Reagent Manual

Author:
Amir Iqbal Manager Quality Control

Reviewed By:
Mr. M. Rafiq CH. General Manager

Approved By:
Dr. M. Akram CH. Chief Executive
 Table of Contents

----------------------------------------------------------------------------------------------------------------

AMMENDMENT 03

1.0 PURPOSE 04

2.0 SCOPE 04

3.0 RESPONSIBILITY 04

4.0 STORAGE 04

5.0 INDICATOR SOLUTION 04

6.0 STANDARD SOLUTION 06

7.0 TEST SOLUTION……………………………………………………...08

8.0 VOLUMETRIC SOLUTION 20

9.0 REFERENCE DOCUMENTS………………………………………..22

 AMENDMENT SHEET

REV. INITIATED PAGE

DATE NATURE OF AMENDMENT DONE BY
# BY #
1.0 PURPOSE

It is established for the preparation of the Laboratory Reagents to ensure

accuracy of Laboratory Analysis of Raw Material and Products.

2.0 SCOPE

It is applicable to the Chemical section of Quality Control Department.

3.0 RESPONSIBILITY

1.3.1 Quality Control Manager

1.3.2 Quality Control Analyst

1.3.3 Quality Assurance Officer

4.0 STORAGE

1.4.1 Paste Reagent identification Slip on the bottle.

1.4.2 This solution is stored in glass bottle.

5.0 INDICATOR SOLUTION

5.1 Mordant Black (II) Triturate

5.1.1 Preparation

A mixture of 1 part of mordant black (II) with 99 parts of

Sodium Chloride.

5.2 Xylenol Orange Triturate

5.2.1 Preparation

Triturate 1 part of Xylenol Orange with 99 parts of Potassium Nitrate.

5.3 Bromothymol Blue Solution R1

5.3.1. Preparation

Dissolve 50 mg of methyl thymol in a mixture of 4 ml of

0.02 M Sodium Hydroxide and 20 ml of 96% Ethanol and
dilute to 100 ml with water. pH range (6.0 - 7.6).

5.4 Bromocresol Green Solution

5.4.1. Preparation
 Dissolve 50 mg of Bromocresol Green in 0.72 ml of 0.1 M
Sodium Hydroxide and 20 ml of 96% ethanol. Add sufficient
water to produce 100 ml. (pH 3.6 yellow - pH 5.2 (blue).

5.5 Crystal Violet Solution

5.5.1 Preparation

Dissolve 0.5 g Crystal Violet in sufficient Anhydrous Acetic Acid

to produce 100 ml.

5.6. Ferroin Sulphate Solution

5.6.1. Preparation

Dissolve 0.7 g of Ferrous Sulphate and 1.76 of 110 Phenanthroline

Hydrochloric in 70 ml of water and add sufficient water to produce 100 ml.

5.7 Litmus Solution

5.7.1. Preparation

Boil 25 g powdered litmus with 100 ml of 90% ethanol water a reflux condenser
for 1 hr. discord the clear liquid and repeat this operation with two 75 ml
quantities of 90% ethanol. Digest or treated litmus with 250 ml of water and filter.

5.8. Methyl Red Solution

5.8.1. Preparation

Dissolve 50 mg of methyl red in a mixture of 1.86 ml of 0.1 M Sodium

Hydroxide and 50 ml of 96% ethanol and add sufficient water to produce 100 ml.

5.9. Methyl Blue Solution 0.1% W/V

5.9.1. Preparation

Dissolve 0.18 of methylene blue in sufficient distilled water to produce 100 ml.

5.10 Methyl Orange Solution

5.10.1 Preparation

Dissolve 1 g of methyl orange in 80 ml of water and add sufficient 96% ethanol to

produce 100 ml.

5.11. Naphthol Benzein Solution

5.11.1. Preparation

Dissolve 0.2 g of 1 - Naphthol Benzein in sufficient anhydrous acetic acid

to produce 100 ml.

