2207/WA/2013/1 Water Analysis (1)Introduction- water for drinking, recreation & others (2) Water quality and standards

(3)Analytical Methodologies: Field and laboratory measurement, Pretreatment and Chemical Analysis (4)Special Methods: Automated Analysis, Flow Injection Analysis (FIA), and Ion Chromatography (IC) (1) INTRODUCTION - Standard or Reference Values set up by government bodies e.g. EPD, Water Works Department, etc - Analytical methods two types (laboratory and field methods) REFERENCE: Standard Methods for the Examination of Water & Waste Water (628.161 S78) jointly issue by i) American Public Health association ii) American Water Works Association iii) Water Environment Federation - Different methods used by different water bodies : i) Clean water - drinking water, fresh water ii)Water for recreation-seawater, beach & river water iii) Dirty water - waste water, water pollution, etc. iv) Processes water - water for industrial use, consideration for cost, corrosion & fit for purpose. Some can be very clean such as DI water. v) Water for specific uses - agriculture/irrigation, cement/concrete mixing, cooling water, etc.

Wastewater, Water, Soil & Sediments Analysis

(2) WATER QUALITY AND STANDARDS Standards are normally set up by government agents or international bodies & variable with different water bodies 2207/WA/2013/2 2.1) Drinking water : i) Microbial aspects, ii) Chemical aspects, iii) Radiological aspects, iv) Acceptable aspects WHO Guideline values - Historically, spread of water borne disease is due to contamination of potable water by dirty water (sewage) with faecal bacteria e.g. London before building deep underground sewers & surface drainage water debate in Bombay, India before link of illness to bacteria has establishedDevelopment of water supply system for cities and associated regulations, practices and control -1914: U.S. Public Health Service set the standards for water for bacteria. -1925 : Cu, Pb & Zn, Pb = 0.1 mg/L (City water supply by leaded pipeline) -1946: 7 additional chemical substances -1962 : Pb 0.05 mg/L (availability of method) Current WHO guideline for Pb : 0.01 mg/L -other chemicals added to water: i) chlorination for disinfecting water to avoid water borne hyphoid & other enteric diseases, ii) fluoridation: start in 1945 to reduce dental cavities. Not too high or too low for best effect. -Drinking water is metered & used to calculate waste water charges in HK.World-wide shortage of water.In California, increase 3x cost to achieve effective water-saving.

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World health Organiszation (WHO): Guidelines for drinking water quality, Third Edition, 2004. - WHO issues guidelines for quality and standards of drinking water, risk assessment, monitoring & analysis methods, sources of pollution, water treatment - Depending on sources of water, water quality varies greatly in different places, some salty water or hard water have to be used in drinking. The most important is fit for drinking with no known health hazards - Following parameters specified in WHO guidelines: i) Bacteriological quality: Faecal bacteria indicator (E. coli) as general standard (individual bacteria if needed) ii) Chemicals of health significance (Guidelines) a)Inorganic constituents - heavy metals & anions b)Organic constituents-chlorinated hydrocarbon, PAH, Nitro-compounds, BTEX (benzene, toluene, ethylbenzene and xylene) c) Pesticides d) Disinfectants and disinfectant by-products iii) Radionuclides for alpha and beta emitters iv) Substances that may give rise to complaints- other metals, hardness, pH, turbidity, dissolved solid, etc. v) Early edition focus on guidelines & standards. From 3rd edition (2004), focus is changed to pollution source control & prevention measures (www.WHO.int)

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Guideline for Bacteriological Quality: [System survey guidelines for water classification] Standard based on E.Coli as indicator organism problem of mixing up portable water with sewer [connect to the wrong pipe]

[Guideline for verification of microbial quality]

Analytical question : Zero detection limit ??

Guidelines for health significant chemicals (86 listed with guideline values)

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Guidelines for Radionuclides

[HK ~3mCi (100MBq) <100mCi]

-Determination of individual radioactive species requires sophisticated and expensive analysis and their concentrations are normally very low in water, a screening procedure for total radioactivity present in the form of alpha and beta radiation is thus used. The screening levels for drinking water below which no further action is required are 0.5 Bq/litre for gross alpha activity and 1 Bq/litre for gross beta activity. [Note : SI unit for radioactivity is Becquerel (Bq), where 1 Bq=1 disintegration per sec.] - To facilitate quick response, HK Observatory hosts the monitoring network operated by chemists from Government Lab - Monitoring air, water and dairy products to check against potential leakage from nearby Daya Bay nuclear power station - High background in HK due to HK granite (igneous rock)

