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1. CHLORINE RESIDUAL COLIFORM RELATIVE STABILITY 1 DISSOLVED OXYGEN TEMPERATURE 9 9 9 PREPARATION OF REAGENTS pH TURBIDITY CHLORINE RESIDUAL COLIFORM T RELATIVE STABILITY 9999 DISSOLVED OXYGEN APPENDICES A APPARATUS AND CHEMICALS B REFERENCES C ALTERNATIVE METHODS 4
2. D DISTILLING AND AUTOCLAVING 1 E CONVERSION TABLE Page f t o 10 10 12 13 14 15 15 16 17 18 19 20 23 24 28 30 INTRODUCTION The field kit is designed to provide quality checks on the water and sewage facilities Seven tests were defined as being appropriate to provide an adequate health index of the water These tests are pH chlorine residual turbidity coliform bacteria relative stability biochemical oxygen demand dissolved oxygen and temperature Objectives of the kit were that it 1 economical and rugged 2 responsive to data needs 3 available for mass distribution 4 usuable by in country skills and 5 that expendables are replace able in country Commercially available test kits such as La Motte or Hach are accurate However they require knowledgeable operators The chemicals had to be ordered out of the country and the cost was well over 250 The kit presented here consists of materials available locally and can be made fresh each day before going into the field Results are adequately accurate and require no special skills or specific knowledge The cost 15 under 100 for the entire kit This manual provides pictures of all operations and equipments needed for each test Measurement in the manual are given in grams and milliliters Howe
3. Fisher Scientific International Division 52 Faden Road Springfield New Jersey 07081 U S Fisher Scientific de Mexico S A Medellin 43 1402 Mexico 7 D F Mexico 22 APPENDIX B REFERENCES Standard Methods for the Examination of Water and Wastewater 13th Ed American Public Health Association 1015 Eighteenth Street N W Washington D C 20036 U S A Shugar Shugar and Bauman Chemical Technicians Ready Reference Handbook McGraw Hill Book Company Manchester Road Manchester Missouri 63011 U S A 23 APPENDIX C ALTERNATIVE METHODS 1 meter 2 Indicators a The following liquid indicators can be used for various titrations INDICATORS Color change Acid Alkaline red yellow red yellow red yellow yellow purple yellow blue yellow red red brown yellow rose brown blue green colorless red colorless blue Name Transformation Interval Thymol Blue 1 2 2 8 Dimethyl yellow 2 9 4 1 Methyl Red 4 4 6 3 Bromcresol purple 5 2 6 8 Bromthymol blue 6 0 7 6 Phenol red 6 4 8 2 Neutral red 6 8 8 0 Naphtholphthalein 7 8 9 0 Phenolphthalein 8 0 9 8 Thymolphthalein 9 3 10 5 Tropeolin 11 1 12 7 yellow orange brown b Cop Cop is the alcoholic blue cabbage extract This extract exhibits the following colors red at pH 2 rose at 4 5 blue at 7 green at 8 yellow at 11 The solution is obtained by extracting blue cabbage cop with 50 60 alcohol The ab
4. nani DM Wed NET p SN agi pe Hance niue py y Water Test Kat I User s Manual APPROPRIATE METHODS OF TREATING WATER AND WASTEWATER IN DEVELOPING COUNTRIES THE UNIVERSITY OF OKLAHOMA BUREAU OF WATER AND ENVIRONMENTAL RESOURCES RESEARCH Sponsored by U S AGENCY FOR INTERNATIONAL DEVELOPMENT WASHINGT 950 3540 51 63 ae WATER TEST KIT I USER S MANUAL o 78 o4 S Bt 63 E 25 WS WA Submitted to Office of Health Agency for International Development Department of State Washington D C 70523 Prepared by George Reid Project Director Regent Professor and Director Bureau of Water and Environmental Resources Research The University of Oklahoma Norman Oklahoma 73069 September 1975 INTRODUCTION TABLE OF CONTENTS SUMMARY OF METHODS STANDARDS AND FREQUENCY OF TESTS ee ee eee eese eo oet nn DATA SHEET OF OU AID WATER amp WASTE WATER TESTING I SET UP OF KIT I 1 CHECKLIST BEFORE GOING TO FIELD TEST METHODS TURBIDITY