5.12. Starch Mucilage

 5.12.1. Preparation

Triturate 0.5 g of starch or soluble starch with 5 ml of water, stirring

continuously and add sufficient water to produce 100 ml. Boil for
a few minutes cool and filter.

5.13. Phenolphthalein Solution Dilute.

5.13.1. Preparation

Dissolve 0.1 g of phenolphthalein in 80 ml of 96% ethanol and add sufficient

distilled water to produce 100 ml.
6.0 STANDARD SOLUTION

6.1. Arsenic Standard Solution (10 ppm)

6.1.1. Preparation

Dilute 1 volume of Arsenic standard solution (10 ppm ) to 10 volumes with water
immediately before use.

6.2. Calcium Standard Solution (100 ppm)

6.2.1 Preparation

Dissolve 2.5 g of dried calcium carbonate in 12 ml of 5 M acetic acid and

dilute to 1000 ml with water. Dilute 1 ml of this solution to 10 ml with
96% ethanol immediately before use.

6.3. Calcium Standard Solution (10 ppm)

6.3.1. Preparation

Dissolve 0.624 g of dried calcium carbonate in distilled water containing

ml of 5 M acetic acid and dilute to 250 ml with water. Dilute 1 volume
of this solution to 100 volume with distilled water immediately before use.

6.4. Chloride Standard Solution (5ppm)

6.4.1. Preparation

Dissolve 82.4 mg of sodium chloride in 100 ml distilled water. Dilute

1 ml volume of this solution to 100 volume immediately before use.

6.5. Iron Standard Solution (1 ppm )

6.5.1. Preparation

Dilute 1 volume of Iron standard solution (20ppm) to 20 volume with water

immediately before use.

6.6. Iron Standard Solution (60 Ppm)

6.6.1. Preparation
 Dissolve 0.863 g of ammonium iron (iii) solution in water containing
25 ml of 1 M sulfuric acid and add sufficient distilled water to produce 500 ml.
Dilute 1 volume immediately before use.

6.7. Lead Standard Solution (100 ppm)

6.7.1. Preparation

Dilute 1 volume of lead standard solution (0.01% Pb) to 10 volume

with water immediately before use. For lead standard solution 0.1% Pb .
dissolve 0.400 g of led (II) nitrate in sufficient water to produce 250 ml.

6.8. Lead standard solution (10ppm)

6.8.1. Preparation

Dilute 1 volume of lead standard (100 ppm) to 10 volume with water immediately
before use. Dilute 159.8 g of lead nitrate in 100 ml of water to which has beach
added 1 ml nitric acid, then dilute with water to 1000 ml.

6.9. Lead Standard Solution (1ppm)

6.9.1. Preparation

Dilute 1 volume of lead standard solution (1 ppm) to 10 volume with water

immediately before use.

6.10. Lead Standard Solution (20 ppm)

6.10.1. Preparation

Dilute 0.80 of lead (II) nitrate in water containing 2 ml of nitric acid and add sufficient
volume to produce 250 ml. Dilute 1 volume to 100 volume with water immediately before
use.

6.11. Lead Standard Solution (2ppm)

6.11.1. Preparation

Dilute 1 volume of lead standard solution (10 Ppm Pb) to 5 volume with water
immediately before use.

6.12. Nitrate Standard Solution (2ppm)

6.12.1. Preparation

Dilute 2 ml of Nitrate Standard (2 ppm) with to 100 ml with distilled water.

6.13. Phosphate Standard Solution (5ppm )

6.13.1. Preparation
 Dissolve 71.6 mg of potassium dihydrogen arthophosphate in sufficient water to
produce 100 ml. Dilute 1 volume of the above solution to 100 volume with water
immediately before use.

6.14. Sulphate Standard Solution (100 ppm)

6.14.1. Preparation

Dissolve 81 g of potassium sulphate in 100 ml of water.Dilute 1 ml of this solution

to 10 ml with water immediately before use.

6.15. Sulphate Standard Solution (10 ppm)

6.15.1. Preparation

Dilute 1 volume of a 0.181% w/v solution of Potassium Sulphate in ethanol (30%)

to 100 volume with ethanol (30%) immediately before use.