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The sievert (Sv) is the SI unit of equivalent or actual radiation dose received by receptor (Note: Bq/Ci is energy emitted from radiation source). Radiation measured in Sv are designed to represent the biological effects of ionization radiation. However, differential absorption exhibits by different biological tissue. Taking differential absorption into consideration, the unmodified absorbed dosage of radiation energy is known as grays (Gy). For biological impact, further calculations must be performed to convert absorbed dose into effective dose, the details of which depend on biological context, far more complicated than just multiplying by a weighting factor. Man-made From previous Sv Natural radiation radiation exposure study, received, Sv/Yr <<Whole 1 Sv carries body CT withit a Cosmo 0.39 scan Crustal 0.46 5.5% chance (once) Food 0.29 of eventually <<World Air- 2.6 General clinical developing Japan >> <<exposure/Yr cancer.
Stomach X-ray <<check (once) Average Sv received per air travel >>

Yearly explosure to natural & manmade radiation

Doses > 1 Sv received over a short time period are likely to cause Lung X-ray radiation poisoning, possibly leading to check (once) death within weeks.

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Table 4 Guidelines for Aesthetic Quality Constituent/Characteristics Unit

Guidelines for recreation use

Guideline values 0.2 250 no guldeline value set Remarks

Alumintum mg/L Chloride mg/L Chlorobenzenes & chlorophenols -

these compounds may affect taste & odour

Colour true colour units (TCU) 15 Copper mg/L 1.0 Detergents no guldeline value set Hardness mg/L (as CaCO3) 500 Hydrogen sulfide not detectable to consumers Iron mg/L 0.3 Manganese mg/L 0.1 Oxygen-dissolved no guldeline value set pH 6.5 - 8.5 Sodium mg/L 200 Solids--tolal dissolved mg/L 1000 Sulfate mg/L 400 Taste and odour inoffensive to most consumers Temperature no guldeline value set ) Turbidity nephelometric turbidity units (NTU) 5 Zinc mg/L 50

no foaming or taste & odour problems

preferably <1 for dislnfection efficiency

2.2) Water for recreation (beach & swimming pool) - less stringent as compared to drinking water 2207/WA/2013/10 -Under Water Pollution Control Ordinance, HK is divided into 10 water control zones. Water quality varies depending on different water control zone, with the most stringent in Tolo Harbour & Channel. The reason is due to the special regional geography with small mouth for holding a large volume of water. -In HK, 36 gazetted bathing beaches, 11 on HK Island & 25 in NT & outlying islands. -Beaches are classified by E.Coli count /100 mL as Good (<24), Fair (25-180), Poor (181-610) & Very Poor (>610). HKEPD issues regular announcement on beach water quality during bathing season. Major water pollution is due to excess sewage discharged with little treatment. - Major problem areas : Typhoon shelter, Kowloon Bay near Kaitak : stagnant water with little circulation

2207/WA/2013/11 2.3) Dirty Water -Polluted water- Three types : i) Sewage water (toilet & kitchen), ii) Drainage water (rainwater) & iii) Industrial effluent waters -Standards set by HKEPD are different for different water zones. Allow mixing of sewage & industrial water in old industrial areas. Mix of drainage & sewage water not allow. -Control water zones with different standards for discharge of wastewater. Most strict at Tolo harbour -parameters includes BOD, COD, grease & oil, metals, etc. -information available from HKEPD website - In Holland, COD & BOD are used to charge wastewater. 2.4) Processes water: ingredients in finished products, buoyant transporting Table 3 Recommended values for water used in mixing concrete media,coolant,steam Threshold value Substance /power generation 25 mg/L free CO2 (variable quality by pH approx. 7 different industries) sulfide not detectable 2.5) Water for 250 mg/L sulfate 1 500 mg/L specific uses : water chloride 100 mg/L for irrigation, mixing ammonium 200 mg/L cement /concrete & magnesium 25 mg/L potassium permanganate for cooling water in consumption air conditioning not detectable humic acids, hydrocarbons For concrete reinforced with iron (water quality 100 mg/L regulated & regularly chloride nitrate 20-50 mg/L monitored )

(3) ANALYTICAL METHODOLOGY 3.1) Field Sampling And Measurement 3.1.1) Sampling and preservation 3.1.2) Direct measurement in field 3.2) Measurement in Laboratory 3.2.1) Water quality testing methods I) Microbiological testing methods II) Radiological testing methods III) Chemical testing methods A) Sample pretreatment B) Chemical analysis a) General properties analysis b) Specific chemical analysis 3.2.2) Automatic Analysis Methods

