Hand Book of Water, Air and Soil Analysis (A Lab Manual)

Transcription

1 Hand Book of Water, Air and Soil Analysis (A Lab Manual) BY SADHANA CHAURASIA & ANAND DEV GUPTA International E Publication

2 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, , India & ANAND DEV GUPTA Department of Energy & Environment MGCGV, Chitrakkot, Satna MP, , India 2014 International E - Publication

3 International E - Publication 427, Palhar Nagar, RAPTC, VIP-Road, Indore (MP) INDIA Phone: , Mobile: contact@isca.co.in, Website: 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:

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

5 CONTENTS S. No. Description Page No. I- Water Analysis 1 Section A (Physico-Chemical test) 1. Temperature 2 2. Transparency 3 3. Turbidity ph 6 5. Conductivity 7 6. Total Solids (TS) 8 7. Total Suspended Solids (TSS) 9 8. Total Dissolved Solids (TDS) Total Volatile Solids (TVS) Volatile Suspended Solids (VSS) Fixed Dissolved Solids (FDS) Alkalinity Acidity Chloride (Cl - ) Sulphate (SO -- 4 ) Fluoride (F - ) Total Hardness Calcium and Magnesium hardness Sodium Potassium Percent Sodium Sodium Absorption Ratio (SAR) Total Iron (Fe) Total Kjeldhal Nitrogen (TKN) Nitrite- Nitrogen (NO 2 -N) Nitrate- Nitrogen (NO 3 -N) Dissolved Oxygen (DO) Bio-Chemical Oxygen Demand (BOD) Chemical Oxygen Demand (COD) Residual Chlorine Chlorine Demand Boron Total Phosphorous Section B (Biological test) 34. Methods of sampling for MPN test Total MPN Feacal MPN Hand Book of Water, Air and Soil Analysis.. iv

6 II- Air Analysis Temperature Minimum and Maximum Relative humidity of Atmosphere Particulate Matter (PM) PM PM SO NO x Ozone Ammonia III- Soil Analysis Texture Bulk Density Specific Gravity Moisture Content Water Holding Capacity ph Electrical Conductivity (EC) Redox Potential Alkalinity Chloride Sulphate Nitrogen (Kjeldhal Nitrogen) Nitrate Total Phosphorus Phosphate Calcium Magnesium Sodium Potassium Iron Organic Matter Hand Book of Water, Air and Soil Analysis.. v

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

8 TEMPERATURE Object- Determine the temperature of given water sample. Requirements- Thermometer and beaker etc. Principle- Relation between Celsius, Fahrenheit and Kelvin scale- 100 = = R 80 =K = 32 = R 9 4 =K 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 Result- The temperature of given sample was observed C. 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. Hand Book of Water, Air and Soil Analysis 2

9 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- 1. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 3

10 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, (NH 2 ) 2, H 2 SO 4 in distilled water and dilute to 100 ml in a volumetric flask. (b) Solution II- Dissolve 10 gm hexamethylenetetramine, (CH 2 ) 6 N 4, 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 25±3 0 C. 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 Hand Book of Water, Air and Soil Analysis 4

11 Observation table- S. No. Sample Description Turbidity (NTU) Average turbidity (NTU) 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. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 5

12 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 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 4±01 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 Result- The ph of given water sample was observed Precautions- 1. All the glassware should be clean. 2. Calibrate the instruments properly. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 6

13 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 1.412±0.01 at 25 0 C. Rinse the electrode and dip in sample and note the reading with temperature 0 C. Rinse the electrode and dip in another sample respectively. Observation table- S. No. Sample description Conductivity (µ mhos cm -1 ) Average conductivity Result- The conductivity of given water sample was obtained...µmhos 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. Hand Book of Water, Air and Soil Analysis 7

14 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 (W 1 ) already kept at C 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 C 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 (W 2 ). Repeat the process of weighing and drying till constant weight is achieved. Calculation- TS (mg/l) = (W2-W1) x 10 6 / 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) Result- Total solids in the given water sample was found----- mg/l at C. Precautions- 1. Glassware should be clean. 2. Cool the dish properly. 3. Note the weight carefully. Hand Book of Water, Air and Soil Analysis 8

15 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 (105 0 C) 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) Result- The total suspended solids in given water sample was observed mg/l. Precautions- 1. Glassware should be clean. 2. Weigh the filter paper carefully. 3. Note the weight carefully. Hand Book of Water, Air and Soil Analysis 9

16 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 (105 0 C) 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) Result- The total dissolved solids in given water sample was observed mg/l. Precautions- 1. Glassware should be clean. 2. Weigh the beaker carefully. 3. Note the weight carefully. Hand Book of Water, Air and Soil Analysis 10

17 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 C 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 C. W2= final weight when TS measured at C. TVS mg/l= (W2-W3) x 10 6 / sample volume (ml) S. No. Crucible No. Initial weight (W1) Final weight at C (W2) Final weight at C (W3) TVS mg/l Average TVS mg/l Result- Total volatile solids of the water sample at C were found mg/l. Precautions- 1. Glassware should be clean. 2. Weigh the beaker carefully. 3. Note the weight carefully. Hand Book of Water, Air and Soil Analysis 11

