QUALITY OF WATER

GENERAL INTRODUCTION

Absolutely pure water is never found in nature and contains number of impurities in varying
amounts. The rainwater which is originally pure, also absorbs various gases, dust and other
impurities while falling. This water when moves on the ground further carries salt, organic and
inorganic impurities. So this water before supplying to the public should be treated and purified
for the safety of public health, economy and protection of various industrial process, it is most
essential for the water work engineer to thoroughly check analyse and do the treatment of the
raw water obtained the sources, before its distribution. The water supplied to the public should
be strictly according to the standards laid down from time to time.

CHARACTERISTICS OF WATER For the purpose of classification, the impurities present in water
may be divided into the following three categories.

Sample collection
Objective: •
Objective of sampling is to collect a portion of material small enough in volume to be
transported comfortably and yet large enough for analytical purposes while still representing
the material being sampled.

• It is to demonstrate whether continuing compliance with specific regulatory requirements has

been achieved

Selection of sample containers

• Selection of sample container is utmost importance in sampling. Containers are generally

made of glass or plastic. Some sample analytes may get absorbed into the walls of plastic
containers and/or some contaminants may leach into samples.

• Trace level of some metals and pesticides may get adsorbed and/or absorbed onto the walls of
the glass container. In the same way, silica, sodium, and boron may be leached from soft glass.

Always use hard glass containers for all organics analyses such as pesticides, volatile organics,
PCBs, and oil & grease.
• Some of the analytes like pesticides, PAH etc. are light sensitive. Hence collect them in amber-
coloured glass containers to minimize photo degradation.

Selection of type of sampling

• Grab

• Composite

• Integrated

Grab sampling
• Grab samples are also called as spot or catch samples. Grab samples are single samples
collected at a specific spot at a site in specified time. Grab samples are to be collected only
when the source is known to be constant in composition for an extended period of time.
Examples are, ground water samples, well mixed surface waters, large lakes, rivers, estuaries,
shorelines, wastewater streams that are expected to be constant in composition over an
extended period of time, like spent wash line in a distillery.

• When the source composition varies from location to location, like upstream and downstream
of a river, then grab samples can be collected from appropriate locations. This helps in finding
out the extent of variation and duration of variation.

Composite sampling
• Composite sampling is carried out when the liquid matrix is expected to be heterogeneous and
varies from time to time or depth or at many sampling locations. This type of sampling provides
a representative sampling for this type of matrix and is carried out by combining portions of
multiple grab samples collected at regular intervals. If the flow is expected to be constant, then
volume based sampling can be carried out. If the flow varies, like sewerage line, then sampling
can be done by flow based composite, i.e., collecting sample that is proportional to the
discharge. Time composite sampling represents a 24hour period, with interval being 1-3 hours.

Use composite samples only for parameters that will remain unchanged under the sampling
conditions, preservation and storage. For parameters like pH, temperature, residual chlorine,
carbon dioxide, alkalinity, sulfide, dissolved oxygen, Oil & Grease etc. avoid composite sampling
and analyse individual samples as soon as possible, preferably in the field itself, except for
sulfide and Oil &Grease.

Integrated sampling
• Integrated sampling is carried out by collecting mixture of grab samples collected from
different points simultaneously. The points may be horizontal or vertical variation. Examples
include river, stream or reservoir or lake that varies in composition across the width and depth.
Also in industries that have different streams and combined treatment is proposed, than
integrated sampling of different streams can be made to understand the significant effect on
treatment.

Selection of sampling points

• Selection of sampling points plays an important role in sampling. The site selection should be
based on the objective of the study. If the monitoring is carried out for judging suitability of
water for drinking purpose, then the sampling point shall be near the intake point. Always
samples must be taken from locations that are representative of the source, treatment plant,
storage facilities, point of discharge, and point of use. Eventhough there is no methodology for
site selection on a cook book basis some general basic rules can be followed to have a sound
sampling programme.

The points chosen should generally yield samples that are representative of the system as a
whole. For this it is important to select a well-mixed zone. When samples are collected from a
river or stream, the observed results may vary with depth, flow and distance from the shore.
Hence, if equipments are available collect an integrated sample from top to bottom in the
middle of the lake or river or from side to side at mid-depth. Otherwise, preferably collect
samples at various points of equal distance across the water body. If only one sample can be
collected, collect it in the middle of the water body at mid depth.