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3.1) Field Sampling and Measurement 3.1.1) Sampling and Preservation - Surface water : Collection by bucket or sample container dipped into water in front of vessel - Drinking water : Running time of 15 to 30 min prior to collection. Sterile glass bottle for bacteria testing - Waste water: Similar to surface water collection. For QC purpose, use 2-hrs pooled or daily pools samples -Processes water: Use processes water to wash bottles - Ground water: Tube wells or shafts with pumps or buckets

Fig.1 Schematic representation of a tube well Sample Conservation - Different methods for different analytes - No HNO3 for COD - No chemical added for BOD -Sample preservation can affect analysis
Table 1 Conservation of substance present in water Parameter trace metals Preservation Method 5 mL HNO3/L

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Maximum storage time several weeks few days few days

o

ammonium, total N2 5 mL HNO3/L mercury nitrite cyanides iron-II sulfide phenols 1 mL 10% K2Cr207- solution/L no stabilization possible, cooling to 4 C alkali to pH = 11 addition of 2,2-bipyridine 2 mL 10% zinc acetate solution/L alkali to pH = 12

1 day 1 day 1 day 1 week 1 week

Domestic Waste Water Sampling

Waste water from sewer Screening & Degritting Remove solids > 6mm Primary Sedimentation

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Water Samples
Effluent Discharged Excess activated sludge digested, de-water & landfill

Remove 50% suspended solid as Primary Sludge

Reduce ammonia-N and total nitrogen by microbes in activated sludge Sedimentation Tank

Secondary Biological Treatment (6-8 hrs)

Activated Sludge Recycled

Aeration with activated sludge

3.1.2) DIRECT FIELD MEASUREMENT 2207/WA/2013/15 - Simple operation and procedure, give immediate results with no problems associated with sample storage and transportation - Specific equipment for each measurement, e.g. glass electrode for pH, Fluoride ISE for F- , oxygen electrode for dissolved oxygen, conductivity meter for conductivity measurement - For spectrophometric measurement, coloured glass comparators are used for quantitation e.g. use of o-toluidine-HCl for residual chlorine. Recent development using optical fibre with computer-attached monochromator - Parameters includes temperature, weather conditions, Redox potential, Dissolved oxygen, Active chlorine, conductivity, pH, etc. - Meter-type instruments with no power requirement or run by battery power
Colorimeter pH meter

Coloured glass as standard

2207/WA/2013/16 3.2) Measurement in Laboratory 3.2.1) Water quality testing methods I) Microbiological Testing Methods -Expensive to detect specific pathogen. Thus, use of indicator organisms are used. Requirements : i) They are always present in contaminated water, absent when faecal contamination not present, ii) generally survive longer than pathogens, and iii) easy to identify. -If indicator organism absent, water is assumed safe. Total coliform group : defined as all of the aerobic & anaerobic, Gram-negative, non-spore-form rod-shaped bacteria which ferment lactose with gas formation within 48 hours at 35oC given by Standard Methods for the Examination of Water & Wastewater. Faecal or thermal tolerant coliform (E.coli): the incubation temperature is 44.5 0.2C and other conditions kept constant coliform from human Method : Pass 100-mL sample through sterile membrane filtration following by incubation in lactose-peptone solution at 35oC and counting confirmed colonies (by morphology, colour and biochemical identification) after 24-48 hours. For individual pathogenic bacteria : tradition method by colony counts in selective media, or new PCR/DNA techniques for detecting specific bacteria

II) Radiological Testing Methods Alpha particle: 1)Positive

Beta particle: 1) Negatively charged particles 2) Longer penetration distance -Unstable nucleus decays by emitting alpha particle, beta particle and Gamma ray (high energy radiation e.g. X-ray) / x neutron
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charged particles, 2) shortest pentration distance

Gamma Ray:1) Low abundance radiation, 2) longest penetration distance

Heavy water

Paper

A) Measurement in field : Thin metal foil Thick Pb block - Measure beta-particle or gamma radiation due to long distance measurement. Alpha particle is too short a distance to measure. - It depends on the penetration power of radiation through given thickness of Aluminum block to distinguish 241Pu, 129I or 90Sr. Thus, one can put detector behind a given thickness of Al block to identify type of radiation. (Qualitative) - The intensity measurement is more a problem for quantitative analysis, as sources & their distance to the detector are needed.