18 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 C. (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 paper no. Initial weight (W1) Final weight (W2) at C Final weight (W3) at C VSS Average VSS Calculation- Volatile suspended solids = (W2-W3) W2= final weight of filter paper at C W3= weight of filter paper at C drying VSS mg/l= (W2-W3) x 10 6 / sample volume (ml) Result- VSS of the water sample was found----- mg/l at C. Precautions- 1. Glassware should be clean. 2. Weigh the beaker carefully. 3. Note the weight carefully. Hand Book of Water, Air and Soil Analysis 12

19 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 C. (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 C 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 10 6 / Volume of sample Observation table- S. No. Crucible No. Initial weight (W1) Final weight (W2) FDS mg/l Average FDS mg/l Result- FDS in the given water sample was found mg/l at C. Precautions- 1. Glassware should be clean. 2. Weigh the beaker carefully. 3. Note the weight carefully. Hand Book of Water, Air and Soil Analysis 13

20 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 H 2 SO 4 by diluting 30 ml conc. H 2 SO 4 to 1 litre distilled water. Standardized it with standard sodium carbonate Na 2 CO 3 0.1N. Dilute appropriate volume of sulphuric acid, approximately 100 ml to 500 ml to obtained standard 0.02N H 2 SO 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 gm methyl orange diluted 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 H 2 SO 4 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 CaCO 3. Phenolphthalein alkalinity (mg/l as CaCO 3 ) = A x N x 1000/ volume of sample Total alkalinity (mg/l as CaCO 3 ) = B x N x 1000/ volume of sample Methyl orange alkalinity (mg/l as CaCO 3 ) = (B-A) x 1000/ volume of sample Hand Book of Water, Air and Soil Analysis 14

21 Observation table- S. No. Sample H 2 SO 4 used H 2 SO 4 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 CaCO 3 ) CaCO 3 ) CaCO 3 ) (ml) B (A+B) Result- The alkalinity of given water sample was observed P alkalinity mg/l as CaCO 3, T alkalinity mg/l as CaCO 3 and M alkalinity mg/l as CaCO 3. Precautions- 1. Glassware should be clean. 2. Prepare solution carefully. 3. Note the end point carefully. Hand Book of Water, Air and Soil Analysis 15

22 ACIDITY (Titrimetric method) Object- Determine the acidity of given water sample. Requirements- Burettes, pipette, volumetric flask, conical flask and beaker. Reagents- (a) Standard sodium hydroxide (0.02N) 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 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 CaCO 3 (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) CaCO 3 ) Result- The acidity of given water sample was observed-----mg/l as CaCO 3. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the end point reading carefully. Hand Book of Water, Air and Soil Analysis 16

23 CHLORIDE (Argentometric method) Object- Determine the chloride in given water sample. Requirements- Burettes, pipette, volumetric flask, conical flask and beaker. Reagents- (a) Potassium chromate indicator solution- 50 gm K 2 Cr 2 O 4 is dissolved in a small quantity of distilled water AgNO 3 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 N AgNO 3 is 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 K 2 CrO 4 indicator, titrate with AgNO 3 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 x 1000/ volume of sample Where, A= ml AgNO 3 used for sample B= ml AgNO 3 used for blank N= Normality of AgNO 3 Observation table- S. No. Sample Normality of AgNO 3 used AgNO 3 used Chloride description AgNO 3 for sample for blank (mg/l) (ml) A (ml) B Hand Book of Water, Air and Soil Analysis 17

24 Result- The chloride in given water sample was observed mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the end point reading carefully. Hand Book of Water, Air and Soil Analysis 18

25 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 BaCl 2 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 SO -2 4 /litre and measure the absorbance. Prepare standard graph between absorbance and concentration of sulphate ions. Calculation- Deduce theso -2 4 concentration in water sample from standard graph. Observation table- S. No. Sample Sample volume Absorbance Sulphate (mg/l) description (420 nm) Result- The sulphate (SO -2 4 ) in given water sample was observed -----mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Measure the absorbance carefully. Hand Book of Water, Air and Soil Analysis 19

26 FLUORIDE (Spands-Zirconium colorimetric method) Object- Determine the fluoride in the given water sample. Requirements- Nessler s cylinder, burette, pipette, reagent bottle, volumetric flask, kjeldalh flask, heating mental and hot plate. Interferences removal- interferences due to higher concentration of Al, Cl - -2, Fe, PO 4, F, SO 4 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 gm (ZrOCl 2.8H 2 O) 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 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 (NaAsO 2 ) in 100 ml distilled water. (e.) Concentrate H 2 SO 4 1liter (2 x 500 ml) (f.) Concentrate HCl 1 liter (2 x 500 ml) (g.) Silver sulphate crystals (Ag 2 SO 4 ) (h.) Fluoride reference solution- Dissolve gm previously dried (105 0 C 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. Hand Book of Water, Air and Soil Analysis 20

27 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. H 2 SO 4 and shake to mix. (2.) Heat on heating device to boil at C, when about 100 ml distillate is collected put off the heating unit and after cooling to about C 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 nessler s cylinder with stopper mark on the cylinder as blank 1, 2, (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 ml ml ml ml ml 1.0 Hand Book of Water, Air and Soil Analysis 21

28 7. 6 ml ml 1.4 S 1 Sample-I 50 ml Sample-I S 2 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 (B-S) x f] If sample is diluted for direct determination then conc. F - ml/l = [absorbance difference Result-The fluoride in given water sample was found------mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Measure the absorbance carefully. Hand Book of Water, Air and Soil Analysis 22