Avoid areas of turbulence and at weirs. Generally collect samples beneath the surface with the
mouth directed towards the current. For oil and grease, collect sample at the surface. In
case of groundwater sampling, select wells that are in continuous use.

Selection of type of filling the container

Depending on the type of analysis to be performed, fill the container full or leave space for
aeration. For most organic compound determination like pesticides, PAH, VOC etc. and for
sulphide fill the container with out any air space. For microbiological and inorganic analyses
leave space for aeration and mixing. The space to be left for aeration and mixing should be
atleast 1% of the container volume. If the bottle is with preservative take care that the
preservative added is not lost or diluted by overflow.

In – situ measurements
 Parameters like pH, conductivity are temperature dependent. Hence, if the temperature varies
significantly, the results of these parameters also vary. Delay in analysis may also lead to loss of
dissolved gases like carbon dioxide, oxygen. Hence some of the parameters like temperature,
ORP, dissolved gases, shall be analysed in situ and parameters like pH, conductivity, alkalinity,
residual chlorine immediately after sample collection.

Sample labeling

• Labeling is an important part in sampling programme. The following information should be

included in the label. Use water proof ink to record all the information.

• Date and time of sampling • Sample field code • Sampling point • Nature of sample: Effluent /
Surface water / Ground water / Others • Type of sample (Grab/Composite/Integrated) • Pre-
treatment or preservation carried out on the sample • Any special notes for the analyst • Name
and sign of sample collector.

ANALYSIS OF WATER

A) PHYSICAL CHARACTERISTICS

The following are the physical characteristics

1. Turbidity

2. Colour and temperature

3. Taste and odour

1. TURBIDITY

Turbidity is caused due to presence of suspended and colloidal matter in the water. The
character and amount of turbidity depends upon the type of soil over which the water has
moved ground waters are less turbed than the surface water. Turbidity is a measure of
resistance of water to the passage of light through it. Turbidity is expressed as NTU
(Nephelometric Turbidity Units) or PPM (parts per million) or Milligrams per litre (mg/l).
Turbidity is measured by 1) Turbidity rod or Tape 2) Jacksons Turbidimeter 3) Bali‟s
Turbidimeter The Sample to be tested is poured into a test tube and placed in the meter and
units of turbidity is read directly on the scale by a needle or by digital display. Drinking water
should not have turbidity more than 10 N.T.U. This test is useful in determining the
detension time in settling for raw water and to dosage of coagulants required to remove
turbidity.

1) Turbidity rod or Tape:

Turbidity rod is used for measuring turbidity of water in the field. It consists of a
graduated aluminium rod, about 20.3 cm in length, at the upper end of which is
attached a graduated non-stretchable tape of about 12.2 cm long. At the lower
end of the aluminium rod, a screw containing a platinum needle and a nickel ring
is inserted. The graduated tape has a mark at its top end specifying the position
of eye during the test. In order to find the turbidity, the lower end of the rod is
gradually immersed in water whose turbidity is to be determined. Eye is kept
constantly at the marked position and the platinum needle is watched. The rod is
moved slowly in water till the platinum needle just disappears from the vision
due to turbidity of water. The reading of the graduated tape near the water
surface directly gives turbidity in p.p.m. the rod gives only rough value of the
turbidity of water.

2. Jacksons Turbidimeter

This is a laboratory apparatus which is used to measure turbidity when it

is more than 100 p.p.m. it consists of a metal stand holding a metal container and
graduated glass tube in it. A standard candle is placed below the stand. The water
sample is poured in the sample and the image of the flame of the standard
candle is seen through the turbid water in the glass tube. The level of water in
the glass tube is gradually till th image of the flame ceases to be seen. The height
of the water column, measured the graduated glass tube provides the measure
of the turbidity of the water The longer the light path of 21.5 cm corresponds to

.
11 JTU while light path of 10 8 cmincreasedcorresponds to 200 JTU where 1 JTU=

1 p.p.m.
2. COLOUR AND TEMPERATURE

Colour in water is usually due to organic matter in colloidal condition but some times it is
also due to mineral and dissolved organic impurities. The colour produced by one milligram
of platinum in a litre of water has been fixed as the unit of colour. The permissible colour for
domestic water is 20ppm on platinum cobalt scale. The colour in water is not harmful but
objectionable. Temperature of water is measured by means of ordinary thermometers. The
temperature of surface water is generally at atmospheric temperature, while that of ground
water may be more or less than atmospheric temperature. The most desirable temperature
for public supply between 4.4 C to 10 C. The temperature above 35 C are unfit for public
supply, because it is not palatable.