7.0 TEST SOLUTION

7.1 Ammonium Iron (III) Sulphate Solution. (1.5 % w/v)

7.1.1 Preparation

Dissolve 1.5 g in sufficient 0.25 M Sulphuric Acid to produce 100 ml. ( For 0.25
Sulphuric Acid add 14 ml of Sulphuric Acid to 25 ml of water and then make the
volume upto 1000 ml.

7.2 Acetate Buffer pH 3.5

7.2.1 Preparation

Dissolve 25 g of ammonium acetate in 25 ml of water add 38 ml of 7 m

hydrochloric acid. Adjust the ph to 3.5 with either 2 M hydrochloric acid or 6 M
ammonia and dilute to 100 ml with water.

7.3 Acetic Acid 2 M

7.3.1 Preparation

Dilute 12 g (11.42 ml) of gracias acetic acid to make upto 100 ml with distilled
water.

7.4. Acetic Acid 5 M

7.4.1 Preparation

Dilute 30 g or (28.57 ml) of glacial acetic acid with sufficient distilled water to
produce 100 ml.
7.5. Acetic Acid 6 M :

7.5.1 Preparation

Dissolve 36 g (34.28 ml) of glacial acetic acid with sufficient water to produce
100 ml.

7.6 Acetic Anhydride Solution R1 :

7.6.1 Preparation

Dissolve 21 ml of acetic anhydride in anhydrous pyridine and dilute to

100 ml with anhydrous pyridine. Protect for light and air.

7.7. Ammonia 10 N.

7.7.1 Preparation

Dilute 60-47 ml of 32% (or 16.535 m) ammonia with make to make upto
100 ml.

7.8 Ammonia Solution 2 M.

7.8.1. Preparation

Dilute 120 ml of 32% (16.53 M) ammonia to 1000 ml with distilled

water.

7.9. Ammonia 5 M.

7.9.1 Preparation

Dilute 30.24 ml of 16.53 (32%) ammonia in sufficient distilled water

to produce 100 ml.

7.10 Ammonium Chloride Solution (10.7% W/V).

7.10.1 Preparation

Dissolve 10.7 g of ammonium chloride in sufficed water to produce 100

ml.

7.11. Ammonium Iron (iii) Sulphate Solution R2 (10% W/V).

7.11.1 Preparation

Dissolve 10 g in sufficient distilled water to produce 100 ml filter before

use.
 7.12 Ammonium Iron (III) Sulphate Solution R6.

7.12.1 Preparation

Dissolve 20 g of ammonium iron sulphate in 75 ml of water add 10 ml of

a 2.8% w/v solution of sulphuric acid (96% w/v) and dilute to 100 ml with
water.

7.13 Ammonium Molybdate T.S (10% W/V).

7.13.1 Preparation

Dissolve 10 g of ammonium molybdate in sufficient distilled water to

produce 100 ml and filter.

7.14 Ammonium Sulphamate Solution (T.S) (5% W/V).

7.14.1 Preparation

Dissolve 5 g Ammonium sulphamate in sufficient distilled water to

produce 100 ml.
.
7.15 Ammonium Thiocyanate (10% W/V).

7.15.1 Preparation

Dissolve 10 g of Ammonium thiocyanate in sufficient distilled water to

produce 100 ml.

7.16 Ammonium Thiocyanate (8% W/V).

7.16.1 Preparation

Dissolve 8.0 g of Ammonium thiocyanate in sufficient distilled water to

produce 100 ml.

7.17 Acetic Acid 1 N

7.17.1 Preparation

Dilute 6 g (5.71 ml) of glacial acetic acid to make upto 100 ml with
distilled water.

