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Standard operating procedure (SOP) to prepare the Reagent Solution for chemical analysis in the quality control laboratory

General Reagent Solution Preparation

1.0   Objective

• • To lay down the procedure for the preparation of the general reagent solution.

2.0   Scope

• • This SOP is applicable in the Analytical / Quality Control Laboratory for the preparation of reagent solutions

3.0   Responsibility

• • Chemist QC: To follow the Procedure

• • Head QC: For system compliance

4.0   Procedure for preparation of reagent solution

• General Reagent Solutions : 

  • Acetic Acid, Dilute:

• • Dilute 5.7 ml of glacial acetic acid to 100 ml with water.

• • Hydrochloric Acid, Dilute:

• • Dilute 24 ml of concentrated Hydrochloric Acid acid to 100 ml with water.

• • Nitric Acid, Dilute:

• • Dilute 10.6 ml of concentrated nitric acid to 100 ml with water.

• • Sulphuric Acid, Dilute:

• • Dilute 5.7 ml of concentrated sulphuric acid to 100 ml with water.

• • Ammonia Solution, Dilute:

• • Dilute 42.5 mI of strong ammonia solution (25 %) to 100 ml with water.

• • Sodium Hydroxide Solution:

• • Weigh and dissolve 20 g of sodium hydroxide in 100 ml of water.

• • Sodium Hydroxide Solution, Dilute:

• • Weigh and dissolve 5 g of sodium hydroxide in 100 ml of water.

• • Reagent Solution: Acetic Acid,

• • 2 M: Dilute 11.4 ml of glacial acetic acid to 100 ml with water.

• • 5M: Dilute 28.5 ml of glacial acetic acid to 100 ml with water.

• • Reagent Solution: Hydrochloric Acid

• • 0.2 M: Dilute 8.5 ml of concentrated Hydrochloric Acid to 500 ml with water.

• • 5 M: Dilute 42.5 ml of concentrated Hydrochloric Acid to 100 ml with water.

• • 2 M: Dilute 17 ml of concentrated Hydrochloric Acid to 100 ml with water.

• • Nitric Acid,2 M:

• • Dilute 12.6 ml of concentrated Nitric Acid to 100 ml with water.

• • Reagent Solution: Sulphuric Acid,

• • 0.5 M: Dilute 2.9 ml of concentrated Sulphuric acid to 100 ml with water.

• • 1 M: Dilute 5.7 ml of concentrated Sulphuric acid to 100 ml with water.

• • 0.1 M: Dilute 0.6 ml of concentrated Sulphuric acid to 100.0 ml with water.

• • Reagent Solution: Sodium Hydroxide

• • 0.1M Ethanolic: Prepared by dissolving 2 g of sodium hydroxide in sufficient ethanol to produce 500.0 ml.

• • 0.5M: Prepared by dissolving 2 g of sodium hydroxide in sufficient water to produce 100.0 ml.

• • Reagent Solution: Ammonium Chloride 2 M:

• • Weigh and dissolve 10.698 g of Ammonium Chloride into 100 ml water.

• • Potassium Permanganate,0.02 M:

• • Dissolve 1.6 g of potassium permanganate in 500 ml of water; heat on a water bath for 1 hour allow standing for 2 days and filter.

• • Ferrous Ammonium Sulphate, 0.1 M;

• • Ammonium Iron (ll) Sulphate, 0.1 M; Dissolve 20 g of ferrous ammonium sulfate in a previously cooled mixture of 20 ml of sulphuric acid and 100 ml of water, dilute with sufficient freshly boiled and cooled water to produce 500 ml.

• • Ceric Ammonium Sulphate, 0.1M Ammonium Ceric Sulphate, 0.1 M:

• • Dissolve 32.5 g of ceric ammonium sulphate, with the aid of gentle heat, in a mixture of 15 ml of sulphuric acid and 250 ml of water. Cool, fIlter the solution, if turbid, and dilute to 500 ml with water.

• • Reagent Solution: Disodium Edetate,

• • 0.1 M: Dissolve 18.6 g of disodium edetate in sufficient water to produce 500 ml.

• • 0.01 M: Dissolve 1.86 g of disodium edentate in sufficient water to produce 500 ml.

• • Reagent Solution: Iodine,

• • 0.05 M: Dissolve about 7 g of iodine in a solution of 18 g of potassium iodide in 50 ml of water, add three drops of hydrochloric acid, and dilute with water to 500 ml.

