Hand Book of Water, Air and Soil Analysis

Hand Book of Water, Air and Soil Analysis
(A Lab Manual)
BY
SADHANA CHAURASIA
&
ANAND DEV GUPTA
International E Publication
www.isca.me , www.isca.co.in
Hand Book of Water, Air and Soil Analysis
(A Lab Manual)
BY
SADHANA CHAURASIA
Head, Department of Energy & Environment
Faculty of Science & Environment
MGCGV, Chitrakkot, Satna MP, 485331, India
&
ANAND DEV GUPTA

Department of Energy & Environment
MGCGV, Chitrakkot, Satna MP, 485331, India
2014
International E - Publication
www.isca.me , www.isca.co.in
International E - Publication
427, Palhar Nagar, RAPTC, VIP-Road, Indore-452005 (MP) INDIA
Phone: +91-731-2616100, Mobile: +91-80570-83382
E-mail: contact@isca.co.in , Website: www.isca.me , www.isca.co.in
Copyright Reserved
2014
All rights reserved. No part of this publication may be reproduced, stored, in

a retrieval system or transmitted, in any form or by any means, electronic,
mechanical, photocopying, reordering or otherwise, without the prior
permission of the publisher.
ISBN: 978-93-83520-91-4
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co, www.isca.net.in
PREFACE
This lab manual Hand Book of Water, Air and Soil Analysis provides
knowledge and understanding for large as well as small class group of the under
graduate and post graduate environmental science students. These analytical
methods form central building pillar in many environmental studies. The manual
emphasizes basic experimental, skill in student which is quintessential of acquiring
laboratory skills. This lab manual serves two purposes: 1- To provide students with
information necessary to conduct activities and experiment that will enhance their
understanding of environmental science through a hand on approach that cannot be
provided in the lecture class room setting. 2- To function as a modified version of
the traditional laboratory. This lab manual contains three sections- Section-1
contains water analysis, Section-2 air analysis and Section-3 soil analysis methods.
As reflected in the title, I am sure that this manual would bring out the fundamental
experimental procedure in environmental studies and experimentation and would
certainly help the new generation environmentalist as the work would be available
on line.
This lab manual provides the current practices in water, air and soil analysis.
The objective is to transfer knowledge of the subject throughout the world to
person who is interested in environmental study and sanitary technology and
engineering and the person interested in maintenance of water and waste water
facility. Based on my experience in education, I believe our readers benefit from
this lab manual in environmental study.
Dr. Sadhana Chaurasia
Head,
Department of Energy & Environment
Faculty of Science & Environment
MGCGV, Chitrakoot, Satna MP, 485331, India
Hand Book of Water, Air and Soil Analysis.. iii

CONTENTS
S. No. Description Page No.
I- Water Analysis 1
Section A (Physico-Chemical test)
1. Temperature 2
2. Transparency 3
3. Turbidity 4-5
4. pH 6
5. Conductivity 7
6. Total Solids (TS) 8
7. Total Suspended Solids (TSS) 9
8. Total Dissolved Solids (TDS) 10
9. Total Volatile Solids (TVS) 11
10. Volatile Suspended Solids (VSS) 12
11. Fixed Dissolved Solids (FDS) 13
12. Alkalinity 14-15
13. Acidity 16
14. Chloride (Cl-) 17-18
15. Sulphate (SO4--) 19
16. Fluoride (F-) 20-22
17. Total Hardness 23-24
18. Calcium and Magnesium hardness 25-26
19. Sodium 27
20. Potassium 28
21. Percent Sodium 29-30
22. Sodium Absorption Ratio (SAR) 31
23. Total Iron (Fe) 32-35
24. Total Kjeldhal Nitrogen (TKN) 36-38
25. Nitrite- Nitrogen (NO2-N) 39-41
26. Nitrate- Nitrogen (NO3-N) 42-44
27. Dissolved Oxygen (DO) 45-46
28. Bio-Chemical Oxygen Demand (BOD) 47-49
29. Chemical Oxygen Demand (COD) 50-51
30. Residual Chlorine 52-53
31. Chlorine Demand 54
32. Boron 55-56
33. Total Phosphorous 57-58
Section B (Biological test)
34. Methods of sampling for MPN test 59-60
35. Total MPN 61-64
36. Feacal MPN 65-66
Hand Book of Water, Air and Soil Analysis.. iv

II- Air Analysis 67

37. Temperature Minimum and Maximum 68
38. Relative humidity of Atmosphere 69
39. Particulate Matter (PM) 70
40. PM10 71-72
41. PM2.5 73-76
42. SO2 77-79
43. NOx 80-82
44. Ozone 83-85
45. Ammonia 86-87
III- Soil Analysis 88
46. Texture 89-93
47. Bulk Density 94
48. Specific Gravity 95
49. Moisture Content 96
50. Water Holding Capacity 97
51. pH 98
52. Electrical Conductivity (EC) 99
53. Redox Potential 100
54. Alkalinity 101
55. Chloride 102
56. Sulphate 103
57. Nitrogen (Kjeldhal Nitrogen) 104-105
58. Nitrate 106-107
59. Total Phosphorus 108
60. Phosphate 109
61. Calcium 110
62. Magnesium 111
63. Sodium 112
64. Potassium 113
65. Iron 114
66. Organic Matter 115-116
Hand Book of Water, Air and Soil Analysis.. v

I- WATER
ANALYSIS
Hand Book of Water, Air and Soil Analysis 1

TEMPERATURE
Object- Determine the temperature of given water sample.
Requirements- Thermometer and beaker etc.
Principle- Relation between Celsius, Fahrenheit and Kelvin scale-
32 R K 273
= = =
100 180 80 100
32 R K 273
= = =
5 9 4 5
Method- Take the water sample in three different beakers and dips the thermometer in the first
beaker for 2 minutes and notes the temperature in the record file. This process is repeated with
second and third beaker and notes the temperature reading in the record. Calculate the average
temperature with this data.
Observation table-
S. No. Sample description Temperature Average temperature
1.
2.
3.
Result- The temperature of given sample was observed --------0C.

Precautions-
1. Beaker should be clean properly.

2. Bulb of the thermometer not to be touched to the body.
3. Thermometer should be put into the water carefully.
4. Note the reading carefully.

TRANSPARENCY
Object- Determine the transparency of the water body.
Requirements-Sacchi disc, string, notebook and inch tape.
Procedure- Lower the sacchi disc in water and note the depth (cm) at which it becomes
disappear. Now raise the disc slowly and note the depth at which it becomes reappear again.
Calculation-
Sacchi disc transparency (SDT) cm = A = B / 2
Where A = depth at which sacchi disc disappears (cm)
B = depth at which sacchi disc reappears (cm)
Euphotic limit (cm) = SDT x 2.5
Vertical attenuation coefficient = 1.9 / SDT
Where SDT = Sacchi disc transparency (cm)
Result- The transparency of water body was found ------ cm.
Precautions-

TURBIDITY (Nephelometric method)
Object- Determine the turbidity of given water sample.
Requirements-Nephelo turbidity meter with sample cells, volumetric flask and amber glass
bottle etc.
Reagents-
(a) Solution I- Dissolve 1.0 gm hydrazine sulphate, (NH2)2, H2SO4 in distilled water and
dilute to 100 ml in a volumetric flask.
(b) Solution II- Dissolve 10 gm hexamethylenetetramine, (CH2)6N4, in distilled water and
dilute to 100 ml in a volumetric flask.
(c) 4000 NTU suspension- In a flask mix 5.0 ml of solution I and 5 ml of solution II. Let
stand for 24 hours at 253 0C. This result in a 4000 NTU suspension. Store in an amber
glass bottle. The suspension is stable for up to 1 year.
(d) Dilute 4000 NTU stock solution with distilled water to prepare dilute standards just
before use and discard after use.
Method- Calibrate the Nephelo turbidity meter with the 100 NTU suspensions and verify the
setting with a reading on the 10 NTU suspensions. Then thoroughly shake sample, wait until air
bubbles disappear and pour sample into turbidity meter tube. Read turbidity directly from
instrument display. In case the turbidity is going beyond the scale dilute the sample and measure
the turbidity. The dilution factor should be taken into account for the final calculation of the
result.
Calculation-
Turbidity (NTU)= NTU measured/proportion of sample in dilution

Observation table-
S. No. Sample Description Turbidity (NTU) Average turbidity

(NTU)
1.
2.
3.
Result- The turbidity of given water sample was observed--------- NTU (Nephelo Turbidity
Unit).
Precautions-
1. Calibrate the instrument properly.

2. Prepare the standard solution carefully.

pH
Object- Determine the pH of given water sample.
Requirement-pH meter, beaker, volumetric flask, buffer tablets (4.0, 7.0 and 9.2 pH), tissue
paper, distilled water and wash bottle.
Principle- pH is a negative logarithm of H+ concentration of a solution.
Method- Switch on the pH meter and leave for 10 minutes to warm up. Rinse the electrode with
distilled water and wipe off with tissue paper. Dip the electrode in standard buffer solution of
pH- 7 for calibration, wait until the displayed reading establish at 7.00. Rinse the electrode with
distilled water and wipe off. Dip the electrode in standard buffer solution of pH- 4 and wait to
establish reading at 401 or adjust with calibration knob. Rinse the electrode and dip in the
sample, wait until reading establish and note the reading.
Observation table-
S. No. Sample description pH value Average pH

1.
2.
3.
Result- The pH of given water sample was observed -----.
Precautions-
1. All the glassware should be clean.

2. Calibrate the instruments properly.

CONDUCTIVITY
Object- Determine the conductivity of given water sample.
Requirements- Conductivity meter, single pan balance, standard solution of KCl, volumetric
flask, funnel, beaker and distilled water.
Method- Switch on the instrument and leave for 10 minutes to warm up. Rinse the electrode
with distilled water and wipe off with tissue paper. After it dip the electrode with probe in the
standard 0.01 M solution of KCl. Press calibration knob and wait for calibration completed and
press continued knob check the displayed reading 1.4120.01 at 250C. Rinse the electrode and
dip in sample and note the reading with temperature 0C. Rinse the electrode and dip in another
sample respectively.
Observation table-
S. No. Sample description Conductivity Average conductivity

( mhos cm-1)
1.
2.
3.
Result- The conductivity of given water sample was obtained........mhos cm-1.
Precautions-
1. Rinse the electrode with distilled water.

2. Beaker should be cleaned with distilled water,
3. Electrode should be put into solution properly.

TOTAL SOLIDS (TS) (Gravimetric method)

Object- Determine the total solid in given water sample.
Requirements- Porcelain dishes, measuring cylinder, balance and oven etc.
Procedure- Weight the previously dried empty porcelain dishes (W1) already kept at 1050C for 2
hours and in desiccator for 30 minutes. Take 100 ml well mixed sample in each porcelain dishes
and place the porcelain dishes in the oven at 1050C carefully for 24 hours. After 24 hours take
the dishes out and keep in the desiccator for 30 minutes for cooling. Now weight the porcelain
dishes and note the final weight (W2). Repeat the process of weighing and drying till constant
weight is achieved.
Calculation-
TS (mg/l) = (W2-W1) x 106 / Sample volume (ml)
Similarly calculate for other two dishes & take average to repeat the results as below-
Observation table-
S. No. Sample no. Initial weight Final weight TS (mg/l) Average TS

(mg/l)
1.
2.
3.
Result- Total solids in the given water sample was found----- mg/l at 1050C.
Precautions-
1. Glassware should be clean.

2. Cool the dish properly.
3. Note the weight carefully.

TOTAL SUSPENDED SOLID (TSS) (Gravimetric method)
Object- Determines the TSS of given water sample.
Requirement- Filter paper (whatman no-41), beaker, hot plate, physical balance, funnel,
measuring cylinder, desiccator, oven etc.
Method- Dry the filter paper in oven (1050C) and cool in desiccator then weight it. Then filter
300 ml sample through filter paper, keep the filter paper in oven for overnight to dry, cool the
filter paper in a desiccator and again weight it. Calculate the TSS using following formula-
Formula- W2-W1 x 1000/V
Where,
W1 = Initial weight of filter paper
W2 = final weight of filter paper
V = Volume of sample
Observation table-
S. No. Sample Initial weight of Final weight of TSS (mg/l)

description filter paper (gm) filter paper (gm)
1.
2.
3.
Result- The total suspended solids in given water sample was observed------- mg/l.
Precautions-

2. Weigh the filter paper carefully.

TOTAL DISSOLVED SOLID (TDS) (Gravimetric method)
Object- Determines the TDS of given water sample.
Requirement- Filter paper (whatman no-41), beaker, hot plate, physical balance, funnel,
measuring cylinder, desiccator, oven etc.
Method- Dry the beaker in oven (1050C) and cool in desiccator then weight it. Then Take 300
ml filtered sample in this beaker and evaporate up to dryness on a hot plate, cool in a desiccator
and again weight it. Calculate the TDS using following formula-
Formula= W2-W1 x 1000/V
Where,
W1 = Initial weight of beaker
W2 = final weight of beaker
V = Volume of sample
Observation table-
S. No. Sample Initial weight of Final weight of TDS (mg/l)

description beaker (gm) beaker (gm)
1.
2.
3.
Result- The total dissolved solids in given water sample was observed ------- mg/l.
Precautions-

2. Weigh the beaker carefully.

TOTAL VOLATILE SOLIDS (TVS) (Gravimetric method)
Object- Determine the TVS in the water sample.
Requirements- Measuring cylinder, nickel crucible, balance, muffle furnace, oven.
Procedure-
(i.) After determining total solids (W2), use the same crucible with residue for TVS
determination carefully.
(ii.) Keep the crucible after determining TS in the muffle furnace at 5500C for 25
minutes.
(iii.) After that keep out the dishes from muffle furnace with the help of tong & keep
in the desiccator for 30 minutes for cooling.
(iv.) Weigh the dishes and note final weight (W3) repeat the process of drying and
weighing till constant weight is achieved.
(v.) W3 will be less than W2.
Calculation-
Volatile Solids = (W2-W3)
W3= weight of dish after keeping in muffle furnace at 5500C.
W2= final weight when TS measured at 1050C.
TVS mg/l= (W2-W3) x 106 / sample volume (ml)
S. No. Crucible Initial Final Final TVS mg/l Average

No. weight weight at weight at TVS mg/l
(W1) 1050C 5500C
(W2) (W3)
Result- Total volatile solids of the water sample at 5500C were found------- mg/l.
Precautions-


VOLATILE SUSPENDED SOLIDS (VSS) (Gravimetric method)
Object- Determine the VSS in water sample.
Requirements- Filter paper, filtration assembly, oven, balance, nickel crucible, tong and
desiccator.
Procedure-
(i.) The filter paper after determining suspended solids may be used otherwise before
doing VSS we have to first TSS (W2).
(ii.) Then keep the filter paper in nickel crucible and keep in the muffle furnace for 30
minutes at 5500C.
(iii.) Take out with the help of tong and put in to desiccator for 30 minutes.
(iv.) After 30 minutes take the weight of filter paper (W3). Repeat the process of drying
and weighing till constant weight is achieved.
Observation table-
S. No. Filter Initial Final Final VSS Average

paper no. weight weight weight VSS
(W1) (W2) at (W3) at
1050C 5500C
Calculation-
Volatile suspended solids = (W2-W3)
W2= final weight of filter paper at 1050C
W3= weight of filter paper at 5500C drying
VSS mg/l= (W2-W3) x 106 / sample volume (ml)
Result- VSS of the water sample was found----- mg/l at 5500C.
Precautions-


FIXED DISSOLVED SOLIDS (FDS) (Gravimetric method)
Object- Determine the FDS in water sample.
Requirements- Whatman filter paper, balance, nickel crucible, measuring cylinder, muffle
furnace, conical flask, desiccator and crow pliers.
Procedure-
(i.) Filter the sample with whatman no. 42 filter paper.