7.18 Acetic Acid - Ammonium Acetate Buffer (pH 6)

7.18.1 Preparation

Dissolve 100 g of ammonium acetate in 300 ml of water, add 4.1 ml of

glacial acetic acid, adjust the pH, if necessary using 10 M ammonia or
5M acetic acid and dilute of 500 ml with water.
 7.19 Ammonia Solution Dilute R1

7.19.1 Preparation

Dilute 41 g ( 46.60 ml) of Concentrated Ammonia (13.5 M) to make

upto 100 ml with distilled water.

7.20 Barium Chloride Solution (25 %w/v)

7.20.1 Preparation

Dissolve 25 g of barium chloride in sufficient water to produce 100 ml.

7.21 Barium Chloride Solution (2% W/V).

7.21.1 Preparation

Dissolve 2 g of barium chloride in sufficient distilled water to produce

100 ml.

7.22 Barium Chloride Solution R1 (6.1% W/V)

7.22.1 Preparation

Dissolve 6.1 g of barium chloride in sufficient water to produce 100 ml.

7.23 Bromine Solution (1 % in Glacial Acetic Acid)

7.23.1. Preparation

Dilute 1 ml of bromine with sufficient Glacial Acetic Acid to produce 100

ml.

7.24 Cetrimide Solution (1% w/v)

7.24.1 Preparation

Dissolve 1 g in sufficient distilled water to make up the volume upto 100

ml.

7.25 Citric Acid Solution 20% w/v

7.25.1 Preparation

Dissolve 20 g in sufficient distilled water to make volume upto 100 ml.

7.26 Cobalt Nitrate (II) Solution 10% w/v

7.26.1 Preparation

Dissolve 10 g in sufficient distilled water to make volume upto 100 ml.

7.27. Cobalt Uranyl Acetate

 7.27.1 Preparation

 Dissolve with warming 40 g of uranyl acetate in a mixture of

30 g of glacial acetic acid and sufficient water to make 500 ml.
 Separately prepare a solution containing 200 g of cobaltous
acetate in mixture of 30 g of glacial acetic and add sufficient
water to make 500 ml.
 Mix the two solution still warm and coot to 20oC. Maintain the
temperature at 20° C for 2 hours to separate the excess salt
from solution and then filter through dry filter.

7.28. Cobalt(II) Acetate Solution (0.2% w/v).

7.28.1 Preparation

Dissolve 0.2 g of Cobalt(II) Acetate in sufficient water to make

volume upto 100 ml.

7.29 Cupric Tartrate Solution Alkaline (Fehling Solution a& B)

7.29.1. Preparation

Solution A:

Dissolve 34.6 g of Copper (II) Sulphate in a mixture of 0.5 ml of Sulphuric Acid &
sufficient water to make volume upto 500 ml.

Solution B:

Dissolve 176 g of Potassium Sodium Tartrate and 77 g of Sodium Hydroxide in

sufficient water to make volume upto 500 ml.

Mix equal volumes of solution A & B immediately before use.

7.30 Disodium Tetraborate Solution 19% w/v

7.30.1. Preparation

Dissolve 19 g in sufficient distilled water to produce 100 ml. Protect for light
and air.

7.31 Diphenyl Carbazide Solution

7.31.1. Preparation

Dissolve 0.2 g of Diphenyl Carbazide in 10 ml of glacial acetic acid and dilute to

100 ml with Absolute Ethanol.
7.32 Disodium Hydrogen Phosphate Solution

7.32.1. Preparation

Dissolve 9 g of Disodium Hydrogen Orthophosphate in sufficient distilled water to

make up the volume upto 100 ml.

7.33. Diphenylamine Solution(0.1 % w/v)

7.33.1 Preparation

Dissolve 0.1 g of Diphenyl amine in sufficient Sulphuric Acid to make up the

volume upto 100 ml.

7.34. Ferric Chloride Solution 0.5% w/v

7.34.1. Preparation

Dissolve 0.5 g of Ferric chloride in sufficient distilled water to make upto the
volume upto 100 ml.