• • 0.02 M: Dissolve about 2.8 g of iodine in a solution of 7.2 g of potassium iodide in 50 ml of water, add three drops of hydrochloric acid, and dilute with water to 500 ml.

• • Reagent Solution: Sodium Arsenite, 0.1 M:

• • Dissolve 4.946 g of arsenic trioxide in 40 ml of 1M sodium hydroxide, dilute to 400 mI with water and add 2 M hydrochloric acids until the solution is neutral to litmus paper. Add 1 g of sodium bicarbonate, dissolve by stirring, and dilute with water to produce 500 mI.

• • Arsenic Standard Solution (10 ppm As):

• • Dissolve 0.33 g of arsenic trioxide in 5 mI of 2M sodium hydroxide and dilute to 250.0 mI with water. Dilute 1 volume of this solution to 100 volumes with water.

• • Arsenic Standard Solution (1 ppm As):

• • Dilute 1 volume of arsenic standard solution (10 ppm As) to 10 volumes with water.

• • Chloride Standard Solution (25 ppm CI):

• • Prepared by dissolving 82.4 mg of Sodium chloride in 100.0 ml of water. Dilute 5 volumes of solution to 100 volumes with water.

• • Chloride Standard Solution (5 ppm CI):

• • Dilute I volume of 0.0824 percent w/v solution of sodium chloride to 100 volumes with water.

• • Iron Standard Solution (20 ppm Fe):

• • Dissolve 172.6 mg of ferric ammonium sulfate in 100 ml of water. Dilute I volume of to 10 volumes with water.

• • Reagent Solution: Lead Standard Solution

• • 0.1 percent Ph:

• • Dissolve 0.4 g of lead nitrate in water containing 2 mI of nitric acid and add sufficient water to produce 250.0 ml.

• • 20 PPM: Dissolve 0.4 g of lead nitrate in water containing 2 mI of nitric acid and add sufficient water to produce 250.0 ml. Dilute 2.0 ml to 100.0 ml with water.

• • 1 ppm: Dilute 5 volumes of lead standard solution (20 ppm Pb) to 100 volumes with water.

• • Sulphate Standard Solution (10ppm S04):

  • Dissolve 181 mg of potassium sulfate in sufficient water and makeup with water to produce 100.0 ml. Dilute 1.0 ml to 100.0 ml with water.

• • Sulphate Standard Solution (10 ppm S04), Ethanolic:

• • Dissolve 181 mg of potassium sulfate in sufficient ethanol (30%) and makeup with ethanol (30%)  to produce 100.0 ml. Dilute 1.0 ml to 100.0 ml with water.

• • Nitrate Standard Solution (0.2 ppm N03):

• • Dilute I volume of nitrate standard solution (10 ppm N03) to 50 volumes with water.

• • Nitrate Standard Solution (10ppm N03):

• • Dissolve 163 mg of potassium nitrate in sufficient water and make up with water to produce 100.0 ml. Dilute 1.0 ml to 100.0 ml with water.

• • Reagent Solution: Mercuric Chloride Paper:

• • Smooth white filter paper, not less than 25 mm in width, soaked in a saturated solution of mercuric chloride, pressed to remove the superfluous solution, and dried at about 60° in the dark.

• • The grade of filter paper is such that the weight is between 65 and 120g per sq.m; the thickness in mm of 400 papers is approximately equal, numerically, to the weight in g per sq.m.

• • Store in a stoppered bottle in the dark. The paper which has been exposed to sunlight or to the vapor of ammonia affords lighter stain or no stain at all when employed in the limit test for arsenic.

• • Lead Acetate Cotton:

• • Immerse absorbent cotton in a mixture of 10 volumes of lead acetate solution and 1 volume of 2 M acetic acid. Drain off the excess liquid by placing it on several layers of filter paper without squeezing the cotton.

• • Allow to dry at room temperature. Store protected from moisture.

• • Reagent Solution: Lead Acetate Solution

• • 10 %: Weigh and dissolve 10 g of Lead Acetate in water to produce 100.0 ml.

• • 20%: Weigh and dissolve 20 g of Sodium Acetate in water to produce 100.0 ml.

• • Sodium Nitrite Solution (2.0%):

• • Weigh and dissolve 2 g of Sodium Nitrite in water to produce 100.0 ml.

• • Sodium Hydroxide Solution (20.0%):

• • Weigh and dissolve 20 g of Sodium Hydroxide in water to produce 100.0 ml.

• • Potassium Iodide Solution (16.6%):

• • Weigh and dissolve 16.6 g of Potassium Iodide in water to produce 100.0 ml.

