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AppendicesWhen a method, test or other matterdescribed in an appendix is invoked ina monograph reproduced from theEuropean Pharmacopoeia, Part III ofthe general notices applies.When amethod, test or other matter describedin an appendix is invoked in any othermonograph, Part II of general noticesapplies.

Contents of the Appendices

APPENDIX I

A. General Reagents A

B. Volumetric Reagents and Solutions A

C. Standard Solutions A

D. Buffer Solutions A

E. Chemical and Biological Reference Materials A

APPENDIX II

A. Infrared Spectrophotometry and Near Infrared Spectrophotometry A

B. Ultraviolet and Visible Spectrophotometry A

C. Nuclear Magnetic Resonance Spectrometry A

D. Atomic Spectrophotometry: Emission and Absorption A

E. Fluorescence Spectrophotometry A

F. X-Ray Fluorescence Spectrometry A

G. Mass Spectometry A

APPENDIX III

Chromatographic Separation Techniques

A. Thin-layer Chromatography A

B. Gas Chromatography A

C. Size-exclusion Chromatography A

D. Liquid Chromatography A

E. Paper Chromatography A

F. Electrophoresis A

G. Capillary Electrophoresis A

H. Supercritical Fluid Chromatography A

APPENDIX IV

A. Clarity of Solution A

B. Colour of Solution A

APPENDIX V

Determination of:

A. Melting Point A

B. Freezing Point A

C. Distillation Range A

D. Boiling Point A

E. Refractive Index A

73-2

F. Optical Rotation and Specific Optical Rotation A

G. Weight per Millilitre, Density, Relative Density and Apparent Density A

H. Viscosity A

J. Circular Dichroism A

K. Reaction of Indicators A

L. pH Values A

M. Thermal Analysis A

N. Osmolality A

O. Conductivity A

P. Total Organic Carbon in Water for Pharmaceutical Use A

Q. Density of Solids A

APPENDIX VI

Qualitative Reactions and Tests A

APPENDIX VII

Tubes for Comparative Tests A

Limit Test for:

Aluminium A

Ammonium A

Arsenic A

Calcium A

Chlorides A

Fluorides A

Heavy Metals A

Iron A

Lead in Sugars A

Magnesium A

Magnesium and Alkaline-earth Metals A

Nickel in Hydrogenated Vegetable Oils A

Nickel in Polyols A

Phosphates A

Potassium A

Sulphates A

APPENDIX VIII

A Non-aqueous Titration A

B. Amperometric and Potentiometric Titrations A

C. Oxygen-flask Combustion A

D. Complexometric Titrations A

E. Ion-selective Potentiometry A

F. Determination of Ethanol A

G. Determination of Methanol and Propan-2-ol A

H. Determination of Nitrogen A

J. Tetrazolium Assay of Steroids A

K. Vacant

L. Residual Solvents A

M. Residual Ethylene Oxide and Dioxan A

N. Determination of N,N-Dimethylaniline A

O. Determination of 2-Ethylhexanoic Acid A

P. Total Protein Assay A

Q. Acetic Acid in Synthetic Peptides A

APPENDIX IX

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Determination of:

A. Sulphated Ash A

B. Sulphur Dioxide A

C. Water A

D. Loss on Drying A

E. Carbon Monoxide in Medicinal Gases A

F. Carbon Dioxide in Medicinal Gases A

G. Nitrogen Monoxide and Nitrogen Dioxide in Medicinal Gases A

H. Oxygen in Medicinal Gases A

J. Water in Medicinal Gases A

K. Impurities in Gases using Gas Detector Tubes A

APPENDIX X

A. Acetyl Value A

B. Acid Value A

C. Ester Value A

D. Hydroxyl Value A

E. Iodine Value A

F. Peroxide Value A

G. Saponification Value A

H. Unsaponifiable Matter A

J. Determination of Cineole A

K. Determination of Aldehydes A

L. Oxidising Substances A

M. Volatile Oils A

N. Fixed Oils A

APPENDIX XI

A. Total Solids A

B. Ethanol-soluble Extractive A

C. Swelling Index A

D. Foreign Matter A

E. Volatile Oil in Drugs A

F. Continuous Extraction of Drugs A

G. Complete Extraction of Alkaloids A

H. Stomata A

J. Ash A

K. Acid-insoluble Ash A

L. Pesticide Residues A

M. Tannins in Herbal Drugs A

APPENDIX XII

A. Disintegration Test for Tablets and Capsules A

B. Disintegration Test for Enteric-coated Tablets A

C. Disintegration Test for Suppositories and Pessaries A

D. Dissolution Test for Tablets and Capsules A

E. Dissolution Test for Transdermal Patches A

F. Aerodynamic Assessment of Fine Particles A

G. Uniformity of Weight A

H. Uniformity of Content A

J. Extractable Volume A

K. Drug Release from Medicated Chewing Gum A

L. Uniformity of Weight (Mass) of Delivered Doses from Multidose Containers A

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M. Deliverable Weight (Mass) or Volume of Liquid and Semi-solid Preparations A

APPENDIX XIII

Particulate Contamination

A. Sub-visible Particles A

B. Visible Particles A

C. Microscope Method A

APPENDIX XIV

Biological Assays and Tests

A. Antibiotics A

B. Immunochemical Methods A

C. Bacterial Endotoxins A

D. Pyrogens A

E. Abnormal Toxicity A

F. Depressor Substances A

G. Histamine A

H. Hormones

1. Corticotropin A

J. Blood and Related Products

1. Factor VII Fraction A

2. Factor VIII Fraction A

3. Factor IX Fraction A

4. Heparin in Coagulation Factor Concentrates A

5. Heparin A

6. Fc Function of Immunoglobulins A

7. Anti-A and Anti-B Haemagglutinins A

8. Prekallikrein Activator A

9. Anticomplementary Activity of Immunoglobulins A

10. Assay of Anti-D Immunoglobulin A

K. Immunological Products

1. Diphtheria Vaccine (Adsorbed) A

2. Pertussis Vaccine A

3. Tetanus Vaccine (Adsorbed) A

4. Hepatitis A Vaccine A

5. Hepatitis B Vaccine (rDNA) A

L. Nucleic Acid Amplification Techniques A

M. Cytokines A

1. Assay of Interferons A

APPENDIX XV

Production and Testing of Vaccines

A. Terminology A

B. Aluminium A

C. Calcium A

D. Free Formaldehyde A

E. Phenol A

F. Neurovirulence

1. Live Viral Vaccines A

2. Poliomyelitis Vaccine (Oral) A

G. Composition of Polysaccharide Vaccines A

H. Specified Pathogen-free Flocks A

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J. Cell Substrates for the Production of Vaccines for Human Use A

