National University of Singapore

Department of Civil and Environmental Engineering

ESE2401 Water Science and Technology

Session 2014/2015

Laboratory Manual

for

Experiment E1 & E2

Venue: EW2 03-14 ESE Teaching Lab.

Name_________________________________________

Group_________________________________________

Date of Experiment ______________________________

 Laboratory Hazards: A Warning

Many reagents require handling with the utmost care, either in their original state or in
solution or both. This is to protect the health and safety of the user. Reagents bearing
labels with the word POISON, DANGER, CAUTION, FLAMMABLE, or comparable
warnings must be handled with special care. Concentrated acids and bases may cause
chemicals burns and are especially hazardous if spilled or splashed into the eyes. Always
handle with extreme care to avoid contact. In diluting concentrated acids, always add acid
to water to prevent local overheating and possible injury; never add water to acid. Students
are advised to use face shields or safety glasses and to familiarize themselves with the
positions of the eye wash fountains. Mercury and its compounds are used to prepare
standards, displace gases and preserve samples. These compounds must always be
handled with extreme caution as they are very toxic. Sodium azide (APHA) is toxic and
reacts with acid to produce more toxic hydrazoic acid. Avoid inhalation, ingestion and
skin exposure. The samples provided may also contain pathogenic organisms. Students
must wash their hands before leaving the laboratory and are advised to leave their
belongings well away from the work benches to avoid possible contamination. Food
should not be brought into the laboratory. Eating, drinking and smoking are not allowed
in the laboratory. Students are required to wear shoes which cover their feet.

Students are expected to clean up all the equipment used during their experiments
before leaving the laboratory.

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Experiment E1 & E2

The purpose of these experiments is to illustrate some of the tests used to assess selected
properties of water and wastewater.

Experiment E1

Purpose

To determine the various categories of solids that are commonly defined in water
and wastewater analyses and to investigate the nature of the materials that these
solids categories define.

1a) Gravimetric Analysis

Introduction

Gravimetric analysis is based on the determination of constituents by measurements of

their weights. In addition to weighing, gravimetric analysis involves three other
techniques. These are filtration which separates suspended or particulate fractions from
the dissolved fractions, evaporation which separates water from the material dissolved or
suspended in it and combustion which separates the inorganic material from organic
matter by destroying the latter.

Total Solids and Total Volatile Solids

This includes both dissolved and suspended material.

Apparatus:

1. Porcelain or nickel dishes

2. Steam Bath
3. Oven
4. Furnace
5. Analytical balance
6. Dessicator

Procedure:

1. Weigh an ignited nickel or porcelain dish, W1.

2. Add 25 mL of sample (more is sample contains little suspended material).
3. Evaporate to dryness on a steam bath.
4. Further drying of samples for 1 hr at 103oC (due to time constraint, students may
dry samples for hr at 103oC) in an oven.
5. Cool samples in a dessicator and weigh, W2.
6. Place dishes into a furnace at 500oC for 20 min.
7. Remove from furnace and allow dishes to cool slightly before further cooling in a
dessicator. Reweigh., W3.

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Calculations

W2 W1
Total solids, (mg/L): TS =
Vol of sample

W2 W3
Total volatile solid, (mg/L): TVS =
Vol of sample

W3 W1
Total fixed solid, (mg/L): TFS =
Vol of sample

Suspended Solids and Volatile Suspended Solids

Apparatus:

1. Filter apparatus
2. Vacuum pump
3. Drying oven
4. Muffle furnace
5. Analytical balance
6. Glass fibre disks
7. Dessicator

Procedure:

1. Weigh pretreated fibre disk (stored in dessicators) and container, W1.

(Warning: do not remove more than one disk for weighing at any given time. Disks
absorb moisture from fingers and the atmosphere very quickly resulting in
erroneous values of W1. Weigh disks must not be marked).
2. Carefully place glass-fibre disk in the filter apparatus.
3. Apply a vacuum and wet the disk with distilled water.
4. Add 25 mL of sample (more if sample contains little suspended material).
5. Remove the disk from the filter apparatus and transfer to its container, dry at
103oC for 1 hr; cool to room temperature in a dessicator then weigh, W2.
6. Place dishes into a furnace at 500oC for 20 min. (Warning: do not use aluminum
dishes).
7. Remove from furnace and allow dishes to cool slightly before further cooling in a
dessicator. Reweigh, W3. (Warning: when loading or removing samples from a
furnace or oven use protective gloves and tongs).