• • Citric Acid Solution (20%):

• • Weigh and dissolve 20 g of Citric Acid in water to produce 100.0 ml.

• • Ammonium Sulphamate Solution (5%):

• • Weigh and dissolve 5 g of Ammonium Sulphamate in water to produce 100.0 ml.

• • Sodium Periodate Solution:

• • Weigh and dissolve 2.14 g of sodium periodate in water and add 5 ml of 1 M sulphuric acid and sufficient water to produce 100 ml.

• • Mercuric Acetate Solution:

• • Weigh and dissolve 5 g of mercuric acetate in glacial acetic acid to produce 100 ml.

• • Potassium Chloride Solution (10%):

• • Weigh and dissolve 10 g of Potassium Chloride in water to produce 100.0 ml.

• • Potassium Bromide Solution (12.5%):

• • Weigh and dissolve 12.5 g of Potassium Bromide in water to produce 100.0 ml.

• • Barium Chloride Solution (10%):

• • Weigh and dissolve 10 g of Barium Chloride in water to produce 100.0 ml.

• • Boric Acid Solution (4%):

• • Weigh and dissolve 4 g of Boric Acid in water to produce 100.0 ml.

• • Aniline Solution (2.5% v/v):

• • Mix 5 ml of Aniline with 200 of Methanol.

• • Borax Acid Solution (1%):

• • Weigh and dissolve 1 g of Borax in water to produce 100.0 ml.

• • Potassium Ferrocyanide Solution:

• • Weigh and dissolve 5 g of Potassium Ferrocyanide in water to produce 100.0 ml.

• • Ammonium Oxalate Solution:

• • Weigh and dissolve 2 g of Ammonium Oxalate in water to produce 100.0 ml.

• • Silver Nitrate Solution:

• • Weigh and dissolve 5 g of Silver Nitrate in water to produce 100.0 ml.

• • Diphenyl Carbazide Solution:

• • Dissolve 0.2 g of 1,5- Diphenyl Carbazide in  10 ml of glacial acetic acid and dilute to 100 ml with ethanol.

• • Potassium Permanganate Solution (4%):

• • Weigh and dissolve 4 g of Potassium Permanganate in water to produce 100.0 ml, heat if necessary.

• • Potassium Chromate Solution (5%):

• • Weigh and dissolve 5 g of Potassium Chromate in water to produce 100.0 ml.

• • P- dimethylamino benzaldehyde:

• • Weigh and dissolve 0.5 g of P- dimethylaminobenzaldehyde dissolve in a dilute sulphuric acid solution and dilute to 100 ml with the dilute sulphuric acid solution.

• • Sulphuric Acid, Dilute (For Menthol):

• • Dilute 61 ml of Sulphuric acid with 39 ml of water to produce 100 ml.

• • Bromocresol Purple Solution:

• • Dissolve 50mg of bromocresol purple in 0.92 ml of 0.1 M sodium hydroxide and 20 ml of ethanol (95 percent). After the solution is effected, add sufficient water to produce 100 ml.

• • Calcon Mixture:

• • A mixture of 1 part of calcon with 99 parts of freshly ignited anhydrous sodium sulfate.

• • Crystal Violet Solution:

• • Weigh and dissolve 0.5 g of crystal violet in anhydrous glacial acetic acid to produce 100 ml.

• • Dimethyl Yellow-Oracet Blue B Solution:

• • Dissolve 15 mg of dimethyl yellow and 15 mg of oracet blue B in chloroform and dilute to 500 ml with chloroform.

• • Eosin Solution:

• • Dissolve 0.5 g of eosin in water to produce 100 mI.

• • Eriochrome Black T Mixture: Mordant Black 11 Mixture:

• • Prepare a mixture of 0.1 g of eriochrome black and 9.9 g of sodium chloride.

• • Eriochrome Black T Solution; Mordant Black 11 Soluton:

• • Dissolve 0.2 g of eriochrome black T and 2 g of hydroxylamine hydrochloride in sufficient methanol to produce 50 ml.

• • Ferroin Solution; Ferroin Sulphate Solution:

• • Dissolve 0.7 g of ferrous sulfate and 1.5 g of 1,10-phenanthroline hydrochloride in 70 ml of water and add sufficient water to produce 100 ml.

• • Methyl Orange Solution:

• • Dissolve 0.1 g of methyl orange in 80 mI of water and add sufficient ethanol (95 %) to produce 100 mI.