I)Geiger Mller (GM) counter Cheap, portable, detect ionizing radiation, e.g. beta particle & gamma ray. Special model detect alpha particle. However, low sensitivity. An inert gas-filled tube (He, Ne or Ar with halogen added) briefly conducts electricity when a particle or high energy radiation makes the gas conductive. The tube amplifies this conduction by a cascade effect with outputs displayed by a needle or lamp and/or audible sound.(Personal monitor, Yes/No indication)
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II) Scintillation counter measures ionizing radiations. The sensor consists of a transparent phosphor, plastic usually containing anthracene that fluoresces when struck by ionizing radiations. A sensitive PMT measures the light from the crystal for counting and possibly quantify the amplitude of the signals from phospor (More sensitive than GM,semi-quantitive) B) Measurement in the Lab. I) Liquid scintillation counting is a standard laboratory method for measuring raditaion from beta-emitting nuclides.(Fixed demension Quantitative)

Samples are dissolved or suspended in a "cocktail/mixed solution containing an aromatic solvent & small amounts of other fluorescence additives (fluors) Beta particles emitted from sample transfer energy to the solvent molecules, which transfer their energy to the fluors. The excited molecules dissipate energy by emitting light for measurement. II) Sealed Xenon Proportional Counter /Argon gas-flow proportional counter for accurate determination of & radiation. A quench gas added to prevent proportional counter acting as a GM counter. Ionization excites inert gas. When coming down to ground state, emit light which is amplified by cascade electrodes like PMT for pulse mode of measurement. 2207/WA/2013/19 Lab procedures : Two common methods for gross alpha & gross beta activities: 1) Evaporation of known sample volume to dryness & measure activity of residue (as alpha absorbs by solid material, not for samples with high total dissolved solid (TDS). 2) Radioactive co-precipitation (precipitation on a carrier precipitate or collector for trace metals. Using a large amount of insoluble K salt to co-precipitate radioactive cations):simple, specific, but time consuming. Lots of foreign materials added to sample, producing contamination problem. Applicable to high TDS samples due to saturated effect. Hard to get 100% recovery.

2207/WA/2013/20 III) CHEMICAL TESTING METHODS A) SAMPLE PRETREATMENT (waste & clean water) Purpose of sample pretreatment : a) cleanup major sample matrix, b) to remove interference & c) to enhance sensitivity (preconcentration) before GC separation & quantitation Types of Pretreatment Methods : i) Preconcentration Evaporation (heat lamp or hot plate): simple, inexpensive, use for most substances, compatible to most methods, no chemical added & no contamination. However, time consuming, not for volatile compounds, adsorpt-ion loss, non-specific, high solid content, only for highly soluble compounds. Maximum ~50x . Ion exchange (cation, anion, chelating): simple, rapid, inexpen-sive setup,regenerated column, very high preconcentration factor. However, suitable detector, specific exchanger & solvent needed ii) Isolation & Phase Separation Precipitation (inorganic & organic reagents):simple, inexpen-sive & quite specific. However, for low soluble compounds with large crystals,lots of foreign ions added & re-dissolution required. iii) Extracting Trace Pollutants from Liquid Phase - Extract trace levels of pollutants from complicated matrixes, remove interferences and concentrate analytes prior to chemical analysis.Commonly used for waste water. - Instrument are often used to speed up the extraction and for automation of procedure

-Three major types of extraction from liquid phase : Purge & Trap, Solid Phase Extraction & Solvent Extraction a) Purge & Trap for Volatile (Purgeable) Organic compounds : Two steps, 1st step is to purge organic compounds out from aqueous solution by gas and this leads to preconcentration and removal of interferents. Second step is to strip organic compounds out from the adsorbents by carbon 2207/WA/2013/21 disulphide or by thermal desorption. - Two types : Off-line and on-line Purge & Trap Off-line Purge & Trap- Removal of moisture by Nafion drier to reduce the effect of water on activated carbon (polymer membrane allowed water to go through but not organic compounds, dried by desiccants packed in Drier) On-line Purge & Trap with cryogenic on-column focusing (capillary column in liquid N2 to trap VOCs) with direct coupling to GC: used to handle a lot of samples at low concentration.
Purge Gas Nafion Drier Activated Carbon Adsorbent Trap

Aqueous solution sample

A hygroscopic ion exchange membrane fabricated in tubular form and made up of a co-polymer of tetra-fluoroethylene and fluorosulfonyl monomer
F F F F