7.35 Fuchsin Solution (Decolorized).

7.35.1. Preparation

Dissolve 0.1 g of basic fuchsin in 60 ml of water and add 1 g of anhydrous

sodium sulfite dissolved in 10 ml of water. Slowly add 2 ml of hydrochloric acid
stirring continuously and dilute to 100 ml with water allow to stand protected for
light for at test 12 hors. Shake with sufficient activated charcoal (0.2-0.3 g) to
remove the colour and then filter immediately. If solution becomes cloudy filter
before use. If on standing solution becomes violet decolorize again by adding
charcoal.

7.36 Formaldehyde T.S

7.36.1. Preparation

Analytical reagent grade of commerce containing not less than 34% w/v and not
more than 37% w/v of formaldehyde.

7.37. Glacial Acetic Acid 2.4% w/v

7.37.1 Preparation

Dilute 2.4 g (2.4 ml) of glacial acetic acid with distilled water to make
up the volume upto 100 ml.

7.38. Hydrochloric Acid Solution 0.02 M

7.38.1. Preparation

Dilute 1.7 ml of 37% (11.76 M) HCl with sufficient distilled water to make
up the volume upto 1000 ml.
 7.39. Hydrochloric Acid Solution (3 M).

7.39.1 Preparation

Dilute 25.5 ml of 37% Hydrochloric Acid (11.5 M) with distilled water to

make up the volume 100 ml with distilled water.

7.40 Hydrochloric Acid Solution 10%.

7.40.1 Preparation

Dilute 27.03 ml of 37% HCl with distilled water to make up the volume
upto 100 ml.

7.41 Hydrochloric Acid Solution 6 M.

7.41.1 Preparation

Dilute 51.0 ml of conc hydrochloric acid with distilled water to make up the
volume upto 100 ml.

7.42. Hydrochloric Acid Solution 7 M.

7.42.1 Preparation

Dilute 59.5 ml of 37% Hydrochloric Acid with distilled water to make

up the volume upto 100 ml.

7.43 Hydrochloric Acid R1 25% w/v.

7.43.1 Preparation

Dilute 70 g (59.32 ml) of concentrated Hydrochloric Acid with distilled water to make
volume upto 100 ml. It contains 25% w/v of Hydrochloric Acid.

7.44 Lead Acetate Solution 9.5% w/v

7.44.1 Preparation

Dissolve the 9.5 g of lead (ii) acetate in sufficient CO 2 free distilled

water to make volume upto 100 ml.

7.45 Magnesia Mixture

7.45.1 Preparation

Dissolve 5.5 g of magnesium chloride and 7.0 g of ammonium chloride in 65 ml

of water and 35 ml of ammonia T.S. set the mixture aside for a few day in well
stopper bottle and filter. The solution is not perfectly clear filter it before using.

7.46 Nitric Acid 2 M.

 7.46.1 Preparation

Dilute 12.6 ml of cone nitric acid (70%) with sufficient distilled water
to make upto volume upto 100 ml.

7.47. Nitric Acid Dilute 12.5% w/v

7.47.1. Preparation

Dissolve 01 g in sufficient distilled water to produce 100 ml.

7.48 Phosphate Buffer pH 4.4

7.48.1 Preparation

Dissolve 5.04 g of disodium hydrogen orthophosphate and 3.01 g

of potassium dihydrogen orthophosphate in sufficient water to
produce 1000 ml and adjust the pH with glacial acetic acid.

7.49 Potassium Ferrocyanide Solution 5.3% w/v

7.49.1 Preparation

Dissolve 5.3 g of potassium hexacyanoferrate (ii) in sufficient distilled

water to make the volume upto 100 ml.

7.50 Potassium Ferricyanide Solution 5% w/v

7.50.1. Preparation

Wash 5 g of potassium hexacyanoferrate with a little water and dilute in sufficient

water to produce 100 ml.

Prepare immediately before use.

7.51. Potassium Hydroxide Solution (Ethanolic)

7.51.1. Preparation

Solution of required molarity may be obtained by dissolving appropriate amount

of potassium Hydroxide in sufficient 96% ethanol to produce 1000 ml.

