Experiment 3: Determination of Chloride, Sulphate, and Nitrate Ions.

Objective

To investigate the concentration of the chloride, sulphate, and nitrate anions in tap water
and drinking water samples from water dispenser by using HACH DR 3900
spectrophotometer.

Introduction

Every chemical compound absorbs, transmits or reflects light (electromagnetic

radiation) over a certain range of wavelength. Therefore, a spectrophotometer, an instrument
that measures the intensity of light absorbed after it passes through the solution can used to
determine the concentration of known chemical.

In this experiment, we have measured the concentration of chloride, sulphate and

nitrate anions in different water samples via this spectrophotometry. A measured amount of
the indicator compound is mixed with measured amount of water sample. After mixing the
solution, amber color will appear and then it is measured in the spectrophotometer.

The chloride, sulphate and nitrate anions are commonly present in water. High
concentration of nitrate levels in water will bring adverse effects to human and animal
health. Low concentration of chloride level in water will not have a significant impact to
human and animal health as well, however it can still jeopardized the health of people who
suffer from heart and kidney diseases. Chloride will increases the electrical conductivity of
water thus increases its corrosivity. For metal pipes that contain drinking water, chloride
reacts with metal ions to form soluble salts hence increases the levels of metal ions in
drinking water. Other than that, the amount of sulphate is one of considerable concern
because it is indirectly responsible for two major problems which associated with the
wastewater treatment process. Sulphate will cause odor and sewer corrosion problem caused
by the reduction of sulphates to hydrogen sulphide under the anaerobic conditions.

1
Materials and Equipment

HACH DR 3900 spectrophotometer, sample cells with stoppers, micropipette, micropipette

tips, tissues, beakers, Nitra Ver 5 pillow powder, Sulfa Ver 4 pillow powder, Ferric Ion
solution, Mercuric Thiocyanate, tap water, deionized water, and drinking water.

Figure 1

Figure 2

Figure 3

2
Procedure

Mercuric Thiocyanate Method for Determination of Chloride Ions

1. The HACH DR 3900 Spectrophotometer was switched on and the program “70
chloride” was selected.
2. The blank was prepared by adding 10mL of deionised water into a sample cell.
3. The sample was prepared by filling the second sample cell with 10mL of tap water.
4. 0.8mL of Mercuric Thiocyanate Solution was added into each sample cell using a
micropipette.
5. Both the solutions were swirled to allow proper mixing.
6. 0.4mL of Ferric Ion Solution was added into each sample cell using a micropipette.
7. Both the sample cells are swirled to allow mixing. An orange colour appeared on the
sample that contained chloride ions.
8. The instrument timer was started to allow a 2-minute reaction.
9. The blank was cleaned once the timer expired. The following five steps were
completed within 5 minutes.
10. The blank was inserted into the cell holder.
11. The “ZERO” button on the screen was pushed and the display showed 0.0mg/L Cl-
and this reading was recorded.
12. The prepared cell sample was cleaned.
13. The prepared cell sample was inserted into the cell holder.
14. The “READ” button on screen was pushed, the results were shown in mg/L Cl- and
recorded.
15. Step 3 to 14 were repeated using water from water dispenser as sample.
16. The experiment was repeated for both sample to obtain average reading.
17. The results were tabulated.

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USEPA SulfaVer 4 Method for Determination of Sulphate Ions (Powder Pillow
Procedure)

1. The HACH DR 3900 Spectrophotometer was restarted and the program “680
Sulfate” was selected.
2. The sample was prepared by filling a sample cell with 10mL of tap water using a
miropipette.
3. The contents of Sulfa Ver 4 pillow powder was added to the sample cell.
4. The sample cell was swirled to allow proper mixing. White turbidity formed as there
were sulphate present in the sample.
5. The instrument timer was started to allow a five-minutes reaction. The cell was not
disturbed within this 5 minutes.
6. The blank was prepared by filling a second sample cell with 10mL of deionized
waster using a micropipette.
7. The blank was cleaned once the timer expired.
8. The blank was inserted in to the cell holder.
9. The “ZERO” button on screen was pushed, the display showed 0mg/L SO42- and this
reading was recorded.
10. The prepared sample was cleaned.
11. The prepared sample was inserted into the cell holder within five minutes after the
timer expired.
12. The “READ” on screen was pushed, the results were shown in mg/L SO42- and
recorded.
13. The sample cell was cleaned with soap and a brush.
14. Step 2 to 13 were repeated using water from water dispenser as sample.
15. The experiment was repeated for both samples to obtain an average reading.
16. The results were tabulated.