F2C=C OCC OCCSO3H n F CF3 F F Du Pont Nafion

2207/WA/2013/22 Procedures in SPE : -C18 SPE (Sorbent) to preconcentrate trace non-polar organic compounds from aqueous medium

Pressure Solid Phase Extraction using cartridge Vacuum manifold for solid phase extraction of multiple cartridges b) Solid Phase Extraction (SPE) -Technique involves the use of a solid sorbent selectively adsorbing the analyte from a large volume of diluted sample solution, and then eluting by a small volume of strong solvent - Ion Exchange Resin for pre-concentration of trace metals from aqueous low-ionic strength solution, then by hydroxide stripping

- Chelating resin (Chelex 100) [styrene-divinyl copolymer with paired iminodiacetate ions as chelating groups to complex with transitional metals] to preconcentrate transition metals from high ionic solution and removal of interferents 2207/WA/2013/23 c) Solvent Extraction Organic solvent extraction Separating funnel more efficient using discrete multiple extraction with several small volumes of extractants Extracting Trace organic pollutants -like-dissolve-like concept for organic compounds with principle based on favourable distribution ratio in the organic phase. K = Corg /Caq. -More complete extraction by multiple extractions using several lots of smaller volumes than a large solvent volume -Choice of solvent: purity, cost, safety (toxicity & fire hazards) and solubility of analyte such as hydrocarbon (hexane) & chlorinated hydrocarbon solvents (1,1,1-trichloroethylene) -Best is CFC on safety & chemical grounds (high purity with only C-F bonds & good solubility of organics) -Problem of long extraction time a few hours and CFC is not environmental friendly Extraction and Preconcentration of Trace Metals -Chelation of metals by EDTA and other complexes before solvent extraction by MIBK (methyl isobutyl ketone) for AAS or ICP determination removal of interference by other ionic compounds, e.g. phosphate -Substantial number of metals can be extracted by chloride complexes in HCl, e.g. Fe (III), Sb(V), Ga(III), Ti(III), Au(III), Mo(VI), As(III) & Ge(IV) - pH is important for extraction & should be optimized

B) CHEMICAL ANALYSIS - Two main types : general and specific analysis a) General properties of water i)General observations:colour, odour & appearance ii) pH or alkalinity/ acidity (acidic/base capacity) iii) Dissolved and undissolved substances Total residue dried at 103-105o C (as mg/L): both dissolved and undissolved solid content Total suspended matter(as mg/L): undissolved solid only Turbidity : cloudiness of solution, related to suspended solid content except for smaller particles. NTU turbidimeter is used for measuring nephelometric turbidity units (NTU) based on scattered light measurement with calibration by a primary standard for turbidity (an aqueous suspension of formazin, an insoluble polymer formed by condensation of hydrazine sulfate and hexamethylenetetramine)

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2207/WA/2013/25 Conductivity (as mS): electrical conductivity is a total parameter for dissolved and dissociated ionic compounds. In seawater, it is related to salinity. It can be used to calculate the total ion content in given samples Dissolved oxygen -important indicators for water quality b) Specific Chemical Analysis: i) Organic matters include Grease & Oil, Hydrocarbons, BOD & COD, Trace organics Grease & Oils: indicate total organic content, measure by dissolving organic matters in CFC (now hexane after CFC ban), evaporate to dryness and weigh Hydrocarbons: direct FTIR on solvent extract for total hydrocarbons. COD (chemical oxygen demand) & BOD (biochemical oxygen demand). -BOD starts as an empirical parameter to assess water pollution by measuring the quantity of dissolved oxygen to oxidize organic compounds found in water with assistance of microorganisms & under defined experimental conditions. It varies with conditions such as temperature, dissolved O2 , levels or type of bacteria & less reproducible compared to COD. However, a lot of historical data using BOD for polluted waters, making it an essential parameter (To set charges for waste water in EU)

BOD - Uptake of oxygen during 5 days or longer by biological reducing system such as activated sludge system. In UK, 5 days adopted during London sewer construction based on average time for water to go through river Thames. For EU, 7 days is adopted to meet weekly work schedule.Dissolved Oxygen (DO) measured by electrode with amperometric detection before and after 5 or 7 days incubation at 20/27oC with or without the seeding of bacteria. Oxygen 2207/WA/2013/26 Electrode

ABS Body BNC Connector Inner glass tubing Noble Metal Cathode (Gold or Platinum) Teflon membrane Anode (silver wire) Ag/AgCl or Calomel KCl Electrolyte Solution