(ii.) Keep washed and marked nickel crucible in muffle furnace for 1 hour at 5500C.
(iii.) Take out the crucible after 1 hour and keep in the desiccator for 30 minutes.
(iv.) Weight the crucible for initial weight (W1).
(v.) Now pour 50 ml filtered sample in the each crucible and carefully keep in the muffle
furnace at 5500C for 2 hours.
(vi.) After that take out from the muffle furnace with help of tong and keep in the
desiccator for 30 minutes.
(vii.) After 30 minutes weight the crucible for final weight (W2).
Calculation-
FDS mg/l= (W2-W1) x 106 / Volume of sample
Observation table-
S. No. Crucible No. Initial weight Final weight FDS mg/l Average FDS
(W1) (W2) mg/l
1.
2.
3.
Result- FDS in the given water sample was found------ mg/l at 5500C.
Precautions-

ALKALINITY (Titrimetric method)
Object- Determine the alkalinity of given water sample.
Requirements- Burettes, pipette, volumetric flask, conical flask and beaker.
Reagents-
1. Standard sulphuric acid (0.02N) - Prepare the 0.1 N H2SO4 by diluting 30 ml conc.
H2SO4 to 1 litre distilled water. Standardized it with standard sodium carbonate Na2CO3
0.1N. Dilute appropriate volume of sulphuric acid, approximately 100 ml to 500 ml to
obtained standard 0.02N H2SO4.
2. Phenolphthalein indicator solution (alcoholic, pH 8.3) - Dissolve 5 gm phenolphthalein in
500 ml 95% ethyl alcohol. Add 500 ml distilled water.
3. Methyl orange indicator- 0.05 gm methyl orange diluted 10 100 ml with distilled water.
Method- Take 100 ml sample in volumetric flask add 2-3 drops of Phenolphthalein indicator. If
pink colour develops titrate with 0.02N H2SO4 till a colour disappears. Note the volume of
sulphuric acid used. Add 2-3 drops of Methyl orange indicator to the same flask and continue
titration till yellow colour changes to orange. Note the volume of sulphuric acid used.
Formula- Calculate total (T), Phenolphthalein (P) and Methyl orange (M) alkalinity as follows
and express in mg/l as CaCO3.
Phenolphthalein alkalinity (mg/l as CaCO3) = A x N x 1000/ volume of sample
Total alkalinity (mg/l as CaCO3) = B x N x 1000/ volume of sample
Methyl orange alkalinity (mg/l as CaCO3) = (B-A) x 1000/ volume of sample

Observation table-
S. No. Sample H2SO4 used H2SO4 used P T M

description with with Methyl alkalinity alkalinity alkalinity
Phenolphthalein orange (mg/l as (mg/l as (mg/l as
indicator (ml) A indicator CaCO3) CaCO3) CaCO3)
(ml) B
(A+B)
1.
2.
3.
Result- The alkalinity of given water sample was observed P alkalinity-------mg/l as CaCO3, T
alkalinity-------mg/l as CaCO3 and M alkalinity-------mg/l as CaCO3.
Precautions-

2. Prepare solution carefully.
3. Note the end point carefully.

ACIDITY (Titrimetric method)
Object- Determine the acidity of given water sample.
Reagents-
(a) Standard sodium hydroxide (0.02N) - 0.8 gm sodium hydroxide is dissolved in distilled
water and dilute to 1000 ml using carbon dioxide free distilled water. Standardized
against standard oxalic acid solution.
(b) Phenolphthalein indicator- 0.05 gm Phenolphthalein is dissolved in 500 ml 95% ethyl
alcohol 500 ml distilled water is added. Add drop wise 0.02N sodium hydroxide till faint
pink colour appears.
Method- Take 100 ml sample in conical flask add 2 drops of Phenolphthalein indicator and
titrate with standard NaOH solution till pink colour appears.
Calculation- Acidity as CaCO3 (mg/l) = ml titrante used (NaOH) x 1000/ ml sample taken
Observation table-
S. No. Sample Normality of NaOH used Acidity (mg/l as

description NaOH (ml) CaCO3)
1.
2.
3.
Result- The acidity of given water sample was observed-----mg/l as CaCO3.
Precautions-

3. Note the end point reading carefully.

CHLORIDE (Argentometric method)
Object- Determine the chloride in given water sample.
Reagents-
(a) Potassium chromate indicator solution- 50 gm K2Cr2O4 is dissolved in a small quantity of

distilled water AgNO3 solution is added drop by drop until a definite red precipitate is
formed. Let stand for 12 hours; filter and dilute to one liter with distilled water.
(b) Standard silver nitrate solution 0.0141N- 2.395 AgNO3is dissolved in distilled water and
dilute to one liter standardized with standard sodium chloride (NaCl). Store in brown
bottle.
Method- Take 100 ml sample in a conical flask and adjust the pH in the range of 7-10.Add 1 ml
K2CrO4 indicator, titrate with AgNO3 end point will be pinkish yellow. Note the reading repeat
the titration with distilled water blank.
Calculation- Cl- (mg/l) = (A-B) x N x 35.45 x 1000/ volume of sample
Where,
A= ml AgNO3 used for sample
B= ml AgNO3 used for blank
N= Normality of AgNO3
Observation table-
S. No. Sample Normality of AgNO3 used AgNO3 used Chloride

description AgNO3 for sample for blank (mg/l)
(ml) A (ml) B
1.
2.
3.

Result- The chloride in given water sample was observed------- mg/l.
Precautions-


SULPHATE (Spectrophotometric method)
Object- Determine the sulphate in given water sample.
Requirements- Spectrophotometer, magnetic stirrer, physical balance, beaker, conical flask,

measuring cylinder, wash bottle etc.
Reagents-
Method- Take 50 ml sample in a conical flask add 20 ml buffer solution and 2 ml conditioning
reagent, mix with magnetic stirrer. While stirrer add a pinch of BaCl2 crystal stir for 1 minute at
constant speed, after stirring pour solution into cell and measure absorbance at 420 nm in
spectrophotometer. Prepare standard from 5 mg to 40 mg SO4-2/litre and measure the absorbance.
Prepare standard graph between absorbance and concentration of sulphate ions.
Calculation- Deduce theSO4-2concentration in water sample from standard graph.
Observation table-
S. No. Sample Sample volume Absorbance Sulphate (mg/l)

description (420 nm)
1.
2.
3.
Result- The sulphate (SO4-2) in given water sample was observed -----mg/l.
Precautions-

3. Measure the absorbance carefully.

FLUORIDE (Spands-Zirconium colorimetric method)
Object- Determine the fluoride in the given water sample.
Requirements- Nesslers cylinder, burette, pipette, reagent bottle, volumetric flask, kjeldalh
flask, heating mental and hot plate.
Interferences removal- interferences due to higher concentration of Al, Cl-, Fe, PO4, F, SO4-2
occur also residual chlorine interference in bleaching action. In this case sample is first
distilled and distillation is taken for determination of fluoride.
Reagents-
(a.) Zirconyl acid reagents- Dissolve 0.133 gm (ZrOCl2.8H2O) zirconium oxichloride in 25

ml distilled water. Transfer in a 500 ml volumetric flask and add 350 ml conc. HCl and
dilute to the mark with distilled water keep in refrigerator.
(b.) Spands Reagent- Dissolve 0.958 gm spands in 500 ml distilled water in a volumetric
flask 0f 500 ml capacity. Keep in refrigerator.
(c.) Reference solution for setting instruments zero- add 10 ml spands solution to 100 ml
distilled water, dilute 7 ml HCl to 10 ml & mix both solutions (total vol. 120 ml). Use
this solution for setting instruments zero before analysis.
(d.) Sodium arsenite 0.5% solution- Dissolve 0.50 gm sodium arsenite (NaAsO2) in 100 ml
distilled water.
(e.) Concentrate H2SO4 1liter (2 x 500 ml)
(f.) Concentrate HCl 1 liter (2 x 500 ml)
(g.) Silver sulphate crystals (Ag2SO4)
(h.) Fluoride reference solution- Dissolve 0.221 gm previously dried (1050C for 2 hours)
sodium fluoride (NaF) in 1 liter volumetric flask. Level as stock reference (100 ppm).
(i.) Fluoride working reference solution- Dilute 10 ml of stock reference F- solution in a 100
ml. volumetric flask dilute to the mark. This will give a solution of 10 ppm fluoride.

Procedure for distillation of sample-
If sample contains interferences in higher concentration than distillation of sample is done,

otherwise clean ground water or surface water samples can be taken for colour development
after filtration. Distillation of the sample is done in the following way:
(1.) Take 400 ml distilled water in a kjeldhal flask and add 200 ml conc. H2SO4 and shake to
mix.
(2.) Heat on heating device to boil at 1800C, when about 100 ml distillate is collected put off
the heating unit and after cooling to about 1000C discard the distillate and add 300 ml
sample in the kjeldhal flask, and keep on hot plate for distillation add a pinch of silver
sulphate (0.5 gm) in the kjeldhal before distillation to avoid interference due to chloride
(Cl-). Collect exactly 300 ml distillate in the receiving conical flask. Take out the flask
while distillation is continued.
Colour Development- Take the sample directly for colour development if sample do not has
interferences in higher concentrations and turbidity otherwise sample is distilled after
distillation proceed as below-
(1.) Take 50 ml capacity nesslers cylinder with stopper mark on the cylinder as blank 1, 2, 3-
--- 10.
(2.) Take reference fluoride solution in the cylinders for colour development for preparation
of calibration curve as below-
S.No. ml of 10 ppm Concentration Absorbance at B-S

reference ppm 510 nm
working solution
1. Blank 0.0
2. 1 ml 0.2
3. 2 ml 0.4
4. 3 ml 0.6
5. 4 ml 0.8
6. 5 ml 1.0

7. 6 ml 1.2
8. 7 ml 1.4
S1 Sample-I 50 ml Sample-I
S2 Sample-II 50 ml Sample-II
B-S= Blank absorbance Sample absorbance.
(3.) After taking the volumes of reference solution make up the volume to 50 ml in each
cylinder.
(4.) Add 5 ml spands solution and 5 ml zirconyl chloride solution and shake well to mix and
let stand for 2 minutes.
(5.) Mean time put on the spectrophotometer for warm up.
(6.) Set instruments 0 with reference solution.
(7.) Now measure the absorbance of blank and note the reading of blank.
(8.) Prepare standard graph (calibration curve) on a graph paper- Taking difference of
absorbance (B-S) at x-axis and concentration (ppm) at y-axis. The curve comes almost
linear.
.
= f (factor)

Calculation- The concentration of fluoride in the sample = absorbance (B-S) x f (factor) mg/l or
ppm
If sample is diluted for direct determination then conc. F- ml/l = [absorbance difference
(B-S) x f]
Result-The fluoride in given water sample was found------mg/l.
Precautions-

3. Measure the absorbance carefully.

TOTAL HARDNESS (EDTA Titrimetric method)

Object- Determine the total hardness of given water sample.
Requirements- Volumetric flask, conical flask, burette, pipette, measuring cylinder, burette
stand, pipette stand and wash bottle etc.
Reagents-
(a) Buffer solution- Dissolve 16.9 gm NH4Cl in 143 ml conc. NH4OH. Add 1.25 gm
magnesium salt of ethylene diamine tetra acetic acid (EDTA) and dilute to 250 ml with
distilled water. Store in a plastic bottle stopper tightly for no longer than one month.
(b) Eriochrome Black T solution (as indicator)- Dissolve 0.5 gm dye in 100 ml
triethanolamine or 2 ethylene glycol monomethyl ether. The salt can also be used in dry
powder form by grinding 0.5 gm dye with 100 gm NaCl.
(c) Standard EDTA titrant (0.01M)- Weight 3.723 gm di-sodium salt of EDTA, dehydrate,
dissolve in distilled water and dilute to 1000 ml. Store in polyethylene bottle.
(d) Standard calcium solution- Weight 1 gm anhydrous CaCO3 in a 500 ml flask. Add 1 + 1
HCl slowly through a funnel till all CaCO3 is dissolved. Add 200 ml distilled water and
boil for a few minutes to expel CO2. Cool and add a few drops of methyl red indicator
and adjusts to the intermediate orange colour by adding 3N NH4OH or 1 + 1 HCl, as
required. Transfer quantitatively and dilute to 1000 ml with distilled water, 1 ml=1mg
CaCO3.
Method- Standardize the EDTA titrant against standard calcium solution using procedure given
below-
Take 50 ml sample in a conical flask add 2 ml buffer solution and 2 drops EBT
indicator solution, wine red colour appears. Titrate with EDTA till the colour changes to blue.
Note the volume of EDTA used. Calculate the total hardness by following formula-
Formula-
Total Hardness (EDTA) mg/l= A x B x 1000/ volume of sample
Where,

A= ml EDTA used
B= mg CaCO3 equivalent to 1 ml EDTA
Observation table-
S. No. Sample Sample volume EDTA used Total hardness

description (ml) (ml) (mg/l) as
CaCO3
1.
2.
3.
Result- The total hardness of given water sample was observed------- mg/l as CaCO3.
Precautions-


Ca & Mg HARDNESS (Titrimetric method)
Object- Determine the Ca and Mg hardness in given water sample.
Requirements- Volumetric flask, conical flask, burette, pipette, measuring cylinder, burette
stand, pipette stand and wash bottle etc.
Reagents-
(a) Buffer solution- Dissolve 16.9 gm NH4Cl in 143 ml conc. NH4OH. Add 1.25 gm
magnesium salt of ethylene diamine tetra acetic acid (EDTA) and dilute to 250 ml with
distilled water. Store in a plastic bottle stoppered tightly for no longer than one month.
(b) Standard EDTA titrant (0.01M)- weight 3.723 gm di-sodium salt of EDTA, dehydrate,
dissolve in distilled water and dilute to 1000 ml. Store in polyethylene bottle.
(c) Standard calcium solution- weight 1 gm anhydrous CaCO3 in a 500 ml flask. Add 1 + 1
HCl slowly through a funnel till all CaCO3 is dissolved. Add 200 ml distilled water and
boil for a few minutes to expel CO2. Cool and add few drops of methyl red indicator and
adjusts to the intermediate orange colour by adding 3N NH4OH or 1 + 1 HCl, as required.
Transfer quantitatively and dilute to 1000 ml with distilled water, 1 ml-1mg CaCO3.
(d) Sodium hydroxide (1N) - 4 gm NaOH dissolved in 100 ml distilled water.
(e) Murexide (ammonium purpurate) indicator- 75 gm of the indicator is dissolved in 50 gm
absolute ethylene glycol.
Method- Take 50 ml sample in conical flask raise the pH to 12-13 by adding 2 ml NaOH. Add
1-2 drops of indicator and titrate with EDTA until solution becomes purple from pink.
Calculation- Calcium hardness as CaCO3 (mg/l) =
A x B x 100 x 1000/ volume of sample
Where,
A= ml EDTA used
B= mg CaCO3 equivalent to 1 ml EDTA

Magnesium hardness
Magnesium hardness is determined by subtraction of Ca hardness from the total hardness
Magnesium hardness (mg/l) = total hardness - calcium hardness
Calcium
Calcium is determined by multiplying the Ca hardness by 0.4 i.e.
calcium as Ca (mg/l) = calcium hardness x 0.40
Magnesium
Magnesium is determined by multiplying the magnesium hardness by 0.243 i.e.
Magnesium as Mg (mg/l) = magnesium hardness x 0.243.
Observation table-
S. No. Sample Sample volume EDTA used Calcium

description (ml) (ml) hardness (mg/l)
1.
2.
3.
Result- The Ca and Mg hardness was observed ------mg/l as CaCO3 and------mg/l asCaCO3
respectively and Ca and Mg was observed -------mg/l and -------mg/l.
Precautions-


SODIUM (Flame photometric method)
Object- Determine the sodium in water sample.
Requirements- Flame photometer, conical flask etc.
Reagents-
(a.) Stock sodium solution- 2.542 gm dried NaCl is dissolved in distilled water and make up
to 1 liter. (1 ml = 1 gm Na)
(b.) Working sodium solution- 10 ml of the stock solution is diluted to a1 liter (1 ml = g
Na).
Procedure-
(1.) Start the electrical supply and switch on the air supply, stabilize the air. The needle
should be steady at the mark.
(2.) Switch on the gas and maintain the gas fuel mixture so that the blue flame is seen through
the viewing window.
(3.) Aspirate distilled water and adjust the flame photometer reading to zero.
(4.) Calibrate the instrument by aspirating the standard and adjusting the flame photometer
reading to desired mark by 0-10 and 0-100 mg/l.
(5.) Aspirate distilled water to bring the reading to zero mark.
(6.) Aspirate sample and note down the flame photometer reading.
(7.) Put off the fuel supply first followed by air and then main switch.
Result- The sodium in given water sample was observed------------ mg/l.
Precautions-

3. Calibrate instrument properly.

POTASSIUM (Flame photometer method)
Object- Determine the potassium in given water sample.
Requirement- Flame photometer, plastic bottle, volumetric flask, desicator and distilled water.
Reagents-
(a.) Stock potassium solution- 1.907 gm KCl, dried at 1100C and cooled in desicator, transfer
to 1 liter volumetric flask and make to 1 liter with water, (1 ml= 1 mg K).
(b.) Intermediate potassium solution- 10 ml stock potassium solution dilute with 100 ml
water, (1 ml=0.1 mg K), prepare calibration curve in the range of 1 to 10 mg/l.
(c.) Standard potassium solution- Dilute 10 ml intermediate solution with 100 ml water, (1
ml= 10 g K), prepare calibration curve in the range of 0.1 to 1 mg/l.
Procedure- Select proper photocell, wavelength, slit width adjustments, fuel gas and air
pressure, step for warm up, correcting for interference and flame back ground, rinsing of burner,
sample ignition and emission intensity of flame photometer. Now prepare a blank and potassium
calibration standards, in any of the application ranges, 0-100, 0-10 or 0-1 mg K/l. Measure
emission at 766.5 nm and prepare calibration curve. Determine potassium concentration of the
sample, or dilute sample from the curve.
Calculation-
Mg K/l = mg K from the calibration curve x Dilution
Where,
Dilution = sample (ml) + distilled water (ml) / Sample (ml)
Result- The potassium in given water sample was observed------------ mg/l.
Precautions-


PERCENT SODIUM (Indirect method)
Object- Determine the percent sodium in given water sample.
Procedure for determination of % Na-
(i.) To determine % Na the following formula is applied
% Na = Na x 100/ Total cations (Ca+2 + Na+ + K+ + Mg+2)

Where concentrations of cations are in meq/l therefore we see that for determining %
Na the concentration of Ca+2, Mg+2 are determined by titrimetric followed by
calculation and Na+, K+ are determined either by flame photometric method or by
atomic absorption spectroscopy.
After determination of concentration of Ca+2, Na+, K+ and Mg+2 is mg/l.
These are then converted to meq/l
Meq/l = concentration mg/l of cation / equivalent weight of cation
Equivalent weight of Ca+2 = 20, Na+ = 23, K+ = 39.1 and Mg+= 12.15
(ii.) After calculating the concentration in mili equivalent per liter (meq/l) % Na is
calculated.
Calculation-
% Na = Na x 100/ Total Cations (Ca+2 + Na+ + K+ + Mg+2)
Where concentrations of cations are in meq/l.