5. Shake well BE CAREFUL NaOH pellets are extremely corrosive avoid contact rinse thoroughly with water and consult a doctor if Burn persists Mineral oil used is commercially available Apparatus APPENDIX A 20 APPARATUS amp CHEMICALS Square bottles 30 ml Test tubes 30 ml Dropper bottle 125 ml Dropper bottle 60 ml Glass stoppered bottle Beaker 100 ml Plastic Syringe 10 m1 with needle Measuring spoon 1 0 g spoon A Measuring spoon 0 05 g Spatula stainless steel Thermometer Brush II Chemicals Bromcresol purple B Phenolphthalein C 50 60 alcohol D Fuller s earth E Starch F Potassium Iodide KI Rice potato corn meal or beef bullion H Powdered milk I Methylene Blue J Ferrous sulfate capsule FeSO K Mineral Oil L Sodium Hydroxide NaOH i D i ro be obtained locally not provided in the kit 21 IIT Companies where equipment and chemicals can be obtained Hach Chemical Company P O Box 907 Ames Iowa 50010 U S A Hach Chemical Company Laivsco Mex S de V Presidenie Masaryk No 17 Apartado Postal 17 519 Mexico 17 D F Mexico Sargent Welch International Division 7300 North Linder Avenue Skokie Illinois 60076 U S A Curtin Scientific Company 4220 Jefferson Avenue Houston Texas 77011 U 5 A Curtin De Mexico S de C V Apartado Postal 13265 Mexico 13 D F Mexico
6. a min B Record result in data sheet 14 PREPARATION OF REAGENTS Important Label all containers that has reagents in them 1 prepare bromcresol purple indicator solution Dissolve 2 spoons use spoon B about 0 05 gm of bromcresol purple indicator in the dropper bottle with distilled water water and fill the bottle to the neck B prepare phenolphthalein indicator solution Dissolve 6 spoons use spoon B of phenolphthalein indicator in the dropper bottle with 50 60 alcohol and fill the bottle to the neck Alcohol 2 Turbidity A prepare stock solution Add 1 spoon use spoon B of Fuller s Earth to 50 ml of distilled water This makes a stock solution with a turbi dity of 1 000 JTU spoon of reagent Fill the spoon with one level spoonful of reagent use a sheet of paper to scrap off the excess from the top and the sides Invert the spoon tap the end of the spoon handle to release the powder 15 3 Starch 16 To prepare 50 JTU solution Shake the stock solution well Take 5 ml of stock solution and dilute to 100 ml with dis tilled water This makes the 50 JTU solution standard Preservation Add mercuric chloride a few specks or bleach a few drops to each standard solution Standards must be prepared fresh each month LABEL ALL SOLUTIONS PREPARED Chlorine residual To prepare starch solution Measure out one spoon of
7. over surrounding s water temperature The influent water either domestic water supply or waste water treat ment plant should be tested once every three months All listed tests should be performed This is to check if the condition of the water enter ing the plant has changed and also to compare and see if the treatment is effectively improving the water condition after treatment If the effluent water water coming out of the treatment plant is not meeting exceeds these standards then the treatment plant is perhaps not functioning properly and adjustments should be made in order that these standards can be achieved See page 13 for determination of reduction 3 1 54591 4 ATddng o2r3seuoq 5158901 y N N es es es ese ec P d doo d od te Tt N N CN CN NN O O ooo o o in LE cn dM e 9 2 y A T 5 ueshxo TWOOT NdX Ini 9SAI3 I N WAOJTTO 950 T LIX dO SAVN juenrija 9 4 211 215 quent 2 CI 99 SHHIVM ONILSAL 40 4204105 03 qorwud 3WIL HIIS ul LIN ONILSAL 9 GI
8. FIELD Bromcresol purple and phenolphthalein indicator solutions One clean bottle 25 JTU and 50 JTU standards be sure they are securely capped and not leaking One clean bottle same as those containing the standards Potassium iodide crystals Starch solution in dropper bottle Two clean bottles Fifteen sterilized screw capped bottles each contains 15 ml of media and five drops of bromcresol purple indicator solution One clean One clean 10 ml syringe 100 ml beaker for sample collection One clean Methylene glass stopper bottle blue indicator solution Four clean test tubes One clean 10 ml syringe with clean needle One clean 100 ml beaker for sample collection Sodium hydroxide solution in dropper bottle Methylene blue indicator solution Ferrous sulfate solution preserved under a layer of mineral oil Thermometer See the section about preparation of reagents 8 TEST METHODS 1 A B TABLE 1 Color of solution pH range Yellow lt 6 0 Blue 6 0 8 5 Purplish blue 8 5 9 5 Red gt 9 0 C D Bromcresol Phenolph purple thalein Use a square bottle take 15 ml of water sample Add 1 drop of bromcresol purple solution and 1 drop of phenolphthalein indicator solution Observe closely the color of colution Comment on pH too low OK Still OK Too high Record result in data sheet If having difficulty in identi fying the red color the follow ing st