3. TASTE AND ODOUR

Taste and odour in water may be due to presence of dead or live micro-organisms, dissolved
gases such as hydrogen sulphide, methane, carbon dioxide or oxygen combined with organic
matter, mineral substances such as sodium chloride, iron compounds and carbonates and
sulphates of other substances. The tests of these are done by sense of smell and taste
because these are present in such small proportions that it is difficult to detect them by
chemical analysis. The water having bad smell and odour is objectionable and should not be
supplied to the public. The intensities of the odours are measured in terms of threshold
number. This number is numerically equal to the amount of sample of water in C.C‟s
required to be added to one litre of fresh odourless water.

B) CHEMICAL CHARACTERISTICS In the chemical analysis of water, these tests are done that
will reveal the sanitary quality of the water. Chemical tests involve the determination of
total solids, PH value, Hardness of water, Chloride content etc.

1) TOTAL SOLIDS AND SUSPENDED SOLIDS

Total solids includes the solids in suspension colloidal and in dissolved form. The quantity of
suspended solids is determined by filtering the sample of water through fine filter, drying
and weighing. The quantity of dissolved and colloidal solids is determined by evaporating
the filtered water obtained from the suspended solid test and weighing the residue. The
total solids in a water sample can be directly determined by evaporating the filtered water
obtained from the suspended solid test and weighing the residue. The total solids in a water
sample can be directly determined by evaporating the water and weighing the residue of
the residue of total solids is fused in a muffle furnace the organic solids will decompose
where as only inorganic solids will remain. By weighing we can determine the inorganic
solids and deducting it from the total solids, we can calculate organic solids.

2) PH VALUE OF WATER PH value denotes the concentration of hydrogen ions in the water
and it is a measure of acidity or alkanity of a substance. PH = - log 10[H+] or 1 / log 10[H+]
Alkalinity Depending upon the nature of dissolved salts and minerals, the PH value ranges
from 0 to 14. For pure water, PH value is 7 and 0 to 7 acidic and 7 to 14 alkaline range. For
public water supply PH value may be 6.5 to 8.5. The lower value may cause tubercolation
and corrosion, where as high value may produce incrustation, sediment deposits and other
bad effects. PH value of water is generally determined by PH papers or by using PH meter.
PH can read directly on scale or by digital display using PH meter.

3) HARDNESS OF WATER

It is a property of water, which prevents the lathering of the soap. Hardness is of two types.
1. Temporary hardness: It is caused due to the presence of carbonates and sulphates of
calcium and magnesium. It is removed by boiling. 2. Permanent hardness: It is caused due to
the presence of chlorides and nitrates of calcium and magnesium. It is removed by zeolite
method. Hardness is usually expressed in gm/litre or p.p.m. of calcium carbonate in water.
Hardness of water is determined by EDTA method. For potable water hardness ranges from
5 to 8 degrees. HARDNESS REMOVABLE Generally a hardness of 100 to 150 mg/litre is
desirable. Excess of hardness leads to the following effects. 1. Large soap consumption in
washing and bathing 2. Fabrics when washed become rough and strained with precipitates.
3. Hard water is not fit for industrial use like textiles, paper making, dye and ice cream
manufactures. 4. The precipitates clog the pores on the skin and makes the skin rough
Precipitates can choke pipe lines and values 6. It forms scales in the boilers tubes and
reduces their efficiency and cause in erustations 7. Very hard water is not palatable When
softening is practices when hardness exceed 300mg/lit. Water hardness more than 600
mg/lit have to rejected for drinking purpose.

METHODS OF REMOVAL OF HARDNESS

1. Boiling

2. Freezing

3. Lime addition

4. Lime soda process

5. Excess Lime treatment

6. Caustic soda process

7. Zeolete process
 8. Dimineralisation or exchange process.

Methods 1,2 and 3 are suitable for removal of temporary hardness and 4 to 8 for both
temperory and permanent hardness. The temporary hardness is removed as follows.