7.52. Potassium Periodate Solution (2.14% w/v)

7.52.1. Preparation

Dissolve 2.14 g in sufficient distilled water to make up a volume of 100 ml.

7.53. Potassium Permanganate Solution 0.32% w/v

7.53.1. Preparation

Dissolve 0.32 of potassium permanganate in sufficient distilled water

to make the volume up to 100 ml.

7.54. Potassium Permanganate Solution Dilute (1% w/v)

7.54.1. Preparation

Dissolve 1 g of potassium permanganate solution in sufficient distilled water

to produce 100 ml.

7.55. Potassium Permanganate Solution (3% w/v)

7.55.1. Preparation

Dissolve 3 g of Potassium Permanganate in sufficient distilled water

to make the volume upto 100 ml.

7.56. Potassium Pyroantimonate Solution

7.56.1. Preparation

Dissolve 2 g of Potassium Pyroantimonate in 95 ml of hot water, cool rapidly

and add a solution containing 2.5 g of potassium hydroxide in 50 ml of water
and 1 ml of 2 M sodium hydroxide. Allow to stand for 24 hours filter and
dilute to 150 ml with water.

7.57. Sodium Nitroprusside Solution (2.0%)

7.57.1. Preparation

Dissolve 2.0 g of sodium Nitroprusside in sufficient distilled water to produce

100 ml.

7.58. Sodium Hydroxide Solution (0.05 M) or (0.2%)

7.58.1. Preparation

Dissolve 2.0 g of sodium hydroxide in sufficient water to produce a total

volume of 100 ml.

7.59. Sodium Hydroxide Solution (10 M) or (40%)

7.59.1. Preparation
 Dissolve 40.0 g of sodium hydroxide in sufficient water to produce a total
volume of 100 ml.

7.60. Sodium Hydroxide Solution (Dilute) 8.5% w/v

7.60.1. Preparation

Dissolve 8.5 g of sodium hydroxide in sufficient water to produce a total

volume of 100 ml.

7.61. Sodium Hydroxide Solution 0.02 M

7.61.1. Preparation

Dissolve 2.0 g of sodium hydroxide in sufficient water to produce a total

volume of 100 ml.

7.62. Sodium Nitroprusside Solution R (2.5%)

7.62.1. Preparation

Dissolve 2.5 g of sodium Nitroprusside solution in sufficient distilled water

to make the volume up to 100 ml.

7.63. Sodium Nitroprusside Solution (1% w/v)

7.63.1. Preparation

Dissolve 1 g of sodium nitroprusside in sufficient distilled water to produce a total volume

of 100 ml.

7.64. Sodium Periodate Solution (2.14% w/v)

7.64.1. Preparation

Dissolve 2.14 g of sodium periodate in water and add 5 ml of

l.

7.65. Sulphomolybdic Reagent R3

7.65.1. Preparation

Dissolve with battery 2.5 g of ammonia molybdate in 20 ml of distilled water

separately add 28 ml of sulphuric acid to 50 ml of water and cool. Mix two
solutions and dilute to 100 ml with water.

7.66. Sulphuric Acid (10% w/v)

7.66.1. Preparation

Dilute 10 g (5.43 ml) of 98% sulphuric acid with sufficient distilled water
to produce a total volume of 100 ml.
7.67. Sulphuric Acid (20% w/v)

7.67.1. Preparation

Dilute 20 g (10.89 ml) of 98% sulphuric acid with sufficient distilled water
to produce a total volume of 100 ml.

7.68. Sulphuric Acid (5% w/v)

7.68.1. Preparation

Dilute 5 g (2.7 ml) of 98% sulphuric acid with sufficient distilled water to
produce a total volume of 100 ml.

7.69. Sulphuric Acid (Dilute)

7.69.1. Preparation

Add 5.5 ml of sulphuric acid (96.0%) to 60 ml of water, allow to cool

add sufficient water to produce 100 ml.

7.70. Sulphuric Acid 0.02 N

7.70.1. Preparation

Dilute 0.06 of 96% sulphuric acid with distilled water to produce a

volume of 100ml.