• • Methyl Red Solution:

• • Dissolve 50 mg of methyl red in a mixture of 1.86 ml of 0.1 M sodium hydroxide and 50 ml of ethanol (95 %). After the solution is affected, add sufficient water to produce 100 ml.

• • Phenolphthalein Solution:

• • Weigh and dissolve 1 g of phenolphthalein in ethanol (95 percent) to produce 100 ml.

• • Starch Solution:

• • Weigh 1 g of soluble starch with 5 ml of water and stir continuously, to 100 ml of boiling water containing 10 mg of mercuric iodide.

• • Acetate Buffer pH 3.5:

• • Weigh and dissolve 25 g of ammonium acetate in 25 ml of water and add 38 ml of 7M hydrochloric acid. Adjust the pH to 3.5 with either 2 M hydrochloric acid or 6 M ammonia and dilute with water to 100 ml.

• • Phosphate Buffer pH 4.0, Mixed:

• • Dissolve 5.04 g of disodium hydrogen phosphate and 3.01 g of potassium dihydrogen phosphate in sufficient water to produce 1000 ml. Adjust the pH with glacial acetic acid.

• • Phosphate Buffer pH 4.0:

• • Dissolve 1.15 g of citric acid and transfer and dissolve in a 250 ml beaker with 20 ml of water. Add 11 ml of 1M Sodium Hydroxide and dilute to 50 ml. Add 45 ml of 0.1 M Hydrochloric acid. Adjust pH 4.0 with 0.1 M Hydrochloric acid or 0.1 M Sodium Hydroxide.

• • Phosphate Buffer pH6.8, Mixed:

• • Dissolve 28.80 g of disodium hydrogen phosphate and 11.45 g of potassium dihydrogen phosphate in sufficient water to produce 1000 ml.

• • Phosphate BufferpH6.8, 0.2 M Mixed:

• • Dissolve 13.872 g of potassium dihydrogen phosphate and 35.084 g of disodium hydrogen phosphate in sufficient water to produce 1000 ml.

• • Sulphanilic Acid Buffer Solution:

• • Dissolve 2.5 g of sulphanilic acid and 10 g of anhydrous Sodium Acetate in 40 ml of water and make volume to 200 ml with water.

• • Ammonia-Ammonium Chloride Buffer:

• • Dissolve 67.5 g of ammonium chloride in about 200 ml of water, add 570 ml of strong ammonia solution, and dilute with water to 1000 ml.

• • Thioacetamide Reagent:

• • Add 1 ml of a mixture of 15 ml of 1M sodium hydroxide, 5 ml of water and 20 ml of glycerin (85 per cent) to 0.2 ml of thioacetamide solution, heat in a water-bath for 20 seconds cool and use immediately.

• • Thioacetamide Solution:

• • Weigh and dissolve 4 g of thioacetamide in 100 ml of water

• • Boric Buffer pH 9.0:

• • Dissolve 6.20 g of boric acid in 500 mI of water, adjust to pH 9.0 with 1M sodium hydroxide (about 41.5 mI) and dilute with water to 1000 mI.

• • Ammonia Solution, Dilute:

• • Dilute 425 mI of strong ammonia solution to 1000m with water.

• • Hydrogen Sulphide Solution:

• • Dissolve hydrogen sulphide to saturated solution in water. (Prepare Fresh).

• • Ferric Chloride Solution:

• • Weigh and dissolve 15 g of ferric chloride hexahydrate in to 100 ml water.

• • Diphenylamine Solution:

• • Weigh and dissolve 0.1 g of Diphenylamine in 100 sulphuric acid.

• • Calcium Chloride Solution:

• • Weigh and dissolve 10 g of Calcium Chloride into 100 ml water.

• • Potassium Permanganate Solution:

• • Weigh and dissolve 3 g of Potassium Permanganate into 100 ml water.

• • 1-10 Phenanthroline Solution:

• • Weigh and dissolve 0.1 g of 1-10 Phenanthroline into 100 ml water.

• • Potassium Ferricyanide Solution:

• • Weigh and dissolve 5 g of Potassium Ferricyanide crystals into 100 ml water.

• • Potassium Carbonate Solution:

• • Weigh and dissolve 15 g of Potassium Carbonate in 100 ml water.

• • Potassium Antimonate Solution:

• • Weigh 2 g of Potassium Antimonate and boil with  95 ml of water until dissolved. Cool rapidly and add 50 ml of potassium hydroxide solution and 5 ml of 1M Sodium hydroxide. Allow to stand for 24 hours, filter, and add sufficient water to produce 150 ml.