Boiling

heat

Ca(HCO3)2 -----------> CaCO3 + CO2 +H2O

Mg(HCO3)2 -----------> MgCO3 + CO2 +H2O

Addition of lime

Ca (HCO3)2 + Ca(OH)2 -----------> 2CaCO3 + 2H2O

Mg(HCO3)2 + Ca(OH)2 -----------> CaCO3 + MgCO3 + 2H2O

Removal of permanent Hardness:

1. Lime soda process :

2. Zeolite process

CHLORIDE CONTENT

The natural waters near the mines and sea dissolve sodium chloride and also presence of
chlorides may be due to mixing of saline water and sewage in the water. Excess of
chlorides is dangerous and unfit for use. The chlorides can be reduced by diluting the
water. Chlorides above 250p.p.m. are not permissible in water.
NITROGEN CONTENT
The presence of nitrogen in the water indicates the presence of organic matters in the
water. The nitrogen may be present in the water may be in one or more of the following
forms. 1. Nitrates 2. Nitrates 3. Free ammonia 4. Albuminoid nitrogen. Excess presence of
nitrogen will cause “MATHEMOGLOBINEMIA” disease to the children.

NITRATE

Nitrate constitutes the final stage in the oxidation of nitrogen compounds, and

normally reaches important concentrations in the final stages of biological oxidation.

The nitrate contained in pure well water derived from an extensive catchment is largely
 the result of biological activity in the surface layers of the soil, enhanced by cultivation
and the application of manures. When the nitrate in in the excessive amounts, t
contributes to the illness known as infant methemoglobinemia. Nitrate is measured
either by reduction to ammonia or by matching the colours produced with
phenoldisulphonic acid.

NITRITES

Nitrite in water is either due to oxidation of ammonium compounds or due to reduction of

nitrate. As an intermediate stage in the nitrogen cycle, it is unstable. A usual concentration
in natural water is in the range of some tenths of mg/L. Higher concentrations are present
in industrial wastes, sewage and in biologically purified effluents and in polluted streams.
In chlorinated supplies, levels of nitrite are often less than the limit of detection, .e.
0.005mg/L NO2-N but high levels may occur in unchlorinated water. Very high nitrite
levels are usually associated with water of unsatisfactory microbiological activity.

Nitrites can be determined by the following

methods

1. Colorimeter or spectro-photometer that can be operated at 543nm

2. Nessler tubes or 100mL capacity volumetric flask.

FREE AMMONIA

Ammonia is produced by the microbiological degradation of organic nitrogenous

matter. It appears, therefore, in many groundwaters as well as surface waters.
Concentrations of ammonia above a certain level in water polluted either due to
sewage or industrial waste is toxic to fish. The proportions of the two forms of
ammonia nitrogen in surface water depend pH. For accurate results, it is
generally preferable to distill off ammonia from the sample, and absorb in boric
acid. It is then determined either by titration or colorimetrically using Nessler
reagent. Direct nesslerisation of the sample is quicker depending upon
 interference. Ammonia produces a yellow coloured compound when reacts with
alkaline Nessler reagent, provided the sample is clarified properly. Pretreatment
with ZnSO4 and NaOH precipitates Ca, Fe, Mg and sulphide and removes
turbidity and apparent colour. Addition of EDTA (Before Nessler reagent) or
Rochelle salt prevents precipitation of residual Ca and Mg in the presence
of alkaline Nessler reagent.

METALS AND OTHER CHEMICAL SUBSTANCES

Water contains various minerals or metal substances such as iron, manganese, copper,
lead, barium, cadmium, selenium, fluoride, arsenic etc. The concentration of iron and
manganese should not allow more than 0.3 ppm . Excess will cause discolouration of
clothes during washing and incrustation in water mains due to deposition of ferric
hydroxide and manganese oxide. Lead and berium are very toxic, low p.p.m of these are
allowed. Arsenic, Selenium are poisonous and may cause totally, therefore they must be
removed totally. Human beings are effected by presence of high quality of copper in the
water. Fewer cavities in the teeth will be formed due to excessive presence of fluoride in
water more than 1 p.p.m. A laxative effect is caused in the human body due to excessive
presence of sulphate in the water.