Quality of irrigation water on the basic of some common parameters-
Quality of Conductivity pH Cl-meq/l SAR % Na

water in mhos/cm
Good 0.5 to 1.5 7.5 to 8.0 2.5 to 5.0 1 to 2 30 to 60
Very good 0.5 6.5 to 7.5 2.5 1.0 30
Poor 3.0 to 5.0 8.5 to 9.0 7.5 to 10.0 4.0 to 8.0 75 to 80
Very poor 5.0 to 6.0 1.0 to 10.0 10.0 to 12.5 8.0 to 15.0 80 to 90
Unsuitable 6.0 10.0 12.5 15 90
Result-The percent sodium in given water was found------%.

SODIUM ABSORPTION RATIO (SAR)
Object- Determine SAR in given water sample.
Procedure-
Ca+2, Mg+2 and Na+ as mg/l are determined. Then these are converted to milli equivalent per liter
by using formula-
Milli equivalent per liter= mg/l (concentration) / equivalent weight
As we know the equivalent weight of Ca+2, Mg+2 and Na+ as below
Element Equivalent weight

+
Na 23
Ca+2 20
Mg+2 12.5
After that calculate milli equivalent per litre as below-
/ / /
Cameq/lit = Mgmeq/lit = Na meq/lit =
. .
Calculate SAR by the formula given below-
Calculation-

SAR=
/
Concentration of Na, Ca and Mg cat ions in meq/lit.
Result-SAR value in the given water sample was found-------.

TOTAL IRON [Fe] (Ferrous & Ferric) (Spectrophotometric method)
Object- Determine the total Fe in water sample.
Apparatus Required-Porceline dishes, beaker, volumetric flask, measuring cylinder, reagents

bottle, glass rod, wash bottle, iron wire, spectrophotometer and nessler cylinder.
Reagents-
(1.) Glacial acetic acid

(2.) Conc. HCl -500 ml
(3.) Hydroxyl amine hydrochloride solution- Dissolve 10 gm (NH2OH) solid in 100 ml
volumetric flask dilute to the mark with distilled water.
(4.) Ammonium acetate buffer solution- Dissolve 250 gm NH2C2O3.3H2O in 1 liter
volumetric flask shake to dissolve and make up the volume with distilled water
transfer the reagent in liter reagent bottle.
(5.) 1,10phenenthroline solution- Dissolve 0.100 gm 1,10 phenenthroline in 100 ml
distilled water in a 100 ml volumetric flask. Heat to dissolve (Do not boil) discard the
solution. If darken no need to heat if 2 drops of conc. HCl are added to dissolve.
(6.) Dissolve 1.404 gm ammonium ferrous sulphate (FAS) is 50 ml distilled water and 20 ml
conc. H2SO4shake to dissolve. Add 0.1 N KMnO4 (potassium permanganate solution) drop
wise to get faint pink colour of solution then dilute to the mark with distilled water in 1 liter
volumetric flask. This will give a solution of 1 ml = 200 g Fe or 200 ppm solution of Fe.
(7.) Reference (working) Fe solution (1 ppm) - Dilute 5.0 ml of stock (200 ppm) solution to 1
liter to get a solution of 1 ppm.
Preparation of Calibration Curve
(i.) Arrange volumetric flask of 100 ml capacity 10 numbers and mark on them. Take the
volumes of reference (working) solution as below & make up the volume to 100 ml
after addition of reagents.

S. No. Flask No. Volume of 1 Volume taken Absorbance at

ppm reference for colour 510 nm
solution development
1. Blank Distilled water 50 ml
100 ml
2. 1 10 ml dilute 100 50 ml
ml
3. 2 20 ml dilute 100 50 ml
ml
4. 3 30 ml dilute 100 50 ml
ml
5. 4 40 ml dilute 100 50 ml
ml
6. 5 50 ml dilute 100 50 ml
ml
7. 6 60 ml dilute 100 50 ml
ml
8. 7 70 ml dilute 100 50 ml
ml
9. 8 80 ml dilute 100 50 ml
ml
10. 9 90 ml dilute 100 50 ml
ml
(ii.) Take reference solution in 100 ml volumetric flask then ad distilled water to make up
the volume to 100 ml. this will give reference solution of 0.1, 0.2, 0.3, 0.4----- 0.9
ppm reference solution.
(iii.) Take 10 numbers of porcelain dishes of 100 ml capacity mark. Blank 1, 2, 3,----9.

(iv.) Take exactly 50 ml of reference solution from the volumetric flask of same numbers.
Then we have reference solution 50 ml in each cylinder having concentration of 0.1,
0.2----0.9 ppm respectively.
(v.) Now add 2 ml conc. HCl and 1 ml hydroxylamine hydrochloride and shake.
Evaporate to remain 10-15 ml. cool and transfer in 100 ml nessler cylinder dilute to
50 ml.
(vi.) Add 10 ml ammonium acetate buffer solution and 4.0 ml 1, 10 phenenthroline
solutions and make up 100 ml and shake to mix properly.
(vii.) Keep for 10 to 15 minutes for colour development mean time put on
spectrophotometer to warm up.
(viii.) Take absorbance at 510 nm.
(ix.) Prepare reference curve on a graph paper taking absorbance at y axis and
concentration at x axis.
Analysis of Sample-
(i.) Take 50 ml water (or an adequate dilute to 50 ml) in a porcelain dish and add 2 ml
conc. HCl & 1 ml hydrochloride and heat on a water bath till volume is left 15-20 ml.
If sample is ashed add 20 ml 1+1 HCl to dissolve the residue cool to room
temperature.
(ii.) Then transfer in 100 ml nessler cylinder wash the porcelain dish with distilled water
approx 10 ml and add washing in cylinder dilute to 50 ml with distilled water.
(iii.) Then add 10 ml ammonium acetate buffer solution, 4.0 ml 1, 10 phenenthroline
solution and shake it well.
(iv.) Let stand for 15-20 minutes mean time switch on spectrophotometer and set at 510
nm.
(v.) Measure the absorbance and compare the reading with calibration curve to obtain
concentration.
Calculation- Results are calculated from calibration curve directly in ppm
Results-The total iron in given water sample was found --------------- mg/l.

Precautions-


TOTAL KJELDHAL NITROGEN (TKN)
Object- To determine the TKN in given water sample
Apparatus Required- ammonia distillation apparatus, hot plate, spectrophotometer, burette,

nesslers cylinders, other laboratory glass ware.
Reagents-
(a) Digestion reagents- Dissolve 134.0 gm potassium sulphate (K2SO4) in 650 ml distilled
water & add 200 ml conc. H2SO4 and add this solution to the solution prepare by
dissolving 2.0 gm HgO (mercuric-Oxide) in 25 ml conc. H2SO4. Dilute the whole
solution to 1 liter in a 1 liter volumetric flask.
(b) Phenolpthaline indicator- Dissolve 0.10 gm solid phenolpthline powder in to 100 ml 96%
isopropyl or ethyl alcohol, add drop wise 0.02 N NaOH to obtain orange red colour,
(c) Mixed indicator- Dissolve 0.20 gm methyl red in to 100 ml 95% ethanol & mix this
solution to the solution prepare by dissolving 0.100 gm methylene blue in 50 ml ethanol.
(d) Boric acid solution (indicating)- Dissolve 20.0 gm H3BO3 (Boric acid) in approximately
600 ml distilled water in 1 liter flask add 10 ml mixed indicator and makeup the volume
of 1 liter with distilled water.
(e) Sodium hydroxide (NaOH) 6N solution- Dissolve 240.0 gm NaOH in 1 liter beaker by
adding distilled water slowly and shaking with glass rod, cools and transfer in 1 liter
volumetric flask and dilute to the mark with distilled water.
(f) Sulphuric acid 0.02 N- Prepare as in alkalinity determination.
(g) Digestion & Distillation of the sample-
1. Take 400 ml sample in a 500 ml capacity kjeldhal flask and add 50 ml digestion mixture.
If suspended organic matter is high adding more digestion mixture to keep salt to acid
ratio 0.8 (8:10) arrange the unit for heating in a digestion chamber.
2. Keep on heating when content of the flask become clean heat for another 30 minutes for
complete decomposition of organic matter present in the sampler reduce volume to 100
ml approx.
3. Cool and dilute to 250 ml and add 0.5 ml phenolphthalein indicator and add 6 N NaOH
till pH rises to about 8.5.

4. Arrange the flask for distillation on heating mental or hot plate and continue distillation.
Keep conical flask blow the receiving end of distillation unit having 50 ml boric acid
solution. Use plain boric acid solution if nesslerization method is to be followed or use
indicating boric acid solution if titrimetric method is followed. Place 0.04N H2SO4 in
place of boric acid. If NH3 selective electrode method is to be followed. Collect
approximately 200 ml distillate.
5. After complete distillation do not off the hot plate or heating mental before taking out of
receiving conical flask otherwise back suctions will occur.
6. Make up the distillate volume 250 ml in volumetric flask, measure the concentration of
ammonical nitrogen in the 50 ml part of the distillate as below-
A. Nesslerisation Method-
(i) Take 50 ml portion of the distillate.
(ii) Add 1 ml nesslers reagent and shake. Keep for 10 minutes for colour development.
(iii) Mean time put on spectrophotometer to warm up.
(iv) Prepare distilled water blank.
(v) Measure absorbance of sample at 420 nm.
(vi) Compare the reading for concentration in mg/l with calibration curve of NH3-N as
prepared in the determination of NH3-N (ammonical nitrogen).
Calculation-
Compare the absorbance reading with calibration curve and take concentration mg/l and
multiply with 5 because only 50 ml of distillate is taken for analysis out of 250 ml total
volume.
B. Titrimetric Method-
(i) Take the conical flask after complete distillation & cool.
(ii) Titrate with 0.02N H2SO4 and note the volume of H2SO4 consumed with blank and
sample.
Calculation-

TKN mg/l =


Where, A= ml of 0.02N H2SO4 for sample titration.
B= ml of 0.02N H2SO4 for blank titration.
N= Normality of H2SO4
Result- The total kjeldhal nitrogen in given water sample was observed ---------mg/l.
Precautions-


NITRITE- NITROGEN (Spectrophotometric method)
Object- Determine the nitrite concentration in given water sample.
Requirements- spectrophotometer, pipette, glass stopper flask, beaker and distilled water.
Reagents-
(a.) Colour reagents- 100 ml 85% phosphoric acid and 10 ml sulphanilamide mix in 800 ml
water. After dissolving add 1 gm N-1- naphthylethylene diamine dihydro chloride. Mix to
dissolve, then dilute to 1000 ml with distilled water. (Solution is stable for one month
when stored in dark in refrigerator).
(b.) Sodium oxalates (0.05N) - Dissolve 3.350 gm Na2C2O4 primary standard grade in water
and dilute to 1000 ml.
(c.) Stock nitrite- Dissolve 1.232 gm NaNO2 in water and dilute to 1000 ml (1 ml= 250 g
N). Preserve with 1 ml chloroform (CHCl3. Standardize by pipetting in order 50 ml 0.01
M KMnO4, 5 ml conc. H2SO4 and 50 ml stock NO2- solution in to a glass stoppered flask.
Shake well and warm to 70-800C. Discharge permanganate colour by adding 10 ml
portions of 0.025 M sodium oxalate. Titrate excess oxalate with 0.01 M (0.05N) KMnO4
to faint pink end point.
Calculation Nitrite content of stock solution-
A= [(B x C) (D x E)] x 7 / F
Where,
A= NO2--N/ml in stock solution (mg)
B= total KMnO4 used (ml)
C= normality of KMnO4
D= total oxalate added (ml)
E= normality of oxalate
F= stock nitrite taken for titration (ml)

(d.) Standard nitrite solution- Dilute 10 ml intermediate NO2- solution to 1000 ml with water
(1 ml = 0.500 g NO2--N, prepare daily).
(e.) Standard potassium permanganate solution (0.05N) - Dissolve 1.6 gm KMnO4 in 1 liter
distilled water. Allow ageing for 1 week then decant supernatant. Standardize this
solution frequently as follows-
Weigh to nearest 0.1 mg several 100 to 200 mg samples for anhydrous sodium oxalate in
beaker. To each beaker add 100 ml distilled water, 10 ml 1+1 H2SO4 and heat rapidly to 90 to
950C. Titrate with permanganate solution to a slight pink end point that persists to at least 1 min.
Do not allow temperature to fall below 850C. Run a blank on distilled water +H2SO4.
Normality KMnO4 = Na2C2O4 (gm) / (A-B) x 0.33505

Where,
A= titrant for sample (ml)
B= titrant for blank (ml)
Average the result of several titrations.
Procedure-
(a.) Add 2 ml colour reagent to 50 ml sample, or to a portion to 50 ml and mix. After this
measure absorbance at 543 nm. Wait between 10 minute and 2 hours after addition of
colour reagent before measurement. Prepare standard curve by diluting 1, 2, 3, 4 and 5 ml
of standard nitrite solution to 100 ml to give 5, 10, 15, 20 and 25 g/l concentration,
respectively.
Calculation-
Deduce sample concentration directly from the curve, taking in consideration dilution of the
sample if applicable.

Result-The NO2 concentration in given water sample was found ---------mg/l NO2-N
Precautions-


NITRATE- NITROGEN (Spectrophotometric method)
Object- Determine the nitrate-nitrogen concentration in given water sample.
Requirements- Volumetric flask, beaker, glass rod, heating mental, reagent bottles, nesslers
cylinder, balance, spectrophotometer, water bath, porcelain dishes, rubber and desicator.
Reagents-
(a.) Phenoldisulphonic acid (PDA) - Dissolve 25 gm white phenol in 150 ml conc. H2SO4 and
add 74 ml fuming H2SO4 (15% free SO3) stir well and heat for 2 hours on a water bath. If
fuming H2SO4 is not available add 86 ml conc. H2SO4 in place of fuming acid in the
solution and shake in a beaker, heat for 2 hours to dissolve, keep in reagent bottle.
(b.) Potassium hydro-oxide solution (12N)- Dissolve 336.5 gram AR grade KOH in 200 ml
distilled water in a beaker stir with glass rod to dissolve transfer in 500 ml volumetric
flask and makeup to volume with distilled water.
(c.) Nitrate reference Solution (100 ppm) - Dissolve 0.7219 gm AR grade anhydrous KNO3
(potassium nitrate) dried at 1050C for 2 hours. Dissolve in 1 liter distilled water.
(d.) EDTA reagent- Mix 50 gm ethylene diamine tetra acitic acid (EDTA) in 20 ml distilled
water and 60 ml NH4OH and mix.
Procedure-
(a.) Take 100 ml sample in a beaker check the pH of the sample and adjust pH 7 by adding
adequate amount of acid or alkali drop wise to adjust the pH do not use nitric acid
(HNO3) for adjusting pH often weak acids like acetic acid or diluted strong acid or alkali
are used for fixing pH and not more than 3 to 5 ml acid or alkali.
(b.) Take neutralized 50 ml sample in a porcelain dish and 50 ml sample in other porcelain
dish and in third porcelain dish 50 ml distilled water always take sample in duplicate.
(c.) Keep the dishes on water bath and evaporate to dryness.
(d.) Then add 2 ml phenoldisulphonic acid reagent and dissolve the residue in it by rubbing
with rubber polish men. Add about 10 ml distilled water and shake to dissolve.
(e.) Transfer the content in 100 ml capacity nesselers cylinder wash the porcelain dish with 2
portions of distilled water 10 ml each and take the washings in the cylinder makeup to 50

ml with distilled water and shake. Then add 10 ml KOH reagent in each cylinder. If
turbidity is developed add drop wise EDTA reagent solution to just clear the solution in
the cylinder. Then makeup to 100 ml with distilled water.
(f.) Filter the contents if necessary. (NH4OH) ammonia solution may be added in the sample
in place of EDTA reagent to avoid turbidity. Let stand the cylinder for 20 minutes for
colour development.
(g.) Put on the spectrophotometer to warm up for 20 minutes.
(h.) Set the absorbance at 410 nm and set instruments 0 with blank treated with sample.
(i.) Take the absorbance of the samples and note the reading for results. The absorbance
readings are compared with calibration curve of NO3-N prepared as below-
Preparation of Calibration Curve of Nitrate-N