BOD bottle BOD Empirical parameter covering organic compounds, reduced substances of nitrogen, sulphur, phosphorus & some metallic species. (Dl-D2)-(Bl-B2)x f BOD (mg O2/L) = P where D1 & D2 are DO (mg/L) of diluted sample immediately after preparation and after 5 or 7 days of incubation at 20/27oC respectively; B1 & B2 are DO seed control before and after incubation respectively; P is dilution factor and f is the ratio of seed in diluted sample in seed control. If no seed is added, the factor [(B1-B2)xf ] will be zero. Several BOD bottles are used (dilutions + seed) with water filled up to top with no air gap left.
O2 O2 O2

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Chemical oxygen demand (COD) is defined as the amount of oxygen in the form of oxidizing agent consumed in the oxidation of organic water components. The degree of oxidation depends upon the type of substance, pH value, temperature, reaction time and concentration of oxidizing agent as well as the type of added accelerators, if any. COD- a few hours measurement -a wet chemical analytical technique -refluxing acid-dichromate with a catalyst to carry out chemical oxidation (catalyst: Ag2 (SO4)) -back-titration with ferrous ammonium sulfate method COD : oxidation of reduced inorganic species + carbonaceous matter -oxidise non-carbonaceous compounds such as reduced states of N & S, some halides, many metals & bound hydrogen inorganic compounds -variable oxidation efficiency e.g. pyridine 0.8% benzene 8%, toluene 22.5%, o-cresol, valine, sodium stearate >90% Trace Organics -Often in complicated matrix require cleanup -Analytical methods: IR, MS, GC, GC-MS, Florescence (PAH) and electrochemical techniques - Organic pollutants e.g. phenols, surfactants, etc

ii) Inorganic compounds 2207/WA/2013/28 Hardness of water : total dissolved divalent cations as measured by EDTA titration. Trace metallic elements i) AAS : flame & graphite furnace, ii) ICP-AES (ppm) and ICP-MS (ppb), iii) Electrochemical stripping techniques, useful for trace metal analysis in seawater. Anions:required ppm/ppb determination in acid rain analysis & high tech industries. 2 methods i) Ion Selective Electrode: F- ISE, ii) Suppressed Ion Chromatography Ion Exchange Column to separate anions using NaOH as eluent.Suppressor column to reduce background conductivity via removing H+ with OH- to form H2O before conductivity detector iii) Non-suppressed Ion Chromatography (Non-suppressed IC) -Same as (ii) except no suppressed column, Use weak organic acid as eluent which absorb UV for indirect UV detection to avoid patent on suppressor (expired now)
Anions (Cl- and SO4=) are monitored in cooling water as indicator for corrosion Active chlorine compounds

-residual chlorine determine by amperometric titration using standard ferrous sulphate or by colorimetric methods using o-toluidine-HCl and coloured glass comparator. Nutrients in water (PO4= and NO3-). - Both are limiting nutrients in soiul for algae and plants. Used as fertilizers to enhance plant growth. Can cause
eutrophication (excessive algae growth known as algae bloom if present in water at high levels under favourable conditions.

Nitrogen Content in Wastewater 2207/WA/2013/29 Total Nitrogen = Ammonia-nitrogen (NH3-N)+Organic-nitrogen (Org-N)+Nitrate-nitrogen (NO3-N)+ Nitrite-nitrogen (NO2-N) -Nitrogen is essential for formation of proteins in cell growth and thus needed to keep activated sludge.However,too much causes algae boom. Ammonia is toxic to fish, and nitrates at high enough dosages in the drinking water cause methemoglobinemia in infants as it can convert to nitrites in the stomach and interfere with the oxygen-carrying capacity of hemoglobin in blood. - Nitrogen exists in NH3, organic-N, nitrate & nitrite in waste- water. Under right conditions, each can convertible to the others. -For NH3-N, Nessler titration or electrode (ISE) method (Fig. 1, a glass electrode mounted inside a gasFig. 1. Construction of permeable membrane to ammonia electrode measure pH) is used. It measures only NH3. The ratio between NH3 & NH4+ depends on solution pH (Fig. 2). Fig. 2. - It operates at 0-50oC, Fraction of working range:0.01-17000 ammonia and ppm (1.0 to 5x10-7M). ammonium ions as - Only operate in aqueous a function of pH. solution & interfere by oil & grease covering up the gas-permeable membrane. -For sample ionic strength >1.0M, requires adjustment

-In field, pH should keep to < 2 by H2SO4 and immediately after sampling to reduce degradation. - In laboratory, pH adjusted to 9.5 using a borate buffer and solution distilled into a receiving solution of either boric acid for Nessler titration or into H2SO4 for ammonia ISE determination. - Nessler reagent (0.09 mol/L solution of K2[HgI4] in 2.5 mol/L KOH) is used to detect small amounts of ammonia. A yellow coloration indicates the presence of ammonia and at higher concentrations, a brown precipitate may form. The sensitivity is about 0.3 g NH3 in 2 L.