DISSOLVED GASES

oxygen and carbondi-oxide are the gases mostly found in the natural water. The surface
water contain large amount of dissolved oxygen because they absorb it from the
atmosphere. Algae and other tiny plant life of water also give oxygen to the water. The
presence of oxygen in the water in dissolved form keep it fresh and sparkling. But more
quantity of oxygen causes corrosion to the pipes material. Water absorbs carbon-dioxide
from the atmosphere. If water comes across calcium and magnesium salts, carbon-dioxide
reacts with the salts and converts them into bicarbonates, causes hardness in the water.
The presence of carbon-dioxide is easily determined by adding lime solution to water gives
milky white colour.

BIO-CHEMICAL OXYGEN DEMAND

If the water is contaminated with sewage, the demand of oxygen by organic matter in
sewage is known as biochemical oxygen demand. The aerobic action continues till the
oxygen is present in sewege. As the oxygen exhausts the anerobic action begins due to
which foul smell starts coming. Therefore indirectly the decomposable matters require
oxygen, which is used by the organisms. The aerobic decomposition of organic matters is
done in two stages. The carbonaceous matters are first oxidized and the oxidation of
nitrogeneous matters takes place in the latter stage.

The Biochemical Oxygen Demand (BOD) is an empirical standardized laboratory test

which measures oxygen requirement for aerobic oxidation of decomposable organic
matter and certain inorganic materials in water, polluted waters and wastewater under
controlled conditions of temperature and incubation period. The quantity of oxygen
required for above oxidation processes is a measure of the test. The test is applied for
fresh water sources (rivers, lakes), wastewater (domestic, industrial), polluted receiving
water bodies, marine water (estuaries, coastal water) and also for finding out the level
of pollution, assimilative capacity of water body and also performance of waste
treatment plants.

This test measures the oxygen utilised for the biochemical degradation of organic
material (carbonaceous demand) and oxidation of inorganic material such as sulphides
and ferrous ions during a specified incubation period. It also measures the oxygen used
to oxidize reduced forms of nitrogen (nitrogenous demand) unless their oxidation is
prevented by an inhibitor. Temperature effects are held constant by performing a test
at fixed temperature. The methodology of BOD test is to compute a difference between
initial and final Do of the samples incubation. Minimum 1.5 L of sample is required for
the test. DO is estimate by iodometric titration.

Since the test is mainly a bio-assay procedure, it is necessary to provide standard

conditions of temperature, nutrient supply, pH (6.5-7.5), adequate population of
microorganisms and absence of microbial-growth-inhibiting substances. The low
solubility of oxygen in water necessitates strong wastes to be diluted to ensure that the
demand does not increase the available oxygen. A mixed group of microorganisms
should be present in the sample; otherwise, the sample has to be seeded Generally
temperature is controlled at 20ºC and the test is conducted for 5 days, as 70 to 80% of
the carbonaceous wastes are oxidized during this period. The test can be performed at
any other temperature provided the correlation between BOD5 20ºC is established
under same experimental condition (for example BOD5, 27ºC) is equivalent to BOD3,
 27ºC) for Indian conditions. While reporting the results, the incubation period in days
and temperature in ºC is essential to be mentioned

Equipment and apparatus

a. BOD bottles 300mL capacity (clean with a detergent, rinse thoroughly and drain

before use) with a water seal.

b. Incubator or water-bath to be controlled at 20ºC or at any desired temperature 1ºC.

Exclude all light to prevent photosynthetic production of DO.

Preparation of dilution water:

a.The source of dilution water may be distilled water, tap or receiving-stream water free

of biodegradable organics and bioinhibitory substances such as chlorine or heavy metals.

b. Aerate the required volume of dilution water in a suitable bottle by bubbling

clean-filtered compressed air for sufficient time to attain DO saturation at room
temperature or at 20ºC/27ºC. Before use stabilise the water at 20ºC/27ºC.

c.Add 1mL each of phosphate buffer, magnesium sulphate, calcium chloride and
ferric chloride solutions in that order for each Litre of dilution water. Mix well.
Quality of dilution water may be checked by incubating a BOD bottle full of
dilution water for 5 days at 20ºC for 3 days at 27ºC. DO uptake of dilution water
should not be more than 0.2mg/L and preferable not more than 0.1mg/L.d.