(1.) Take 10 Nos of 100 ml capacity necesslers cylinders and clean, then rise with
distilled water.
(2.) Arrange & number the cylinder in increasing order, take the volume of reference
(working) solution as below-
Cylinder No. Volume of reference Concentration in Absorbance at 410

solution 10 ppm to ppm nm
makeup 50 ml
1. 0.0 ml blank 0.0 ppm
2. 2.5 ml 5 ppm
3. 5.0 ml 10 ppm
4. 7.5 ml 15 ppm
5. 10 ml 20 ppm
6. 12.5 ml 25 ppm
7. 15 ml 30 ppm

8. 17.5 ml 35 ppm
9. 20 ml 40 ppm
(3.) After taking reference solution in the 100 ml capacity nessler cylinder makeup
the volume to 50 ml
(4.) Add 2 ml PDA reagent 10 ml KOH solution and 10 ml NH4 OH solution and
shake to mix. Let stand for 20 minutes for colour development.
(5.) Mean time put on spectrophotometer.
(6.) After 20 minutes measure absorbance by setting instruments 0 with distilled
water reagent blank.
(7.) Plot a calibration curve absorbance verses concentration (ppm) on a graph paper.
(8.) Deduce the nitrate concentration from standard graph.
Results-the nitrate-nitrogen in given water sample was found ------- NO3-N mg/l.
Precautions-


D.O. (DISSOLVED OXYGEN) (Modified Winklers method)
Object- Determine the concentration of dissolved oxygen in given water sample.
Requirements- BOD bottle, pipette, burette, conical flask, volumetric flask, beaker, measuring
cylinder, burette stand, and pipette stand, etc.
Reagents-
(i) Sodium thiosulphate [Na2S2O3.5H2O] (0.025 N) - dissolve 6.205 gm of solution

thiosulphate in previously boiled distilled water add 0.4 gm solid NaOH and make up
the volume to one litre.
(ii) Manganous sulphate solution [MnSO4]- 480 gm MnSO4.4H2O / 400 gm
MnSO4.2H2O / 364 gmMnSO4.H2O is dissolved in distilled and dilute to 1000 ml.
(iii) Alkali-iodide azide reagent - 500 gm NaOH and 135 gm NaI are dissolved in distilled
water and dilute to 1000 ml. Add 10 gm NaN3 dissolved in 40 ml distilled water.
Or
700 gm KOH and 150 gm KI are dissolved in distilled water and dilute to 1000 ml.
Add 10 gm NaN3 dissolved in 40 ml distilled water.
(iv) Starch solution (use an aqueous solution or soluble starch powder)- 2 gm soluble
starch and 0.2 gm salicylic acid as preservative in 100 ml hot distilled water.
Method- Take sample in BOD bottle add 2 ml MnSO4 solution followed by 2 ml alkali-
iodide-azide reagent, yellow colour precipitate appears (if white precipitate appears it
indicates that there is no DO). Put stopper carefully to exclude air bubbles and mix by
inverting bottle 40-50 times. Allow to settle the precipitate. When precipitation has settled
sufficiently add 2 ml conc. H2SO4. Mix by inverting bottle several times until precipitate
dissolved completely.
Titrate 203 ml sample with standard 0.025N sodium thiosulphate solution to a

pale yellow colour, add 1 ml starch indicator and continue titration until the disappearance of
blue colour. Note the reading and calculate DO by following formula-


Formula-

Where,
V= Volume of sodium thiosulphate used in ml
N= Normality of sodium thiosulphate
Observation table-
S. No. Sample Normality of Sodium DO (mg/l)

description sodium thiosulphate
thiosulphate used (ml)
1.
2.
3.
Result- The dissolved oxygen in given water sample was observed -------mg/l.
Precautions-

2. Avoid bubbling during sampling.
4. Note end point carefully.

BOD (BIO-CHEMICAL OXYGEN DEMAND)
Object- Determine the BOD of given water sample.
Requirements- BOD incubator, BOD bottle, pipette, burette, conical flask, volumetric flask,
beaker, measuring cylinder, burette stand, and pipette stand, wash bottle etc.
Reagents-
(i) Phosphate buffer solution- 8.5 gm potassium dihydrogen phosphate (KH2PO4), 21.75
gm dipotassium hydrogen phosphate (K2HPO4) 33.4 gm. Disodium hydrogen
phosphate (Na2HPO4.7H2O) and 1.7 gm ammonium chloride NH4Cl are dissolved in
about 500 ml distilled water and dilute to 1000 ml. pH of the solution should be 7.2.
(ii) Magnesium sulphate solution [MgSO4.7H2O] - 22.5 gm MgSO4.7H2O is dissolved in
distilled water and dilute to 1000 ml.
(iii) Calcium chloride solution (CaCl2) - 27.5 gm CaCl2 is dissolved in distilled water and
dilute to 1 litre.
(iv) Ferric chloride solution [FeCl3.6H2O] - 0.25 g. FeCl3.6H2O is dissolved in 1000ml
distilled water.
(v) Sulphuric acid [H2SO4] (1N)- Slowly and while stirring 28 ml conc. H2SO4 is added
in distilled water and dilute to 1000 ml.
(vi) Alkali sodium hydroxide [NaOH] (1N) - 40 gm NaOH is distilled water and dilute to
1000 ml.
(vii) Glucose (Glutamic acid solution) - 150 gm glucose and 150 gm glutamic acid are
dissolved in distilled water and dilute to 1 litre. Prepare fresh immediately before use.
Method-Preparation of dilution water.: Place desired volume of water in a suitable vessel

and 1 ml each of phosphate buffer, MgSO4, CaCl2, FeCl3, solution per litre of water are
added. Seed dilution water, if desired prepare 2-3 percentage of the sample by dilution water
and fill the 3 BOD bottles of each dilution. Also prepare 2% dilution of glucose-glutamic
acid by dilution water and fill the 3 BOD bottles. Fill 3 BOD bottles of dilution water as
blank.

Measure the dissolved oxygen of one of the three bottles of each dilution, of
2% glucose-glutamic acid and blank, on starting day (0 day) by iodometric or electrometric
method. Keep the remaining bottles (2 bottles of each sample and blank) in the incubator for
5 days if temperature is kept 200C or for 3 days if temperature is maintained at 270C. After 5
or 3 days as the case may be measure the DO of the remaining bottles (2 bottles of each
sample and blank). Calculate the BOD using following formula-
(a) When dilution water is not seeded:

BOD (mg/l) =

(b) When dilution water is seeded:

BOD (mg/l) =

Where,
D1= DO of diluted sample initially
D2 = DO of diluted sample after incubation.
P = Decimal volumetric fraction of sample used.
B1 = DO of seed control initially
B2 = DO of seed control after incubation
F = Ratio of % seed in diluted sample to % seed in seed control
Observation table-
S. No. Description Volume of Initial DO Final DO BOD (mg/l)

of sample sample/dilution (mg/l) (mg/l)
factor
1.
2.
3.

Result- The BOD in given water sample was observed------- mg/l.
Precautions-

2. Avoid bubbling during sampling.

COD (CHEMICAL OXYGEN DEMAND) (Open reflux method)
Object- Determine the COD of given water sample.
Requirements- COD reflux assembly, hot plate, glass beads, conical flask, beaker, measuring
cylinder, wash bottle, burette stand, pipette stand, burette, pipette etc.
Reagents-
(i) Standard potassium dichromate solution (0.25N) - 12.259 gm K2Cr2O7 is dissolved,

previously dried at 1030C for 24 hours, in 1000 ml distilled water.
(ii) Sulphuric acid reagents (0.1N)- Add 22 gm silver sulphate (AgSO4) to a 9 lbs (4 kg)
bottle of concentrated H2SO4. Keep overnight for dissolution and after that shake well
to it.
(iii) Standard ferrous ammonium sulphate solution (0.1N) - Dissolve 39.2 gm of Fe
(NH4)2 (SO4)2.6H2O in 500 ml of distilled water. Add 20 ml of concentrated H2SO4,
and dilute to 1 litre.
Standardization- Dilute 10 ml standard K2Cr2O7 solution to about 100 ml. Add 30 ml conc.
H2SO4 and cool. Add 3-4 drops of ferroin indicator and titrate with the ferrous ammonium
sulphate solution till the colour changes to wine red. The normality of the Fe(NH4)2 (SO4)2
solution is given by the following formula-
.
Normality of Fe(NH4)2 (SO4)2 solution =

(iv) Ferroin indicator solution- 1.485 gm. 1-10 phenanthroline monohydrate and 695 mg
ferrous sulphate FeSO4.7H2O are added in distilled water and dilute to 1000 ml.
(v) Mercury sulphate (HgSO4) - Crystal or powder.
Method- 50 ml of sample is taken into a 500 ml refluxing flask. 1 gm HgSO4, glass beads
and very slowly added 5 ml sulphuric acid reagent with mixing to dissolve HgSO4 cool while
mixing to avoid possible loss of volatile materials. 25 ml K2Cr2O7solution is added and mix.
Flask is attached to condenser and turn on cooling water. Remaining sulphuric acid reagents
(70 ml) is added through open end of condenser. Reflux for 2 hours, cool, and wash down
condenser with distilled water.

Cool to room temperature and titrate excess K2Cr2O7 with F.A.S. using 0.1-0.15 ml (2-3
drops) ferroin indicator.

Formula- COD (mg/l) =

Where,
A= F.A.S. used for blank in ml
B= F.A.S. used for sample in ml
N= Normality of F.A.S.
8000= Milli equivalent weight of oxygen x 1000 ml/lit.
Observation table-
S. No. Sample Normality of F.A.S. used (ml) COD (mg/l)

description F.A.S.
1.
2.
3.
Precautions-

3. Note end point carefully.

RESIDUAL CHLORINE
Object- Determine the free chlorine in given water sample.
Requirements-burette, pipette, beaker, conical flask, measuring cylinder, wash bottle etc.
Reagents-
(a.) Acetic acid (Glacial)

(b.) Potassium iodide KI crystal
(c.) Standard sodium thiosulphate (0.1N)- 25 gm Na2S2O3.5H2O is dissolved in one litre
distilled water and standardize with standard potassium dichromate.
70-80 ml distilled water is taken in a 250 ml flask, 1 ml conc. H2SO4 is added followed
by 10 ml K2Cr2O7 solution and KI. Allow the reaction-mixture to stand for 5-7 minute in
dark. Titrate with sodium thiosulphate solution until the yellow colour of iodine librated
is discharged starch indicator is added and titrate, until the blue colour disappeared.
(d.) Sulphuric acid H2SO4 conc.
(e.) Standard sodium thiosulhate (0.1B) - 100 ml of solution (c) is taken and diluted to one
litre by distilled water.
(f.) Starch indicator solution- 5 gm starch is taken in a little cold water make the paste and
pour into one litre boiling distilled water, stir and let settle down for overnight, use clear
supernatant preserve with 1.25 gm salicylic acid and 4.0 gm zinc chloride.
Procedure- In a sample 5 ml acetic acid is added to reduce the pH four or less than four, add
about one gm KI, mix and titrate with standard sodium thiosulphate titrant, until the yellow
colour of the librated iodine is discharged. Add 1 ml starch solution is added and titrate until
blue colour is disappeared.
Calculation-
Free chlorine as Cl2mg/lit =

A Bx N x 35450
ml sample
Where

A= ml titration for sample
B= ml titration for blank (Positive or negative)
N= normality of Na2S2O3
Result- The free chlorine in given water sample was observed -----mg/l.
Precautions-


CHLORINE DEMAND
Object- Determine the chlorine demand in given water sample.
Reagents-
(a.) Standard sodium thiosulphate (0.025 N)

(b.) Glacial acetic acid
(c.) Potassium Iodide (KI)
(d.) Starch indicator solution
(e.) Standard chlorine solution- Dilute commercially available 5-7% sodium hypochlorite,
standardize as residual chlorine. Keep in dark or in a brown, glass stoppered bottles.
A suitable strength of chlorine usually will be between 100 and 1000 mg/liter.
(f.) Ortho-toludine reagent
Method- Measure 5-10 equal sample portions of 100 ml each into glass Stoppard bottles or
flask. Add increasing amounts of standard chlorine to successive portion in the series. Stir the
mixture well and withdraw 0.25 ml of the solution with a pipette on a spot plate and immediately
test for residual chlorine with a drop of 0-toludine reagents, continue this process until there is
slight, excess of chlorine which is indicated by light yellow colour. The quantity of chlorine
required is regarded as the immediate chlorine demand. Find out the residual chlorine in the last
solution by iodometric titration.
Calculation-
Mg chlorine demand/litre = chlorine added (mg/l) residual chlorine (mg/l)
Result- The chlorine demand in given water sample was observed -----mg/l.
Precautions-


BORAN
Object- Determine the boron in given water sample.
Reagents-
(a.) Stock boron solution- 571.6 mg anhydrous boric acid H3BO3 is dissolved in distilled
water and dilute to one liter. (1 ml = 100 g B)
(b.) Standard boron solution- 10 ml stock boron solution is diluted to one liter with distilled
water. (1 ml = 1 g B)
(c.) Curcumin reagent- 40 mg finally ground curcumin and 5 gm oxalic acid are dissolved in
80 ml 95% ethyl alcohol, 4.2 ml concentration HCl is added and make up to 100 ml with
ethyl alcohol in a 100 ml volumetric flask.
(d.) Ethyl or isopropyl alcohol (95%)
Method- Pipette 0.2, 0.5, 0.8 and 1.0g boron into evaporating dish of the same type, size and
shape add distilled water to make the volume 1 ml of each. In a dish take 1 ml water as blank.
Add 4 ml curcumin reagent to each and swirl gently to mix contents thoroughly. Float dishes on
water bath set at 5520C and let them remain for 80 minute which is usually sufficient for
complete drying and removal of HCl. Keep drying time constant for standard and sample, after
dishes cool to room temperature add 10 ml 95% ethyl alcohol to each dish and stir gently with a
polyethylene rod to ensure complete dissolution of the red colour produced.
Transfer contents of the dish into a 25 ml volumetric flask, make up the mark
with 95% ethyl alcohol and mix thoroughly measure the absorbance of standard and samples at
540 nm and prepare a calibration graph.
Calculation-
Boran (mg/l) = A2 x C/A1 x S
Where,
A1 = absorbance of standard

A2 = absorbance of sample
C = g Boron in standard taken
S = ml sample
Result- The Boron in given water sample was observed -----mg/l.
Precautions-

2. Prepare the solution carefully.
3. Note the reading carefully in record.

TOTAL PHOSPHORUS (Spectophotometric method)
Object- Determine the total phosphorus in water sample.
Requirements- Hot plate, spectrophotometer, glass ware etc.
Reagents-
(a.) Perchloric acid (70%)

(b.) Phenolphthalein indicator- Dissolve 1.0 gm of phenolphthalein in 100 ml of ethyl alcohol
and add 100 ml of distilled water.
(c.) Sodium hydroxide solution (1N) - Dissolve 4 gm of sodium hydroxide in distilled water
to prepare 100 ml of solution.
(d.) Ammonium molybdate solution- Add 62ml of sulphuric acid (conc.) slowly to 80 ml of
distilled water and let cool. Dissolve 5gm of ammonium molybdate in 35 ml of distilled
water and mix it with sulphuric acid solution 200 ml.
(e.) Stannous chloride solution- Dissolves 0.5gm of stannous chloride in 2ml of conc. HCl
and dilute to 20 ml distilled water.
(f.) Standard phosphate solution- Dissolve 4.388gm of dried anhydrous potassium hydrogen
phosphate in distilled water to make the volume 1liter. Take 10 ml of this solution and
add distilled water to make 1 liter of stock solution containing 1 mg P/l. Prepare standard
phosphorous solution of various strengths (preferably in the range of 0.0 to 1.0 mg P/l at
intervals of 0.1mg P/l) by diluting the stock solution with distilled water.
Procedure
Take 25 ml of sample in an Erlenmeyer and evaporate to dryness. Cool and dissolve the
residue in 1 ml of perchloric acid. Heat the flask gently so that the contents become
colorless. Cool and add 10 ml of distilled water and 2 drops of phenolphthalein indicator.
Titrate against sodium hydroxide solution until the appearance of slight pink color. Make
the volume to 25 ml by addition distilled water. Add 1 ml of ammonium molybdate
solution and three drops of stannous chloride solution. A blue color will appear. Wait for
10 minutes (never more than 15 minutes) and record absorbance in spectrophotometer
meter at 690 nm. Run simultaneously distilled water blank in similar manner.