4
Cadmium Reduction Method for Determination of Nitrate Ions (Powder Pillow
Procedure)

1. The HACH DR 3900 Spectrophotometer was restarted and the program “353 N,
Nitrate MR PP” was selected.
2. The sample was prepared by filling a sample cell with 10mL of tap water using a
micropipette.
3. The contents of Nitra Ver 5 pillow powder was added to the sample cell
4. The instrument timer was started to allow a 1-minute reaction.
5. The stopper was placed on the sample cell followed by a vigorous shaking until the
timer expired.
6. The instrument timer was started to allow a 5-minute reaction. An amber colour was
shown as there were nitrate ions present in the sample.
7. The blank was prepared by filling a second sample cell with 10mL of deionized
water using a micropipette.
8. The blank sample cell was cleaned when the timer expired.
9. The blank was inserted into the cell holder.
10. The “ZERO” button on screen was pushed, the display showed 0.0mg/L NO3- -N
and this reading was recorded.
11. The prepared sample cell was cleaned.
12. The prepared sample was inserted into the cell holder within 2 minutes after the
timer expired.
13. The “READ” button on screen was pushed, the results were shown in mg/L NO3- -
N and recorded.
14. Step 2 to 13 were repeated using water from water dispenser as sample.
15. The experiment was repeated for both samples to obtain an average reading.
16. The results were tabulated.

5
Results

Table 1: Concentration of chloride ions (mg/L) in water samples

Water type Concentration of chloride ions (mg / L)

Blank Reading 1 Reading 2 Average
Tap water 0.0 2.8 3.7 3.25
Water from 0.0 3.7 4.6 4.15
water dispenser

Table 2: Concentration of sulphate ions (mg/L) in water samples

Water type Concentration of sulphate ions (mg / L)

Blank Reading 1 Reading 2 Average
Tap water 0.0 14 14 14
Water from 0.0 13 11 12
water dispenser

Table 3: Concentration of nitrate ions (mg/L) in water samples

Water type Concentration of nitrate ions (mg / L)

Blank Reading 1 Reading 2 Average
Tap water 0.0 0.8 0.7 0.75
Water from 0.0 0.6 0.5 0.55
water dispenser

6
DISCUSSION

Mercuric Thiosulphate Method

Anthropogenic sources of 𝐶𝑙 − , including sewage, water conditioning salts, and

synthetic fertilizer(primarily KCl), brine disposal pits associated with oil fields, chemical
and other industries, and, in snowy climates, road salt runoff. Naturally, 𝐶𝑙 −
concentrations in shallow aquifers ranged from less than 1 to 15 mg/L. Discharge from
wastewater treatment plant which had 𝐶𝑙 − concentrations that ranged from 113 to 291
mg/L. In-home water treatment, specifically water softening, typically uses NaCl to
recharge ion exchange columns in order to reduce hardness (Ca + Mg) by replacement
with Na. For a family of three or four with moderately hard water, the recommended
amount of NaCl for water softening is between 1.8 and 2.7 kilograms per day, or 600 to
nearly 1000 kg of NaCl per year.1 KCl is the most commonly available potassium (K)
fertilizer and usually the cheapest. Municipal landfill leachate has 𝐶𝑙 − as high as 6,170
mg/L and the greatest 𝐶𝑙 − concentration recorded in municipal landfill leachate to be
27,100 mg/L. Potential sources of 𝐶𝑙 − in landfills include food scraps and pet wastes.

When chloride is mixed with a solution of undissociated mercuric thiocyanate, the

chloride preferentially combines with the mercury to form mercuric
chloride/chloromercurate(Il) complex ion. The thiocyanate that is released combines with
ferric ions present in the reagent to form ferric thiocyanate which can be measured
spectrophotometrically by measuring the intensity of red colour. There is possible
formation of the higher complexes, such as Fe(SCN)2+ and Fe(SCN)3 .2

FeSCN2+ + SCN- <-> Fe(SCN)2+

Fe(SCN)2+ + SCN- <-> Fe(SCN)3
Fe(SCN)3 + SCN- <-> Fe(SCN)4-
Fe(SCN)4- + SCN- <-> Fe(SCN)52-
Fe(SCN)52- + SCN- <-> Fe(SCN)63-