• • Methoxyphenylacetic acid Reagent:

• • Weigh 2.7 g of methoxyphenylacetic acid in 6 ml of tetramethylammonium hydroxide solution and add 20 ml of ethanol.

• • Barium Chloride Solution:

• • Weigh and dissolve 10 g of Barium Chloride into 100 ml water.

• • Stannous Chloride Solution:

• • Weigh and dissolve 33 g of Stannous Chloride into 10 ml hydrochloric acid and add sufficient water to produce 100 ml.

• • Iodine Solution:

• • Weigh and dissolve 2 g of iodine and 3 g of potassium iodide in water to produce 100 ml

• • Ammonium Acetate Solution:

• • Dissolve 15 g of ammonium acetate in 20 mI of water, add 0.3 ml of glacial acetic acid, and dilute to 100 ml with water.

• • Appearance of Solution

• • Clarity of Solution

• • Standard Suspension:

• • Dissolve 1.0 g of hydrazine sulfate in sufficient water to produce 100.0 ml and set aside for about 6 hours.

• • To 25.0 ml of this solution add 25.0 ml of a 10.0 percent w/v solution of hexamine, mix well, and allow to stand for 24 hours.

• • Keep in a glass container with a smooth internal surface in which the suspension does not adhere to the glass. Store in this manner, the suspension is stable for about 2 months.

• • Prepare the standard suspension by diluting 15 ml of the well-mixed suspension to 1000 ml with water. The standard suspension should be used within 24 hours of preparation.

• • Opalescence Standards:

• • Prepare opalescence standards by mixing aliquots of the standard suspension with water as indicated in Table 1. Each opalescence standard should be shaken well before use.

Table 1

Opalescence Standard	Standard Suspension (ml)	Water (ml)
OS1	5.0	95.0
OS2	10.0	90.0
OS3	30.0	70.0
OS4	50.0	50.0

• • Method of Preparation :

• • Transfer to a flat-bottom test tube of neutral glass, 15 to 25 mm in diameter, a suitable volume of the solution under examination such that the test tube is filled to a depth of 40 mm. Into another matched test tube add the same volume of the freshly prepared opalescence standard. After 5 minutes, compare the contents of the test tubes against a black background by viewing under diffused light down the vertical axis of the tubes.

• • Clarity or opalescence:

• • Express the degree of opalescence in terms of the opalescence standard. A liquid is considered clear if its clarity is the same as that of water or of the solvent used for preparing the solution under examination or if its opalescence is not more than that of opalescence standard OS1.

• • Color of Solution

• • Special Reagents

• • Ferric Chloride Colorimetric Solution (FCS):

• • Dissolve about 46 g of ferric chloride hexahydrate in enough of a mixture of 25 ml of hydrochloric acid and 975 ml of water to produce 1000 ml.

• • Standardization procedure of Opalescence:

• • Pipette 10 ml of this solution into a 250 ml iodine flask, add 15 ml of water, 3 g of potassium iodide, and 5 ml of hydrochloric acid, and allow the mixture to stand for 15 minutes.

• • Dilute with 100 ml of water and titrate the liberated iodine with 0.1 M sodium thiosulphate using 0.5 ml of starch solution, added towards the end of the titration, as the indicator.

• • Carry out a blank titration. I ml of 0.1 M sodium thiosulphate is equivalent to 0.02703 g of FeCI3,6H₂0. Adjust the final volume of the solution by the addition of enough of the mixture of hydrochloric acid and water so that each ml contains 0.045 g of FeCI3, 6H20.

• • The solutions should be stored and protected from light.

• • Cobaltous Chloride Colorimetric Reagent Solution (CCS):

• • Dissolve about 60 g of cobaltous chloride in enough of a mixture of 25 ml of hydrochloric acid and 975 ml of water to produce 1000 ml.

• • Standardization procedure:

• • Pipette 5 ml of this solution into a 250 ml iodine flask, add 5ml of hydrogen peroxide solution (10 volume) and 15 ml of sodium hydroxide solution, boil for 10 minutes, cool, and add 2 g of potassium iodide and 60 ml of dilute sulphuric acid.

• • Dissolve the precipitate by gentle shaking, if necessary, and titrate the liberated iodine with 0.1 M sodium thiosulphate using 0.5 ml of starch solution, added towards the pink endpoint, as the indicator.

• • Carry out a blank titration: 1 ml of 0.1 M sodium thiosulphate is equivalent to 0.02379 g of CoCh,6H20.

