TOTAL DISSOLVED SOLIDS

Procedure:

1. Preparation of glass-fiber filter with wrinkled side up into filtration apparatus. Apply vacuum and
wash disk with three times with 20-mL volumes of reagent-grade water. Continue suction
toremove all traces of water. Discard washings.
2. Preparation of evaporating dish: Heat clean dish to 180 ± 2°C for 1 h in an oven. Store in
desiccator until needed. Weigh immediately before use.
3. Selection of filter and sample sizes: Choose sample volume to yield between 2.5 and 200mg
dried residue.
4. Sample analysis: Stir sample with a magnetic stirrer and add measured volume onto aglass-fiber
filter with applied vacuum. Wash with 3 times with 10-mL volumes ofreagent-grade waterand
continue suction forabout 3 min after filtration is complete. Transfer total filtrate (with
washings) to a weighedevaporating dish and evaporate to dryness on a steam bath or in a drying
ovenfor at least 1 h inan oven at 180 ± 2°C, cool in a desiccator to balance temperature, and
weigh. Repeat dryingcycle of drying, cooling, desiccating, and weighing until a constant weight is
obtained or untilweight change is less than 4% of previous weight.Analyze at least10% of all
samples in duplicate. Duplicate determinations should agree within 5% of theiraverage weight.

Calculations

TURBIDITY

Procedure:

Gently agitate sample. Wait until air bubbles disappear and pour sample into cell. When possible, pour
well-mixed sample into cell and immerse it in an ultrasonic bath for 1 to 2 s or apply vacuum degassing,
causing complete bubble release. Read turbidity directly from instrument display.

CONDUCTIVITY

Procedure:

1. Determination of cell constant: Rinse conductivity cell with at least three portions of 0.01M KCl
solution. Adjust temperature of a fourth portion to 25.0 ± 0.1°C. If a conductivity meter displays
resistance, R, ohms, measure resistance of this portion and note temperature.Compute cell
constant, C:
C, cm–1 = (0.001412)(R(KCl))[1 + 0.0191(t − 25)]
where: R(KCl) = measured resistance, ohms, and t = observed temperature, °C.
 2. Conductivity measurement: Thoroughly rinse cell with one or more portions of sample. Adjust
temperature of a final portion to about 25°C. Measure sample resistance or conductivity and
note temperature to ±0.1°C.

Calculations:

pH

The electromotive force (emf) produced in the glass electrode system varies linearly with pH. This linear
relationship is described by plotting the measured emf against the pH of different buffers. Sample pH is
determined by extrapolation.

Procedure:

Establish equilibrium between electrodes and sample by stirring sample to ensure homogeneity; stir
gently to minimize carbon dioxide entrainment. For buffered samples or those of high ionic strength,
condition electrodes after cleaning by dipping them into sample for 1 min. Blot dry, immerse in a fresh
portion of the same sample, and read pH. With dilute, poorly buffered solutions, equilibrate electrodes
by immersing in three or four successive portions of sample. Take a fresh sample to measure pH.
BOD FOR 5 DAYS AT 20 DEGREE CENTIGRADE

Preparations

Preparation of Dilution Water:1ml each of(PO4 buffer + MgSO4 +CaCl2 + FeCl3) /L of water.
Temperature: 20 ± 1℃. Saturate with DO

Seeding :

➢ Seed source: Domestic wastewater, unchlorinated effluents from biological waste treatment plant

➢ Seed control: BOD of seeding material

➢ Series of dilution:

1. DO depletion = Initial DO - Final DO

2. DO depletion/(volume of seed in ml)

➢ Required criterion: DO depletion - 2mg/L; Residual DO - 1mg/L (minimum)

Procedure

Sample taken in 300ml BOD bottle + 1ml conc. sulfuric acid + 1ml Manganese sulfate solution + 1ml
Alkali-iodide. Starch indicator (solution turns to blue color. Titrate with 0.025M sodium thiosulfate (pale
straw color solution). Endpoint: Blue to colorless
Calculations:

COD

Procedure:

Place sample in culture tube/ ampule and add digestion solution. Carefullyrun sulfuric acid reagent
down inside of vessel so an acid layer is formedunder the sample-digestion solution layer. Tightly cap
tubes or seal ampules,and mix.Place tubes or ampules in block digester preheated to 150°C and reflux
for 2 h. Cool to room temperature. Remove culture tube caps and add small TFE-covered magnetic
stirring bar.Add 1to 2 drops ferroin indicator and stir rapidly on magnetic stirrer while titratingwith
standardized 0.10M FAS. The end point is a sharp color change fromblue-green to reddish brown,
although the blue-green may reappear withinminutes. Titrate a blank containing the reagents and
avolume of distilled water equal to that of the sample.
TOTAL ALKALINITY AS CaCO3

Procedure

Color change: Select sample size and normality of titrant. Adjust sample to room temperature, if
necessary, and with a pipet discharge sample into an erlenmeyer flask, while keeping pipet tip near flask
bottom. If free residual chlorine is present add 0.05 mL (1 drop) 0.1M Na2S2O3 solution, or destroy with
ultraviolet radiation. Add 0.2 mL (5 drops) indicator solution and titrate over a white surface to a
persistent color change characteristic of the equivalence point. Commercial indicator solutions or solids
designated for the appropriate pH range (3.7 or 8.3) may be used. Check color at end point by adding
the same concentration of indicator used with sample to a buffer solution at the designated pH.

c. Potentiometric titration curve:

1) Rinse electrodes and titration vessel with distilled water and drain. Adjust sample to room
temperature, if necessary, and with a pipet discharge sample while keeping pipet tip near the titration
vessel bottom.

2) Measure sample pH: Add standard alkali in increments of 0.5 mL or less, such that a change of less
than 0.2 pH units occurs per increment. After each addition, mix thoroughly but gently with a magnetic
stirrer. Avoid splashing. Record pH when a constant reading is obtained. Continue adding titrant and
measure pH until pH 9 is reached. Titrate to theend-point pH without recording intermediate pH values
and without undue delay. As the endpoint is approached make smaller additions of acid and be sure that
pH equilibrium is reachedbefore adding more titrant.

d. Potentiometric titration of low alkalinity: For alkalinities less than 20 mg/L titrate 100 to200 mL
according to the above procedure, using a 10-mL microburet and 0.02N standardacid solution. Stop the
titration at a pH in the range 4.3 to 4.7 and record volume and exact pH.Carefully add additional titrant
to reduce the pH exactly 0.30 pH unit and again record volume.

Calculations:
Alkalinity Relationships

TOTAL HARDNESS AS CaCO3

Procedure:

Pretreatment of polluted water and wastewater samples: Use nitric acid-sulfuric acid or nitric acid-
perchloric acid digestion.

Titration of sample: Dilute 25.0 mL sample to about 50 mL with distilled water. Add 1 to 2 mL buffer
solution (pH of10.0 to 10.1). Add 1 to 2 drops indicator solution or an appropriate amount of dry-
powder indicatorformulation. Add standard EDTA titrant slowly, with continuous stirring, until the
lastreddish tinge disappears. Add the last few drops at 3- to 5-s intervals. At the end point thesolution
normally is blue. Daylight or a daylight fluorescent lamp is recommended highlybecause ordinary
incandescent lights tend to produce a reddish tinge in the blue at the end point.
c. Low-hardness sample: For ion-exchanger effluent or other softened water and for naturalwaters of
low hardness (less than 5 mg/L), take a larger sample, 100 to 1000 mL, for titrationand add
proportionately larger amounts of buffer, inhibitor, and indicator. Add standard EDTAtitrant slowly from
a microburet and run a blankofthe same volume as the sample, to which identical amounts of buffer,
inhibitor, and indicatorhave been added. Subtract volume of EDTA used for blank from volume of EDTA
used forsample.