29 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 NH 4 Cl in 143 ml conc. NH 4 OH. 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 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 CaCO 3 in a 500 ml flask. Add HCl slowly through a funnel till all CaCO 3 is dissolved. Add 200 ml distilled water and boil for a few minutes to expel CO 2. Cool and add a few drops of methyl red indicator and adjusts to the intermediate orange colour by adding 3N NH 4 OH or HCl, as required. Transfer quantitatively and dilute to 1000 ml with distilled water, 1 ml=1mg CaCO 3. 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, Hand Book of Water, Air and Soil Analysis 23

30 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 CaCO Result- The total hardness of given water sample was observed mg/l as CaCO 3. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the end point reading carefully. Hand Book of Water, Air and Soil Analysis 24

31 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 NH 4 Cl in 143 ml conc. NH 4 OH. 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 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 CaCO 3 in a 500 ml flask. Add HCl slowly through a funnel till all CaCO 3 is dissolved. Add 200 ml distilled water and boil for a few minutes to expel CO 2. Cool and add few drops of methyl red indicator and adjusts to the intermediate orange colour by adding 3N NH 4 OH or HCl, as required. Transfer quantitatively and dilute to 1000 ml with distilled water, 1 ml-1mg CaCO 3. (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 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 Hand Book of Water, Air and Soil Analysis 25

32 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 i.e. Magnesium as Mg (mg/l) = magnesium hardness x Observation table- S. No. Sample Sample volume EDTA used Calcium description (ml) (ml) hardness (mg/l) Result- The Ca and Mg hardness was observed mg/l as CaCO 3 and------mg/l ascaco 3 respectively and Ca and Mg was observed mg/l and mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. Hand Book of Water, Air and Soil Analysis 26

33 SODIUM (Flame photometric method) Object- Determine the sodium in water sample. Requirements- Flame photometer, conical flask etc. Reagents- (a.) Stock sodium solution 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 = µg 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 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- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Calibrate instrument properly. Hand Book of Water, Air and Soil Analysis 27

34 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 gm KCl, dried at C 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 µg 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 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- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Calibrate instrument properly. Hand Book of Water, Air and Soil Analysis 28

35 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 (ii.) Equivalent weight of Ca +2 = 20, Na + = 23, K + = 39.1 and Mg + = 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. Hand Book of Water, Air and Soil Analysis 29

36 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 to to to 2 30 to 60 Very good to Poor 3.0 to to to to to 80 Very poor 5.0 to to to to to 90 Unsuitable Result-The percent sodium in given water was found------%. Hand Book of Water, Air and Soil Analysis 30

37 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 Mg 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 Hand Book of Water, Air and Soil Analysis 31

38 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 (NH 2 OH) solid in 100 ml volumetric flask dilute to the mark with distilled water. (4.) Ammonium acetate buffer solution- Dissolve 250 gm NH 2 C 2 O 3.3H 2 O 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 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 gm ammonium ferrous sulphate (FAS) is 50 ml distilled water and 20 ml conc. H 2 SO 4 shake to dissolve. Add 0.1 N KMnO 4 (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 µg 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. Hand Book of Water, Air and Soil Analysis 32

39 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 ml dilute ml ml ml dilute ml ml ml dilute ml ml ml dilute ml ml ml dilute ml ml ml dilute ml ml ml dilute ml ml ml dilute ml ml ml dilute 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, ppm reference solution. (iii.) Take 10 numbers of porcelain dishes of 100 ml capacity mark. Blank 1, 2, 3, Hand Book of Water, Air and Soil Analysis 33

40 (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, ppm respectively. (v.) Now add 2 ml conc. HCl and 1 ml hydroxylamine hydrochloride and shake. Evaporate to remain 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 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 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. Hand Book of Water, Air and Soil Analysis 34

41 Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 35

42 TOTAL KJELDHAL NITROGEN (TKN) Object- To determine the TKN in given water sample Apparatus Required- ammonia distillation apparatus, hot plate, spectrophotometer, burette, nessler s cylinders, other laboratory glass ware. Reagents- (a) Digestion reagents- Dissolve gm potassium sulphate (K 2 SO 4 ) in 650 ml distilled water & add 200 ml conc. H 2 SO 4 and add this solution to the solution prepare by dissolving 2.0 gm HgO (mercuric-oxide) in 25 ml conc. H 2 SO 4. 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 gm methylene blue in 50 ml ethanol. (d) Boric acid solution (indicating)- Dissolve 20.0 gm H 3 BO 3 (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 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. Hand Book of Water, Air and Soil Analysis 36

43 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 H 2 SO 4 in place of boric acid. If NH 3 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 NH 3 -N as prepared in the determination of NH 3 -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 H 2 SO 4 and note the volume of H 2 SO 4 consumed with blank and sample. Calculation- TKN mg/l = Hand Book of Water, Air and Soil Analysis 37

44 Where, A= ml of 0.02N H 2 SO 4 for sample titration. B= ml of 0.02N H 2 SO 4 for blank titration. N= Normality of H 2 SO 4 Result- The total kjeldhal nitrogen in given water sample was observed mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 38

45 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 gm Na 2 C 2 O 4 primary standard grade in water and dilute to 1000 ml. (c.) Stock nitrite- Dissolve gm NaNO 2 in water and dilute to 1000 ml (1 ml= 250 µg N). Preserve with 1 ml chloroform (CHCl 3. Standardize by pipetting in order 50 ml 0.01 M KMnO 4, 5 ml conc. H 2 SO 4 and 50 ml stock NO - 2 solution in to a glass stoppered flask. Shake well and warm to C. Discharge permanganate colour by adding 10 ml portions of M sodium oxalate. Titrate excess oxalate with 0.01 M (0.05N) KMnO 4 to faint pink end point. Calculation Nitrite content of stock solution- A= [(B x C) (D x E)] x 7 / F Where, A= NO - 2 -N/ml in stock solution (mg) B= total KMnO 4 used (ml) C= normality of KMnO 4 D= total oxalate added (ml) E= normality of oxalate F= stock nitrite taken for titration (ml) Hand Book of Water, Air and Soil Analysis 39