APPENDIX XVI

A. Test for Sterility A

B. Test for Microbial Contamination A

1. Tests for Specified Micro-organisms A

2. Total Viable Aerobic Count A

3. Mycoplasmas A

4. Mycobacteria A

5. Extraneous Agents in Viral Vaccines A

C. Efficacy of Antimicrobial Preservatives A

D. Microbiological Quality of Pharmaceutical Preparations A

APPENDIX XVII

A. Particle Size of Powders

1. Classification A

2. Limit Test, Microscopy A

B. Sieves and Filters

1. Sieves A

2. Filters A

C. Specific Surface Area by Air Permeability A

D. Apparent Volume A

E. Flowability A

F. Consistency by Penetrometry A

G. Friability of Uncoated Tablets A

H. Resistance to Crushing of Tablets A

J. Softening Time of Lipophilic Suppositories A

K. Pycnomeric Density of Solids A

L. Resistance to Rupture of Suppositories and Pessaries A

M. Specific Surface Area by Gas Adsorption A

APPENDIX XVIII

Methods of Sterilisation A

APPENDIX XIX

Containers

A. Introduction A

B. Glass Containers for Pharmaceutical Preparations A

C. Plastic Containers and Closures

1. For Aqueous Intravenous Infusions A

D. Containers for Blood and Blood Components

1. Sterile Plastic Containers A

2. Empty Sterile Containers of Plasticised PVC A

3. Empty Sterile Containers of Plasticised PVC with an Anticoagulant Solution A

E. Rubber Closures for Containers for Aqueous Parenteral Preparations A

F. Sets for the Transfusion of Blood and Blood Components A

G. Sterile Single-use Plastic Syringes A

APPENDIX XX

Materials for the Manufacture of Containers

A. Poly(vinyl chloride)

1. For Blood, Blood Components and Aqueous Intravenous Infusions A

2. Tubing for Transfusion Sets A

3. For Non-injectable, Aqueous Solutions A

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4. For Dry Dosage Forms A

5. For Containers for Aqueous Solutions for Intravenous Infusion A

B. Polyolefines A

C. Polyethylene

1. without Additives A

2. with Additives A

D. Polypropylene A

E. Poly(EthyleneVinyl Acetate) A

F. Silicone

1. Oil A

2. Elastomer A

G. Plastic Additives A

H. Rubber for Closures

APPENDIX XXI

A. Abbreviated Titles A

B. Approved Synonyms A

C. Codes for Eye Drops in Single-dose Containers A

APPENDIX XXII

Names, Symbols and Atomic Weights of Elements A

APPENDIX XXIII

Weights and Measures: SI Units A

APPENDIX XXIV

Abbreviations A

73-7

European Pharmacopoeia Equivalent TextsIn monographs reproduced from the European Pharmacopoeia the analytical

methods, tests and other supporting texts are invoked by means of the referencenumber of the text in the general Chapters of the European Pharmacopoeia.The table below lists the contents of the General Chapters of the EuropeanPharmacopoeia and gives the British Pharmacopoeia or British Pharmacopoeia(Veterinary) equivalents. It is provided for information but it is emphasised that fortexts of the European Pharmacopoeia, in cases of doubt or dispute the textpublished by the Council of Europe is authoritative. Appendices of the BritishPharmacopoeia (Veterinary) are identified by inclusion of (Vet) after theAppendix letter, for example, Appendix XV J(Vet) 1.