9. V O VIVA KIT SET UP The kit is divided into three levels The following figures show the set up of top m ddle and bottom levels 1 Top Level A Test tube Square bottle for turbidity test B Square bottle G 50 JTU standard storage bottle C Phenolphthalein solution 25 JTU standard storage bottle D Spoon A I Hole for holding test tube when E Spoon B doing the test 2 Middle Level MAMAS i A Thermometer D Spatula B Square bottle E Syringe C Bottle brush 3 Bottom Level gt s GIO e Gg sj A Methylene blue J Potassium Iodide B FeSO capsules K Dry Milk C Sodium Hydroxide NaOH L Methylene blue solution D Fuller s earth M Mineral oil E Starch N FeSO solution Bromcresol purple O Glass bottle for relative stability Square bottle P Sodium hydroxide solution H 100 ml beaker Q Starch solution I Phenolphthalein R Bromcresol purple solution For ease of identification of apparatus and chemicals a color code system is used as follows Color Code Blue Yellow Green Orange Brown Red Test involved pH Turbidity Chlorine Residual Coliform Bacteria Relative Stability Dissolved Oxygen CHECKLIST pH A B Turbidity Chlorine Residual A B C Coliform A B C Relative Stability A B Dissolved Oxygen Temperature BEFORE GOING TO
10. andard red solution may be prepared Repeat step A but in addition add 5 drops of NaOH solution This should give a standard red color solution for comparison D D 2 Turbidity A B C Fill the sample bottle completely full with water sample Shake the standards by inverting them Compare the sample with the two standards and determine if the sample is less than 25 JTU between 25 50 JTU or greater than 50 JTU Observe movement of particles in solution Record result in data sheet 3 ocr iced 22 10 KI e 3 Starcb 5 min 10 Fill a clean bottle with water sample up to the bottle neck If water sample is turbid with color fill a second bottle as with the first bottle This is for comparison of color in later steps Let the water sample s stand for ten minutes Add two crystals of potassium iodide DO NOT ADD THIS TO THE SECOND BOTTLE Add five drops of starch solution TO BOTH BOTTLES Shake the sample s vigorously and let it stand for five minutes Observe the solut on for change of color change of color intensity upon longer standing should be disregarded Record result in data sheet No color absence of chlorine 0 ppm Faint blue color correct amount of chlorine 0 15 ppm Dark blue color too much gt 0 2 ppm To get better results water sample should be thoroug
11. clean starch with spoon A Add enough cold water and stir to produce a thin paste Add approximately 100 ml of boiling water and keep stirring Boil for 2 3 minutes Add a few drops of chloroform or formal dehyde to preserve the solution Fresh solution should be prepared as often as possible two weeks or less 4 Coliform prepare media Any of the following four methods may be used i Rice Broth Boil 25 grams or fill 1 square bottle full of rice and add 4 spoons use spoon A about 1 gram of powdered milk in 450 ml of water for 5 minutes stir occasionally Decant carefully the rice broth into a glass bottle and discard the rice residue ii Potato Broth Peeled or sliced potatoes or sweet potatoes may be used zm Boil 50 grams of potato in C ov place of the 25 grams of o rice and 4 spoons spoon A Potato Milk 15 one of powdered milk for 15 min then follow the same steps as with the rice broth iii Corn Meal Broth Heat 400 ml of water to 70 C 158 F Add 1 square bottle full of 2 Ten D corn meal and 4 spoons EET use spoon of powdered 0 milk stir frequently Gace Milk Decant carefully the broth meal into a glass bottle and discard the residue 17 iv Lactose Broth Dissolve k of a beef bullion bar 0 approximately 1 gram Ss 4 us 250 ml and 4 spoons spoon A 0 of powdered milk in ee Milk 250 ml of di