46 (d.) Standard nitrite solution- Dilute 10 ml intermediate NO - 2 solution to 1000 ml with water (1 ml = µg NO - 2 -N, prepare daily). (e.) Standard potassium permanganate solution (0.05N) - Dissolve 1.6 gm KMnO 4 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 H 2 SO 4 and heat rapidly to 90 to 95 0 C. Titrate with permanganate solution to a slight pink end point that persists to at least 1 min. Do not allow temperature to fall below 85 0 C. Run a blank on distilled water +H 2 SO 4. Normality KMnO 4 = Na 2 C 2 O 4 (gm) / (A-B) x 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 µg/l concentration, respectively. Calculation- Deduce sample concentration directly from the curve, taking in consideration dilution of the sample if applicable. Hand Book of Water, Air and Soil Analysis 40

47 Result-The NO 2 concentration in given water sample was found mg/l NO 2 -N Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 41

48 NITRATE- NITROGEN (Spectrophotometric method) Object- Determine the nitrate-nitrogen concentration in given water sample. Requirements- Volumetric flask, beaker, glass rod, heating mental, reagent bottles, nessler s cylinder, balance, spectrophotometer, water bath, porcelain dishes, rubber and desicator. Reagents- (a.) Phenoldisulphonic acid (PDA) - Dissolve 25 gm white phenol in 150 ml conc. H 2 SO 4 and add 74 ml fuming H 2 SO 4 (15% free SO 3 ) stir well and heat for 2 hours on a water bath. If fuming H 2 SO 4 is not available add 86 ml conc. H 2 SO 4 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 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 gm AR grade anhydrous KNO 3 (potassium nitrate) dried at C 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 NH 4 OH 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 (HNO 3 ) 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 nesseler s cylinder wash the porcelain dish with 2 portions of distilled water 10 ml each and take the washings in the cylinder makeup to 50 Hand Book of Water, Air and Soil Analysis 42

49 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. (NH 4 OH) 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 NO - 3 N prepared as below- Preparation of Calibration Curve of Nitrate-N (1.) Take 10 Nos of 100 ml capacity necessler s 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 solution 10 ppm to makeup 50 ml Concentration ppm ml blank 0.0 ppm ml 5 ppm in Absorbance at 410 nm ml 10 ppm ml 15 ppm ml 20 ppm ml 25 ppm ml 30 ppm Hand Book of Water, Air and Soil Analysis 43

50 ml 35 ppm 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 NH 4 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 NO - 3 N mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 44

51 D.O. (DISSOLVED OXYGEN) (Modified Winkler s 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) (ii) (iii) Sodium thiosulphate [Na 2 S 2 O 3.5H 2 O] (0.025 N) - dissolve gm of solution thiosulphate in previously boiled distilled water add 0.4 gm solid NaOH and make up the volume to one litre. Manganous sulphate solution [MnSO 4 ]- 480 gm MnSO 4.4H 2 O / 400 gm MnSO 4.2H 2 O / 364 gmmnso 4.H 2 O is dissolved in distilled and dilute to 1000 ml. Alkali-iodide azide reagent gm NaOH and 135 gm NaI are dissolved in distilled water and dilute to 1000 ml. Add 10 gm NaN 3 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 NaN 3 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 MnSO 4 solution followed by 2 ml alkaliiodide-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 times. Allow to settle the precipitate. When precipitation has settled sufficiently add 2 ml conc. H 2 SO 4. 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- Hand Book of Water, Air and Soil Analysis 45

52 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) Result- The dissolved oxygen in given water sample was observed mg/l. Precautions- 1. Glassware should be clean. 2. Avoid bubbling during sampling. 3. Prepare solution carefully. 4. Note end point carefully. Hand Book of Water, Air and Soil Analysis 46

53 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 (KH 2 PO 4 ), gm dipotassium hydrogen phosphate (K 2 HPO 4 ) 33.4 gm. Disodium hydrogen phosphate (Na 2 HPO 4.7H 2 O) and 1.7 gm ammonium chloride NH 4 Cl 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 [MgSO 4.7H 2 O] gm MgSO 4.7H 2 O is dissolved in distilled water and dilute to 1000 ml. (iii) Calcium chloride solution (CaCl 2 ) gm CaCl 2 is dissolved in distilled water and dilute to 1 litre. (iv) Ferric chloride solution [FeCl 3.6H 2 O] g. FeCl 3.6H 2 O is dissolved in 1000ml distilled water. (v) Sulphuric acid [H 2 SO 4 ] (1N)- Slowly and while stirring 28 ml conc. H 2 SO 4 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) 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, MgSO 4, CaCl 2, FeCl 3, 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. Hand Book of Water, Air and Soil Analysis 47

54 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 20 0 C or for 3 days if temperature is maintained at 27 0 C. 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) (b) When dilution water is not seeded: BOD (mg/l) = 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 Hand Book of Water, Air and Soil Analysis 48