Ph. Eur. Subject of text British Pharmacopoeiareferencereference

1 General Notices

2.1.1 Droppers Appendix I A

2.1.2 Sintered-glass filters Appendix XVII B2

2.1.3 UV lamps Appendix III A

2.1.4 Sieves Appendix XVII B

2.1.5 Tubes for comparative tests Appendix VII

2.1.6 Gas detector tubes Appendix IX K

2.2.1 Clarity and degree of opalescence of liquids Appendix IV A

2.2.2 Degree of coloration of liquids Appendix IV B

2.2.3 Potentiometric determination of pH Appendix V L

2.2.4 Reaction of solution, pH and indicator colour Appendix V K

2.2.5 Relative density Appendix V G

2.2.6 Refractive index Appendix V E

2.2.7 Optical rotation Appendix V F

2.2.8 Viscosity Appendix V H

2.2.9 Capillary viscometer method Appendix V H, Method II

2.2.10 Rotating viscometer method Appendix V H, Method III

2.2.11 Distillation range Appendix V C

2.2.12 Boiling point Appendix V D

2.2.13 Water by distillation range Appendix IX C, Method II

2.2.14 Melting point: Capillary method Appendix V A, Method I

2.2.15 Melting point: Open capillary method Appendix V A, Method IV

2.2.16 Melting point: Instantaneous method Appendix V A, Method V

2.2.17 Drop point Appendix V A, Method III

2.2.18 Freezing point Appendix V B

2.2.19 Amperometric titrations Appendix VIII B

2.2.20 Potentiometric titrations Appendix VIII B

2.2.21 Fluorimetry Appendix II E

2.2.22 Atomic emission spectrometry Appendix II D

2.2.23 Atomic absorption spectrometry Appendix II D

2.2.24 Infrared spectrophotometry Appendix II A

2.2.25 Visible and ultraviolet spectrophotometry Appendix II B

2.2.26 Paper chromatography Appendix III E

2.2.27 Thin-layer chromatography Appendix III A

2.2.28 Gas chromatography Appendix III B

2.2.29 Liquid chromatography Appendix III D

73-8

2.2.30 Size-exclusion chromatography Appendix III C

2.2.31 Electrophoresis Appendix III F

2.2.32 Loss on drying Appendix IX D

2.2.33 Nuclear magnetic resonance spectrometry Appendix II C

2.2.34 Thermogravimetry Appendix V M

2.2.35 Osmolality Appendix V N

2.2.36 Ion-selective potentiometry Appendix VIII E

2.2.37 X-ray fluorescent spectrophotometry Appendix II F

2.2.38 Conductivity Appendix V O

2.2.39 Molecular mass distribution in dextrans Appendix III C

2.2.40 Near infrared spectrometry Appendix II A

2.2.41 Circular dichroism Appendix V J

2.2.42 Density of Solids Appendix V Q

2.2.43 Mass Spectrometry Appendix II G

2.2.44 Total Organic Carbon in Water Appendix V P

2.2.45 Supercritical Fluid Chromatography Appendix III H

2.2.46 Chromatographic Separation Techniquies Appendix III

2.2.47 Capillary Electrophoresis Appendix III G

2.3.1 Identification reactions Appendix VI

2.3.2 Identification of fatty oils by TLC Appendix X N

2.3.3 Identification of phenothiazines by TLC Appendix III A

2.3.4 Odour Appendix VI

Limit test for:-

2.4.1 ammonium Appendix VII

2.4.2 arsenic Appendix VII

2.4.3 calcium Appendix VII

2.4.4 chloride Appendix VII

2.4.5 fluorides Appendix VII

2.4.6 magnesium Appendix VII

2.4.7 magnesium and alkaline-earth metals Appendix VII

2.4.8 heavy metals Appendix VII

2.4.9 iron Appendix VII

2.4.10 lead in sugars Appendix VII

2.4.11 phosphates Appendix VII

2.4.12 potassium Appendix VII

2.4.13 sulphate Appendix VII

2.4.14 Sulphated ash Appendix IX A, Method II

2.4.15 Limit Test for nickel in polyols Appendix VII

2.4.16 Total ash Appendix XI J, Method II

2.4.17 Limit test for aluminium Appendix VII

2.4.18 Free formaldehyde Appendix XV D

2.4.19 Alkaline impurities in fatty oils Appendix X N

2.4.20 Antioxidants in fatty oils Suppressed 1/1/99

2.4.21 Foreign oils in fatty oils by TLC Appendix X N

2.4.22 Foreign oils by gas chromatography Appendix X N

2.4.23 Sterols in fatty oils Appendix X N

2.4.24 Residual solvents Appendix VIII L

2.4.25 Residual ethylene oxide and dioxan Appendix VIII M

2.4.26 N,N-Dimethylaniline Appendix VIII N

2.4.27 Nickel in hydrogenated vegetable oils Appendix VII

2.4.28 2-Ethylhexanoic Acid Appendix VIII O

73-9

2.5.1 Acid value Appendix X B

2.5.2 Ester value Appendix X C

2.5.3 Hydroxyl value Appendix X D

2.5.4 Iodine value Appendix X E

2.5.5 Peroxide value Appendix X F

2.5.6 Saponification value Appendix X G, Method II

2.5.7 Unsaponifiable matter Appendix X H, Method II

2.5.8 Assay of primary aromatic amino nitrogen Appendix VIII B

2.5.9 Semi-micro determination of nitrogenby sulphuric acid digestion Appendix VIII H

2.5.10 Oxygen-flask method Appendix VIII C

2.5.11 Complexometric titrations Appendix VIII D

2.5.12 Semi-micro determination of water Appendix IX C, Method I

2.5.13 Aluminium in adsorbed vaccines Appendix XV B

2.5.14 Calcium in adsorbed vaccines Appendix XV C

2.5.15 Phenol in immunosera and vaccines Appendix XV E

Polysaccharide vaccines:-

2.5.16 protein Appendix XV G

2.5.17 nucleic acids Appendix XV G

2.5.18 phosphorus Appendix XV G

2.5.19 O-acetyl Appendix XV G

2.5.20 hexosamines Appendix XV G

2.5.21 methylpentoses Appendix XV G

2.5.22 uronic acids Appendix XV G

2.5.23 sialic acid Appendix XV G

2.5.24 Carbon dioxide in medicinal gases Appendix IX G

2.5.25 Carbon monoxide in medicinal gases Appendix IX E

2.5.26 Nitrogen monoxide and nitrogen dioxide inmedicinal gases Appendix IX H

2.5.27 Oxygen in medicinal gases Appendix IX J

2.5.28 Water in medicinal gases Appendix IX K

2.5.29 Sulphur dioxide Appendix IX B

2.5.30 Oxidising substances Appendix X L

2.5.31 Ribose in polysaccharide vaccines Appendix XV G

2.5.32 Coulometric titration of water Appendix IX C

2.5.33 Total protein assay Appendix VIII P