7.71. Sulphuric Acid 0.5 M (Methanolic)

7.71.1. Preparation

Dilute 2.7 ml of sulphuric acid with sufficient methanol to produce a volume of

100 ml.

7.72. Sulphuric Acid 13% w/v

7.72.1. Preparation

Dilute 13 ml of Concentrated sulphuric acid with sufficient distilled water

to make a volume of 100 ml.

7.73. Sulphuric Acid 40% w/v

7.73.1. Preparation

Dilute 21.7 ml (40 g) of 96% sulphuric acid with sufficient distilled water
to produce a total volume of 100 ml.

7.74. Sulphuric Acid 50% w/v

7.74.1. Preparation

Dilute 50 ml of Concentrated (96%) sulphuric acid with sufficient distilled

water to produce a total volume of 100 ml.
7.75. Sulphuric Acid 96% w/v

7.75.1. Preparation

Use Concentrated sulphuric acid.

7.76. Thioacetamide Reagent

7.76.1. Preparation

Add 1 ml of a mixture of 15 ml of 1 M sodium hydroxide 5 ml of water and

20 ml of glacial (85%) to 0.2 ml of thioacetamide solution heat in water
bath for 20 seconds cool and use immediately.

7.77. Tin (II) Chloride Solution

7.77.1. Preparation

Heat 20 g of tin with 85 ml of HCl until no more hydrogen in enclosed.

Store over an excess of tin protected from air.

7.78. Tin (II) Chloride Solution (AST)

7.78.1. Preparation

Prepared by adding on equal volume of HCl, reducing the original volume

by boiling and filtering through a fine grain filter paper. Pass dry CO 2 free
nitrogen gas through the solution for 5 minutes.

7.79. Xanthydrol 0.02% w/v Solution

7.79.1 Preparation

To 0.2 ml of a 10.0% w/v solution of Xanthydrol in methanol, add 100 ml

of anhydrous acetic acid, and 1 ml of HCl allow to stand for 24 hors.

7.80. Ammonia - Ammonium Chloride Buffer (pH 10)

7.80.1. Preparation

Dissolve 5.4 g of ammonium chloride in 20 ml of water, add 35 ml of

10 M ammonia and dilute to 100 ml with water.

7.81. Sodium Hydroxide (20% Solution)

7.81.1 Preparation

Dissolve 20 g in sufficient distilled water to produce a volume of 100 ml.

7.82. Ethanol (50%)

7.82.1 Preparation
 To make 100 ml of 50% alcohol take 52 ml of 96% ethanol and dilute
with water to make a total volume of 100 ml.

7.83. Phosphate Buffer pH 7

7.83.1. Preparation

Dissolve 6.805 g of potassium dihydrogen orthophosphate in 29.63 if in sodium

hydroxide and dilute with distilled water upto 1000 ml.

7.84. Phosphate Buffer pH 8

7.84.1. Preparation

Dissolve 6.805 g of potassium dihydrogen orthophosphate in 46.80 ml of 1 N

Sodium Hydroxide and dilute the solution upto 1000 ml with distilled water.

7.85. Phosphate Buffer pH 7.6

7.85.1 Preparation

Dissolve 6.805 in 42.8 ml of 1 N sodium hydroxide VS and dilute the resulting

solution up to 1000 ml with distilled water.
8.0 VOLUMETRIC SOLUTION

8.1. 0.1 N Potassium Permanganate Solution

8.1.1 Preparation

Take 3.3 g of potassium permanganate dissolve in 1000 ml of distilled water and

heat on a water bath for 1 hour, cool & filter through sintered glass filter.

Factor:
Each ml of 0.1 N potassium permanganate is equivalent to 5.7 mg of
sodium oxalate

8.2 0.05 N Potassium Permanganate Solution

8.2.1. Preparation

Take 8 g of potassium permanganate dissolve in 1000 ml of distilled water and

heat on a water bath for 1 hour, cool & filter through sintered glass filter.