Calculations:

NITRATE (NO3-)

Procedure:

25mL sample or a portion diluted to 25mL + 75mL NH4Cl-EDTA solution and mix. Pour mixed sample into
column andcollect at a rate of 7 to 10 mL/min. Discard first 25mL. Collect the restin original sample flask.
If to be used again in a few hours, pour 50 mLdilute NH4Cl-EDTA solution on to the top and let it pass
through thesystem. Store Cu-Cd column in this solution and never let it dry.

Not more than 15 min afterreduction, 2mL color reagent + 50mL sample and mix. Between 10min

and 2h afterward, measure absorbance at 543 nm against a distilled

water-reagent blank.

Using the intermediate NO3¯-N solution, preparestandards in the range 0.05 to 1.0mg NO3¯-N/L by
diluting thefollowing volumes to 100 mL in volumetric flasks: 0.5, 1.0, 2.0, 5.0,and 10.0 mL. Carry out
reduction of standards exactly as describedfor samples.Reactivate Cu-Cd granules when efficiency of
reductionfalls below about 75%.

Calculation:

Obtain a standard curve by plotting absorbance of standards againstNO3¯-N concentration. Compute

sample concentrations directly fromstandard curve.

NITRITE (NO2-)

Reagents:

1. Nitrite-free water : 1L DW + KMnO4 +Ca(OH)2. Re-distillall-borosilicate-glass apparatus. Initial 50ml is

discarded. Collect distillate fractionthat is permanganate free.DPD reagentis red incolour,indicates
presence of permanganate
2. Colour reagent: 800ml NO2-free-water + 85% H3PO4 + 10g sulfanilamide+ 1g NED
dihydrochloridedilute to 1L

3. Stock Solution: To determine NaNO2 (Sodium nitrate) content: 0.01M KMnO4 titrate (std. Reductant)
+ 0.025M Na2C2O4back titrate permanganate solution

Preparation of stock soln: 1.232g NaNO2 + 1000ml DW1mL = 250mg N

Standardisation of stock soln: 50ml KMnO4 + 5ml H2SO4 + 50ml stock solution.Shake gently & warm to
70 – 80 degree centigrade on hot plate.Titarate0.025M Na2C2O4 soln. Against 0.01M (0.05N) KMnO4.
Faint pink end point.

Procedure:

50ml Standard solutions + 2ml color reagent. Standard curve_absorbance vs NO2- N. 50ml sample + 2ml
color reagent. Read absorbance at 543nm.

Calculation:

Prepare a standard curve by plotting absorbance of standards against NO2 –-N concentration. Compute
sample concentration directly from curve.

AMMONIA-N

Procedure:

Prilimnary Distillation: 500ml water + 20ml of borate buffer solution to a distillationflask (pH to 9.5 with
6N NaOH solution) + few glass beads anduse this mixture to steam out the distillation apparatus
untildistillate shows no trace of ammonia.500ml of dechlorinated sample or a portion diluted to 500 ml+
water +dechlorinatingagent (remove residual chlorine) +25ml of borate buffer (pH to 9.5 with 6N NaOH
solution).

To minimize contamination, leave distillation apparatus assembled after steaming out and until just
before starting the sample distillation. Disconnect steaming out flask and immediately transfer sample
flask to distillation apparatus. Distil at the rate of 6 to 10ml/minute with the tip of the delivery tube
below the surface of acid receiving solution.Use 50 ml indicating boric acid solution fortitrimetric
method.

Titrate ammonia in distillate against standard sulphuric acid until indicator turns a pale lavender. Carry a
blank through all steps of the procedure and apply the necessary correction to the results.

Calculation:

TOTAL NITROGEN

Procedure:

1. Selection of sample volume and sample preparation:

2. Ammonia removal: Sample + 25 mL borate buffer + 6N NaOH(until pH 9.5 reached) +few glass beads
or boiling chips and boil off 300 mL. Ifdesired, distill this fraction and determine ammonia nitrogen. For
sludge and sediment samples, weigh wet sample in a crucible or weighing bottle, transfer contents to a
kjeldahl flask, and determine kjeldahl nitrogen.