The formation of higher complexes presumably does not occur at great dilutions;
two other complicating reactions are:
(i) hydrolysis of Fe3+ to FeOH2+, Fe2(OH)24+ , Fe(OH)2+ , Fe(OH)30, and Fe(OH)4- .
(ii) formation of thiocyanic acid molecules.
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 An estimate can be made of the extent to which the two reactions interfere, and
conditions chosen so that this interference is not serious and can be allowed for. Results
also provide some evidence on the possible existence of the ion Fe(OH) (SCN)+ . In
almost every investigation dealing with Fe(SCN)2+, deviations have been found at higher
thiocyanate concentrations which have been ascribed to higher complexes.

In conclusion, the chemical equation for this reaction is:

2 𝐶𝑙 − + 2Fe3+ + Hg(SCN)2 HgCl2 + 2Fe(SCN)2+ +( FeOH2+, Fe2(OH)24+ , Fe(OH)2+ ,

Fe(OH)30, and Fe(OH)4- + Fe(SCN)2+ + Fe(SCN)3+ Fe(SCN)4-+ Fe(SCN)52-+ Fe(SCN)52-+
Fe(SCN)63-+…..)

USEPA SulfaVer 4 method

On the other hand, sulphate ions are formed through a combination of sulphur and
oxygen and are a part of naturally occurring minerals in some soil and rock formations that
contain groundwater. The mineral dissolves over time and is released into groundwater.
As groundwater moves through these, some of the sulphate is dissolved into the water.
Some minerals that contain sulphate are sodium sulphate (Glauber's salt), magnesium
sulphate (Epsom salt), and calcium suphate (gypsum).3 Therefore, sulphate are found in
appreciable quantity in all natural waters, particularly high in arid and semi-arid regions
where natural waters in general have high salt content.

Sulphate is commonly use in the industrial. Sulphates and sulphuric acid products
are used in the production of chemicals, dyes, glass, paper, soaps, textiles, fungicides,
insecticides, astringents and emetics. They are also used in the mining, wood pulp, metal
and plating industries. Sulphate salts are mostly soluble in water and impart hardness.
Sulphate minerals can cause scale buildup in water pipes similar to other minerals and
may be associated with a bitter taste in water that can have a laxative effect on humans and
young livestock.

Through this SulfaVer 4 method, the concentration of sulphate ions in the sample
(tap water and reversed osmosis water) can be determined in order to maintain the
concentration of sulphate anions within safe level. The reagent used in this method is
SulfaVer 4 Reagent Powder Pillows. This reagent used in this experiment contains barium

8
chloride. The chemical equation below shows the reaction between SulfaVer 4 Reagent
Powder Pillows and sample water.

𝐵𝑎2+ (𝑠) + 𝑆𝑂4 2− (𝑎𝑞) → 𝐵𝑎𝑆𝑂4 (𝑎𝑞)

Hence, when the SulfaVer 4 Reagent Powder Pillows is added into the sample
contains sulphate ions, barium ions will reacts with the sulphate ions, and thus barium
sulphate precipitate is produced. 5 minutes instrument time was set for reaction time
between barium ions and sulphate ions. Barium sulphate is white crystalline solid and
poorly soluble in water. The observation of white turbidity produced after reagent is added
indicates presence of sulphate ions in the sample. The turbidity is proportional to the
concentration of sulphate ions. When the turbidity increases, the spectrophotometer rays
pass through the sample decreases, and this makes the reading of spectrophotometer
increases, which means concentration of sulphate ions increase.

Cadmium Reduction Method

Unlike sulphate, Nitrate and nitrite are naturally occurring ions that are part of the
nitrogen cycle. The nitrate ion (NO3−) is the stable form of combined nitrogen for
oxygenated systems. Although chemically unreactive, it can be reduced by microbial
action. The nitrite ion (NO2−) contains nitrogen in a relatively unstable oxidation state.
Chemical and biological processes can further reduce nitrite to various compounds or
oxidize it to nitrate.4

Nitrate can reach both surface water and groundwater as a consequence of

agricultural activity (including excess application of inorganic nitrogenous fertilizers and
manures), from wastewater treatment and from oxidation of nitrogenous waste products in
human and animal excreta, including septic tanks. Nitrite can also be formed chemically in
distribution pipes by Nitrosomonas bacteria during stagnation of nitrate-containing and
oxygen-poor drinking-water in galvanized steel pipes or if chloramination is used to
provide a residual disinfectant and the process is not sufficiently well controlled.