55 Result- The BOD in given water sample was observed mg/l. Precautions- 1. Glassware should be clean. 2. Avoid bubbling during sampling. 1. Prepare solution carefully. 2. Note the end point carefully. Hand Book of Water, Air and Soil Analysis 49

56 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) (ii) (iii) Standard potassium dichromate solution (0.25N) gm K 2 Cr 2 O 7 is dissolved, previously dried at C for 24 hours, in 1000 ml distilled water. Sulphuric acid reagents (0.1N)- Add 22 gm silver sulphate (AgSO 4 ) to a 9 lbs (4 kg) bottle of concentrated H 2 SO 4. Keep overnight for dissolution and after that shake well to it. Standard ferrous ammonium sulphate solution (0.1N) - Dissolve 39.2 gm of Fe (NH 4 ) 2 (SO 4 ) 2.6H 2 O in 500 ml of distilled water. Add 20 ml of concentrated H 2 SO 4, and dilute to 1 litre. Standardization- Dilute 10 ml standard K 2 Cr 2 O 7 solution to about 100 ml. Add 30 ml conc. H 2 SO 4 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(NH 4 ) 2 (SO 4 ) 2 solution is given by the following formula- Normality of Fe(NH 4 ) 2 (SO 4 ) 2 solution =. (iv) (v) Ferroin indicator solution gm phenanthroline monohydrate and 695 mg ferrous sulphate FeSO 4.7H 2 O are added in distilled water and dilute to 1000 ml. Mercury sulphate (HgSO 4 ) - Crystal or powder. Method- 50 ml of sample is taken into a 500 ml refluxing flask. 1 gm HgSO 4, glass beads and very slowly added 5 ml sulphuric acid reagent with mixing to dissolve HgSO 4 cool while mixing to avoid possible loss of volatile materials. 25 ml K 2 Cr 2 O 7 solution 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. Hand Book of Water, Air and Soil Analysis 50

57 Where, Cool to room temperature and titrate excess K 2 Cr 2 O 7 with F.A.S. using ml (2-3 drops) ferroin indicator. Formula- COD (mg/l) = 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 Precautions- 1. Glassware should be clean. 2. Prepare solution carefully. 3. Note end point carefully. Hand Book of Water, Air and Soil Analysis 51

58 RESIDUAL CHLORINE Object- Determine the free chlorine in given water sample. Requirements-burette, pipette, beaker, conical flask, measuring cylinder, wash bottle etc. Reagents- Where (a.) Acetic acid (Glacial) (b.) Potassium iodide KI crystal (c.) Standard sodium thiosulphate (0.1N)- 25 gm Na 2 S 2 O 3.5H2O is dissolved in one litre distilled water and standardize with standard potassium dichromate ml distilled water is taken in a 250 ml flask, 1 ml conc. H 2 SO 4 is added followed by 10 ml K 2 Cr 2 O 7 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 H 2 SO 4 conc. (e.) Standard sodium thiosulhate (0.1B) 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 Cl 2 mg/lit = A±B x N x ml sample Hand Book of Water, Air and Soil Analysis 52

59 A= ml titration for sample B= ml titration for blank (Positive or negative) N= normality of Na 2 S 2 O 3 Result- The free chlorine in given water sample was observed -----mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. Hand Book of Water, Air and Soil Analysis 53

60 CHLORINE DEMAND Object- Determine the chlorine demand in given water sample. Requirements-burette, pipette, beaker, conical flask, measuring cylinder, wash bottle etc. 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- 1. Glassware should be clean. 2. Prepare the standard solution carefully. Hand Book of Water, Air and Soil Analysis 54

61 BORAN Object- Determine the boron in given water sample. Requirements-burette, pipette, beaker, conical flask, measuring cylinder, wash bottle etc. Reagents- (a.) Stock boron solution mg anhydrous boric acid H 3 BO 3 is dissolved in distilled water and dilute to one liter. (1 ml = 100 µg B) (b.) Standard boron solution- 10 ml stock boron solution is diluted to one liter with distilled water. (1 ml = 1 µg 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 1.0µg 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 55±2 0 C 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 Hand Book of Water, Air and Soil Analysis 55

62 A2 = absorbance of sample C = µg Boron in standard taken S = ml sample Result- The Boron in given water sample was observed -----mg/l. Precautions- 1. Glassware should be clean. 2. Prepare the solution carefully. 3. Note the reading carefully in record. Hand Book of Water, Air and Soil Analysis 56

63 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. Hand Book of Water, Air and Soil Analysis 57

64 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- 1. Glassware should be clean. 2. Prepare the standard solution carefully. Hand Book of Water, Air and Soil Analysis 58

65 SECTION-B BIOLOGICAL TEST Method of sampling for bacteriological test Apparatus Required- BOD bottle, incubator, duraham s 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 C 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. Hand Book of Water, Air and Soil Analysis 59

66 (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 4 0 C in ice but do not delay more than 6 hours. Hand Book of Water, Air and Soil Analysis 60

67 TOTAL COLIFORM (MPN) TEST (Multiple tube fermentation technique) Object- Determine total MPN in water sample. Apparatus- Laminar flow, auto clave, sprit lamp, durham s 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 Lactose Dipotassium hydrogen phosphate (K 2 HPO 4 ) Potassium dihydrogen phosphate (KH 2 PO 4 ) NaCl Sodium lauryle sulphate Distilled water 20.0 gm 05.0 gm gm gm 5.0 gm 0.1 gm 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 durham s 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. Hand Book of Water, Air and Soil Analysis 61