For wastes which are not expected to have sufficient microbial population, seed
is essential. Preferred seed is effluent from a biological treatment system. Where
this is not available, supernatant from domestic wastewater (domestic sewage)
settled at room temperature for at least 1h but not longer than 36hours is
considered sufficient in the proportion 1-2mL/L of dilution water. Adopted
microbial population can be obtained from the receiving water microbial
population can be obtained from the water body preferably 3-8 km below the
 point of discharge. In the absence of such situation develop an adapted seed in
the laboratory.

e.Determine BOD of the seeding material. This is seed control From the value of
seed control determine seed DO uptake. The DO uptake of seeded dilution water
should be between 0.6mg/L and 1mg/L.

Sample preparation:

a.Neutralise the sample to pH 7, if it is highly acidic or alkaline

b.The sample should be free from residual chlorine. If it contains residual chlorine

remove it by using Na2S2O3 as described below.

c.Take 50mL of the sample and acidify with addition of 10mL 1 + 1 acetic acid.
Add about 1g Kl. Titrate with 0.025N Na2S2O3, using starch indicator. Calculate
the volume of Na2S2O3 required per Litre of the sample and accordingly add to
the sample to be tested for BOD.

d.Certain industrial wastes contain toxic metals, e.g. planting wastes. Such

samples often require special study and treatment.

e.Bring samples to 20 ± 1ºC before making dilutions

f.If nitrification inhibition is desired, add 3mg 2-chloro-6-(trichloromethyl)

pyridine (TCMP) to each 300mL bottle before capping or add sufficient amount to
the dilution water to make a final concentration of 30mg/L. Note the use of
nitrogen inhibition in reporting results.

g.Samples having high DO contents, DO ≥ 9mg/L should be treated to reduce the

DO content to saturation at 20ºC. Agitate or aerate with clean, filtered
compressed air.

Dilution of sample: Dilutions that result in a residual DO of at least 1mg/L and

DO uptake of at least 2mg/L produce reliable results. Make several dilutions of the
pre- treated sample so as to obtain about 50% depletion of DO or DO uptake of
2mg/L

Siphon out half the required volume of seeded dilution water in a graduated

volumetric flask without entraining air. Add the desired quantity of mixed
sample and dilute to the appropriate volume by siphoning dilution water. Mix
well with plunger type mixing rod to avoid entraining air.

General guidelines for dilution range are as follows:

0.1% to 1% : Strong trade waste

1% to 5% : Raw or settled sewage

5% to 25% : Treated effluent

25% to 100% : River water

Sample processing:

a.Siphon the diluted or undiluted sample in three labeled bottles and stopper
immediately.

b.Keep 1 bottle for determination of the initial DO and incubate 2 bottles at 20ºC

for 3days. See that the bottles have a water seal.

c.Prepare a blank in triplicate by siphoning plain dilution water (without seed) to

measure the O2 consumption in dilution water.

d.Also prepare a seed blank in triplicate to measures BOD of seed for correction

of actual BOD.

E.Determine DO in a BOD test can in the blank initial day and end of incubation

period by Winkler method as described for DO measurement.

f.DO estimation in a BOD test can also be done by membrane electrodes. A DO probe
with a stirrer is used to determine initial and final DO after incubation in BOD
samples. The semi-permeable membrane provided in the DO probe acts as a
diffusion barrier against impurities between sensing element and sample.
Calculate BOD of the sample as follows:
a. When dilution water is not seeded

BOD as O2 mg/L = (D1 –D2) x 100/% dilution

b.When dilution is seeded

BOD O2 mg/L =(D1 – D2) – (B1 –B2) x 100/%

dilution c.When material is added to sample or to

seed control

BOD O2 mg/L = (D1 – D2) – (B'1 x B'2) x F x 100/% dilution

where,

D1 = DO of sample immediately after preparation, mg/L

D2 = DO of sample after incubation period, mg/L

B1 = DO of blank (seeded dilution water) before incubation, mg/L

B2 = DO of blank (seeded dilution water) after incubation, mg/L

F = ration of seed in diluted sample to seed in seed control (Vol. Of seed in diluted

sample / Vol. of seed in seed control)