Calculations

W2 W1
Suspended solids, (mg/L): SS =
Vol of sample

W2 W3
Volatile suspended solid, (mg/L): VSS =
Vol of sample

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 W3 W1
Fixed suspended solid or residue, (mg/L): FSS =
Vol of sample

1b) Turbidity

Introduction

The technique used is based on nephelometry (i.e., the measurement of the forward
scattering of light at 90o to the path of an incandescent light beam). Suspended particles in
a water sample reflect a portion of the incident light. A photoelectric detector measures
the light reflected at 90o and compares this against light reflected by a reference standard.
The higher the turbidity the higher is the intensity of scattered light. Formazin polymer is
used as the reference turbidity standard suspension. Turbidimeters are usually calibrated
to read directly in nephelometric turbidity units (NTU).

Apparatus:

1. Turbidimeter
2. Formazin reference standards

Procedure:

1. Switch machine on to warm-up.

2. Place reference standard into sample well and follow calibration instructions
(Hold reference standard tubes only by the top. Glass surfaces must be kept clean.
Wipe surfaces with the lens tissue provided.).
3. Thoroughly mix samples provided and collect approximately 200 mL of each.
4. Rinse a clean sample tube with the sample; then fill the tube full.
5. Tap the tube to remove any air bubbles. Wipe glass surface clean.
(Hold reference standard tubes only by the top).
6. Place sample tube into sample well and cover with shield. Read the scale and
report turbidity in NTU.

1c) Conductivity

Introduction

The ability of an aqueous solution to carry an electric current depends on the presence of
ions and on the temperature of measurement. A numerical expression of this ability is
conductivity. Solutions of most inorganic acids, bases and salts will have relatively high
conductivities but solutions or organic compounds will not conduct a current well. The
conductance of a solution is the reciprocal of its resistance and is given the units of mhos
(i.e. reciprocal ohms). The specific conductance, K, of a solution is the conductance of 1
cc of solution between electrodes of 1 sq cm area which are 1 cm apart and has units
mho/cm. In a conductivity meter, resistance measurements are made and converted to
conductance on the instrument read out. Since conductivity cells used do not have
electrodes exactly 1 sq cm. in area and 1 cm apart, readings must be corrected to standard
conditions by using a cell constant. The conductivity of a particular sample can usually be
related to the dissolved solids content.

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Apparatus:

1. Conductivity meter

Procedure:

1. Rinse cell with sample to be tested.

2. Collect about 100 mL of sample in a beaker.
3. Measure the conductivity of the sample with the cell. Also record the temperature
of the sample.

Calculation (if standard conditions do not apply to the electrodes)

Conductivity = mq

where m = specific conductance

q = cell constant
(Note: The instrument gives a direct reading from the scale and does not require the above
calculation)

1d) pH

Introduction

pH or hydrogen ion activity indicates the intensity of the acidic or basic character of a
solution. Measurement of pH is important because almost every phase of water and
wastewater treatment is pH dependant. The pH of a sample is measured electrometrically
by potentiometric measurement using a glass electrode and a reference electrode
(calomel).

Apparatus:

1. Buffer solution pH 4.0

2. Buffer solution pH 7.0
3. pH meter

Procedure:

1. Check pH meter calibration with Buffer solutions provided. (Note: Always rinse
probe with distilled water before immersing it into a solution. On completion, place
probe back into a beaker of distilled water.)
2. Collect samples in beakers and measure pH with probe. To provide gentle mixing
during pH measurement, drop a magnetic bar into the beaker. Adjust magnetic
stirrer so that there is only gentle mixing.
3. Read the meter and record results.
4. On completion leave probe in a beaker of water.

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1e) Chloride

Introduction

In costal areas, the presence of large amounts of chloride in wastewater indicates salt
water infiltration of the collection system. Zeolite and ion exchange water softeners add
chloride from regeneration. Human excretions also add chlorides. The chloride content
should remain constant through the plant and have no significance in interpreting process
changes or efficiencies. With a low pH they indicate industrial waste containing muriatic
acid.

Volumetric Techniques

This depends on the measurement of volumes of liquid reagents of known strength. The
requirements of this mode of analysis are:

(i) Equipment to measure the sample volume accurately e.g. pipette.

(ii) A standard solution of suitable strength.
(iii) An indicator to show when stoichiometric end point has been reached.
(iv) A graduated burette for accurate measurement of the volume of standard solution
necessary to reach end point.