68 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 % % % influent effluent 4. Bathing Ghats % % % 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 35±0.5 0 C. 14. After 24 hrs check the tubes if there is air in the durham s tube also the colour of the media get changed light red if coliform is present due to fermentation of the media and production of CO 2 gas which gets filled in the durham s tube. These called positive tubes. Count and note the number of positive tubes. Hand Book of Water, Air and Soil Analysis 62

69 Reference Table- Combination of (+) tubes dilution % MPN index per 100 ml Combination of (+) tubes dilution % MPN index per 100 ml Reference Table- Combination of (+) tubes dilution 10% MPN index per 100 ml 1.0% 0.1% Hand Book of Water, Air and Soil Analysis 63

70 Result-The total coliform was found MPN/100ml in given water sample. Precautions- 1. Sample should be taking carefully. 2. Sterilize carefully. Hand Book of Water, Air and Soil Analysis 64

71 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, durham s 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 Lactose Bile salts K 2 HPO 4 KH 2 PO 4 NaCl 20.0 gm 6.0 gm 1.5 gm 4.0 gm 1.5 gm 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 45±0.5 0 C for 40 hours. Calculate MPN fecal coliform following the reference table. Hand Book of Water, Air and Soil Analysis 65

72 Result-The total coliform was found MPN/100ml in given water sample. Precautions- 3. Sample should be taking carefully. 4. Sterilize carefully. Hand Book of Water, Air and Soil Analysis 66

73 AIR ANALYSIS Hand Book of Water, Air and Soil Analysis 67

74 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 C and C during a 24 hours dial cycle. Precaution- 1. Set the index properly. 2. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 68

75 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. 2. Note the reading carefully. Table -1. Relative humidity (%) as measured by Psychrometer Dipression in wet bulb Temperature in dry bulb thermometer ( 0 C) thermometer( 0 C) Hand Book of Water, Air and Soil Analysis 69

76 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 µg/m 3. 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. Hand Book of Water, Air and Soil Analysis 70

77 PARTICULATE MATTER (PM 10 ) (Gravimetric method) Object- Determine the particulate matter (PM 10 ) 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 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 C and 40-50% relative humidity or in an airtight desiccator for 24 hours. Take final weight of the filter paper (Wf). Calculation C PM 10 μg/m 3 = (Wf Wi) x 10 6 / V Where, C PM 10 = Concentration of Nitrogen dioxide (μg/m 3 ) Wf = Initial weight of filter in (gm) Wi = Initial weight of filter in (gm) 10 6 = Conversion of g to (μg) V = Volume of air sampled (m 3 ) Hand Book of Water, Air and Soil Analysis 71

78 Result- The PM 10 in ambient air was found µg/m 3. 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. Hand Book of Water, Air and Soil Analysis 72

79 PARTICULATE MATTER (PM2.5) (Gravimetric method) Object-Determine the particulate matter (PM 2.5 ) in ambient air. Requirements- Respirable dust sampler, filter paper, balance and weight box. Definition PM 2.5 refers to fine particles that are 2.5 micrometers (μm) or smaller in diameter. Ambient air is defined as any unconfined part of the Earth s 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 PM 2.5 size ranges divided by the actual volume of air sampled, and is expressed in μg/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 room s Hand Book of Water, Air and Soil Analysis 73

80 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 ± ºC. 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 PM 2.5 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 l pm(1 m 3 /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 5 0 C 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 to50 0 C. The supporting run-time (interval) data, which are stored in detailed 5-minute intervals in the sampler s 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 μg/m 3 and approximately ± 2% at 100μg/m 3. 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 Hand Book of Water, Air and Soil Analysis 74

81 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: M 2.5 = (Mf Mi) mg x 10 3 μg Where, M2.5 = total mass of fine particulate collected during sampling period (μg) Mf = final mass of the conditioned filter after sample collection (mg) Mi = initial mass of the conditioned filter before sample collection (mg) 10 3 = unit conversion factor for milligrams (mg) to micrograms (μg) Field records of PM 2.5 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 m 3 Where, V = total sample value (m 3 ) Q avg = average flow rate over the entire duration of the sampling period (L/min) t = duration of sampling period (min) 10 3 = unit conversion factor for liters (L) into cubic meters (m 3 ) The equation given below can be used to determine PM 2.5 mass concentration: PM 2.5 = M 2.5 / V Where, PM 2.5 = mass concentration of PM2.5 particulates (μg/m 3 ) Hand Book of Water, Air and Soil Analysis 75

82 PM 2.5 = total mass of fine particulate collected during sampling period (μg) V = total volume of air sampled (m 3 ) Result- The PM 2.5 in ambient air was found µg/m 3. 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. Hand Book of Water, Air and Soil Analysis 76

83 SULPHUR DIOXIDE (Improved West and Gaeke method) Object- Determine the SO 2 in ambient air. Requirements- High volume sampler and spetrophotometer. Reagents/Chemicals- Absorbing Reagent- Potassium tetrachloromercurate (TCM 0.04M)- Dissolve gm mercuric chloride 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 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 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 C, 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 Hand Book of Water, Air and Soil Analysis 77

84 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 Hand Book of Water, Air and Soil Analysis 78