B'1 = DO of seed control before incubation, mg/L

B'2 = DO of seed control after incubation, mg/L

CHEMICAL OXYGEN DEMAND (COD)

Chemical Oxygen Demand (COD) test determines the oxygen requirement

equivalent of organic matter that is susceptible to oxidation with the help of a
strong chemical oxidant. It is an important, rapidly measured parameter as
means of measuring organic strength for streams and polluted water bodies. The
test can be related empirically to BOD, organic carbon or organic matter in
samples from specific source taking into account its limitations. The test is useful
in studying performance evaluation of wastewater treatment plants and
 monitoring relatively polluted water bodies. COD determination has advantage
over BOD determination. COD results can be 4 hrs as compared to 3-5days
required for BOD test. Further, the test is relatively easy, precise, and is
unaffected by interferences as in the BOD test. The intrinsic limitation of the test
lies in its inability to differentiate between the biologically oxidisable and
biologically inert material and to find out the system rate constant of aerobic
biological stabilization.

Principle

The open reflux method is suitable for a wide range of wastes with a
large sample size. The dichromate reflux method is preferred over procedures
using other oxidants (e.g. potassium permanganate) because of its superior
oxidizing ability, applicability to a wide variety of samples and ease of
manipulation. Oxidation of most organic compounds is up to 95-100% of the
theoretical value.

The organic matter gets oxidised completely by potassium dichromate (K2Cr2O7)

with silver sulphate as catalyst in the presence of concentrated H2SO4 to
produce CO2 and H2O. The excess K2Cr2O7 remaining after the reaction is titrated
with ferrous ammonium sulphate [Fe (NH4)2(SO4)2]. The dichromate consumed
gives the oxygen (O2) required for oxidation of the organic matter. The chemical
reactions involved in the method are as under:

a. 2K2Cr2O7 + 8 H2SO4 ® 2 K2 SO4 + 2Cr2(SO4)3 + 8 H2O + 3O

C6H12O6 + 6O2 ® 6CO2 + 6H2O

. Cr2O7-- + 6Fe++ + 14H+ ® 6Fe+++ + 2Cr3+ + 7H2O

Procedure

Sample preparation: All samples high in solids should be blended for 2

minutes at high speed and stirred when an aliquot is taken for analysis. Select the
appropriate volume of sample based expected COD range, e.g. for COD range of 50-
500 mg/L take 25-50mL of sample. Sample volumeless than 25mL should not be
pipetted directly, but serially diluted and then a portion of the diluted sample taken.
Dilution factor should be incorporated in calculations.

a.500mL of sample diluted to 1000mL = 0.5mL sample/mL of diluent, 50mL = 25mL

of sample.

b.100mL of sample diluted to 1,000mL = 0.1mL sample/mL diluent, 50mL of diluent =

5mL of sample.

Reflux of samples:

a.Place 0.4g HgSO4 in a 250mL reflux sample

b.Add 20mL sample or an aliquot of sample diluted to 20mL with distilled water. Mix

well.

c.Add clean pumic stones or glass beads.

d.Add 10mL 0.25N (0.04167M) K2Cr2O7 solution and mix.

e.Add slowly 30mL concentrated H2SO4 containing Ag2SO4 mixing thoroughly. This
slow addition along with swirling prevents fatty acids to escape due to generation of
high temperature. Alternatively attach flask to condenser with water flowing and
then add H2SO4 slowly through condenser to avoid escape of volatile organic
substance due to generation of heat.

f.Mix well. If the colour turns green, either take fresh sample with lesser aliquot or

add more potassium dichromate and acid.

g.Connect the flask to condenser. Mix the contents before heating. Improper mixing

will result in bumping and blow out of flask content.

h.Reflux for a minimum of 2 hours. Cool and then wash down condenser with

distilled water.
 .Disconnect reflux condenser and dilute the mixture to about twice its volume with
distilled water. Cool to room temperature and titrate excess K2Cr2O7 with0.1M FAS
using 2-3 drops of ferroin indicator. The sharp colour change from blue green to
reddish brown indicates end-point or completion of the titration. After a small time
gap, the blue-green colour may reappear. Use the same quantity of ferroin indicator
for all titrations.

j.Reflux blank in the same manner using distilled water instead of sample.