Apparatus:

1. Burette, 25 mL
2. Beakers
3. 5 mL pipette
4. Silver nitrate, 0.0141 N
5. Sodium Chloride, 0.0141 N
6. Potassium Chromate indicator

Procedure:

1. Standardize silver nitrate solution.

Add exactly 20 mL of sodium chloride to 1 mL of potassium chromate in a beaker.
Titrate as in step 4. Then,

(N of NaCl) (mL NaCl)

Normality Constant, C N =
mL AgNO3

2. Place 100 mL sample or a smaller amount diluted to 100 mL with distilled water
into a beaker.
3. Add 1 mL of potassium chromate.
4. Add silver nitrate solution from burette agitating the sample constantly until a
uniform pinkish-yellow end point is reached. Record volume of silver nitrate used,
M1 .
5. Repeat steps 2, 3 and 4 using 100 mL distilled water as sample. Record volume of
AgNO3 used, M2.

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Calculations

(M1 M 2 ) x 35.45 x 1000 x C N

mg/L Cl =
ml sample

7
Experiment E2

Measurement of Organic Strength

The standard methods used for the measurement of the organic strength of samples
(eg wastewaters and effluents) are COD, BOD and TOC. To enable the tests to be
completed during the laboratory period the reaction times for the COD test have
been reduced from the standard method. All of the determinations (except TOC)
involve the measurement of the difference between initial and final amounts of an
oxidizing agent and thus the sample must not exert such a high oxygen demand
that no oxidizing agent remains at the end of the test.

3a) Chemical Oxygen Demand (COD)

(Closed Reflux Titrimetric Method)

Overall objective

Determine the oxygen equivalent required to completely oxidize organic matter contained
in a sample of interest.

Background

Most types of organic matter are oxidized by a boiling mixture of chromic and sulfuric
acids. The sample is refluxed in strongly acid solution with a known excess of potassium
dichromate. After the digestion, the unreduced potassium dichromate is titrated with
ferrous ammonium sulfate to determine the amount of potassium dichromate consumed
and the oxidizable organic matter is calculated in terms of oxygen equivalent. COD is used
as a measure of the concentration of organic matter in a sample of interest. It is a common
means of expressing the strength of wastewater streams, especially as it relates to
biological treatment.

Typical applications

Measuring COD of feed solutions or reactor contents for biological systems. Typically
used for carbon or oxygen mass balances.

Equipment and reagents required:

Block Digester or oven at 150 C

Reflux Tubes, 16X100 mm
Tube Rack
Small TFE-covered magnetic stirring bar
Magnetic Stir Bar Plate
0.0167 M Standard Potassium Dichromate Digestion
Sulfuric Acid Reagent
Ferroin Indicator Solution
Standard Ferrous Ammounium Sulfate Titrant (FAS)

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Method

1. Add 2.5 ml of sample to reflux tube. Use distilled water for the blank.
2. Add 1.5 ml of digestion solution to samples.
3. Add 3.5 ml of sulfuric acid reagent to samples.
4. Tightly cap the tubes and shake. Be sure to wear face shield, gloves, and mitts
when shaking tubes, which become hot.
5. Digest tubes for 2 hours
6. Remove test tubes and cool to room temperature in a test tube rack
7. Add 2 drop of ferroin indicator and a magnetic stir bar
8. On a magnetic stir bar plate, titrate to red/orange endpoint with 0.2M FAS. The
endpoint is a sharp endpoint.
9. Dump waste into a hazardous waste container. Rinse the tubes with small amount
of distilled water, and pour into hazardous waste container.
10. Calculate COD as follows:
COD as mg O2 per L = (A - B) x M x 8000 / ml sample where:
A = ml FAS used for blank
B = ml FAS used for sample
M = molarity of FAS

Notes, tips and caveats:

Interferences may result from the presence of straight-chain aliphatic compounds, or
nitrite.

3b) Biochemical Oxygen Demand: 5 day-BOD

A water sample is incubated for 5 days at 20C. The reduction in dissolved

oxygen content after the incubation period yields a measure of the biochemical
oxygen demand. To obtain a reliable result the DO in the incubated sample should
be a residual of at least 0.5 mg/l. Thus if a sample has a BOD higher than 7 mg/l,
it will require dilution to prevent excessive oxygen consumption in the BOD bottle.
It is usual to set up 3 dilutions to straddle a samples estimated BOD and the table
below details the dilutions suitable for various BOD ranges.

PERCENT MIXTURE BOD COVERED

100 2-7 mg/l

50 4-14
20 10-35
10 20-70
5 40-140
2 100-350
1 200-700
0.5 400-1400
0.2 1000-3500
0.1 2000-7000

Apparatus and Reagents

1. DO meter + BOD self-stirring probe

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2. BOD incubator
3. Pipettes, beakers and 1 l measuring cylinders
4. Buffer solution (i.e. distilled water containing trace elements and phosphate
buffer).