2.5.34 Acetic acid in synthetic peptides Appendix VIII Q

2.6.1 Sterility Appendix XVI A

2.6.2 Mycobacteria Appendix XVI B4

2.6.3 Extraneous viruses using fertilised eggs Appendix XVI B(Vet) 4

2.6.4 Leucosis virus Appendix XVI B(Vet) 6

2.6.5 Extraneous viruses using cell cultures Appendix XVI BVet) 7

2.6.6 Extraneous agents using chicks Appendix XVI B(Vet) 8

2.6.7 Mycoplasmas Appendix XVI B3 and Appendix XVI B(Vet) 3

2.6.8 Pyrogens Appendix XIV D

2.6.9 Abnormal toxicity Appendix XIV E

2.6.10 Histamine Appendix XIV G

2.6.11 Depressor substances Appendix XIV F

2.6.12 Total viable aerobic count Appendix XVI B2

2.6.13 Specified micro-organisms Appendix XVI B1

73-10

2.6.14 Bacterial endotoxins Appendix XIV C

2.6.15 Prekallikrein activator Appendix XIV J8

2.6.16 Extraneous agents in viral vaccines Appendix XVI B5

2.6.17 Anticomplementary activity of immunoglobulins Appendix XIV J9

2.6.18 Neurovirulence of live viral vaccines Appendix XV F1

2.6.19 Neurovirulence of poliomyelitis vaccine (oral) Appendix XV F2

2.6.20 Anti-A and anti-B haemagglutinins (indirect method) Appendix XIV J7

2.6.21 Nucleic acid amplification Appendix XIV L

2.7.1 Immunochemical methods Appendix XIV B

2.7.2 Microbiological assay of antibiotics Appendix XIV A

2.7.3 Assay of corticotrophin Appendix XIV H1

2.7.4 Assay of coagulation factor VIII Appendix XIV J2

2.7.5 Assay of heparin Appendix XIV J5

2.7.6 Assay of diphtheria vaccine (adsorbed) Appendix XIV K1

2.7.7 Assay of pertussis vaccine Appendix XIV K2

2.7.8 Assay of tetanus vaccine (adsorbed) Appendix XIV K3

2.7.9 Fc function of immunoglobulins Appendix XIV J6

2.7.10 Assay of coagulation factor VII Appendix XIV J1

2.7.11 Assay of coagulation factor IX Appendix XIV J3

2.7.12 Heparin in coagulation factor concentrates Appendix XIV J4

2.7.13 Assay of anti-D immunoglobulin Appendix XIV J10

2.7.14 Assay of hepatitis A vaccine Appendix XIV K4

2.7.15 Assay of hepatitis B (rDNA) vaccine Appendix XIV K5

2.8.1 Ash insoluble in hydrochloric acid Appendix XI K, Method II

2.8.2 Foreign matter Appendix XI D, Test B

2.8.3 Stomata and stomatal index Appendix XI H

2.8.4 Swelling index Appendix XI C

2.8.5 Water in essential oils Appendix X M

2.8.6 Foreign esters Appendix X M

2.8.7 Fatty oils and resinified volatile oils Appendix X M

2.8.8 Odour and taste of volatile oils Appendix X M

2.8.9 Residue on evaporation Appendix X M

2.8.10 Solubility in ethanol (volatile oils) Appendix X M

2.8.11 Determination of cineole Appendix X J

2.8.12 Essential oil content of crude drugs Appendix XI E

2.8.13 Pesticide residues Appendix XI L

2.8.14 Determination of tannins in herbal drugs Appendix XI M

2.9.1 Disintegration: tablets and capsules Appendix XII A

2.9.2 Disintegration: Suppositories and pessaries Appendix XII C

2.9.3 Dissolution: Solid oral dosage forms Appendix XII D

2.9.4 Dissolution: Transdermal patches Appendix XII E

2.9.5 Uniformity of mass Appendix XII G

2.9.6 Uniformity of content Appendix XII H

2.9.7 Friability of uncoated tablets Appendix XVII G

2.9.8 Resistance to crushing of tablets Appendix XVII H

2.9.9 Measurement of consistency by penetrometry Appendix XVII F

2.9.10 Ethanol content and alcoholimetric tablets Appendix VIII F, Method III

2.9.11 Methanol and 2-propanol Appendix VIII G

2.9.12 Sieve test Appendix XVII A

2.9.13 Particle size by microscopy (limit test) Appendix XIII

2.9.14 Specific surface area by gas permeability Appendix XVII C

73-11

2.9.15 Apparent volume Appendix XVII D

2.9.16 Flowability Appendix XVII E

2.9.17 Extractable volume Appendix XII J

2.9.18 Aerodynamic assessment of fine particles Appendix XII F

2.9.19 Particulate contamination: Sub-visible particles Appendix XIII A

2.9.20 Particulate contamination: Visible particles Appendix XIII B

2.9.21 Particulate contamination: Microscope method Appendix XIII C

2.9.22 Softening time of lipophilic suppositories Appendix XVII J

2.9.23 Pycnometric density of solids Appendix XVII K

2.9.24 Resistance to rupture of suppositores and pessaries Appendix XVII L

2.9.25 Drug Release from Medicated Chewing Gum Appendix XII K

2.9.26 Specific Surface Area by Gas Adsorption Appendix XVII M

2.9.27 Uniformity of mass of delivered doses frommultidose containers Appendix XII L

2.9.28 Test for deliverable mass or volume of liquidand semi-soilid preparations Appendix XII M

3.1.1 Material used for containers for human bloodand blood components Appendix XX A

3.1.1.1 Materials based on PVC for blood and bloodcomponents Appendix XX A1

3.1.1.2 Materials based on PVC for tubing used in sets forthe transfusion of blood and blood components Appendix XX A2

3.1.2 Superseded by 3.1.1.2

3.1.3 Polyolefines Appendix XX B

3.1.4 Polyethylene - without additives Appendix XX C1

3.1.5 Polyethylene - with additives Appendix XX C2

3.1.6 Polypropylene Appendix XX D

3.1.7 Poly(Ethylenevinyl acetate) Appendix XX E

3.1.8 Silicone oil used as a lubricant Appendix XX F1

3.1.9 Silicone elastomer for closures and tubing Appendix XX F2

3.1.10 Materials based on non-plasticised PVC forcontainers for non-injectable, aqueous solutions Appendix XX A3

3.1.11 Materials based on non-plasticised PVC forcontainers for dry dosage forms for oraladministration Appendix XX A4

3.1.12 Rubber for closures for containers for aqueousparenteral preparations and for powders forfreeze-dried products Appendix XX H

3.1.13 Plastic additives Appendix XX G

3.1.14 Materials based on plasticised poly(vinyl chloride)for containers for aqueous solutionsfor intravenous infusion Appendix XX A5