85 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 = SO 2 concentration in mg/ml V1 = Volume of thiosulphate for blank (ml) V2 = Volume of thiosulphate for sample (ml) N = Normality of thiosulphate K = (Milli equivalent weight SO 2 /μg) V = Volume of standard sulphite solution (ml) C (SO 2 μg/m 3 )= (As Ab) x CF x Vs/ Va x Vt Where, C SO 2 = Concentration of sulphur dioxide (μg/m 3 ) As = Absorbance of sample Ab = Absorbance of reagent blank CF = Calibration factor Va= Volume of air sampled (m 3 ) Vs= Volume of sample (ml) Vt = Volume of aliquot taken for analysis (ml) Result- SO 2 in ambient air was observed µg/m 3. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 79

86 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 C N-(1-Naphthyl)-ethylenediamine Di-hydrochloride (NEDA) - A 1%aqueous solution should have only one absorption peak at 320 nm over the range of 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% NaNO 2 or greater Sodium nitrite stock solution (1000 μg NO 2 /ml) Sodium nitrite solution (10 μg NO 2 /ml) Sodium nitrite working solution (1 μg NO 2 /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. Hand Book of Water, Air and Soil Analysis 80

87 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 Preparation of Standards 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 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 C (NO 2 μg/m 3 ) = (As Ab) x CF x Vs/ Va x Vt x 0.82 Where, C NO 2 = Concentration of Nitrogen dioxide (μg/m 3 ) As = Absorbance of sample Ab = Absorbance of reagent blank CF = Calibration factor Va= Volume of air sampled (m 3 ) Hand Book of Water, Air and Soil Analysis 81

88 Vs= Volume of sample (ml) Vt = Volume of aliquot taken for analysis (ml) 0.82 = Sampling efficiency Result-NO x in ambient air was observed µg/m 3. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 82

89 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 The iodine is determined spectrophotometrically by measuring the absorption of tri-iodide ion at 352 nm. The stoichiometry is approximated by the following reaction: O KI + H 2 O --> KI KOH + O 2 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 Reagents / Chemicals All the chemicals should meet specifications of ACS Analytical Reagent grade Distilled water Absorbing Solution (1% KI in 0.1 m Phosphate Buffer) - Dissolve 13.6 gm of potassium dihydrogen phosphate (KH 2 PO 4 ) 14.2 gm of disodium hydrogen phosphate (Na 2 HPO 4 ) or 35.8 gm of the dodecahydrate salt (Na 2 HPO H 2 O) 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 with NaOH orkh 2 PO 4. 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 M I 2 (0.05N) - Dissolve 16 gm of potassium iodide and3.173 gm of resublimed 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 M Na 2 S 2 O 3. Hand Book of Water, Air and Soil Analysis 83

90 The sodium thiosulphate is standardized against primary standard bi-iodate [KH(IO 3 ) 2 ] or potassium dichromate (K 2 Cr 2 O 7 ). M I 2 Solution - Pipette exactly 4.00 ml of the 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 Preparation of Standards Calibrating Iodine Solution - For calibration purposes exactly 5.11 ml of the 0.001M I 2 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 μl of O 3 /ml. Discard this solution after use. Obtain a range of calibration points containing from 1 μl to 10 μl 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 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 C (O 3 μg/m 3 ) = (As Ab) x CF x 1.962/ Va Hand Book of Water, Air and Soil Analysis 84

91 Where, C O 3 = Concentration of Ozone in μg/m 3 As = Absorbance of sample Ab = Absorbance of reagent blank CF = Calibration factor Va = Volume of air sampled in m = Conversion factor, μl to μg Result-The ozone in ambient air was found------µg/m 3. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 85

92 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 Reagents / Chemicals All the chemicals should meet specifications of ACS Analytical Reagent grade 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 NH 3 /ml) Ammonia working solution (10 μg NH 3 /ml) (Prepare fresh daily) Sampling Hand Book of Water, Air and Soil Analysis 86

93 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 Preparation of Standards 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 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 C (NH 3 μg/m 3 ) = (As Ab) x CF / Va Where, C NH 3 = Concentration of Ammonia in μg/m 3 As = Absorbance of sample Ab = Absorbance of reagent blank CF = Calibration factor Va = Volume of air sampled in m 3 Result- The ammonia in ambient air was found------µg/m 3. Precautions- 1. Glassware should be clean. 2. Prepare the standard solution carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 87

94 SOIL ANALYSIS Hand Book of Water, Air and Soil Analysis 88

95 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 20 0 C in 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 Hand Book of Water, Air and Soil Analysis 89

96 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 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 pipette sample Fine sand decantation Depth 28 cm Depth 22 cm Depth 28 cm Depth 10 cm 0 C Minimum Hours Hours Minimum Second /4 24 1/ / / / /2 21 3/4-6 0 Hand Book of Water, Air and Soil Analysis 90

97 /4 21 1/ /4 20 1/ / /2 24 3/ / / /4 18 1/2 23 1/ / /2 17 1/2 22 1/ /4 17 1/ /4 21 1/ / /4-20 1/ /4-19 1/ / /4-28 1/ / /4-17 3/ /2-17 1/ / / 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 Hand Book of Water, Air and Soil Analysis 91

98 oven at C 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 C 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 C 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 C, cool in a desiccator and note the increase in weight (W4) which corresponds to the weight of oven dry fine sand. Hand Book of Water, Air and Soil Analysis 92

99 Observation table- S.No. Weight of 1 st Weight of 2 nd 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. Hand Book of Water, Air and Soil Analysis 93