3. Digestion: Cool, + 50 mL digestion reagent (or substitute 6.7 mL concH2SO4, 6.7 g K2SO4, and 0.365 g
CuSO4) to distillation flask + a few glass beads and mix. Boil brisklyuntil the volume is greatly reduced (to
about 25 to 50 mL) and copious white fumes are observed. Then continue to digest for 30 min. As
digestion continues, colored or turbid samples will become transparent andpale green. After digestion,
dilute to 300 mL with water, and mix + 50 mL sodium hydroxide-thiosulfate reagent (forms alkalinelayer
at flask bottom). Connect flask to a steamed-out distillation apparatus and swirl flask toinsure complete
mixing. The pH of the solution should exceed 11.0.

4. Distillation: Distill and collect 200 mL distillate. Use 50 mL indicating boric acid asabsorbent solution
when ammonia is to be determined by titration

5. Final ammonia measurement: Use the titration method.

6. Standards: Carry a reagent blank and standards through all steps of the procedure.

Calculation:
Prepare standard curves by plotting the absorbance of standards processed through the manifold versus
ammonia concentration. The calibration curve is linear.

PHOSPHATES& TOTAL PHOSPHORUS

Procedure:

1. Preliminary Filtration

2. Preliminary Acid Hydrolysis: 100-mL sample or a portion diluted to 100 mL+ 0.05 mL (1
drop)phenolphthalein indicator solution. If a red color develops, add strong acid solution dropwise,
tojust discharge the color. Then add 1 mL more.Boil for at least 90 min + distilled water to keep the
volume between 25 and 50mL. Cool,neutralize to a faint pink color with NaOH solution, and restore to
the original 100mL volumewith distilled water.

3. Sulfuric Acid-Nitric Acid Digestion: Into a micro-kjeldahl flask + sample + 1 mL conc H2SO4 + 5 mL conc
HNO3. Digest to a volume of 1 mL and then continue until solution becomes colorless to
removeHNO3.Cool, + approx. 20 mL distilled water + 0.05 mL (1 drop) phenolphthalein indicator + 1N
NaOH solution as required to produce a faint pink tinge. Transfer neutralized solution, filtering if
necessary to remove particulate material or turbidity, into a 100-mL volumetric flask. Add filter washings
to flask and adjust sample volume to 100 mL with distilled water.

FOR PHOSPHATE

4. Vanadomolybdophosphoric Acid Colorimetric Method: If sample pH > 10, 50.0 mL sample + 0.05 mL (1
drop) phenolphthalein indicator to and discharge the red color with 1 + 1 HCl before diluting to 100 mL.
Remove excessive color in sample by shaking about 50mL + 200 mg activated carbon, for 5 min and filter
to remove carbon. 35ml sample + 10 mL vanadate-molybdate reagent and dilute to 50ml with DW.
Prepare a blank in which 35mL distilled water is substituted for the sample. After 10 min or more,
measure absorbance of sample versus a blank at a wavelength of 400 to 490 nm. Prepare a calibration
curve by using suitable volumes of standard phosphate solution.

FOR TOTAL PHOSPHORUS

4. Ascorbic Acid Method: 50mL sample + 0.05 mL (1 drop) phenolphthalein indicator. If a red color
develops + 5N H2SO4 solution dropwise (discharge the color) + 8mL combined reagent and mix
thoroughly. After at least 10-30 min, measure absorbance of each sample at 880 nm, using reagent
blank as the reference solution. The absorbance is proportional to conc. of total phosphorus.