3.2 Containers Appendix XIX A

3.2.1 Glass containers for pharmaceutical use Appendix XIX B

3.2.2 Plastic containers and closures Appendix XIX C

3.2.3 Sterile plastic containers for blood andblood components Appendix XIX D1

3.2.4 Empty sterile plastic containers for blood andblood components Appendix XIX D2

3.2.5 Sterile plastic containers with anticoagulantfor blood and blood components Appendix XIX D3

3.2.6 Sets for the transfusion of blood andblood components Appendix XIX F

3.2.7 Plastic containers for aqueous intravenous infusions Appendix XIX H

3.2.8 Sterile single-use plastic syringes Appendix XIX G

73-12

3.2.9 Rubber closures for containers for aqueouspreparations for parenteral use Appendix XIX E

4.1.1 Reagents Appendix I A

4.1.2 Standard solutions Appendix I C

4.1.3 Buffer solutions Appendix I D

4.2.1 Reference substances for volumetric solutions Appendix I B

4.2.2 Volumetric solution Appendix I B

5.1.1 Methods of preparation of sterile products Appendix XVIII

5.1.2 Biological indicators (sterilisation) Appendix XVIII

5.1.3 Efficacy of antimicrobial preservation Appendix XVI C

5.1.4 Microbial quality of pharmaceutical preparations Appendix XVI D

5.1.5 F0 concept (sterilisation) Appendix XVIII

5.2.1 Terminology: Vaccines Appendix XV A andAppendix XV A(Vet)

5.2.2 SPF Chicken flocks Appendix XV H andAppendix XV H(Vet)

5.2.3 Cell substrates for the production of Appendix XV Jvaccines for human use

5.2.4 Cell cultures (veterinary vaccine production) Appendix XV J(Vet) 1

5.2.5 Substances of animal origin (veterinary vaccineproduction) Appendix XV J(Vet) 2

5.2.6 Safety: Veterinary vaccines Appendix XV K(Vet) 1

5.2.7 Efficacy: Veterinary vaccines Appendix XV K(Vet) 2

5.2.8 Minimising the Risk of Transmitting AnimalSpongiform Encephalopathy Agents viaMedicinal Products Supplementary Chapter IV F.

5.3 Statistical analysis of biological assays and tests Supplementary Chapter IV G

5.4 Residual solvents Supplementary Chapter IV D

5.5 Alcoholimetric tables Supplementary Chapter IV E

5.6 Assay of Interferons Appendix XIV M1

5.7 Physical Characteristics of radionuclides RadiopharmaceuticalPreparations

74-1

Appendix IThe specifications given below are strictly for the use of the materials as reagents. The inclusion ofa material in this Appendix does not imply that it is suitable for use in medicines.

Appendix I A. General Reagents In a monograph from the European Pharmacopoeia, the name of a substance or solution followedby the letter R (the whole in italics) indicates a reagent in Appendix I. Where a reagent is describedusing an expression such as hydrochloric acid (10 g/l HCl) the solution is prepared by an appropriatedilution with water of a more concentrated solution specified in this section.

Where the name of the solvent is not stated, an aqueous solution is intended. Reagents in aqueoussolutions are prepared using water. Reagent solutions used in the limit tests for barium, calcium andsulphates are prepared using distilled water.

The description of a reagent may include a Chemical Abstracts Service Registry number (CASnumber) recognisable by its typical format (for example, 9002-93-1).

Some of the reagents included may be injurious to health unless adequate precautions are taken.They should be handled in accordance with good laboratory practice and any relevant regulationssuch as those issued in the United Kingdom in accordance with the Health and Safety at Work etc.Act (1974). The labelling should comply with relevant national legislation and internationalagreements. Reagents and reagent solutions should be kept in well-closed containers.

Droppers (Ph Eur text 2.1.1)

The term drops means standard drops delivered from a standard dropper as described below.

Standard droppers (Fig. 11) are constructed of practically colourless glass. The lower extremity hasa circular orifice in a flat surface at right angles to the axis. Other droppers may be used provided theycomply with the following test.

Fig. 11 Standard dropper

Dimensions in millimetres

74-2

Twenty drops of water at 20 1 flowing freely from the dropper held in the vertical position at aconstant rate of one drop per second weighs 1000 50 mg. The dropper must be carefully cleanedbefore use. Carry out three determinations on any given dropper. No result may deviate by morethan 5% from the mean of the three determinations

Acacia Of the British Pharmacopoeia.

Acacia Solution Dissolve 100 g of acacia in 1000 ml of water, stir mechanically for 2 hours,centrifuge at 2000 g for 30 minutes to obtain a clear solution.Store in polyethylene containers of about 250 ml capacity at 0 to 20.

Acetal 1,1-Diethoxyethane C6H14O2 = 118.2 (105-57-7)General reagent grade of commerce.d2020 , about 0.824; nD20 , about 1.382; boiling point, about 103.Acetaldehyde C2H4O = 44.1 (75-07-0)General reagent grade of commerce.A colourless volatile liquid with an acrid, penetrating odour; d2020 , about 0.788; nD20 , about 1.332;boiling point, about 21.

Acetamide C2H5NO = 59.07 (60-35-5)General reagent grade of commerce.Melting point, about 78.

Acetic Acid Dilute 30 g of glacial acetic acid to 100 ml with water. It contains not less than 29.0%and not more than 31.0% w/v of C2H4O2 (5M).

Acetic Acid, Anhydrous Anhydrous glacial acetic acid; C2H4O2 = 60.1 (64-19-7)Glacial acetic acid of commerce for use in non-aqueous titrations containing not less than 99.6% w/wof C2H4O2.d2020 , 1.0152 to 1.053; boiling point, 117 to 119.Complies with the following test.WATER Not more than 0.4% w/w, Appendix IX C. If the water content is greater than 0.4% it maybe adjusted by adding the calculated amount of acetic anhydride.Store protected from light.

Acetic Acid, Dilute Dilute 12 g of glacial acetic acid to 100 ml with water. It contains not less than11.5% and not more than 12.5% w/v of C2H4O2 (2M).

Acetic Acid, Glacial Analytical reagent grade of commerce.A colourless liquid with a pungent odour, about 17.5M in strength; freezing point, about 16; weightper ml, about 1.05 g. It contains not less than 98.0% w/w of C2H4O2.

Solutions of molarity xM should be prepared by diluting 57x ml (60x g) of glacial acetic acid to1000 ml with water.

Acetic Anhydride C4H6O3 = 102.1 (108-24-7)Analytical reagent grade of commerce containing not less than 97.0% w/v of C4H6O3.A colourless liquid; boiling point, 136 to 142.

Acetic Anhydride Solution R1Dissolve 25 ml of acetic anhydride in anhydrous pyridine and dilute to 100 ml with anhydrous pyridine.Store protected from light and air.

Acetic AnhydrideDioxan Solution Add 1 ml of acetic anhydride to 50 ml of 1,4-dioxan.

Acetic AnhydrideSulphuric Acid Solution Carefully mix 5 ml of acetic anhydride with 5 ml ofsulphuric acid. Add, dropwise and with cooling, to 50 ml of absolute ethanol.Prepare immediately before use.