Process the standard phosphorous solution of different strength (reagent E) in similar

manner and plot a standard curve between absorbance and concentration of standard
phosphorous solutions. Deduce the total phosphorous content of sample by comparing its
absorbance (S) with standard curve and express the result of total phosphorous in mg/l.
The total particulate phosphorous can be estimated as a difference between the concentration of
total phosphorous in unfiltered and filtered sample
Result- The total phosphorous in given water sample was observed -------mg/l.
Precautions-


SECTION-B
BIOLOGICAL TEST
Method of sampling for bacteriological test
Apparatus Required- BOD bottle, incubator, durahams tube, test tube, test tube stand, test tube
basket, tray, pipette, wash bottle, inoculation loop, laminar flow, sprit lamp etc.
Miscellaneous items required-
(1.) Cotton roll (non-absorbant)

(2.) Sprit rectified
(3.) Aluminum foil
(4.) Leucoplast tape/stickers, marker pen
Procedure-
(1.) Sterilize the 125 ml BOD bottle before rapping its mouth with aluminum foil while cap is
fitted to prevent contamination of air at mouth.
(2.) Keep the bottle in the autoclave at 15 psi 1210C for 20 minutes. Cool & take out for
sampling.
(3.) Take sprit in a bottle and little cotton and an iron rod.
(4.) Wrap cotton on iron rod at one end and wet it with rectified sprit and ignite in under the
tap from which sample is to be taken.
(5.) Then open the tap and allow running water for sometime then take sample promptly by
opening cap of sample bottle and immediately closing it. Label on the sample bottle
location, date, time etc.
(6.) Sample from lake can be taken by dipping bottle at a 6 inch depth so that no atmospheric
air enters inside.
(7.) From river take sample by keeping the mouth of the bottle in the direction of flow at this
portion no atmospheric air will enter, also keep the bottle dipped in water at a 6 inch
depth.
(8.) Immediately keep the sample in ice.

(9.) Analysis as soon as possible, because bacteria if present will grow fast and erroneous
results will obtain.
(10.) Take care during analysis from contamination of sample from atmospheric air and
hands etc.
(11.) Sample is treated in a laminar air flow, if laminar air flow is not available sprit
lamps at both side of laboratory bench near the analysis area that keep sterilizing the
working environment.
(12.) Analyze the sample within 4 hours after collection
(13.) Preparation for analysis should be made before sample collection to avoid delay
in analysis.
(14.) Testing of bacteria is very sensitive parameter hence cleanness is very necessary
in the bio monitoring laboratory.
Sample Collection- sample of drinking water are collected in sterilized glass bottle. When
sample is collected from any hand pump or tap first of all the mouth of hand pump or tap is
sterilized by burning of cotton dipped in rectified sprit for approximately two minutes. After
that flush off some water before taking sample do not rinse the sampling bottle with sample.
Close immediately the sample bottle after taking sample and perform the test immediately as
soon as possible. If test is not possible immediately preserve at 40C in ice but do not delay
more than 6 hours.

TOTAL COLIFORM (MPN) TEST (Multiple tube fermentation technique)
Object- Determine total MPN in water sample.
Apparatus- Laminar flow, auto clave, sprit lamp, durhams tubes, test tubes, cotton bundle, test
tube stand, pipette, incubator etc.
Reagents- Chemicals for preparation of culture medium.
Constituents Quantity required for

1 liter medium
Tryptose 20.0 gm
Lactose 05.0 gm
Dipotassium hydrogen phosphate (K2HPO4) 02.75 gm
Potassium dihydrogen phosphate (KH2PO4) 02.75 gm
NaCl 5.0 gm
Sodium lauryle sulphate 0.1 gm
Distilled water 1 liter
For preparation of 1 liter medium dissolve the quantity of all above chemicals by heating
do not boil.
Procedure- Multiple tube fermentation (read the procedure carefully before analysis)
1. Prepare medium as per the procedure given above.

2. Fill 9 ml medium in washed test tubes carefully keep the tubes in test tube basket.
3. Fill the durham tube with medium and put it in the test tube filled with medium.
4. Cap the test tube after putting durhams tube by cotton plug.
5. Cover test tube basket with aluminum foil (food grade) and also put required number of
10 ml pipettes for sterilization in the autoclave.
6. Tighten autoclave cover put on autoclave when pressure is reached at 15 psi, keep on for
about 20 minutes at 15 psi keep the pressure at 15 psi by releasing steam (if required) by
steam release valve.

7. Put off autoclave after complete sterilization and open steam value for releasing pressure.
8. After 30 minutes open autoclave and take out carefully the sterilized test tubes filled with
medium and other glass ware like pipettes.
9. Now test tubes when at normal temp keep in the test tube stand keeping 15 test tubes for
one sample in three groups of 5 test tubes each in the test tube stand with dilution tubes.
10. Arrange test tubes so that for each sample 15 test tubes in 3 groups of 5 test tubes and
label the test tube stands as per sample codes.
11. Now take spirit lamps two numbers and put on the lamps at the table where test is being
performed if laminar air flow is available use that. Then there is no need for spirit lamps.
12. Dilution of sample- There are different type of samples, which have different ranges of
coli forms dilution for some samples are give below-
S. No. Category (Source) Dilutions

I II II
1. Ground water 1% 0.1% 0.01 %
2. River 1% 0.1% 0.01 %
3. Sewage treatment plant 0.0001% 0.00001% 0.000001%
influent effluent
4. Bathing Ghats 0.0001% 0.00001% 0.000001%
5. Recreation at waters 1% 0.1% 0.01 %
6. Swimming pool 1% 0.1% 0.01 %
13. After making proper dilution keep the sample tubes in bacteriological incubator for 24
hrs. at 350.50C.
14. After 24 hrs check the tubes if there is air in the durhams tube also the colour of the
media get changed light red if coliform is present due to fermentation of the media and
production of CO2 gas which gets filled in the durhams tube. These called positive tubes.
Count and note the number of positive tubes.

Reference Table-
Combination of (+) MPN index per 100 Combination of (+) MPN index per 100
tubes dilution % ml tubes dilution % ml
10-1-0.1 10-1-0.1
0-0-0 2 3-0-0 8
0-0-1 2 3-0-1 11
0-1-0 2 3-1-0 11
0-2-0 4 3-1-1 14
1-0-0 2 3-2-0 14
1-0-1 4 3-2-1 17
1-1-0 4 4-0-0 13
1-1-1 6 4-0-1 17
1-2-0 6 4-1-0 17
2-0-0 4 4-1-1 21
2-0-1 7 4-1-2 26
2-1-0 7 4-2-0 22
2-1-1 9 4-2-1 26
2-2-0 9 4-3-0 27
2-3-0 12 4-3-1 33
4-4-0 34
Reference Table-
Combination of (+) MPN index per 100

tubes dilution 10% ml
1.0% 0.1%
5-0-0 23
5-0-1 30
5-0-2 40
5-1-0 30

5-1-1 50
5-1-2 60
5-2-0 50
5-2-1 70
5-2-2 90
5-3-0 80
5-3-1 110
5-3-2 140
5-3-3 170
5-4-0 130
5-4-1 170
5-4-2 220
5-4-3 280
5-4-4 350
5-5-0 240
5-5-1 300
5-5-2 500
5-5-3 900
5-5-4 1600
5-5-5 1600
Result-The total coliform was found-------- MPN/100ml in given water sample.
Precautions-
1. Sample should be taking carefully.

2. Sterilize carefully.

FECAL COLIFORM MPN/100 ml (Multiple tube fermentation techniques)
Object- Determine the fecal coliform in given water sample.
Apparatus Required- Laminar flow, auto clave, sprit lamp, durhams tubes, test tubes,
cotton bundle, test tube stand, pipette, incubator etc.
The fecal coliform test differentiates between coliform of fecal origin (intestines of warm
blooded animals) and colifom from other sources, EC medium is used and prepared as
below-
Tryptose 20.0 gm
Lactose 6.0 gm
Bile salts 1.5 gm
K2HPO4 4.0 gm
KH2PO4 1.5 gm
NaCl 5.0 gm
Dissolve all above in gradients in 1 liter distilled water in a beaker heat for complete
dissolution. Fill the fermentation tubes 9 ml (test tubes) with medium and sterilize the tubes
by keeping in autoclave at 15 psi for 20 minutes.
Inoculation of sample:
(1.) Take positive tubes of total coliform.

(2.) Shake & dip inoculation loop in the medium of positive tube & immediately dip in the
tube to be fixed for fecal coliform test. Equal numbers of tubes are inoculated.
(3.) The exercise is done in the sterilized atmosphere by using in the laminar air flow or keep
sprit lamps near working area.
Keep the sample inoculated tubes in the bacteriological incubator at 450.50C for
40 hours.
Calculate MPN fecal coliform following the reference table.

Result-The total coliform was found-------- MPN/100ml in given water sample.
Precautions-
3. Sample should be taking carefully.

4. Sterilize carefully.

AIR
ANALYSIS

TEMPERATURE (MINIMUM AND MAXIMUM)
Object- Determine the maximum and minimum temperature of atmosphere.
Requirement- Minimum maximum thermometer and magnet.
Method- Bring a horse shoe magnet close to capillary tube of thermometer and with the down
word strokes of it set the index in each limb to rest on mercury column. Preferably set the indices
during morning hours. Place the thermometer in shady atmosphere. After a laps of 24 hours
record the temperature in two limbs of the instruments. The temperature recorded on left limbs
represents the minimum temperature while that on right limbs the maximum.
Result- The minimum and maximum temperature was observed -----0C and ----- 0C during a 24
hours dial cycle.
Precaution-
1. Set the index properly.

RELATIVE HUMIDITY OF ATMOSPHERE
Object- Determine the relative humidity of atmosphere.
Requirement-Psychrometer (Dry & Wet bulb thermometer)
Method- Soak the cotton wick of wet bulb thermometer in water and swing the set of
thermometer in air with the help of handle provided at the top of the instruments. Until the
reading in two thermometers are constant. Record the temperature in both temperature and find
the difference in two reading referred to as depression in wet bulb thermometer. Find the RH (%)
following table-1 which involves temperature in wet bulb thermometer. If both the thermometer
read same temperature then it is indicative of 100% relative humidity.
Result- The relative humidity of atmosphere was observed ------- %.
Precaution-
1. Cotton wick should not dry.

Table -1. Relative humidity (%) as measured by Psychrometer
Dipression in wet bulb Temperature in dry bulb thermometer (0C)

0
thermometer( C) 0 3 5 6 9 10 12 15 18 20 21 24 25 27 30 33 35 36 39 40
0.5 90 92 94 94 94 95 96 96 96 96 96 97 97
1 81 84 87 87 88 88 89 90 90 90 91 92 92 93 93 93 93 93 94 94
1.5 71 76 80 82 83 84 85 86 87 88 90 90 90 90 91
2 64 79 72 73 76 76 78 80 82 82 83 85 86 86 86 87 88
2.5 55 62 66 70 73 76 78 79 81 85 82 82 83 87 84 85 88
3 46 44 60 65 68 71 73 75 77 79 79 80 81 85
3.5 38 46 54 59 63 66 69 71 74 76 76 77 78 79
4 29 40 47 53 58 62 65 67 70 72 73 76 75 76
4.5 21 32 41 48 53 58 61 64 66 68 70 71 72 74
5 13 25 33 35 42 48 53 47 59 60 63 63 65 67 68 70 70 71 72
6 12 21 23 32 34 38 44 52 52 53 56 57 59 61 63 64 64 66 66
7 9 11 22 25 30 36 42 45 46 49 50 53 55 57 59 59 61 61
8 12 15 21 28 35 38 39 43 44 47 50 52 54 54 56 56
9 3 6 12 20 27 30 42 37 38 41 44 47 50 50 62 52
10 4 13 20 24 26 31 33 36 39 42 44 49 47 48
11 4 13 19 26 31 35 37 41 43
12 6 13 21 26 30 33 36 39

PARTICULATE MATTER (Gravitation method)
Object- Determine the particulate matter in atmosphere.
Requirement- Glass jar, balance, weight box.
Method- Take 3 glass jars of known weight and put it in open area where dust fall is to be
estimated. After sufficient gap of time (4 hours) collect the dust settled in each jar and weight it.
Take an average weight of dust in each jar and calculate the result. Dust fall to be expressed as
weight of dust per unit area per unit time.
Particulate matter = final weight of jar initial weight of jar
Result- The particulate matter observed was--------- g/m3.
Precautions-
1. Beaker should be dried properly.

2. Weight the beaker paper carefully.
3. Keep the beaker at the height of approximately two meters.
4. Label the beaker properly.

PARTICULATE MATTER (PM10) (Gravimetric method)

Object- Determine the particulate matter (PM10) in ambient air.
Requirement- Respirable dust sampler, glass fiber filter paper 8 X 10 inch, balance, weight box,
automatic volumetric flow control, flow-measuring device and top loading orifice kit.
Sampling
Field Sampling - Tilt back the inlet and secure it according to manufacturer's instructions.
Loosen the faceplate wing nuts and remove the face plate. Remove the filter from its jacket and
centre it on the support screen with the rough side of the filter facing upwards. Replace the
faceplate and tighten the wing nuts to secure the rubber gasket against the filter edge. Gently
lower the inlet. For automatically flow-controlled units, record the designated flow rate on the
data sheet. Record the reading of the elapsed time meter. The specified length of sampling is
commonly 8 hours or 24 hours. During this period, several reading (hourly) of flow rate should
be taken. After the required time of sampling, record the flow meter reading, take out the filter
media from the sampler, and put in a container or envelope.
Analysis
Filter inspection: Inspect the filter for pin holes using a light table. Loose particles should be
removed with a soft brush. Apply the filter identification number or a code to the filter if it is not
a numbered. Condition the filter in conditioning room maintained within 20-30 C and 40-50%
relative humidity or in an airtight desiccator for 24 hours. Take initial weight of the filter paper
(Wi) before sampling. Condition the filter after sampling in conditioning room maintained within
20-30 C and 40-50% relative humidity or in an airtight desiccator for 24 hours. Take final
weight of the filter paper (Wf).
Calculation
C PM10g/m3 = (Wf Wi) x 106 / V
Where,
C PM10 = Concentration of Nitrogen dioxide (g/m3)
Wf = Initial weight of filter in (gm)
Wi = Initial weight of filter in (gm)
106 = Conversion of g to (g)
V = Volume of air sampled (m3)

Result- The PM10in ambient air was found-------g/m3.

Precautions-
1. Filter paper should be dried properly.
2. Weight the filter paper carefully.
3. Keep the sampler at the height of approximately two meters.
4. Label the filter paper properly.

PARTICULATE MATTER (PM2.5) (Gravimetric method)

Object-Determine the particulate matter (PM2.5) in ambient air.
Requirements- Respirable dust sampler, filter paper, balance and weight box.
Definition
PM2.5 refers to fine particles that are 2.5 micrometers (m) or smaller in diameter.
Ambient air is defined as any unconfined part of the Earths atmosphere, that the surrounding
outdoor air in which humans and other organisms live and breathe.
FRM Federal Reference Method
FEM Federal Equivalent Method
Principle
An electrically powered air sampler draws ambient air at a constant volumetric flow rate (16.7 l
pm) maintained by a mass flow / volumetric flow controller coupled to a microprocessor into
specially designed inertial particle-size separator (i.e. cyclones or impactors) where the
suspended particulate matter in the PM2.5 size ranges is separated for collection on a 47 mm
polytetrafluoroethylene (PTFE) filter over a specified sampling period. Each filter is weighed
before and after sample collection to determine the net gain due to the particulate matter. The
mass concentration in the ambient air is computed as the total mass of collected particles in the
PM2.5 size ranges divided by the actual volume of air sampled, and is expressed in g/m3. The
microprocessor reads averages and stores five-minute averages of ambient temperature, ambient
pressure, filter temperature and volumetric flow rate. In addition, the microprocessor calculates
the average temperatures and pressure, total volumetric flow for the entire sample run time and
the coefficient of variation of the flow rate.
Interferences and Artifacts
The potential effect of body moisture or oils contacting the filters is minimized by using
non-serrated forceps to handle the filters at all times. This measure also moderates interference
due to static electricity. Teflon filters accumulate a surface electrical charge, which may affect
filter weight. Static electricity is controlled by treating filters with a Static Master static charge
neutralizer prior to weighing. Placement of filters on a Static Master unit is required for a
minimum of 30 seconds before any filter can be weighed.
Moisture content can affect filter weight. Filters must be equilibrated for a minimum of
24 hours in a controlled environment prior to pre- and post weighing. The balance rooms

relative humidity must be maintained at a mean value range of 45 5 % and its air temperature
must be maintained at a mean value range of 25.0 C. Airborne particulate can adversely affect
accurate mass measurement of the filter. Filters undergoing conditioning should not be placed
within an air flow path created by air conditioning ductwork, computer printers, or frequently
opened doorways. Cleaning laboratory bench-tops and weighing areas daily, installing sticky
floor mats at doorway entrances to the balance room and wearing clean lab coats over regular
clothing can further minimize dust contamination.
Precision and Accuracy
The performance segment of the PM2.5 FRM specifies strict guidelines for controls that must be
observed, as well as the range of precision and accuracy of control. The flow rate through the
instrument is specified as 16.67 l pm(1 m3/hr). This flow must be volumetrically controlled to a
precision of 5% and an accuracy of 2%. The flow control must be upgraded at least every 30
seconds and recorded (logged) every five minutes. Barometric pressure, ambient temperature and
filter temperature should be measured on the same schedule. Filter temperature, it must not
exceed the ambient temperature by more than 50C for more than 30 minutes. A fan blowing
filtered ambient air through the enclosure provides the necessary cooling effect. It is necessary
for the entire apparatus to provide accurate performance over a temperature range of 20 to500C.
The supporting run-time (interval) data, which are stored in detailed 5-minute intervals in the
samplers microprocessor, as well as 24-hour integrated performance (filter) data, must be
capable of being extracted at the completion of a 24-hour run. The FRM mandates the provision
of an RS232 port for this purpose. Data may be extracted to a portable computer.
Mass of the filter deposit, flow rate through the filter, and sampling time have typical
precision of 0.2 mg, 5%, and 30 seconds, respectively. These uncertainties combine to
yield a propagated precision of approximately 5 %at 10 g/m3 and approximately 2% at
100g/m3.
Sitting Requirements
Samplers should be sited to meet the goals of the specific monitoring project. For routine
sampling to determine compliance with the National Ambient Air Quality Standards (NAAQS),
sampler sitting is described in CPCB guide lines shall apply. The monitoring should be done at
outside the zone of influence of sources located within the designated zone of representation for
the monitoring site. Height of the inlet must be 3 10 m above the ground level and at a suitable

distance from any direct pollution source including traffic. Large nearby buildings and trees
extending above the height of the monitor may present barriers or deposition surfaces for PM.
Distance of the sampler to any air flow obstacle i.e. buildings, must be more than two times the
height of the obstacle above the sampler.
There should be unrestricted airflow in three of four quadrants. Certain trees may also be
sources of PM in the form of detritus, pollen, or insect parts. These can be avoided by locating
samplers by placing them > 20 m from nearby trees. If collocated sampling has to be performed
the minimum distance between two samplers should be 2 meters.
Calculation:
The equation to calculate the mass of fine particulate matter collected on a Teflon filter is as
below:
M2.5 = (Mf Mi) mg x 103g
Where,
M2.5 = total mass of fine particulate collected during sampling period (g)
Mf = final mass of the conditioned filter after sample collection (mg)
Mi = initial mass of the conditioned filter before sample collection (mg)
103 = unit conversion factor for milligrams (mg) to micrograms (g)
Field records of PM2.5 samplers are required to provide measurements of the total volume
of ambient air passing through the sampler (V) in cubic meters at the actual temperatures and
pressures measured during sampling. Use the following formula if V is not available directly
from the sampler-
V = Q avg x t x 10-3 m3
Where,
V = total sample value (m3)
Q avg = average flow rate over the entire duration of the sampling period (L/min)
t = duration of sampling period (min)
103 = unit conversion factor for liters (L) into cubic meters (m3)
The equation given below can be used to determine PM2.5 mass concentration:
PM2.5 = M2.5 / V
Where,
PM2.5 = mass concentration of PM2.5 particulates (g/m3)