12. he rod shall be graduated as follows mark of 100 shall be placed on the rod at a distance of 100 mm from the cen ter of the wire and other graduations made so that 25 Turbidity ppm Vanishing depth of wire mm 10 794 20 426 40 228 60 158 80 122 100 100 150 72 200 57 4 300 43 2 400 35 4 500 30 9 1 000 20 9 2 000 14 8 Chlorine Residual 1 Orthotolidine method refer to Standard Methods Coliform The technique is the same however certain materials can be substituted 1 use durham tubes inverted vials to detect gas production instead of bromcresol purple 2 use pH paper instead of bromcresol purple should end up with a pH of about 5 3 instead of plastic syringe glass syringes or glass pipettes can be used 4 instead of powdered milk and bullion the following normally used media can be applied 8 grams of peptone and beef extract nutrient broth 5 grams of Lactose To be more accurate you can use the confirmed test or selective media refer to Standard Methods 26 E Dissolved Oxygen 1 Winkler method or its various modifications Standard Methods F Relative Stability 1 BOD test refer to Standard Methods or to some source of information on water analysis 27 APPENDIX D DISTILLING amp AUTOCLAVING CLEAR CLEAN SUNLIGHT 1 INVERTED FUNNEL Cox RD TUBING 4 WATER DISTILLED WATER B HEAT n SouRcE B C D E 28 Distilling the bas
13. hly swirled before use To the first group of five bottles with correct amount of media and indicator solution introduce 10 ml of water sample into each by using the syringe BE SURE to record the amount of water sample introduced into each bottle To the second group of five bottles introduce 1 ml of water sample into each To the third group of five introduce 0 1 ml of water sample into each Incubate the bottles at 35 or 95 F for 48 hours Chlorine Residual A Coliform A F After 48 hours observe for color change in the bottles Bottles that have changed from purple to yellow color indicate a positive test Record the number of bottles in each concentration that give positive results G MPN Index most probable number and most positive results i Domestic water supply see Table 2 ii Waste water see Table 3 H Record result in data sheet If MPN is greater than 2 for domestic water supply rerun the test the next day or sooner if possible Table 2 MPN Index for Various Combination of Positive Results For Domestic Water Supply Number of bottles giving positive results MPN Index 10 ml water 1 ml water 0 1 ml water per 100 ml sample sample sample 0 0 0 2 0 0 1 2 0 1 0 2 1 0 0 2 1 0 1 gt 2 1 1 0 gt 2 11 Table 3 MPN Index for Various Combination of Positive Results for wastewater Number of bottles giving positive results MPN Index 10 mi water 1 ml
14. ic idea is to heat the water to its gaseous state through evaporation then condense it and collect it in liquid state This causes the water collected to be free of impurities This can be done in the following ways Evaporation and collection Heating the water to produce steam and then cool the steam to obtain purified water Use of commercially available glass apparatus Use of electric or gas model distilling apparatus Use of de ionizing devices also give water of equivalent quality as distilled water Autoclaving or sterilization The idea here is to heat the mater ial to a high enough temperature using wet or dry heat to kill or inactivate the micro organisms Glassware can be heated in an oven at 1709C for sixty minutes or put in an autoclave or pressure cooker as shown in diagram with fifteen pounds of Wiess re at 1209C for fif teen minutes Media can be steri lized in an autoclave or pressure cooker as above or it can be boiled for fifteen minutes Important 29 Be sure to read the operating manual for the autoclaving and sterilizing devices carefully before any attempt is made to use them APPENDIX E CONVERSION TABLES 16 tablespoons 1 cup or 8 ounces 1 teaspoon 5 ml 1 tablespoon 15 ml 1 cup 1 4 liter l pint 4732 liters u 1 gram 0 0352 ounces 1 ounce avoir 28 35 grams 1 kilogram 2 2046 pounds 1 pound 453 6 grams 1 fluid ounce 29 57 ml 1 q