Acetone Propan-2-one; C3H6O = 58.08 (67-64-1)Analytical reagent grade of commerce.A volatile, flammable liquid; boiling point, about 56; weight per ml, about 0.79 g.Complies with the following test.WATER Not more than 0.3% w/w, Appendix IX C, using anhydrous pyridine as the solvent.

Acetonitrile Methyl cyanide; C2H3N = 41.05 (75-05-8)General reagent grade of commerce.A colourless liquid; d2020 , about 0.78; nD20 , about 1.344. Not less than 95% distils between 80 and82.Acetonitrile used in spectrophotometry complies with the following requirement.TRANSMITTANCE Not less than 98% in the range 255 to 420 nm using water in the reference cell,Appendix II B.

Acetonitrile for Chromatography Chromatographic grade of acetonitrile containing not less than

74-399.8% of C2H3N that complies with the following test.TRANSMITTANCE Not less than 98% from 240 nm using water in the reference cell.

Acetonitrile R1 Acetonitrile containing not less than 99.9% of C2H3N and complying with thefollowing requirement.LIGHT ABSORPTION The absorbance of the reagent being examined at 200 nm, Appendix II B, is notmore than 0.10. Use water in the reference cell.

Acetyl Chloride C2H3ClO = 78.5 (75-36-5)Analytical reagent grade of commerce.d2020 , about 1.10. Not less than 95% distils between 49 and 53.Acetylacetamide 3-Oxobutanamide; C4H7NO2 = 101.1 (5977-14-0)General reagent grade of commerce.Melting point, 53 to 56.

Acetylacetone Pentane-2,4-dione; C5H8O2 = 100.1 (123-54-6)Analytical reagent grade of commerce.A colourless or slightly yellow, easily flammable liquid.nD

20 , 1.452 to 1.453; boiling point, 138 to 140.

Acetylacetone Reagent R1To 100 ml of ammonium acetate solution add 0.2 ml of acetylacetone.

4-Acetylbiphenyl 4-Phenylacetophenone; C14H12O = 196.2 (51-42-3)General reagent grade of commerce.Melting point, about 117.

N-Acetyl-eeeee-caprolactam C8H13NO2 = 155.2 (1888-91-1)General reagent grade of commerce.d2020 , about 1.100; nD20 , about 1.489; boiling point, about 135.Acetylcholine Chloride C7H16ClNO2 = 181.7 (60-31-1)General reagent grade of commerce.Store at 20.

N-Acetyl-L-cysteine C5H9NO3S = 163.2 (616-91-1)General reagent grade of commerce.Melting point, about 110; [ ]D20 , about +4.6.Acetyleugenol C12H14O3 = 206.2 (93-28-7)General reagent grade of commerce.A yellow, oily liquid; nD

20 , about 1.521; boiling point, 281 to 282.Acetyleugenol used in gas chromatography complies with the following test.ASSAY Carry out the test for Chromatographic profile described under Clove Oil using the reagentbeing examined as the test solution. The area of the principal peak is not less than 98.0% of the totalarea of the peaks.

N-Acetylneuraminic Acid O-Sialic acid; C11H19NO9 = 309.3 (131-48-6)Melting point, about 186, with decomposition; [ ]D20 , about 36 (1% w/v in water).N-Acetyltryptophan C13H14N2O3 = 246.3 (1218-34-4)General reagent grade of commerce.Melting point, about 205.ASSAY Dissolve 10 mg in sufficient of a mixture of 10 volumes of acetonitrile and 90 volumes of waterto produce 100 ml. Examine as prescribed in the monograph for Tryptophan under 1,1-Ethylidene-bis(tryptophan) and other related substances. The area of the principal peak in the chromatogramobtained is not less than 99.0% of the areas of all the peaks.

Acetyltyrosine Ethyl Ester Ethyl N-acetyl-L-tyrosinate; C13H17NO4,H2O = 269.3 (36546-50-6)General reagent grade of commerce.A white, crystalline powder; [ ]D20 , +21 to +25 (1% w/v in ethanol); A(1%, 1 cm) at 278 nm, 60 to68 determined in ethanol.

Acetyltyrosine Ethyl Ester, 0.2M 0.2M Ethyl acetyltyrosinateDissolve 0.54 g of acetyltyrosine ethyl ester in sufficient ethanol (96%) to produce 10 ml.

Acid Blue 83 CI 42660; Coomassie brilliant blue R250; brilliant blue R; C45H44N3NaO7S = 826(6104-59-2)General reagent grade of commerce.A brown powder.

Acid Blue 90 CI 42655; Coomassie brilliant blue G; brilliant blue G; C47H48N3NaO7S2 = 854(6104-58-1)A dark brown powder with a violet sheen; some particles have a metallic lustre.Complies with the following tests.LIGHT ABSORPTION A(1%, 1 cm) in a 0.001% w/v solution in phosphate buffer pH 7.0 at 577 nm,

74-4greater than 500, Appendix II B, calculated with reference to the dried substance.LOSS ON DRYING When dried to constant weight at 100 to 105, loses not more than 5.0% of itsweight.

Acid Blue 92 Coomassie blue; CI 13390; trisodium 8-hydroxy-4-phenylaminoazonaphthalene-3,5,6-trisulphonate; C26H16N3Na3O10S3 = 696 (3861-73-2)General reagent grade of commerce.Dark blue crystals.

Acid Blue 92 Solution Coomassie blue solutionDissolve 0.5 g of acid blue 92 in a mixture of 10 ml of glacial acetic acid, 45 ml of ethanol (96%) and45 ml of water.

Acrylamide C3H5NO = 71.08 (79-06-1)General reagent grade of commerce.Melting point, about 84.

Acrylamide/bisacrylamide (29:1) Solution, 30 per cent Prepare a solution containing 290 g ofacrylamide and 10 g of methylenebisacrylamide per litre of water and filter.

Acrylamide/bisacrylamide (36.5:1) Solution, 30 per cent Prepare a solution containing 292 g ofacrylamide and 8 g of methylenebisacrylamide per litre of water and filter.