PM2.5 = total mass of fine particulate collected during sampling period (g)
V = total volume of air sampled (m3)
Result- The PM2.5in ambient air was found-------g/m3.
Precautions-
1. Filter paper should be dried properly.
2. Weight the filter paper carefully.
3. Keep the sampler at the height of approximately two meters.
4. Label the filter paper properly.

SULPHUR DIOXIDE (Improved West and Gaeke method)

Object- Determine the SO2 in ambient air.
Requirements- High volume sampler and spetrophotometer.
Reagents/Chemicals-
Absorbing Reagent- Potassium tetrachloromercurate (TCM 0.04M)- Dissolve 10.86 gm
mercuric chloride 0.066 gm EDTA and 6.0 gm potassium chloride or sodium chloride 4.68 gm in
water and bring to the mark in a 1 liter volumetric flask. The pH of this reagent should be
approximately 4.0. The absorbing reagent is normally stable for six months.
Sulphamic Acid (0.6%) - Dissolve 0.6 gm sulphamic acid in 100 ml distilled water. Prepare
fresh daily.
Formaldehyde (0.2%) - Dilute 5 ml formaldehyde solution (36-38%) to 1 liter with distilled
water. Prepare fresh daily.
Purified Pararosaniline Stock Solution (0.2% Nominal)- Dissolve 0.500 gm of specially
purified pararosaniline (PRA) in 100 ml of distilled water and keep for 2 days (48 hours).
Pararosaniline Working Solution - 10 ml of stock PRA is taken in a 250 ml volumetric flask.
Add 15 ml conc. HCL and make up to volume with distilled water.
Stock Iodine Solution (0.1 N) - Place 12.7 gm iodine in a 250 ml beaker, add 40 gm potassium
iodide and 25 ml water. Stir until all is dissolved then dilute to 1 litre with distilled water.
Iodine Solution (0.01 N) - Prepare approximately 0.01 N iodine solution by diluting 50 ml of
stock solution to 500 ml with distilled water.
Starch Indicator Solution - Triturate 0.4 gm soluble starch and 0.002 gm mercuric iodide
preservative with a little water and add the paste slowly to 200 ml boiling water. Continue
boiling until the solution is clear, cool, and transfer to a glass-Stoppard bottle.
Stock Sodium Thiosulfate Solution (0.1 N) - Prepare a stock solution by placing 25 gm sodium
thiosulphate pentahydrate in a beaker add 0.1 gm sodium carbonate and dissolve using boiled,
cooled distilled water making the solution up to a final volume of 1 liter. Allow the solution to
stand one day before standardizing.
To standardize, accurately weigh to the nearest 0.1 mg, 1.5 gm primary standard potassium
iodate dried at 1800C, dissolve and dilute to volume in a 500 ml volumetric flask. Into a 500 ml
Iodine flask, transfer 50 ml of iodate solution by pipette. Add 2 gm potassium iodide and 10 ml
of N- hydrochloric acid and stopper the flask. After 5 min, titrate with stock thiosulphate solution

to a pale yellow. Add 5 ml starch indicator solution and continue the titration until the blue
colour disappears. Calculate the normality of the stock solution.
Sodium Thiosulphate Titrant (0.01 N) - Dilute 100 ml of the stock thiosulphate solution to 1
litre with freshly boiled and cooled distilled water.
Sampling
Place 30 ml of absorbing solution in an impinger and sample for four hours at the flow rate of 1
L/min. After sampling measure the volume of sample and transfer to a sample storage bottle.
Analysis
Replace any water lost by evaporation during sampling by adding distilled water up to the
calibration mark on the absorber. Mix thoroughly, pipette out 10 ml of the collected sample into
a 25 ml volumetric flask. Add 1 ml 0.6% sulphamic acid and allow reacting for 10 minutes to
destroy the nitrite resulting from oxides of nitrogen. Add 2 ml of 0.2% formaldehyde solution
and 2 ml pararosaniline solution and make up to 25 ml with distilled water. Prepare a blank in
the same manner using 10 ml of unexposed absorbing reagent. After a 30 min colour
development interval and before 60 minutes, measure and record the absorbance of samples and
reagent blank at 560 nm. Use distilled water; not the reagent blank, as the optical reference.
Calibration
The actual concentration of the sulphite solution is determined by adding excess iodine and back
titrating with standard sodium thiosulphate solution. To back-titrate, measure, by pipette, 50 ml
of the 0.01 N iodine solutions into each of two 500 ml iodine flasks A and B. To flask a (blank)
add 25 ml distilled water and into flask B (sample) measure 25 ml sulphite solution by pipette.
Stopper the flasks and allow reacting for 5 minutes. Prepare the working sulphite-TCM solution
at the same time iodine solution is added to the flasks. By means of a burette containing
standardized 0.01N thiosulphate, titrate each flask in turn to a pale yellow. Then add 5 ml starch
solution and continue the titration until the blue colour disappears.
Preparation of Standards
Measure 0.5 ml, 1.0 ml, 1.5 ml, 2.0 ml, 2.5 ml, 3.0 ml, 3.5 ml and 4.0 ml of working
sulphiteTCM solution in 25 ml volumetric flask. Add sufficient TCM solution to each flask to
bring the volume to approximately 10 ml. Then add the remaining reagents as described in the
procedure for analysis. A reagent blank with 10 ml absorbing solution is also prepared. Read the
absorbance of each standard and reagent blank

Standard Curve
Plot a curve absorbance (Y axis) versus concentration (X axis). Draw a line of best fit and
determine the slope. The reciprocal of slope gives the calibration factor (CF).
Calculation
Concentration of sulphite solution:

C=
Where,
C = SO2 concentration in mg/ml
V1 = Volume of thiosulphate for blank (ml)
V2 = Volume of thiosulphate for sample (ml)
N = Normality of thiosulphate
K = 32000 (Milli equivalent weight SO2/g)
V = Volume of standard sulphite solution (ml)
C (SO2 g/m3)= (As Ab) x CF x Vs/ Va x Vt
Where,
C SO2 = Concentration of sulphur dioxide (g/m3)
As = Absorbance of sample
Ab = Absorbance of reagent blank
CF = Calibration factor
Va= Volume of air sampled (m3)
Vs= Volume of sample (ml)
Vt = Volume of aliquot taken for analysis (ml)
Result- SO2 in ambient air was observed--------g/m3.
Precautions-

NITROGEN DIOXIDE (Modified Jacob and Hochheiser method)

Object- Determine the oxide of nitrogen (NOx) in ambient air.
Requirements- High volume sampler and spetrophotometer.
Reagents / Chemicals
All the chemicals should meet specifications of ACS Analytical Reagent grade
Distilled water
Sodium hydroxide
Sodium arsenite
Absorbing solution- Dissolve 4.0 gm of sodium hydroxide in distilled water add 1.0 gm of
sodium arsenite, and dilute to 1000 ml with distilled water.
Sulphanilamide - Melting point 165 to 1670C
N-(1-Naphthyl)-ethylenediamine Di-hydrochloride (NEDA) - A 1%aqueous solution should
have only one absorption peak at 320 nm over the range of 260-400 nm. NEDA showing more
than one absorption peak over this range is impure and should not be used
Hydrogen Peroxide - 30%
Phosphoric Acid - 85%
Sulphanilamide Solution - Dissolve 20 gm of sulphanilamide in 700 ml of distilled water. Add
with mixing, 50 ml of 85% phosphoric acid and dilute to 1000 ml. This solution is stable for one
month, if refrigerated.
NEDA Solution - Dissolve 0.5 gm of NEDA in 500 ml of distilled water. This solution is stable
for one month, if refrigerated and protected from light.
Hydrogen Peroxide Solution - Dilute 0.2 ml of 30% hydrogen peroxide to250 ml with distilled
water. This solution may be used for one month, if refrigerated and protected from light
Sodium nitrite - Assay of 97% NaNO2or greater
Sodium nitrite stock solution (1000 g NO2/ml)
Sodium nitrite solution (10 g NO2/ml)
Sodium nitrite working solution (1 g NO2/ml)
(Dilute with absorbing reagent prepare fresh daily)
Sampling
Place 30 ml of absorbing solution in an impinger and sample for four hour at the flow rate of 0.2
to 1 L/min. After sampling measure the volume of sample and transfer to a sample storage bottle.

Analysis
Replace any water lost by evaporation during sampling by adding distilled water up to the
calibration mark on the absorber, mix thoroughly. Pipette out 10 ml of the collected sample into
a 50 ml volumetric flask. Pipette in 1 ml of hydrogen peroxide solution, 10 ml of sulphanilamide
solution, and1.4 ml of NEDA solution, with thorough mixing after the addition of each reagent
and make up to 50 ml with distilled water. Prepare a blank in the same manner using 10 ml of
unexposed absorbing reagent.
After 10 min colour development interval, measure and record the absorbance of samples
and reagent blank at 540 nm. Use distilled water; not the reagent blank, as the optical reference
samples with an absorbance greater than 1.0 must be re-analyzed after diluting an aliquot of the
collected samples with an equal quantity of un exposed absorbing reagent.
A randomly selected 5-10% of the samples should be re-analyzed as a part of an internal
quality assurance program.
Calibration
Pipette 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15 and 20 ml of working standard solution into 50 ml
volumetric flask. Fill to 20 ml mark with absorbing solution. A reagent blank with 10 ml
absorbing solution is also prepared. Add reagents to each volumetric flask as in the procedure for
analysis. Read the absorbance of each standard and reagent blank against distilled water
reference.
Standard Curve
Calculation
C (NO2g/m3) = (As Ab) x CF x Vs/ Va x Vt x 0.82
Where,
C NO2 = Concentration of Nitrogen dioxide (g/m3)
Va= Volume of air sampled (m3)

Vs= Volume of sample (ml)

Vt = Volume of aliquot taken for analysis (ml)
0.82 = Sampling efficiency
Result-NOx in ambient air was observed--------g/m3.
Precautions-

OZONE (Chemical method)

Object- Determine the ozone in ambient air.
Principle of the method
Micro-amounts of ozone and the oxidants liberate iodine when absorbed in a1% solution of
potassium iodine buffered at pH 6.8 + 0.2. The iodine is determined spectrophotometrically by
measuring the absorption of tri-iodide ion at 352 nm. The stoichiometry is approximated by the
following reaction:
O3 + 3 KI + H2O --> KI3 + 2 KOH + O2
Instrument/Equipment
The following items are necessary to perform the monitoring and analysis of ammonia in
ambient air:
Analytical balance:
Vacuum pump: Any suction pump capable of drawing the required sample flow rate of 1 to 2
liter per minute
Calibrated flow measuring device to control the air flow from 1 to2 l/min.
Absorber: All glass midget impinger
Spectrophotometer: Capable of measuring absorbance at 352 nm.
Glass wares: low actinic glassware must be used for analysis
Distilled water
Absorbing Solution (1% KI in 0.1 m Phosphate Buffer) - Dissolve 13.6 gm of potassium
dihydrogen phosphate (KH2PO4) 14.2 gm of disodium hydrogen phosphate (Na2HPO4) or 35.8
gm of the dodecahydrate salt (Na2HPO2. 12 H2O) and 10.0 gm of potassium iodide in sequence
and dilute the mixture to 1 litre with water. Keep at room temperature for at least 1 day before
use. Measure pH and adjust to 6.8 + 0.2 with NaOH orKH2PO4. This solution can be stored for
several months in a glass Stoppard brown bottle at room temperature without deterioration. It
should not be exposed to direct sunlight.
Stock Solution 0.025 M I2 (0.05N) - Dissolve 16 gm of potassium iodide and3.173 gm of re-
sublimed iodine successively and dilute the mixture to exactly 500 ml with water. Keep at room
temperature at least 1 day before use. Standardize shortly before use, against 0.025 M Na2S2O3.

The sodium thiosulphate is standardized against primary standard bi-iodate [KH(IO3)2] or

potassium dichromate (K2Cr2O7).
M I2 Solution - Pipette exactly 4.00 ml of the 0.025 M Stock solution into a100 ml volumetric
flask and dilute to the mark with absorbing solution. Protect from strong light. Discard after use.
Sampling
Place 10 ml of absorbing solution in a standard impinger and sample for one hour at the flow rate
of 1 L/min. Do not expose the absorbing reagent to direct sunlight. After sampling measure the
volume of sample and transfer to a sample storage bottle.
Analysis
If, appreciable evaporation of the absorbing solution occurs during sampling, add water to bring
the liquid volume to 10 ml. within 30 to 60 minutes after sample collection, read the absorbance
in a cuvette at 352 nm against a reference cuvette containing distilled water.
Measure the absorbance of the unexposed reagent and subtract the value from the absorbance of
the sample.
Calibration
Calibrating Iodine Solution - For calibration purposes exactly 5.11 ml of the 0.001M I2
solution (or equivalent volume for other molarity) is diluted with absorbing solution just before
use to 100 ml (final volume) to make the final concentration equivalent to 1 l of O3/ml. Discard
this solution after use.
Obtain a range of calibration points containing from 1 l to 10 l of ozone equivalent per 10.0 ml
of solution. Prepare by individually adding 1.0, 2.0, 4.0, 6.0, 8.0 and 10.0 ml of the calibrating
iodine solution to 10.0 ml volumetric flasks. Bring each to the calibration mark with absorbing
reagent.
Read the absorbance of each of the prepared calibration solutions at 352 nm against distilled
water reference
Standard Curve
Calculation
C (O3g/m3) = (As Ab) x CF x 1.962/ Va

Where,
C O3 = Concentration of Ozone in g/m3
Va = Volume of air sampled in m3
1.962 = Conversion factor, l to g
Result-The ozone in ambient air was found------g/m3.
Precautions-

AMMONIA (Indophenol blue method)

Object- Determine the ammonia in ambient air.
Principle of the method
Indophenol method (Method 401, Air Sampling and Analysis, 3rd Edition) Ammonia in the
atmosphere is collected by bubbling a measured volume of air through a dilute solution of
sulphuric acid to form ammonium sulphate. The ammonium sulphate formed in the sample is
analyzed colorimetrically by reaction with phenol and alkaline sodium hypochlorite to produce
indophenol. The reaction is accelerated by the addition of sodium nitropruside as catalyst.
Instrument/Equipment
The following items are necessary to perform the monitoring and analysis of ammonia in
ambient air:
Analytical balance
Vacuum pump to maintain a flow rate up to 5 litre per minute
Calibrated flow measuring device to control the air flow from 1 to 2 litre/min.
Absorber: a midget impinger or a fritted bubbler
Spectrophotometer capable of measuring absorbance at 630 nm.
Glass ware: low actinic glass wares must be used for analysis
Distilled water
N Sulphuric Acid (Absorbing solution)
Sodium Nitroprusside
6.75 M sodium hydroxide
Sodium hypochlorite solution
Phenol solution 45% v/v
Sodium phosphate
Ammonium chloride or Ammonium Sulfate
Hydrochloric acid
Ammonia stock solution (1 mg NH3/ml)
Ammonia working solution (10 g NH3/ml) (Prepare fresh daily)
Sampling