15. ly in water Fill the four test tubes by pouring from the beaker 15 all water sample along the slanted side of the tube to avoid getting oxygen into the water All possible means must be exercised to avoid trapping excess air To each test tube add five drops of methylene blue indicator solution and five drops of sodium hydroxide solution D Swirl the iron solution gently to mix it evenly E With the needle tip placed under neath the oil layer fill the syringe with the iron ferrous solution Wipe any oil droplet off the needle tip F Discharge the iron solution Slowly down the side of the test tube So as not to introduce additional oxygen into the solu tion 4 ml of the iron solution into the first bottle 5 ml of TO the iron solution into the second bottle 6 ml into the third and 7 ml into the fourth Recap the test tube securely invert each test tube gently to mix the solution H After two minutes observe the color change of these test tubes color change from blue to strav I Refer to the Table 5 for approxi mate dissolved oxygen reading Record result in data sheet Table 5 Dissolved Oxygen Concentration and Color Change Change of Color from Blue to Straw Dissolved Oxygen Comment ppm lst lt 4 Slightly low lst and 2nd 4 OK lst 2nd and 3rd 24 Good lst 2nd 3rd and 4th gt 4 Good 7 Temperature Submerge 1 3 of thermometer stem into the water sample for
16. ove solution should be stored in orange colored bottles c Universal Indicators i methyl red x naphtolphthalein phenolphthalein brom thymol blue and thymolphthalein Mix equal volumes of 24 0 14 solutions of the above five This solution is red at pH of 4 0 yellow at 6 green yellow at 7 green at 8 blue violet at 10 ii 100 mg phenolphthalein 200 mg of methyl red 300 mg of dimethyl yellow 400 mg bromthymol blue 500 mg of thymol blue in 500 ml of alcohol methyl or ethyl with sufficient 0 1 N alkali added to produce a yellow colour pH 6 orange red at pH 4 yellow at 6 yellow green at 7 green at 8 blue at 10 d paper B Turbidity 1 2 Jackson candle refer to Standard Methods St Louis or Baylis Turbidimeter refer to Standard Methods Turbidity Rod calibrated rod attached to one end is a 1 mm diameter platinum wire which is at right angle to the rod About 1 2 meters from this wire a wire ring is attached through which the fine wire is observed when the rod is lowered into the water To obtain the turbidity of a water with this instrument the rod is lowered vertically into the water being tested The observer places an eye immediately above the ring and watches the platinum wire as the rod is being lowered into the water When the wire disappears from sight the position of the surface of the water is read on the calibrated scale of the rod this is read in parts per million T
17. stilled ullion water Heat if necessary B prepare sterilized culture bottles Take 15 clean screw capped bottles Introduce 15 ml of media into each bottle Add 5 drops of bromcresol purple indicator solution to each bottle Sterilize with the cap loosely placed on the mouth of the bottle Let cool slightly tighten the cap 5 Relative Stability To prepare methylene blue indicator solution Place 4 spoons use spoon B of methylene blue indicator in the dropper bottle Fill the bottle with distilled water to the neck Shake to water dissolve the indicator See appendix D for methods of sterilizing or autoclaving 18 6 Dissolved Oxygen A 2 Water Mineral 40 509C FeSO oil B C p E 3 12 pellets D 19 To prepare ferrous sulfate solution Dissolve one capsule of ferrous sulfate sold under the name of Feosol Spansule in 125 ml of distilled water Keep water temperature at 40 509C 100 1209F for effective dissolving Fill the bottle to the neck with warm water then pull the capsule apart to let the pellets inside the capsule to go into the water Cover the prepared solution with a layer of mineral oil To prepare methylene blue indica tor solution See relative stability To prepare sodium hydroxide NaOH solution Dissolve 12 NaOH pellets in a dropper bottle with distilled water Fill the dropper bottle to the neck with distilled water