Alternate procedure for low COD samples less than 50mg/L: Follow similar
procedure with two exceptions (i) use standard 0.025N (0.004167M) K2Cr2O7 and
(ii) titrate with standardize 0.025M FAS. The sample volume should be 5.mL.
Exercise extreme care

with this procedure because even a trace of organic matter the glassware or from the

atmosphere may cause gross errors. Compute amount of HgSO4 to be added based

on chloride concentrations. Carry blank reagent through the same procedure.

Calculations

COD as mg/L = (a –b) x N x 8000 / mL sample

Where, a = mL FAS used for blank

b = mL FAS used for sample

N = normality of FAS

8000 = Milieq. wt. of O2 x 1000

BACTERIAL AND MICROSCOPICAL CHARACTERISTICS

The examination of water for the presence of bacteria is important for the water supply
engineer from the viewpoint of public health. The bacteria may be harmless to mankind or
harmful to mankind. The former category is known as non-pathogenic bacteria and the
later category is known as pathogenic bacteria.
 Some of these are able to multiply and continue their existence while the remaining die
out in due course of time. The selective medium that promote the growth of particular
bacteria and inbuilt the growth of other organisms is used in the lab to detect the presence
of the required bacteria, usually coliform bacteria. For bacteriological analysis the
following tests are done.

(a) PLATE COUNT TEST

In this method total number of bacteria presents in a millitre of water is counted. 1 ml

of sample water is diluted in 99ml of sterilized water and 1ml of dilute water is mixed
with 10ml of agar of gelatine. This mixture is then kept in incubator at 37 C for 24
hours or 20 C for 48 hours. After the sample will be taken out from the incubator and
colonies of bacteria are counted by means of microscope. Drinking water should not
have more than 10 coliforms/100ml.

(b) M.P.N. TEST (MOST PROBABLE NUMBER) The detection of bacteria by mixing different
dilutions of a sample of water with fructose broth and keeping it in the incubator at
37 C for 48hours. The presence of acid or carbon-dioxide gas in the test tube will
indicate the presence of B-coli. After this the standard statistical tables (Maccardy‟s)
are reffered and the “MOST PROBABLE NUMBER” (MPN) of B-coli per 100ml of water
are determined. For drinking water, the M.P.N. should not be more than 2.

In the multiple-tube method, a series of tubes containing a suitable selective broth culture
medium (lactose-containing broth, such as MacConkey broth) is inoculated with test
portions of a water sample. After a specified incubation time at a given temperature each
tube showing gas formation is regarded as “presumptive positive the gas indicates the
possible presence of coliforms. However, gas may also be produced by other organisms, and
a subsequent confirmatory test is essential. The two tests are known respectively as the
presumptive test and the confirmatory test.

For the confirmatory test, a more selective culture medium (brilliant green bile broth) is
inoculated with material taken from the positive tubes. After an appropriate incubation
time, the tubes are examined for gas formation as before. The most probable number
(MPN) of bacteria present can then be estimated from the number of tubes inoculated and
the number of positive tubes obtained in the confirmatory test. Using specially devised
statistical tables. This technique is known as the MPN method

WATER BORNE DISEASES

 World health organization has observes that 80% of communicable diseases that are
transmitted through water. The diseases like cholera, gastroenteritis, typhoid, amoebia,
diarrhoea, polio, hepatitis (Jaundice), Leptospirosis, Dracontiasis are caused by bacteria. Excess
of fluorides present in water [ above 1.5 mg/litre] cause diseases like dental flurosis, sketetal
flurosis. This is a permanent irresible disease that weakens the bone structure. The patient
becomes immobile and bedridden. Excess of nitrates in water causes Mathaemoglobinaemia or
blue baby symptoms in infants. It effects the hemoglobin in the blood and reduces its capacity
to transport oxygen to the cells. Nitrates in water are caused by industrial effluents, agricultural
runoff. Toxic ions of chromium, lead, arsenic and pesticides in water cause diseases affecting the
kidney, liver and high blood pressure, paralysis, cancer etc. These toxic substances are due to
industrial effluents reaching the surface and ground water sources.