Nitrate is used mainly in inorganic fertilizers. It is also used as an oxidizing agent

and in the production of explosives, and purified potassium nitrate is used for glass
making. Sodium nitrite is used as a food preservative, especially in cured meats. Nitrate is
sometimes also added to food to serve as a reservoir for nitrite. Nitrates occur naturally in

9
plants, for which it is a key nutrient. Nitrate and nitrite are also formed endogenously in
mammals, including humans. Nitrate is secreted in saliva and then converted to nitrite by
oral microflora.

This Cadmium Reduction Method is used to determine the concentration of nitrate

ions in sample water in order to maintain the concentration of nitrate ions is within the
safety range. The reagent used in this experiment was NitraVer Nitrate 5 Reagent Powder
Pillow. This reagent contains Cadmium. The equation below shows the reaction between
NitraVer Nitrate 5 Reagent Powder Pillow and sample water.

Hence, when the NitraVer Nitrate 5 Reagent Powder Pillow is added into the
sample contains nitrate ions, cadmium metal reduces nitrate in the sample to nitrite. The
nitrite ion reacts in an acidic medium with sulfanilic acid to form an intermediate
diazonium salt. The salt couples with gentisic acid to form an amber colored solution. The
measurement wavelength is 500 nm for spectrophotometers or 520 nm for colorimeters.

Shaking time and technique influence the color development. For most accurate
results, use a standard solution that is within the test range and run the test several times.
Increase or decrease the shaking time to get the expected result. The adjusted shaking time
is used for sample measurements. For this experiment, one minute shaking time and five
minutes reaction time are used to get optimal result.

Based on the results, it can be observed that there’s no big difference between
concentration of the tested ions in both tap water and reverse osmosis water, while
concentration of chloride ions in both samples deviates highly from the expected
concentration of chloride ions in mg/L Cl- , this might be due to human error that occurs
during the experiment. During the experiment, a wrong amount of Mercuric Thiocyanate
solution and Feric ion solution was added to the solution due to over pressing while
obtaining solution by using pipette, which causes excess amount of mentioned solution to
be added into the mixture, which then further affects the results since chloride ions in the

10
sample will react with mercuric thiocyanate, forming mercuric chloride and liberate
thiocyanate ions. The liberated thiocyanate ions will the react with ferric ions to form
ferric thiocyanate complex, which contributes to the orange color of the solution. Since the
concentration of thiocyanate complex was said to be proportional to the concentration of
chloride concentration, it will be used as an indicator to determine the concentration of
chloride ions. Thus, concentration of chloride ions obtained will be affected if the wrong
amount of mercuric thiocyanate and ferric ion solution was added.

On the other hand, to prevent errors such as mistakes as mentioned above to

occurs, several precaution should be taken. For example, while using pipette to transfer
solutions, the pipette shouldn’t be over pressed so that no excess solution will be added
into the mixture solution. Besides, air bubbles should be checked so that there’s no air
bubbles exist in the pipette to ensure that the exact amount of solution will be added.

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Conclusion

In this experiment, different methods were used to determine the concentration of

chloride, sulphate and nitrate in water from tap and water dispenser. The concentration of
chloride, sulphate, and nitrate in tap water are 3.25, 14, 0.75mg/L respectively, while
concentrations in water from water dispenser (Reverse Osmosis Water) are 4.15, 12, and
0.55 respectively. It is observed that there’s no big difference between concentrations of
all tested ions in both water samples.

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References

1. Kelly WR, Panno S V, Hackley K, Kelly WR. The Sources , Distribution , and
Trends of Chloride in the Waters of Illinois Illinois State Geological Survey.
2012;(March).

2. Stefánsson A. Iron(III) Hydrolysis and Solubility at 25 °C. Environ Sci Technol.

2007;41(17):6117-6123. doi:10.1021/es070174h.

3. Quality D. Sulfate in Drinking-water Background document for development of

WHO Guidelines for Drinking-water Quality.

4. Guidelines WHO, Quality D. Nitrate and nitrite in drinking-water.

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