2207/WA/2013/30 temp. at 4oC

NH4+ + 2[HgI4]2 + 4OH HgOHg(NH2)I + 7I + 3H2O

NO3- & NO2- Analysis (Inorganic nitrogen, ISE + Cd) -Typically, < 1 % N in form of nitrates (NO3-) or nitrites (NO2-), the ratio reflects microbial balance -NO3- converted to NO2- by a Cd reduction column or using hydrazine sulfate for the determination of NO2- (Total nitrate after nitrate reduction). - NO2- is determined by diazotizing with sulfanilamide and coupling with N-(1-naphthyl)-ethylenediamine dihydrochloride to form a highly colored azo dye which is measured colorimetrically. - NO3-= total NO2- (after nitrate reduction)-NO2- (without nitrate reduction). - Total Kjeldahl Nitrogen (TKN) is obtained by digesting sample for two and a half hours in H2SO4 and collecting NH3 distilled for analysis. Result consists of (organic + inorganic) nitrogen plus NH3-N. Thus, organic nitrogen can be calculated by subtracting NH3-N and inorganic nitrogen from TKN in the same sample.

 Phosphorus compound analysis include Total phosphorus, o-phosphate, hydrolyzable phosphate (total inorganic phosphate) & organically bound phosphate in water, wastewater or soil. - Natural total phosphate in water < 0.1 mg/L, normally fixed in soil and non-toxic. Requires. frequent monitoring in water analysis, leading to use of FIA for automatic determination.

-The intermediate product (NH4)3PO4-12MoO3 is a yellow colored Mo(VI) complex. When it is treated with ascorbic acid, the blue colored Mo(V) complex formed is detected spectrophotometrically.
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Injection of phosphate standards (~100 ug/mL) and samples

(4) SPECIAL METHODS: A) Automation in Water Analysis. 4.1) A lot of samples for analysis automation (different degrees). Normally first for time consuming step in sample preparation such as dilution or pH adjustment. 4.2) 3 essential elements in automation : brain (decision making by computer), eye (sensor) & hand (actuator). 4.3) The most simple system is automated titration with indicator electrode (eye), generator electrode or autopipette (hand), and microprocessor/computer /electronic circuitry (brain) for end-point detection. I) Discrete Automatic titration - electrochemical 2207/WA/2013/32 titrant generation and detection

Indicator electrodes - Two Pt flags -Amperometrically detect Br2/Br- ratio at both Pt electrodes

Generator electrodes -Pt coil - Coulometrically generate Br2 from Br-

- Ascorbic acid is back titrated with Br2 generated by electrolysis at anode reaction - Iodine is added as its redox reaction (I-/I ) with ascorbic acid is more reversible & it can be regenerated by reaction with Br2. - Operation : i) Conditioning of electrolyte by generating a fixed amount of Br2 (indicated by constant current i) ii) Add sample to reduce current, iii) Generate Br2 till current reaches constant current i again, iv) measure coulomb passed and calculate the amount of Br2 generated assuming 100% coulombic efficiency. II) Continuous Flow Segmented Autoanalyser: Commercial autoanalyser incorporating sample preparation & analysis available for COD, trace metals & nutrients (nitrate, nitrite, ammonia and phosphorus) - First autoanalyser for water analysis with segmented flow and air bubbles separating different samples
Dialyzer Sampler

Titration of Ascorbic Acid With Bromine Generated 2207/WA/2013/33 Ascorbic acid + Br2 2Br- + 2H+

Proportional Pump & Manifold

Note:Bubbles causes disturbance in detector baseline.Devices added to remove bubbles before detector

a single-channel Technicon AutoAnalyzer system

2207/WA/2013/34 III) Continuous Flow Non-segmented Analyser - Analyser based on newly deveoped Flow Injection Analysis (FIA). No air bubble to isolate samples. Use only segments of carrier liquid to separate samples.Require accurate control in flowrate & volume of sample injection to achieve repeatable dispersion during each run for sharp peaks at detector. FIA resemble HPLC with an empty column. a) Representation of a simple flow injection analyser; b) Sample flow in tube containing a plug of sample; c) typical response over time for samples and standards