Calculation:

FOR PHOSPHATE:

RESIDUAL CHLORINE
Procedure:

1. Standardize 0.1N Sodium thiosulfate by Dichromate method: 4.904 g K2Cr2O7 + 1000mL DW. 80 mL
DW + 1 mL conc H2SO4 + 10 mL 0.1N K2Cr2O7 + 1kg KI and let reaction mixture stand 6 min in the dark
before titrating with 0.1N Na2S2O3 titrant.

5 mL acetic acid in + 1 g KI (pH to between 3.0 and 4.0) + Sample (mix) + 0.01N Na2S2O3 from a buret
until the yellow color of the liberated iodine almost is discharged. Add 1 mL starch solution and titrate
until blue color is discharged. For blank: DW + 5 mL acetic acid, 1 g KI, and 1 mL starch solution. Perform
blank titration whichever applies:

1) If a blue color develops, titrate with 0.01N Na2S2O3 to disappearance of blue color and record result.
B is negative.

2) If no blue color occurs, titrate with 0.0282N iodine solution until a blue color appears. Back-titrate
with 0.01N or 0.025N Na2S2O3 and record the difference. B is positive.

Before calculating the chlorine concentration, subtract the blank titration of 1) from the sample
titration; or, if necessary, add the net equivalent value of the blank titration of 2).

Calculations:

ANIONIC SURFACTANTS
Procedure

Prepare an initial calibration curve in range of 10 to 200 μg of MBAS standard. If necessary to avoid
decolorization of methylene blue by sulfides, + a few drops of 30% H2O2. Prepare a series of separatory
funnels for a reagent blank and selected standards. Standard LAS solution + water to make 100 mL
volume in each separatory funnel. Make alkaline by dropwise addition of 1N NaOH, using
phenolphthalein indicator. Discharge pink color by dropwise addition of 1N H2SO4 + 10 mL CHCl3 + 25
mL methylene blue reagent. Rock funnel vigorously for 30 s and let phases separate.

To break persistent emulsions + a small volume of isopropyl alcohol (<10 mL) + same volume of
isopropyl alcohol to all standards. Before draining CHCl3 layer, swirl gently, then let settle. Draw off
CHCl3 layer into a second separatory funnel. Repeat extraction two additional times, using 10 mL CHCl3
each time. If blue color in water phase becomes faint or disappears, discard and repeat, using a smaller
sample.

Combine all CHCl3 extracts in the second separatory funnel + 50 mL wash solution and shake vigorously
for 30 s. Let settle, swirl, and draw off CHCl3 layer through a funnel containing a plug of glass wool into a
100-mL volumetric flask; filtrate must be clear. Wash solution twice with 10 mL CHCl3 each and add to
flask through the glass wool. Collect washings in volumetric flask + CHCl3, and mix well. Determine
absorbance at 652 nm against a blank of CHCl3 and plot a calibration curve of absorbance vs.
micrograms LAS taken, specifying the molecular weight of the LAS used.

METALS – ICP/MS METHOD:

Procedure:

1. Sample Preparation: Nitric Acid sample digestion technique for all analytes except silver and
antimony. Perform all sample manipulations in a Class 100 clean hood.

1. Sample Analysis: Instrument operating conditions, tuning, optimization and calibrations are defined
according to manufacturer’s standards. Ensure that all vessels and reagents are free from
contamination. Internal standard recoveries must be between 70% and 125% of internal standard
response in the laboratory-fortified blank; otherwise, dilute sample, add internal standard mix, and
reanalyze. Make known-addition analyses for each separate matrix in a digestion or filtration batch.

Calculations and Corrections:

For correcting solid or sediment sample results for dilution during digestion and moisture content:

Use instrument software to correct for interferences. Establish appropriate reporting limits for method
analytes based on instrument detection limits and the laboratory blank. Maintain documentation for
instrument tuning, mass calibration, calibration verification, analyses of blanks, analyses of samples and
duplicates with known additions, laboratory and field duplicate information, serial dilutions, internal
standard recoveries, and any relevant quality control charts. Also maintain, and keep available for
review, all raw data generated in support of the method.