Procedure

1. Decide on dilution required

2. Using a 1 l cylinder prepare the desired dilutions for each sample. Make up 750 ml.
3. Carefully fill 2 BOD bottles to the brim (avoid the introduction of air bubbles
while pouring sample into the bottle)
4. Immediately determine the DO using the probe (DOSI)
5. On removal of the probe use a little distilled water to fill the bottle again and then
gently stopper it. Ensure that no air bubbles are trapped beneath the stopper. Also
fill flare tops of bottle with distilled water.
6. Also incubate two bottles of dilution water after determining initial DO, DOBI.
These are your blanks.
7. Determine final DO values in bottles after 5 days of incubation i.e. DOSF and
DOBF.

Calculations

(DOSI - DO SF ) - (DO BI - DO BF )
BOD mg/l =
p, decimal volumetric fraction of sample used

Eg. p = 0.1 if a 10% mixture is used.

(3c) Total Organic Carbon: - Combustion-Infrared Method

The organic carbon in water and wastewater represents many compounds and
oxidation states. Biological or chemical processes may oxidize some of these
further but because there are organic compounds that do not respond to either COD
or BOD, these tests may not be a satisfactory measure of total organic carbon
(TOC). TOC, however, does not provide the same kind of information as either
BOD or COD although it can be related to the latter two parameters if a
sufficiently constant empirical relationship can be established. TOC analyzers
offer a means of measuring total organic carbon in the range normally found in
water and wastewater. Organic carbon is oxidized to CO2 by heat and oxygen and
the CO2 is measured directly by a nondispersive infrared analyzer. Inorganic
carbon must be compensated for since it usually forms a significant portion of the
total carbon. Total organic carbon is commonly estimated by subtracting from
total carbon the total inorganic portion.

Apparatus and Reagents:

1. Sample blender or homogenizer

2. Magnetic stirrer
3. Hypodermic Syringe

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4. Total Organic Carbon Analyzer
5. Redistilled water for preparation of blanks and standard solutions
6. Standard carbon solution
7. Air, dry and CO2 free

Procedure:

Warning Students should consult the technician in attendance before they operate the
Total Carbon Analyzer

1. If sample contains gross solids or other insoluble matter, homogenize.

2. Continuously stir sample with a magnetic stirrer following 1.
3. Withdraw a portion using the hypodermic syringe.
4. Inject into the analyzer and obtain peak height reading.
Repeat until 3 consecutive peaks are obtained that are reproducible to within
10%.
5. From calibration curve provided obtain TOC readings.

References

1. Standard Methods for the Examination of Water and Wastewater, 20th edition,
American Public Health Association, 1998.
2. Chemistry for Environmental Engineering, Sawyer, C.N., McCarty, P.L., 4th
edition, McGraw-Hill.
3. Environmental Chemistry, De, A.K., Wiley Eastern Ltd., 1987.

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Report for E1 and E2

Each student undertaking the two laboratory classes will have to submit two separate
detailed formal reports one on each experiment. Each report is due exactly two weeks
after your laboratory class. Reports are to be submitted to the Water and Science
Laboratory (WS2 02-32).

Each report should include the following:

(i) Results neatly tabulated

(ii) All calculations performed clearly set out
(iii) A brief discussion on the sources of errors and ways to reduce them
(iv) A brief discussion of the parameters investigated
outlining their relationship with each other and
their importance in water quality analysis
(v) Conclusion

The following questions may be found useful when preparing the formal report. The
report, however, must not be seen to consist of a series of answers to the questions below:

(i) Discuss the importance of the two temperatures used in gravimetric analysis.
(ii) Tabulate TSS, TVS and TFS. Comment on their relative sources abundance with
reference to the possible of sources of the three samples provided.
(iii) Discuss the relationship between turbidity and the gravimetric results and that
between conductivity and TFS.
(iv) With reference to the possible sample sources, comment on your pH data.
(v) From your chloride test, is there any sample which contains enough to affect the
COD test?
Are you adding sufficient mercuric sulphate?
(vi) Discuss the significance of reflux time on COD yield using the dichromate
oxidation method.
(vii) Tabulate BOD, COD and TOC values determined and calculate the ratios of
BOD/COD, BOD/TOC and COD/TOC, comment on the samples relative
biodegradability.
(viii) When considering the analysis of industrial wastes, it is imperative to evaluate the
BOD, COD and TOC values for various classes of compounds and to attempt to
correlate BOD and COD with TOC.
Discuss the factors which might constrain or negate these correlations.
(ix) Discuss the effect of high dissolved solids on the results of COD tests.
(x) Identify the most likely sources of the given samples.

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