier host
of guinea
worm.
Polluted and
algae laden
waters.
Feed on algae
Euglena. Phacus
f) Miscellaneous organisms
Sponges. Hydra
Fresh water
Plumatella
Polluted
waters
Dreissena, Asellus
Polluted
waters
Harbourpathogenic organisns
Virological Examination
1.
2
302
3.
4.
Bacteriological Examinatio~
1. Water ill distributioll system
Ideally, all samples taken from distribution system including consumer's premises,
should be free from coliform organisms. In practice, this is not always attainable,
and the following standard for water collected in distribution system is therefore
recommended:
a)
b)
c)
d)
Throughout any year, 95percentage of samples should not contain any coliform
organisms in 100 mL.
No sample should contain E. coli in 100 mL.
No sample should contain more than 10 colifonn organisms per 100 mL.
Coliform organisms should not be detectable in 100 mL of any two consecutive
samples.
If any coliform organisms are found the minimum action required is immediate resampling. The repeated or the appearance of higher numbers in individual samples
suggests that undesirable material is gaining access to the water and measures
should at once be taken to discover and remove the source of the polIution.
1.
2.
Where it is impracticable to supply water to consumer's through a piped distribution network and where untreated sources, such as wells, bore-holes and
springs which may not be naturalIy pure, have to be used, the requirements of
piped supplies may not be attainable. In such circumstances, disinfection although is not always practicable, and considerable reliance has to be placed
on sanitary inspection and not exclusively on the results of bacteriological
examination.
Everything possible should be done to prevent pollution of water. Obvious
sources of contamination should be removed from the immediate catchment
area, special attention being given to the safe disposal of excrement.
Appendices
3.
4.
5.
6.
303
Wells and storage tanks should be protected by lining and covering, surface
drainage should be diverted, erosion prevented and the surrounding area paved.
Access of man and animals should be restricted by fencing, and should be so
designed that fouling is prevented when drawing water. Although not supplied through pipes, water from such sources is likely to undergo further deterioration in quality during transport or storage before drinking.
Containers used for water should be kept clean, covered and clean of the floor.
The most important factor in achieving these objectives is to ensure the cooperation of the local community, and the importance of education in simple
sanitary hygiene should be strongly stressed. In hospitals or medical clinics
with such supplies, the value of some form of treatment is stressed.
Bacteriologically, the objective should be to reduce the coliform count to less
than 10 per 100 mL, but more importantly, to ensure the absence offecal
coliform organisms. If these organisms are repeatedly found, or if sanitary
inspection reveals obvious sources of pollution which cannot be avoided,
then an alternative source of drinking water would be sought whenever possible. Greater use should be made of protected ground-water sources and
rainwater catchment which are more likely to meet requirements for potable
water quality.
Although private sources of drinking water may be outside the jurisdiction of
public health and water supply authorities, such supplies should still be of
potable quality. The results of bacteriological tests and those of sanitary surveys should therefore be used to encourage improvement. Partial treatment
may be necessary to remove turbidity even when coliform counts are low; and
other quality criteria may dictate the need for treatment processes.
Index
Acidity, 30
Measurement of, 31
Standards of, 30
Alkalinity, 33
Environmental significance, 34
Estimation of, 34
Estimation of, 63
Standards of, 33
Chlorine demand, 70
Aluminium, 181
Estimation of, 181
Standards of, 181
Ammonia nitrogen (see nitrogen
ammonical), 108
Anionic detergents (see methylene
blue active substances), 100
Arsenic, 182
Estimation of, 182
Standards of, 182
Artificial substrate sampler, 246
Atomic absorption spectrophotometer, 178
Benthic macroinvertebrates, 243
Counting of, 244
Sampling method, 245
Biochemical oxygen demand, 37
Application of BOD data, 37
BOD standards, 37
Procedure of, 40
Biological monitoring of waters, 242
Significance of, 242
Boron, 48
Estimation of, 71
Significance and use, 70
Chlorine (residual), 66
Estimation of, 67
Standards of, 66
Chlorophyll, 261
Estimation of, 261
Chromium, 184
Estimation of, 184
Standards of, 184
Coagulant aids, 1 99
Coagulation-flocculation jar test, 197
Application of, 200
Procedure of, 201
Cold vapour atomic absorption
spectroscopy, 189
Coliform group of bacteria, 214
Estimation of, 215
Presence-absence coliform
test, 223
Coliform standards, 20
Colour, 57
Environmental significance, 58
Estimation of, 48
Estimation of, 58
Standards of, 48
Standards of, 57
Cadmium, 183
Conductivity, 52
Environmental significance, 53
Estimation of, 53
Carbon dioxide, 50
Estimation of free CO 2 , 50
Standards of free CO 2 , 50
Standards of, 52
Copper, 185
Estimation of, 185
305
Index
Standards of, 185
Cyanide, 74
Estimation of cyanide:
Colorimetric method, 77
lon-selective electrode, 78
Flame photometer, 1 52
Fluoride, 85
Estimation of, 87, 89
Removal of fluoride from water, 90
Standards of, 85
Titration, 76
Total cyanide, 74
Hardness (Calcium), 94
Standards of, 74
Description of monitoring area, 2
Digestion of metals, 177
Diseases caused by bacteria, 213
Diseases caused by protozoa, 214
Diseases caused by viruses, 214
Dissolved oxygen, 79
Alum flocculation method, 84
Environmental significance, 80
Estimation of, 95
Standards of, 94
Hardness (Magnesium), 96
Estimation of, 97
Standards of, 96
Hardness (Total), 91
Environmental significance, 92
Estimation of, 92
Standards of, 91
Estimation of, 80
Standard of, 79
Iron, 185
Manganese, 187
306
a radioactivity, 277
~
radioactivity, 275
Removal of hardness, 98
Selenium, 192
Estimation of, 1 92
Standards of, 192
Settleable solids (see solids), 142
Silver, 193
Index
307
Solids, 142
Estimation of:
Total solids, 143
Objectives of, 1
Zooplankton, 253
Counting of, 253, 254