18. uart 946 milliliters 1 liter 1 06 quarts 1 liter 2 113 pints 1 gallon 3 785 liters Temperature Conversion kelvin C 273 Farenheit 9 5 C 32 Centigrade 5 9 32 30
19. ver all weighings are substituted by the measuring spoons provided The appendices nrovide alterative methods for performing each test along with information on references sterilization and distilling and conversion tables SUMMARY OF METHODS pH Simple application of two indicators bromcresol purple and phenolphthalein Turbidity Basically a comparison of water sample with prepared standards made up of precipitated Fuller s Earth Chlorine Residual Potassium iodide and starch solution are added to the water sample Clear sample reflects absence of chlorine When chlorine is present water sample will turn to a blue color the intensity of which is proportional to the amount of chlorine in the sample Coliform Appl cation of normal MPN most probable number coliform _test series of culture bottles are prepared using media that can be made up by locally available ingredients An indicator bromcresol purple 15 then added to the bottles When coliforms are present pH decreased which induces change of indicator color from purple to yellow will occur after 48 hours of incubation The num ber of coliform present coliform 100 ml can be correlated by the use of MPN table provided Biochemical Oxygen Demand B 0 D is measured by the relative sta bility method which replaces the overly complicated standard B O D test The number of days that are required to use up the oxygen is visually shown by the use of meth
20. water 0 1 ml water per 100 ml sample sample sample lt 240 5 4 2 lt 240 5 4 3 gt 240 5 4 4 gt 240 5 0 240 5 5 1 gt 240 Relative Stability as A Fill the glass stoppered bottle or BOD bottle with water sample and avoid trapped air bubbles by letting water in along one side of the bottle and tap gently when it is full to get rid of any trapped air bubbles B Add 4 drops of methylene blue indicator solution stopper the 4 drops bottle and mix by inverting the Methylene blue bottle C Let the bottle stand at room temperature 20 265 64 D Observe the solution twice day morning and evening until the blue color disappears E Record the number of days required for the color to disappear and then read off the corresponding relative stability percentage shown in Table 4 12 Table 4 Relative Stability Percentage and Days Required for Color Disappearance Relative Stability Percentage e n HO OOoOOug oOuocuotuoocucocuoguocuQgundQgwun o 12 To determine percentage reduction he following equation can be used Effluent Influent X 100 Effluent reduction Record result in data sheet 6 Dissolved Oxygen Methylene blue NaOH 13 Fill the beaker with water sample by carefully and slowly Submerging it complete
21. ylene blue indicator which change from blue to clear upon depletion of oxygen Concen tration of oxygen is determined by using a K rate of 0 171 per day Dissolved Oxygen A ferrous sulfate method is being introduced in this kit Commercially available ferrous sulfate capsules iron anemis are dissolved in water Methylene blue and sodium hydro xide are added to each bottle containing the water sample Differ ent amounts of iron solution are then introduced into each bottle The color of the indicator will turn from blue to straw upon con sumption of oxygen Temperature Thermometer 1 STANDARDS AND FREQUENCY OF TESTS Two lists of tests for testing water from the domestic water supply and from the waste water treatment plant with their respective recom mended standards and suggested frequency for testing are compiled in the following charts Domestic Water Supply Test pH Turbidity Chlorine Residual Coliform Bacteria Waste Water Effluent Test pH Turbidity Coliform Bacteria Relative Stability Dissolved Oxygen Temperature Effluent Frequency once week once week once week once week Frequency once week once week once week once week once week once week Standards 7 9 5 JTU most desir able 25 JTU max permissible 0 15 0 2 ppm 2 MPN 100 ml Standards 6 10 Less than 50 JTU 240 MPN 100 ml 30 reduction No less than 4 ppm Not more than 109

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