• • Adjust the final volume of the solution by the addition of enough of the above mixture of hydrochloric acid and water so that each ml contains 0.0595 g of CoCI2, 6H20.

• • Cupric Sulphate Colorimetric Reagent Solution (CSS):

• • Dissolve about 63 g of cupric sulfate in enough of a mixture of 25 ml of hydrochloric acid and 975 ml of water to produce 1000 ml.

• • Standardization procedure:

• • Pipette 10 ml of this solution into a 250 ml iodine flask, add 40 ml of water, 4 ml of acetic acid, 3 g of potassium iodide, and 5 ml of hydrochloric acid, and titrate the liberated iodine with 0.1 M sodium thiosulphate using 0.5 ml of starch solution, added towards the pale brown end-point, as an indicator.

• • Carry out a blank titration. 1 ml of 0.1 M sodium thiosulphate is equivalent to 0.02497g of CuS04,5H20.

• • Adjust the final volume of the solution by the addition of enough of the above mixture of hydrochloric acid and water so that each ml contains 0.0624 g of CuS04,5H20.

• • Reference Solution:

• • Prepare by mixing the volumes of colorimetric solutions and hydrochloric acid (1 percent w/v HCl) as indicated in Table 2.

• • NOTE – Reference solutions must be prepared immediately before use from the Colorimetric solutions which may be stored in the refrigerator. The solutions should be stored and protected from light.

• • Method:

• • Transfer to a flat bottom test tube of neutral glass 15 to 25 mm in diameter, a suitable volume of a liquid been examined such that the test tube is filled to a depth of 40 mm. Into another matched test tube add the same volume of water or of the solvent used for preparing the solution being examined or of the reference solution stated in the individual monograph.

• • Compare the colors in diffused light viewing vertically against a white background.

• • Colorless Solution:

• •  A solution is considered colorless if it has the same appearance as water or the solvent used for preparing the solution or is not more intensely colored than reference solution BS8.

Table 2

Colour of reference  solution	Reference solution	FCS (ml)	CCS (ml)	CSS (ml)	Hydrochloric Acid (l per cent w/v HCl) (ml)
Yellow	YSI	24.0	6.0	0	70.0
	YS2	18.0	4.5	0	77.5
	YS3	12.0	3.0	0	85.0
	YS4	6.0	1.5	0	92.5
	YS5	3.2	0.8	0	96.0
	YS6	1.6	0.4	0	98.0
	YS7	0.8	0.2	0	99.0
Greenish Yellow	GYS1	24.0	0.5	0.5	75.0
	GYS2	14.0	0.1	0.1	85.8
	GYS3	8.5	0.05	0.05	91.4
	GYS4	5.0	0.05	0.05	94.9
	GYS5	5.8	0.05	0.05	194.1
	GYS6	2.9	0.05	0.05	197.0
	GYS7	2.9	0.05	0.05	397.0
Brownish Yellow	BYS1	24.0	10.0	4.0	62.0
	BYS2	18.0	7.5	3.0	71.5
	BYS3	12.0	5.0	2.0	81.0
	BYS4	6.0	2.5	1.0	90.5
	BYS5	3.0	1.5	0.5	95.0
	BYS6	1.5	0.8	0.2	97.5
	BYS7	1.0	0.4	0.1	98.5
Brown	BS1	22.5	22.5	18.0	37.0
	BS2	15.0	15.0	12.0	58.0
	BS3	11.2	11.2	9.0	68.6
	BS4	7.5	7.5	6.0	79.0
	BS5	3.7	3.7	3.0	89.6
	BS6	1.5	1.5	1.2	95.8
	BS7	0.8	0.8	0.6	97.8
	BS8	0.4	0.4	0.2	99.0
Red	RS1	1.0	20.0	0	70.0
	RS2	7.5	15.0	0	77.5
	RS3	5.0	10.0	0	85.0
	RS4	3.8	7.6	0	88.5
	RS5	2.5	5.0	0	92.5
	RS6	1.3	2.6	0	96.1
	RS7	0.5	1.0	0	98.5

• •  Reagent solution shall be used for up to 3 months or as per the shelf life defined in a pharmacopeia.

• • The reagent solution which is not defined in the SOP shall be prepared as per the procedure defined in the Pharmacopoeia.

Annexure – Reagent Solution Preparation

Annexure I:  General format for the procedure of reagent solution preparation.

General format for procedure of reagents preparation

Reagent Name:

Batch  No:

Reference:

Procedure:

Prepared By                                                                                      Checked By

Date                                                                                                   Date

• Reference (S)
• Indian Pharmacopeia.