Place 10 ml of absorbing solution in an impinger and sample for one hour at the flow rate of 1 to
2 L/min. After sampling measure the volume of sample and transfer to a sample storage bottle.
Analysis
Transfer contents of the sample bottle to a 25 ml glass stopper graduated cylinder. Maintain all
the solutions and sample at 25 C. Add 2 ml buffer. Add5 ml of working phenol solution, mix,
and fill to about 22 ml. Add 2.5 ml of working hypochlorite solution and rapidly mix. Dilute to
25 ml, mix and store in the dark for 30 minutes to develop colour. Measure the absorbance of the
solution at 630 nm on a spectrophotometer using 1 cm cells. Prepare a reagent blank and field
blank and measure the absorbance as done in the analysis of samples.
Calibration
Pipet 0.5, 1.0, 1.5, 2.0 ml of working standard solution in to 25 ml glass Stoppard graduated
cylinders. Fill to 10 ml mark with absorbing solution. A reagent blank with 10 ml absorbing
solution is also prepared. Add reagents to each cylinder as in the procedure for analysis. Read the
absorbance of each standard against reagent blank.
Standard Curve
Calculation
C (NH3 g/m3) = (As Ab) x CF / Va
Where,
C NH3 = Concentration of Ammonia in g/m3
Va = Volume of air sampled in m3
Result- The ammonia in ambient air was found------g/m3.
Precautions-

SOIL
ANALYSIS

TEXTURE
Object- Determine the texture of given soil.
Requirements- Shaking machine, motor description unit (this consists of a 0.5 H.P. motor with a
propeller shaft and double-blanded propeller of stainless steel. The base of the unit contains a
600 ml beaker with the baffle grid fitted snugly in it), Sieve (made from 70 mesh brass or copper
gauze), Sedimentation cylinder (40 cm height about 6.5 cm in diameter and graduated to contain
1.25 l.), Pipettes (to deliver 20 ml at 200C in 25-30 second. One such pipette should have its
lower stem 39 cm long and ring attached around it at a distance of 28 cm from lower tip. One
other pipette should have its cover stem 32 cm, long and a ring attached around it at a distance of
22 cm from the lower tip), Beaker (one of 500 ml marked on outside to indicate the height of 10
cm above bottom and other of 800 ml., rubber pestle (it consists of a rubber stopper of 25 mm
height and 20 mm diameter having a 25 cm long glass rod fitted in the centre of narrower end),
Stirring Paddle (it is made up of a brass disc of 6 cm diameter and 3 mm thickness, attached to a
50 cm long spindle. The disk has 8 holes in it, each of about 6 mm diameter), Hot water both,
Buchner funnel, Filter paper (whatman no-50), camel hair brush and reagents as given below-
A. Hydrogen peroxide
B. Hydro-chloric acid (2N): Dilute 175 ml of concentrated hydro-chloric acid (reagent B)
and dilute with 1 litre distilled water.
C. Hydro-chloric acid (0.2N): Take 100 ml of 2N hydro-chloric acid (reagent B) and dilute
with 1 litre distilled water.
D. Sodium Hydroxide Solution (1N): Dissolve 40 gm of sodium hydroxide in distilled water
to make the volume 1 litre.
Method- Take 25 gm of air-dry soil in a beaker of 800 ml, add 50 ml of hydrogen peroxide
(reagents A) and allow standing overnight. Then heat the beaker on boiling water bath until the
vigorous reaction is ceased. It is soil is extra rich in organic matter further add 30 ml of hydrogen
peroxide and heat the beaker on water bath for another 10 minutes. Cool the beaker clean its
sides with a rubber pestle and add 25 ml of 2N hydrochloric acid (reagents B). Dilute the
contents with distilled water to about 250 ml and thoroughly rub the soil with rubber pestle. Wait
for 1 hour and then test the solution with a blue litmus paper to ensure the presence of an excess

of acid. Filter the contents through a Buchner funnel fitted with filter paper. Wash the soil with
200 ml of 0.2N hydrochloric acid (reagents C) and then with excess of water until the filtrate is
almost neutral to litmus.
Transfer the soil from Buchner funnel to 600 ml beaker of dispersion until.
Clean the last tracers of soil from filter paper and funnel with the help of a camel hair brush and
rinsing with water into the same beaker. The volume of suspension should not exceed 250 ml.
Add 10 ml of 1N sodium hydroxide solution (reagent D) and dip the propeller of dispersion unit
in to beaker. Set the propeller in motion and control the speed to have vigorous stirring without
the loss from splashing. Continue the mechanical dispersion for about 15 minutes, then take out
the propeller and baffle grid and rinse the two into beaker.
Filter the suspension in a sedimentation cylinder through a 70 mess sieve.

Wash maximum possible material through the sieve with a stream of water until cylinder is about
one-half full. Transfer sieve to a tray and dry for sand content estimation.
Cover the cylinder with a brass cap and shake it for 12-16 hours in a shaking
machine for complete dispersion. Then stand the cylinder rinse the brass cap in to it and dilute
the suspension with distilled water to 1.25 litres. Record the temperature of the suspension.
According to which the time of pipetting and decantation is determined as shown in table-1-
Table-1. Time the sedimentation at different temperatures.
Temperature First pipette sample Second Fine sand decantation

pipette
sample
Depth 28 cm Depth 22 cm Depth 28 cm Depth 10 cm
0
C Minimum Hours Hours Minimum Second
8 183/4 241/2 - 6 40
9 18 233/4 - 6 30
10 171/2 23 - 6 20
11 17 241/4 - 6 10
1/2 3/4
12 16 21 - 6 0

13 161/4 211/4 - 5 50
14 153/4 201/2 - 5 40
15 151/4 20 - 5 30
16 15 191/2 243/4 5 20
17 141/2 19 241/4 5 10
18 141/4 181/2 231/2 5 0
3/4
19 13 18 23 5 0
20 131/2 171/2 221/2 4 48
21 131/4 171/4 22 4 40
22 13 163/4 211/2 4 30
23 121/2 - 21 4 30
24 121/4 - 201/2 4 20
25 12 - 20 4 15
3/4 1/2
26 11 - 19 4 10
27 111/2 - 19 4 5
28 111/4 - 281/2 4 0
29 11 - 181/4 3 55
30 103/4 - 173/4 3 50
31 101/2 - 171/2 3 45
1/4
32 10 - 17 3 40
33 10 - 163/4 3 35
Stir the suspension vigorously by up and down.
Strokes of stirring paddle. Take out the paddle and note the time of commencement of
sedimentation on completion of sedimentation time for first pippetting (see Table-1) take 39 cm
stemmed pipette, close the upper stem of it with the fore finger and introduce gently the lower
stem into suspension so that 28 cm mark on the pipette corresponds with the surface of
suspension. Fill the pipette to 20 ml by gentle suction and deliver the sample into pre-weighed
silica crucible. Evaporate the sample to dryness on a hot water bath; please the crucible in an

oven at 1050C for about 13 hours, cool in the desiccator and weight. The increase in the weight
(W1) indicates the weight of oven dry silt+clay.
Let the sedimentation cylinder stand until completion of sedimentation time for
second pipetting (see Table-1). Take 32 cm stemmed pipette, aloes its upper stem with forefinger
and introduce gently the lower stem into suspension so that 22 cm mark of the pipette
corresponds with the new surface. Because at higher temperature the pipetting time becomes
inconveniently short the second pipetting may be made by using a 39 cm stemmed pipette
lowered to 28 cm mark after the time period indicated in Table-1. Fill the pipette to 20 ml by
gentle suction, transfer the sample to pre-weighted silica crucible, evaporate and oven dry the
sample at 1050C and record the increase in weight (W2) which indicates the weight of oven dry
clay.
When the sandy residue on 70 mesh sieve has dried rub it lightly until fine
material ceases to pass through. Transfer the coarse sand left in sieve to a pre-weighted silica
crucible, oven dry it at 1050C and find the increase in weight (W3) which corresponds to the
weight oven dry coarse sand.
After second pipetting remove the suspension gently by means of a siphon tube
leaving behind about 4 cm height undisturbed sediment at the bottom of the sedimentation
cylinder. Bring this sediment into suspension by agitating with a stream of water and transfer the
content to a 500 ml beaker marked at 10 cm above the bottom. Add whatever fine material
passed out of 70 mesh sieve by rubbing. Let the beaker stand for 15 minutes. Decant the
supernatant with no or minimum disturbance to the sediment, fill the beaker again with water to
the 10 cm mark stir the sediment wait for 15 minutes and again decant the supernatant. Record
the temperature and find the corresponding time of decantation for fine sand from Table-1. Now
fill the beaker again to 10 cm mark with water and ensure through mixing of sediment. Allow to
stand and on completion of period for decantation as note for Table-1 decant the supernatant as
completely as possible without losing any sediment. Again fill the beaker with water to 10 cm
mark and repeat the exercise until supernatant becomes almost devoid of any suspended matter.
Now transfer the residue in the beaker to pre-weighted silica crucible, evaporate and oven dry the
sample at 1050C, cool in a desiccator and note the increase in weight (W4) which corresponds to
the weight of oven dry fine sand.

Observation table-
S.No. Weight of 1st Weight of 2nd Weight of Weight of Texture

pipet sample pipet sample sandy residue sandy residue
after
decantation
Calculation- Percent fractions in air-dry soil-
Coarse sand (%) = W3x4
Fine sand (%) = W4x4
Silt (%) = (W1-W2) x 250
Clay (%) = (W2 x 250) 1.6
Where, W1= weight of first pipette sample (gm)
W2= weight of second pipette sample (gm)
W3= weight of sandy residue obtained on 70 mesh sieve (gm) and
W4= weight of sandy residue obtained by decantation (gm)
Result- The texture of given soil was observed ---------- gm.
Precautions-
1. Note the value carefully.

2. Weight the soil exactly.
3. Dry the soil properly.

BULK DENSITY
Object- Determine the bulk density of given soil.
Requirements- Oven, measuring cylinder and chemical balance.
Method- Dry the soil sample in oven at 105 0C until a constant weight is attained, transfer a little
dried soil to a measuring cylinder and note the volume. Record the weight of this volume of soil
on a balance.
Observation table-
S. No. Weight of soil (gm) Volume of soil (ml) Bulk density

(gm/cm3)
1.
2.
3.
Calculation-
Bulk density (gm/cm3) = Weight of soil (gm)/Volume of soil (cm3)
Result- The bulk density of given soil was observed ------- gm/cm3.
Precautions-


SPECIFIC GRAVITY
Object- Determine the specific gravity of given soil.
Requirements- Oven, glass bottles, chemical balance.
Method- Dry the soil in an oven at 1050C until a constant weight is attained. Fill a pre-weighted
glass bottle of known volume with dried soil and records its weight. Fill another pre-weighted
glass bottle of same volume with distilled water and records its weight.
Observation table-
S. No. Initial Weight of Initial Weight of Specific

weight of bottle with weight of bottle with gravity
bottle-1(gm) soil (gm) bottle-2 distilled
(gm) water (gm)
1.
2.
3.
Calculation- Formula is given below:
Specific gravity = A2-A1/B2-B1

Where, A2= weight of bottle with soil
A1= weight of empty bottle used for soil
B2= weight of bottle with distilled water
B1= weight of empty bottle used for distilled water
Result- The specific gravity of given soil was observed--------.

Precautions-


MOISTURE CONTENT
Object- Determine the moisture content of given soil.
Requirements- Oven and chemical balance.
Method- Take a fresh homogenized sample of soil and weight it. Now dry it in an oven at 1050C
until a constant weight is attained. Cool in a desiccator and record the final weight of sample.
Observation table-
S. No. Initial weight of soil Weight of dried soil Moisture content

(gm) (gm) (%)
1.
2.
3.
Calculation- Moisture content (%) = I-F/I*100
Where, I= initial weight of soil
F= final weight of soil after drying
Result- The moisture content of given soil was observed-----------%.
Precautions-


WATER HOLDING CAPACITY
Object- Determine the water holding capacity of given soil.
Requirements- Oven, perforated circular soil boxes (5.6 cm diameter, 1.6 cm height bottom
perforated with hales of 0.75 mm diameter), Filter paper (whatman no.1), petridish and chemical
balance.
Method- Dry the crushed soil sample in an oven at 1050C. Place a filter paper inside the
perforated bottom of the circular soil box, weight the box and fill it with dried soil sample. Note
the weight of box filled with dried soil. Place the box in petridish of 10 cm diameter containing
water for about 12 hours. So that water enters the box and saturates the soil. Take the box out of
water wipe it dry on the outside and records its weight.
Observation table-
S. No. Initial weight of Weight of soil Weight of soil Water holding

soil box (gm) box with soil box with soil capacity (%)
(gm) after observing
water (gm)
1.
2.
3.
Calculation-
Water holding capacity (%)= (W2-W1)-(W1-W0)/(W1-W0)*100
Result- The water holding capacity of given soil was observed -----------%.
Precautions-
1. Weight the soil and particulars properly.

2. Dry the soil box from outside properly.

pH
Object- Determine the hydrogen ion concentration (pH) of given soil.
Requirements- Distilled water and pH meter.
Method- Take 10 gm of air-dry soil/sediment and add 100 ml of distilled water to make a
suspension of 1:10 w/v dilution. Determine the pH of suspension.
Observation table-
S. No. Sample description pH

1.
2.
3.
Calculation- (No calculation required. Note the reading directly from pH meter screen).
Result- The hydrogen ion concentration (pH) of given soil was observed---------.
Precaution-
1. Standardize the instruments properly.

2. Prepare soil solution properly.

ELECTRICAL CONDUCTIVITY (EC)
Object- Determine the electrical conductivity of given soil sample.
Requirements- Conductivity meter, distilled water, beaker.
Method- Take 10 gm of air-dry soil/sediment and add 100 ml of distilled water to prepare
suspension (1:10 w/v). Then record the EC (Electrical Conductivity).
Observation table-
S. No. Sample description Conductivity (mho/cm)

1.
2.
3.
Calculation-
Conductivity (mho/cm) = Selector knob reading x dial reading x cell constant.
Result- The electrical conductivity of given soil was observed---------mho/cm.
Precautions-
1. Prepare soil suspension properly.

2. Calibrate the conductivity meter.
3. Note the reading properly.

REDOX POTENTIAL
Object- Determine the redox potential of given soil.
Requirements- Distilled water
Method- Take 10 gm of air-dry soil/sediment and add 100 ml of distilled water to make a
suspension (1:10 w/v). Determine the redox potential of suspension as that of water.
Observation table-
S. No. Sample description Redox potential

1.
2.
3.
Calculation- Digital instrumental method (no calculation).
Result- The redox potential of given soil was observed----------.
Precautions-


ALKALINITY
Object- Determine the alkalinity of given soil.
Requirements- Distilled water, filter paper (whatman no- 44), oven and chemical balance.
Method- Take a 10 gm of air-dry soil/sediment and add 100 ml of distilled water to prepare a
suspension (1:10 w/v) filter it through a filter paper and determine the alkalinity of filtrate.
(Procedure same as in water alkalinity testing) in the fraction of same soil/sediments sample
estimate the moisture contents.
Observation table-
S. No. Sample Initial Final Normality of Alkalinity

description reading of reading of H2SO4 (mg/g)
burette (ml) burette (ml)
1.
2.
3.
Calculation-
Alkalinity (mg/g) = F/10 * V/W * 1/(100-M)
Where, F= Alkalinity of filtrate (mg/l)

V= Total volume of suspension (ml)
W= weight of soil used in suspension (g)
and M= Moisture content of soil/sediment (%)
Result- The alkalinity of given soil was observed----------mg/g.
Precautions-


CHLORIDE
Object- Determine the chloride in given soil sample.
Requirements- Filter paper (Whatman no. 44), weight box, physical balance, distilled water etc.
Procedure- Take 10 gm of soil/sediment and add 100 ml of distilled water to prepare a 1:10 w/v
suspension. Filter it through a filter paper and estimate the chloride in filtrate following the
method given for water.
Calculation-
Chloride (mg/l) = F/10 x V/W x 1/(100-M)
Where, F= chloride estimated in filtrate (mg/l)
V = total volume of suspension (ml)
W = weight of soil/sediment used in suspension (gm)
And M = moisture content of soil/sediment (%)
Result- Chloride in given soil sample was observed--------mg/l.
Precautions-
1. Weight carefully.
2. Collect soil sample carefully.