Factor:
Each ml of 0.05 N potassium permanganate is equivalent to 5.7 mg of sodium
oxalate

STANDARDIZATION OF POTASSIUM PERMANGANATE SOLUTION

Weigh 200 mg Sodium oxalate accurately, previously dried at 110°C to constant

weight and dissolve in 250 ml of water. Add 7 ml of sulphuric acid, heat to 70°C
and titrate against potassium permanganate solution taken in burette, with constant
stirring until a pale pink color, which persists for 15 seconds is produce. The
temperature at the conclusion of titration should be not less then 60°C. Calculate
the normality.

8.3. 0.1 N Hydrochloric Acid Solution

8.3.1. Preparation

Dilute 8.5 ml of conc. Hydrochloric acid (37%) with sufficient distilled water to
produce a volume of 1000 ml.

Factor:
Each ml of 0.1 N Hydrochloric acid is equivalent to 5.30 mg of sodium carbonate.

8.4. 0.5 N Hydrochloric Acid Solution

8.4.1. Preparation

Dilute 42.5 ml of conc. Hydrochloric acid (37%) with sufficient distilled water to
produce a volume of 1000 ml.

Factor:
Each ml of 0.5 N Hydrochloric acid is equivalent to 25.5 mg of sodium carbonate.

STANDARDIZATION OF HYDROCHLORIC ACID SOLUTION

Dissolve 0.1 g of anhydrous sodium carbonate in 20 ml of water, add 0.1 ml

 of methyl orange solution and titrate with hydrochloric acid until the solution
becomes reddish yellow. Boil for 2 minutes and continue titration until reddish
yellow colour is restored. Calculate the normality.

8.5. 0.1 N Sulphuric Acid Solution

8.5.1 Preparation

Dilute 8.8 ml of conc. Sulphuric acid with sufficient distilled water to produce a
volume of 1000 ml.

Factor:
Each ml of 0.1 N Sulphuric acid is equivalent to 5.3 mg of sodium carbonate.

8.6. 2 N Sulphuric Acid Solution

8.6.1 Preparation

Dilute 112 ml of conc. Sulphuric acid with sufficient distilled water to produce a
volume of 1000 ml.

8.7. 0.005 N Sulphuric Acid Solution

8.7.1 Preparation

Dilute 0.28 ml of conc. Sulphuric acid with sufficient distilled water to produce a
volume of 1000 ml.

STANDARDIZATION OF SULPHURIC ACID SOLUTION

Dissolve 0.1 g of anhydrous sodium carbonate in 50 ml of water, add 0.1 ml

of methyl orange solution and titrate with hydrochloric acid until the solution
becomes reddish yellow. Boil for 2 minutes and continue titration until reddish
yellow colour is restored. Calculate the normality.

8.8. 0.1 N Sodium Hydroxide Solution

8.8.1 Preparation

Dissolve 4.2 g of Sodium Hydroxide in sufficient distilled water to produce a

volume of 1000 ml.

Factor:
Each ml of 0.1 N Sodium Hydroxide is equivalent to 0.02042 g of potassium
Hydrogen Phthalate.

8.9. 5 N Sodium Hydroxide Solution

8.9.1 Preparation

Dissolve 21 g of Sodium Hydroxide in sufficient distilled water to produce a volume

of 100 ml.
 8.10. 0.5 N Sodium Hydroxide Solution

8.10.1 Preparation

Dissolve 21 g of Sodium Hydroxide in sufficient distilled water to produce a

volume of 1000 ml.
Factor:
Each ml of 0.5 N Sodium Hydroxide is equivalent to 0.102 g of potassium
Hydrogen Phthalate.

STANDARDIZATION OF SODIUM HYDROXIDE SOLUTION

Dissolve 0.65 g of potassium hydrogen phthalate in 75 ml of carbon dioxide

free water to make up the volume upto 100 ml. Calculate the Normality.

9.0 REFERENCE DOCUMENTS

U.S.P. Vol. I & II

B.P. Vol. I & II (1988)
B.P. Vol. I &II (1998)