Note: Asymmetric peak with tailing

- Successful development of HPLC makes suitable valves and low pressure pump commercially available at affordable price, leading to the development of FIA for non-segmented flow auto-analyzer. Due to its flexible design using parts from HPLC, FIA is now largely replaced the dedicated Continuous Flow Segmented Analyzer in automation for water analysis. The simplest FIA system is using Ion Selective Electrode for continuous flow direct analyte detection.FIA system with the widest applicationis using UV-Visible detector. - Simple Ion Selective Electrode combination electrode with built-in reference electrode for monitoring specific ion. As no separation before detection, the detector system (Fluoride ISE) must be very selective towards the analyte to avoid sample matrix interference.

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- The flowrate and response time of the ISE or UV detector affect the sample throughput (no. of analysis per hour) of the auto-analyzer. -Symmetrical peak gives the least error in measuring peak height for concentration. Asymmetric peak require peak area for quantitation, more complicated and less accurate measurement. -The most efficient sample throughput is when the first peak reaching baseline, the second peak will start with minimal time between samples. Thus, FIA can handle hundreds of samples everyday. This can be done by optimizing the flowrate and mixing rate of the FIA system.

2207/WA/2013/36 B) ION CHROMATOGRAPHY To meet the problem for determination of anions at ppm level, Two chemists working at from Dow Chemicals have patented a suppressor technology in 1975 to enable the use of conductivity detector to detect non-UV active anions like Chloride and Sulphate after their separation by HPLC using an Ion Exchange Column. They left Dow Chemicals to set up a new company (Dionex) to expand the suppressor technology to determine anions at ppm level for acid rain monitoring, taste in food, impurities in DI water and for various industrial monitoring, leading to the birth of a billion dollar industry for Ion Chromatography at California.

NaOH used as eluent (1) OH- + H+ H2O Low Background Conductivity SO4= + H+ H2SO4 (2) Strong acid, high conductivity (1) + (2) High Detection Sensitivity

Suppressor

-Patent by DIONEX is based on suppressor technology which is expired in 2005.

Ion Chromatogram using Suppressor Column for ppm determination of anions in water
Fung & Dao, Anal. Chim Acta, 1995, 315, 347.

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-Non-suppressed Ion Chromatography (Non-suppressed IC) -Same as IC except no suppressed column used, Use weak organic acid as eluent which absorb UV for indirect UV detection of non-UV absorbing anions to avoid patent on suppressor.

-After suppressor patent is expired, other companies enter the IC market, producing cheaper & better design with automatic/onsite regeneration of suppressor column -Dual Cycle with one 1) Running IC Column For Anions Separation & Quantitation Suppressor in use and the other regenerated
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2)On-column Suppressor Regeneration

-To stay ahead in IC technology, DIONEX developed Membrane Suppressor (patent still in force) to reduce postcolumn band broadening, enable continuous insitu suppressor regeneration & achieve ppb detection level.

Flowchart For MMS III Membrane Suppressor

- Membrane suppressor operated by applying +/ voltages across membrane to remove H+ and OH-Another direction taken by DIONEX is coupling IC

Regenerant (H2SO4 or electrolytic H2O) Membrane Eluent (NaOH) Sample (NaF, NaCl, Na2SO4) Membrane Regenerant (H2SO4 or electrolytic H2O)

with MS detector to enable ppt detection of anion and confirmation of anion (ClO4-) in milk & water

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i)High ClO4reduce thyroid hormone production ii) USEPA Max level at 1 ppb in drinking water in 2002 iii)IC2ppb QL at 100 L iv)IC/MS/MSDL 4ppt(ng/L) for 99/83 MRM at 100 L v) MRM confirmation for 37Cl/35Cl at 0.323 ratio

SUMMARY

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1) For water analysis, it is always under government control based on standards, guidelines or objectives. 2) Analytical method should have detection limits at 10 % or lower of the quantitation limit at guideline values. Do not need extreme sensitivity. Method should be fit-for-purpose. 3) Cost is always important for water analysis, as a lot of data are collected on daily basis. There is a trend towards methods capable of analyzing multi-parameters. 4) Validity of method is important. Thus, standard methods are often used with calibration by reliable standards. 5) High degree of automation in analytical method in water analysis. Future method development is towards integration with field sampling and laboratory analysis, as results should represent condition in field and many reactive pollutants can not be stored and required onsite monitoring.