Acrylic Acid Prop-2-enoic acid; C3H4O2 = 72.1 (79-10-7)General reagent grade of commerce containing not less than 99% of C3H4O2 and stabilised with0.02% of hydroquinone monomethyl ether.It polymerises readily in the presence of oxygen.d2020 , about 1.05; nD20 , about 1.421; boiling point, about 141.Adenosine 6-Amino-9-b-D-ribofuranosyl-9H-purine; C10H13N5O4 = 267.2 (58-61-7)General reagent grade of commerce.Melting point, about 234.

Adipic acid C6H10O4 = 146.1 (124-04-9)General reagent grade of commence.Melting point, about 152.

Adrenaline Acid Tartrate L-Epinephrine D-hydrogen tartrate; C13H19NO9 = 333 (51-42-3)General reagent grade of commerce.

Aescin Escin; C54H84O23,2H2O = 1138 (11072-93-8)A mixture of related saponins obtained from the seeds of Aesculus hippocastum L.General reagent grade of commerce complying with the following test.HOMOGENEITY Carry out the test for Identification described under Senega Root applying 20 l ofsolution (2) to the plate. After spraying with anisaldehyde solution the chromatogram exhibits aprincipal spot with an Rf value of about 0.4.

Agar (9002-18-0) The dried extract from Gelidium sp. and other algae belonging to the classRhodophyceae.Microbiological reagent grade of commerce.

Agarose for Chromatography (9012-36-6)Chromatographic reagent grade of commerce.Swollen beads 60 m to 140 m in diameter presented as a 4% suspension in water. It is used in size-exclusion chromatography for the separation of proteins with relative molecular masses of 6 104 to20 106 and of polysaccharides with relative molecular masses of 3 103 to 5 106.

Agarose for Chromatography, Cross-linked (61970-08-9)Chromatographic reagent grade of commerce.Prepared from agarose by reaction with 2,3-dibromopropanol in strongly alkaline conditions.Swollen beads 60 m to 140 m in diameter presented as a 4% suspension in water. It is used in size-exclusion chromatography for the separation of proteins with relative molecular masses of 6 104 to20 106 and of polysaccharides with relative molecular masses of 3 103 to 5 106.

Agarose for Chromatography R1, Cross-linked (65099-79-8)Chromatographic reagent grade of commerce.Prepared from agarose by reaction with 2,3-dibromopropanol in strongly alkaline conditions.Swollen beads 60 m to 140 m in diameter presented as a 4% suspension in water. It is used in size-exclusion chromatography for the separation of proteins with relative molecular masses of 7 104 to40 106 and of polysaccharides with relative molecular masses of 1 105 to 2 107.

Agarose for Electrophoresis (9012-36-6)Electrophoretic reagent grade of commerce.A neutral, linear polysaccharide, the main component of which is derived from agar.

Agarose-DEAE for Ion Exchange ChromatographyChromatographic reagent grade of commerce.Cross-linked agarose substituted with diethylaminoethyl groups, presented as beads.

74-5Agarose/Cross-linked PolyacrylamideAgarose trapped within a cross-linked polyacrylamide network. It is used for the separation ofglobular proteins with relative molecular masses of 2 104 to 35 104.

Alanine L-Alanine; C3H7NO2 = 89.1 (56-41-7)General reagent grade of commerce.Melting point, about 315, with decomposition.

bbbbb-Alanine See 3-aminopropionic acid.

Albumin, Bovine Bovine serum albumin (9048-46-8)Bovine albumin containing about 96% protein.A white to light yellowish-brown powder complying with the following test.WATER Not more than 3.0% w/w, Appendix IX C. Use 0.8 g.Bovine albumin used in the assay of tetracosactide should be pyrogen-free, free from proteolyticactivity when examined by a suitable means, for example using chromogenic substrate, and free fromcorticosteroid activity by measurement of fluorescence as described in that assay.

Albumin Solution, Human Albumin (9048-46-8)Albumin Solution of the British Pharmacopoeia.

Albumin Solution R1, Human Albumin solutionDilute human albumin solution with sufficient sodium chloride injection to produce a proteinconcentration of 0.1% w/v and adjust the pH to between 3.5 and 4.5 with glacial acetic acid.

Alcohol See Ethanol (96%).

Alcohol, Aldehyde-free See aldehyde-free ethanol (96%).

Alcohol (x% v/v) See ethanol (96%).Aldehyde Dehydrogenase Enzyme obtained from bakers yeast which oxidises acetaldehyde toacetic acid in the presence of nicotinamide-adenine dinucleotide, potassium salts and thiols at pH8.0.General reagent grade of commerce.

Aldehyde Dehydrogenase Solution Dissolve in water a quantity of aldehyde dehydrogenaseequivalent to 70 units and dilute to 10 ml with the same solvent. This solution is stable for 8 hours at4.

Aldrin C12H8Cl6 =364.9 (309-00-2)Use a grade suitable for pesticide residue analysis. A suitable certified reference solution (10 ng/l incyclohexane) may be used.boiling point, about 145 C.; melting point, about 104 C.

Aleuritic Acid (9RS,10SR)-9,10,16-Trihydroxyhexadecanoic acid; C16H32O5 = 304.4 (533-87-9)General reagent grade of commerce.Melting point, about 101.

Alizarin S CI 58005; alizarin red S; C14H7NaO7S,H2O = 360.3 (130-22-3)General reagent grade of commerce.A yellowish brown or orange-yellow powder.

Alizarin S Solution A 0.1% w/v solution of alizarin S.Complies with the following test.SENSITIVITY TO BARIUM To 5 ml of 0.05M sulphuric acid add 5 ml of water, 50 ml of acetate buffer pH3.7 and 0.5 ml of the solution being examined. Add, dropwise, 0.05M barium perchlorate VS. Thecolour changes from yellow to orange-red.Colour change: pH 3.7 (yellow) to pH 5.2 (violet).

Aluminium Al = 26.98 (7429-90-5)Analytical reagent grade of commerce.A white, malleable, flexible, bluish metal available as bars, sheets, powder, strips or wire. In moist airan oxide film forms which protects the metal from corrosion.

Aluminium Chloride Aluminium chloride hexahydrate; AlCl3,6H2O = 241.4 (7784-13-6)General reagent grade of commerce containing not less than 98.0% of AlCl3,6H2O.Store in an airtight container.