SULPHATE
Object- Determine the sulphate in given soil sample.
Requirements- Filter paper (Whatman no. 44), weight box, physical balance, distilled water etc.
Procedure- Take 10 gm of soil/sediment and add 100 ml of distilled water to prepare a 1:10 w/v
suspension. Filter it through a filter paper and estimate the sulphate in it following the method
described for water, preferably the gravimetric method. Estimate in a fraction of same
soil/sediment sample the moisture content following the method given water.
Calculation-
Chloride (mg/l) = F/10 x V/W x 1/(100-M)
Where, F= sulphate estimated in filtrate (mg/l)
W = weight of soil/sediment used in suspension (gm)
And M = moisture content of soil/sediment (%)
Result- Chloride in given soil sample was observed--------mg/l.
Precautions-
1. Weight the soil carefully.


NITROGEN (KJELDHAL NITROGEN)
Object- Determine the nitrogen in give soil sample.
Requirements- Kjeldhal distillation assembly, laboratory glassware etc.
Reagents-
(a.) Catalyst mixture- Mix 20 gm of copper sulphate, 3 gm of mercuric oxide, and 1 gm of

selenium powder and grind. Mix 2 gm of this mixture with 40 gm of sodium sulphate to
prepare the catalyst mixture.
(b.) Sulphuric acid (concentrated)
(c.) Sodium hydroxide solution (40%)- Dissolve 40 gm of sodium hydroxide in 100 ml of
distilled water.
(d.) Zink granules.
(e.) Boric acid cum indicator solution-
(f.) Hydrochloric acid (0.1N)- Take 8.34 ml of concentrated hydrochloric acid (12N) and
dilute with distilled water to prepare 100 ml of 0.1N HCl. Dilute this 100 ml for 1.0N
HCl to 1 liter by adding distilled water and get 0.1N HCl.
Procedure- Take 10 gm of air-dry soil/sediment in a 300 ml round bottom flask and add 20 gm
of catalyst mixture and 35 ml of sulphuric acid. Heat the contents from bottom of the flask for
about 2 hours. Cool the contents (digest), add about 100 ml of distilled water, wait for about 5
minutes, and deliver the supernatant into 1 liter distillation flask A of Kjeldhal distillation
assembly. Wash the residue with a little of distilled water several times and transfer the
supernatant each time to the same distillation flask. Add 100 ml of sodium hydroxide solution
and a few granules of zinc. Take 25 ml of boric acid cum indicator solution in a 500 ml
Erlenmeyer flask B and place it below distillation assembly so that the lower open ends of the
condenser is dipped in solution. Heat the distillation flask on a hot plate and collect about 150 ml
of distillate in flask B. Remove the flask with distillate and titrate the distillate (which has turned
blue due to dissolution of ammonia) against 0.1N hydrochloric acid. Turning of blue colour to
light brown-pink indicates the end point. Run distilled water blank in the same manner.

Calculation-
Kjeldhal Nitrogen (mg/l) = T1 T2 x N x 14 / W
Kjeldhal Nitrogen (%) = T1 T2 x N x 1.4 / W
Where, T1 = volume of titrant used against sample (ml)
T2 = volume of titrant used against distilled water blank (ml)
N = normality of titrant (0.1) and
W = weight of soil/sediment used (gm)
Result- The Kjeldhal nitrogen in given soil sample was observed--------mg/l.
Precautions-
1. Weight carefully.

NITRATE
Object- Determine the nitrate of given soil sample.
Requirements- Laboratory glass ware, filter paper (whatman no. 50), chemical balance, flask
etc.
Reagents-
(a.) Extraction reagent- (i.) Dissolve 12.5 gm of copper sulphate in distilled water to make
100 ml of solution.
(ii.) Dissolve 0.6 gm of silver sulphate in distilled water to make 100 ml of solution. Mix
20 ml and 100 ml of solution (i) and (ii) respectively and dilute with distilled water to
make 1 liter of nitrate extraction reagent.
(b.) Calcium hydroxide- Dry powdered.
(c.) Magnesium carbonate-Dry powdered.
Procedure- Dry the soil/sediment in air and take 50 gm of it in an Erlenmeyer flask (500 ml).
Add 250 ml of extraction reagent, shake for 15 minutes, add 0.4 gm of calcium hydroxide, shake
for 5 minutes, and then add 1 gm of magnesium carbonate. Filter the contents through filter
paper and measure the total volume of filtrate. Determine the nitrate content in filtrate following
phenodisulphonic acid method described for water.
Calculation-
NO3- N (mg/l) = F x V / 1000 x W
Where, F = NO3- N determine in filtrate (mg/l)
V = total volume of filtrate (ml) and
W = weight of dried soil/sediment used (gm)/

Result- The nitrate in given soil sample was observed--------mg/l.
Precautions-
1. Weight soil carefully.


TOTAL PHOSPHORUS
Object- Determine the total phosphorus in given soil sample.
Requirements- Filter paper (Whatman no.44), hot plate, distilled water, flask and reagent bottle,
weigh box, physical balance etc.
Reagents-
(a.) Nitric acid.

(b.) Perchloric acid (concentrated)
(c.) Sulphuric acid (dilute)- Take 5 ml of concentrated sulphuric acid and dilute with distilled
water to 100 ml.
Procedure- Air-dry the soil/sediment grinds to fine powder, and take 0.5 gm of it in a round
bottom flask. Add a few drops of distilled water, 2 ml of nitric acid and 2 ml of perchloric acid.
Heat gently on hot plate up to dryness. Cool, add 21 ml of sulphuric acid and boil for 15 minutes.
Cool the flask; filter the digest through a filter paper (Whatman no. 44) and makeup the volume
of it to 250 ml with distilled water in a volumetric flask. Determine the phosphate content in
solution following the method described for estimation of inorganic phosphorus in water.
Calculation-
Total phosphorus (mg/l) = Pd x V / 1000 x W
Where, Pd = PO4-P in digest (mg/l)
V = total volume of solution (ml) and
W = weight of air dry soil/sediment taken (gm)
Result- The total phosphorus in given soil sample was observed--------mg/l.
Precautions-


PHOSPHATE
Object- Determine the phosphate in given soil sample.
Requirements- Filter paper (Whatman no. 50), flask, distilled water etc.
Reagents-
(a.) Sulphuric acid (0.002N)- Dilute 2.8 ml of concentrated sulphuric acid to 1 liter by adding
distilled water. Take 20 ml of it and dilute to 1 liter with distilled water to get 0.002N
suphuric acid.
Procedure- Air-dry the soil/sediment sample and take 1 gm of it in 500 ml flask. Add 200 ml
sulphuric acid (0.002N) to it and shake for about half an hour. Filter the suspension through a
filter paper (Whatman no. 50). Determine the phosphate content in the filtrate following the
method described for phosphorus in water.
Calculation-
PO4-P (mg/l) = Ps x V / 1000 x W
Where, Ps= PO4-P estimated in suspension (mg/l)
V = total volume of suspension (ml) and
W = weight of air dry soil/sediment taken.
Result- The total phosphate in given soil sample was observed--------mg/l.
Precautions-


CALCIUM
Object- Determine the calcium in given soil sample.
Requirements- Filter paper (Whatman no. 50), flask, distilled water, beaker etc.
Reagents-
(a.) Ethyl alcohol- absolute and 40 %.

(b.) Ammonium acetate solution (1N) - Take 57 ml of glacial acetate acid and dilute it to 800
ml with distilled water. Add gradually the concentration ammonium hydroxide solution
till pH 7.0 is attained. Further add distilled water to make the final volume 1 liter.
Procedure- Preparation of soil extract: Air dries the soil/sediment and takes 50 gm of it in a
flask. Add 100 ml of 40% of ethyl alcohol, shake well, wait for about 10 minutes, and filter the
suspension through filter paper. Further wash the soil residue on filter paper with 40% ethyl
alcohol and finally with absolute ethyl alcohol. Transfer the residue to a beaker, add 100 ml of
ammonium acetate solution, stir and allow standing overnight. Filter the supernatant through
filter paper and collect the filtrate (soil extract). Note the total volume of soil extract.
Calculation-
Calcium (mg/l) = T x 400.4 x V1 / V2 x W 10000
Where, T = volume of EDTA titrant used (ml)
V1 = total volume of soil extract (ml)
V2 = volume of soil extract titrated (ml) and
W = weight of air-dry soil/sediment taken for extraction.
Result- The total calcium in given soil sample was observed--------mg/l.
Precautions-


MAGNESIUM
Object- Determine the magnesium in given soil sample.
Requirements- Filter paper (Whatman no. 50), all material including reagents, required for
determination of magnesium in water etc.
Reagents-
(a.) Ethyl alcohol- Absolute and 40%.

(b.) Ammonium acetate solution- Take 57 ml of glacial acetate acid and dilute it to 800 ml
with distilled water. Add gradually the concentration ammonium hydroxide solution till
pH 7.0 is attained. Further add distilled water to make the final volume 1 liter.
Procedure- Prepare the soil extract as in case of determination of calcium. Find out magnesium
content in the extract following the method given for determination of magnesium in water.
Calculation-
Magnesium (mg/l) = T2 T1 x 400.4 x V1 / V2 x 1.645x W x 10000
Where, T2 = volume of EDTA titrant used for determination of calcium and magnesium (ml)
T1 = volume of EDTA titrant used for determination of calcium (ml)
V1 = total volume of soil extract (ml)
V2 = volume of soil extract titrated (ml) and
W = weight of air-dry soil/sediment taken for extraction (gm)
Result- The total magnesium in given soil sample was observed--------mg/l.
Precautions-


SODIUM
Object- Determine the sodium in given soil sample.
determination of sodium in water etc.
Reagents-

Procedure- Prepare the soil extract as described for calcium. Determine the sodium content in
extract following the method of sodium determination in water.
Calculation-
Sodium (mg/l) = A x V/ W x 10000
Where, A = sodium content of soil extract (mg/l)
V = total volume of soil extract (ml) and
Result- The total sodium in given soil sample was observed -------mg/l.
Precautions-


POTASSIUM
Object- Determine the potassium in given soil sample.
determination of potassium in water etc.
Reagents-

Procedure- Prepare the soil extract as described for determination of calcium. Find out the
potassium in extract as it is determined in water.
Calculation-
Potassium (mg/l) = A x V / W x 10000
Where, A = potassium content of soil extract (mg/l)
V = total volume of soil extract (ml) and
Result- The total potassium in given soil sample was observed--------mg/l.
Precautions-


IRON
Object- Determine the iron in given soil sample.
determination of iron in water etc.
Procedure- Take 10 gm of air dried soil/sediment and add 100 ml of distilled water to make
1:10 w/v suspension. Filter it through a filter paper and determine the iron in filtrate following
the method used for iron estimation in water. Analyze a fraction of soil/sediment sample for
moisture content as described moisture content of soil.
Calculation-
Iron (mg/l) = F/10 x V/W x 1/ (100-M)
Where, F = iron determined in filtrate (mg/l)
W = weight of soil/sediment used in suspension (gm) and
M = moisture content of soil/sediment (%)
Result- The total iron in given soil sample was observed--------mg/l.
Precautions-


ORGANIC MATTER
Object- Determine the organic matter in given soil sample.
Requirements- Laboratory glassware, flask etc.
Reagents-
(a.) Potassium dichromeate solution (1N)- Dissolve 49.04 gm of potassium dichromate in

distilled water to prepare 1liter of solution.
(b.) Sulphuric acid (concentrated)
(c.) Phosphoric acid (concentration)
(d.) Sodium fluoride- Dry, powdered
(e.) Diphenylamine indicator- Dissolve 0.25 gm of diphenylamine in 10 ml of distilled water
and add gradually 50 ml of concentrated sulphuric acid.
(f.) Ferrous ammonium sulphate solution (0.5N)- Add 20 ml of concentrated suphuric acid to
800 ml of distilled water and dissolve in it 196.1 gm of ferrous ammonium sulphate.
Further add distilled water to make the volume 1 liter.
Procedure- Air-dry the soil/sediment and take 0.5 gm of it in 500 ml Erlenmeyer flask. Add10
ml of potassium dichromate solution and gradually 20 ml of sulphuric acid. Wait for about half
an hour, and the add 200 ml of distilled water, 10 ml of phosphoric acid, 0.2 gm of sodium
fluoride and 1 ml of diphenylamine indicator. Titrate the contents against ferrous ammonium
sulphate solution, at the end point the dull green colour changes through turbid blue to the
brilliant green.
Calculation-
O M (mg/l) = 6.791/ W [1- T1/T2] x 10
O M (%) = 6.791/ W [1- T1/T2]
Carbon (%) = 6.791/ W x 1.724 [1- T1/T2]
Where, OM = Organic matter
W = weight of soil/sediment taken (gm)

T1 = volume of titrant used against sample (ml) and
T2 = volume of titrant used against distilled water blank (ml)
Result- The total organic matter in given soil sample was found--------mg/l.
Precautions-


ABOUT THE AUTHERS
Dr. Sadhana Chaurasia had his M.Sc. in Botany from Govt.

Science Collage Raipur in 1984, and was awarded with PhD in Bioscience
by Pt. Ravi Shankar University, Raipur in 1990. Presently she is working as
Associate Professor and Head Dept. of Energy & Environment, Faculty of
Science & Environment at MGCGV, Chitrakoot Satna MP, since 1992. She
has over 22 years of teaching experience of UG & PG students of
Environmental Science and handle projects for the individual students in the field of
Environmental Pollution. She has guided around 150 students to obtain their Masters degree. She
is currently guiding Six Ph.D. students and six students have been awarded with Ph.D. degree.
Several M.Phil students were also guided by her. She has published more than 73 research
papers in national and international journals. She has presented various lectures in national and
international seminars/conferences. She has attended several conferences, trainings and
workshops. She was awarded Best Science Research Award (Teacher) by MPCST, University
Cell, 2013.
Mr. Anand Dev Gupta completed his M.Sc. in Environmental Science, 2011
and M.Phil-Environmental Science, 2012 from MGCGV, Chitrakoot Satna
MP. Presently he is pursuing Ph.D. in Environmental Science from the same
university. He has 1 year of teaching experience of UG & PG classes. He has
published more than 19 research paper in national and international Journals.
He has presented 04 research papers in various national seminars/conferences.
He has attended several conferences, trainings and workshops. He has received Chancellor
Gold Medal for standing first in M.Sc. He was awarded Best Science Research Award
(Research Scholar) by MPCST, University Cell; 2013. He is playing the role as editorial board
member of many international journals.

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Hand Book of Water, Air and Soil Analysis

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Dr. Sadhana Chaurasia

Hand Book of Water, Air and Soil Analysis.. iii

Hand Book of Water, Air and Soil Analysis.. iv

II- Air Analysis 67

Hand Book of Water, Air and Soil Analysis.. v

Hand Book of Water, Air and Soil Analysis 1

Object- Determine the temperature of given water sample.

Requirements- Thermometer and beaker etc.

Principle- Relation between Celsius, Fahrenheit and Kelvin scale-

Result- The temperature of given sample was observed --------0C.

1. Beaker should be clean properly.

Hand Book of Water, Air and Soil Analysis 2

Object- Determine the transparency of the water body.

Requirements-Sacchi disc, string, notebook and inch tape.

Sacchi disc transparency (SDT) cm = A = B / 2

Where A = depth at which sacchi disc disappears (cm)

B = depth at which sacchi disc reappears (cm)

Euphotic limit (cm) = SDT x 2.5

Vertical attenuation coefficient = 1.9 / SDT

Where SDT = Sacchi disc transparency (cm)

Result- The transparency of water body was found ------ cm.

1. Note the reading carefully.

Hand Book of Water, Air and Soil Analysis 3

TURBIDITY (Nephelometric method)

Object- Determine the turbidity of given water sample.

Turbidity (NTU)= NTU measured/proportion of sample in dilution

Hand Book of Water, Air and Soil Analysis 4

S. No. Sample Description Turbidity (NTU) Average turbidity

1. Calibrate the instrument properly.

Hand Book of Water, Air and Soil Analysis 5

Object- Determine the pH of given water sample.

Principle- pH is a negative logarithm of H+ concentration of a solution.

S. No. Sample description pH value Average pH

Result- The pH of given water sample was observed -----.

1. All the glassware should be clean.

Hand Book of Water, Air and Soil Analysis 6

Object- Determine the conductivity of given water sample.

S. No. Sample description Conductivity Average conductivity

Result- The conductivity of given water sample was obtained........mhos cm-1.

1. Rinse the electrode with distilled water.

Hand Book of Water, Air and Soil Analysis 7

TOTAL SOLIDS (TS) (Gravimetric method)

Requirements- Porcelain dishes, measuring cylinder, balance and oven etc.

TS (mg/l) = (W2-W1) x 106 / Sample volume (ml)

S. No. Sample no. Initial weight Final weight TS (mg/l) Average TS

1. Glassware should be clean.

Hand Book of Water, Air and Soil Analysis 8

TOTAL SUSPENDED SOLID (TSS) (Gravimetric method)

Object- Determines the TSS of given water sample.

Formula- W2-W1 x 1000/V

W1 = Initial weight of filter paper

W2 = final weight of filter paper

S. No. Sample Initial weight of Final weight of TSS (mg/l)

1. Glassware should be clean.

Hand Book of Water, Air and Soil Analysis 9

TOTAL DISSOLVED SOLID (TDS) (Gravimetric method)

Object- Determines the TDS of given water sample.

Formula= W2-W1 x 1000/V

W1 = Initial weight of beaker

W2 = final weight of beaker