100 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 C 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/cm 3 ) Calculation- Bulk density (gm/cm 3 ) = Weight of soil (gm)/volume of soil (cm 3 ) Result- The bulk density of given soil was observed gm/cm 3. Precautions- 1. Note the value carefully. 2. Weight the soil exactly. 3. Dry the soil properly. Hand Book of Water, Air and Soil Analysis 94

101 SPECIFIC GRAVITY Object- Determine the specific gravity of given soil. Requirements- Oven, glass bottles, chemical balance. Method- Dry the soil in an oven at C 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) 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- 1. Note the value carefully. 2. Weight the soil exactly. 3. Dry the soil properly. Hand Book of Water, Air and Soil Analysis 95

102 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 C 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) (%) 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- 1. Note the value carefully. 2. Weight the soil exactly. 3. Dry the soil properly. Hand Book of Water, Air and Soil Analysis 96

103 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 C. 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) 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. Hand Book of Water, Air and Soil Analysis 97

104 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 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. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 98

105 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 (µmho/cm) Calculation- Conductivity (µmho/cm) = Selector knob reading x dial reading x cell constant. Result- The electrical conductivity of given soil was observed µmho/cm. Precautions- 1. Prepare soil suspension properly. 2. Calibrate the conductivity meter. 3. Note the reading properly. Hand Book of Water, Air and Soil Analysis 99

106 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 Calculation- Digital instrumental method (no calculation). Result- The redox potential of given soil was observed Precautions- 1. Calibrate instrument properly. 2. Prepare soil solution properly. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 100

107 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 H 2 SO4 (mg/g) burette (ml) burette (ml) 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- 1. Note the reading carefully. 2. Prepare soil solution properly. Hand Book of Water, Air and Soil Analysis 101

108 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. 3. Note the end point carefully. Hand Book of Water, Air and Soil Analysis 102

109 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) 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 the soil carefully. 2. Collect soil sample carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 103

110 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. Hand Book of Water, Air and Soil Analysis 104

111 Calculation- Kjeldhal Nitrogen (mg/l) = T 1 T 2 x N x 14 / W Kjeldhal Nitrogen (%) = T 1 T 2 x N x 1.4 / W Where, T 1 = volume of titrant used against sample (ml) T 2 = 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. 2. Collect soil sample carefully. 3. Note the end point carefully. 4. Prepare solution carefully. Hand Book of Water, Air and Soil Analysis 105

112 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- NO 3 - N (mg/l) = F x V / 1000 x W Where, F = NO 3 - N determine in filtrate (mg/l) V = total volume of filtrate (ml) and W = weight of dried soil/sediment used (gm)/ Hand Book of Water, Air and Soil Analysis 106

113 Result- The nitrate in given soil sample was observed mg/l. Precautions- 1. Weight soil carefully. 2. Collect soil sample carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 107

114 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 = PO 4 -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- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 108

115 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- PO 4 -P (mg/l) = Ps x V / 1000 x W Where, Ps= PO 4 -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- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 109

116 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 x V 1 / V 2 x W Where, T = volume of EDTA titrant used (ml) V 1 = total volume of soil extract (ml) V 2 = 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- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 110

117 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) = T 2 T 1 x x V 1 / V 2 x 1.645x W x Where, T 2 = volume of EDTA titrant used for determination of calcium and magnesium (ml) T 1 = volume of EDTA titrant used for determination of calcium (ml) V 1 = total volume of soil extract (ml) V 2 = 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- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 111

118 SODIUM Object- Determine the sodium in given soil sample. Requirements- Filter paper (Whatman no. 50), all material including reagents, required for determination of sodium 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 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 Where, A = sodium content of soil extract (mg/l) V = total volume of soil extract (ml) and W = weight of air-dry soil/sediment taken for extraction (gm) Result- The total sodium in given soil sample was observed mg/l. Precautions- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 112

119 POTASSIUM Object- Determine the potassium in given soil sample. Requirements- Filter paper (Whatman no. 50), all material including reagents, required for determination of potassium 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 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 Where, A = potassium content of soil extract (mg/l) V = total volume of soil extract (ml) and W = weight of air-dry soil/sediment taken for extraction (gm) Result- The total potassium in given soil sample was observed mg/l. Precautions- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 113

120 IRON Object- Determine the iron in given soil sample. Requirements- Filter paper (Whatman no. 50), all material including reagents, required for 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) V = total volume of suspension (ml) 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- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 114

121 ORGANIC MATTER Object- Determine the organic matter in given soil sample. Requirements- Laboratory glassware, flask etc. Reagents- (a.) Potassium dichromeate solution (1N)- Dissolve 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 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- T 1 /T 2 ] x 10 O M (%) = 6.791/ W [1- T 1 /T 2 ] Carbon (%) = 6.791/ W x [1- T 1 /T 2 ] Where, OM = Organic matter W = weight of soil/sediment taken (gm) Hand Book of Water, Air and Soil Analysis 115

122 T 1 = volume of titrant used against sample (ml) and T 2 = volume of titrant used against distilled water blank (ml) Result- The total organic matter in given soil sample was found mg/l. Precautions- 1. Collect soil sample carefully. 2. Weight soil carefully. 3. Prepare solution carefully. 4. Note the reading carefully. Hand Book of Water, Air and Soil Analysis 116

123 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 Presently she is working as Associate Professor and Head Dept. of Energy & Environment, Faculty of Science & Environment at MGCGV, Chitrakoot Satna MP, since 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, 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; He is playing the role as editorial board member of many international journals. Hand Book of Water, Air and Soil Analysis 117