Aluminium Chloride Reagent Dissolve 2.0 g of aluminium chloride in 100 ml of a 5% v/v solutionof glacial acetic acid in methanol.

Aluminium Chloride Solution Dissolve 65.0 g of aluminium chloride in sufficient water to produce100 ml, add 0.5 g of activated charcoal, stir for 10 minutes, filter and add to the filtrate, stirringcontinuously, sufficient of a 1% w/v solution of sodium hydroxide to adjust the pH to about 1.5(approximately 60 ml is required).

Aluminium Hydroxide Gel Al(OH)3 + aq (21645-51-2)A hydrated grade of aluminium hydroxide of commerce.

74-6Aluminium Nitrate Al(NO3)3,9H2O = 375.1 (7784-27-2)Analytical reagent grade of commerce.Deliquescent crystals.Store in an airtight container.

Aluminium Oxide, Activated Acid Al2O3 = 102.1An almost white, fine, granular powder, very hygroscopic, activated by heating at 200 to 250 for3 hours.Mean particle size, 50 to 200 m.

Aluminium Oxide, Anhydrous Al2O3 = 102.0 (1344-28-1)Use a grade of commerce consisting of g-Al2O3 dehydrated and activated by heat treatment. Theparticle size is such that it passes through a 150-m sieve but is retained on a 75-m sieve.

Aluminium Oxide, Basic A basic grade of aluminium oxide of commerce suitable for columnchromatography.Complies with the following test.ALKALINITY pH of a 10% w/v suspension in carbon dioxide-free water, shaken for 5 minutes, 9 to 10,Appendix V L.When used in monographs other than those of the European Pharmacopoeia, the following test applies.ACTIVITY Pack into a chromatographic tube (25 cm 10 mm) sufficient of the basic aluminiumoxide to form a column 50 mm deep. Apply to the column a mixture of 5 ml of sudan yellow solutionand 5 ml of sudan red solution and elute with 20 ml of a mixture of 1 volume of benzene and 4 volumesof petroleum spirit (boiling range, 60 to 80). Sudan red forms a zone about 10 mm in depth on theupper part of the column separated by a colourless zone from a yellow zone of sudan yellow below.

Aluminium Oxide, Deactivated To a suitable basic alumina add 1.5 to 2% of water, mix well andallow to stand overnight in a stoppered bottle. The product complies with the following test.

Prepare a column (20 cm 10 mm) using the alumina and hexane. Add a solution of 0.25 g ofcalciferol in 10 ml of hexane. When the level of the solution falls just to the top of the column, begineluting with a 17.5% v/v solution of ether in hexane adjusting the rate of flow, if necessary, to between1 and 2 ml per minute. Collect 200 ml of eluate; no calciferol is present. Collect a further 100 ml ofeluate; it contains not less than 95% of the calciferol used in the test, when determined by the Assaydescribed under Calciferol Oral Solution.

Aluminium Oxide G A fine, white, homogeneous powder of an average particle size between 10and 40 m containing about 10% w/w of calcium sulphate hemihydrate.CONTENT OF CALCIUM SULPHATE Carry out the test described under silica gel G.ACIDITY OR ALKALINITY pH of a suspension prepared by shaking 1 g with 10 ml of carbon dioxide-free water, about 7.5, Appendix V L.

Aluminium Oxide, Neutral Al2O3 = 102.0General reagent grade of commerce complying with the monograph for Dried AluminiumHydroxide.

Aluminium Potassium Sulphate Alum; aluminium potassium sulphate dodecahydrate;AlK(SO4)2,12H2O = 474.4 (7784-24-9)Analytical reagent grade of commerce.

Aluminium Sulphate Al2(SO4)3,16H2O = 630.4 (10043-01-3)Analytical reagent grade of commerce.

Amaranth S CI 16185; acid red 27; C20H11N2Na3O10S3 = 604 (915-67-3)General reagent grade of commerce.A deep brown or deep reddish brown, fine powder.When used for the titration of iodine and iodides with potassium iodate, the colour changes fromorange-red to yellow.

Amaranth Solution A 0.2% w/v solution of amaranth S.

Amido Black 10B See naphthalene black 12B

Amido Black 10B Solution A 0.5% w/v solution of naphthalene black 12B in a mixture of 10volumes of acetic acid and 90 volumes of methanol.

Aminoazobenzene a-Aminoazobenzene; 4-Aminoazobenzene; 4-phenylazoaniline; CI 11000;C12H11N3 = 197.2 (60-09-3)General reagent grade of commerce.Brownish yellow needles with a bluish tinge; melting point, about 128.

2-Aminobenzoic Acid Anthranilic acid; C7H7NO2 = 137.1 (118-92-3)General reagent grade of commerce.Melting point, about 145.

74-74-Aminobenzoic Acid C7H7NO2 = 137.1 (150-13-0)General reagent grade of commerce.Melting point, about 188.Complies with the following test.HOMOGENEITY Carry out the test for Related substances described under ProcaineHydrochloride. The chromatogram obtained shows only one principal spot.Store protected from light.

4-Aminobenzoic Acid Solution Dissolve 1 g of 4-aminobenzoic acid in a mixture of 18 ml ofanhydrous acetic acid, 20 ml of water and 1 ml of orthophosphoric acid. Immediately before use, mix 2volumes of the solution with 3 volumes of acetone.

(4-Aminobenzoyl)-L-glutamic Acid C12H14N2O5 = 266.2 (4271-30-1)General reagent grade of commerce.Melting point, about 173.

4-Aminobutanoic Acid See 4-amino-n-butyric acid.

Aminobutanol See 2-Aminobutan-1-ol.

2-Aminobutan-1-ol Aminobutanol; C4H11NO = 89.14 (5856-63-3)General reagent grade of commerce.An oily liquid; boiling point, about 180; d2020 , about 0.94; nD20 , about 1.453.4-Amino-n-butyric Acid 4-Aminobutanoic acid; gamma amino butyric acid; GABA; C4H9NO2 =103.1 (2835-81-6)General reagent grade of commerce containing not less than 97% of C4H9NO2.

Aminochlorobenzophenone See 2-Amino-5-chlorobenzophenone.

2-Amino-5-chlorobenzophenone Aminochlorobenzophenone; C13H10ClNO = 231.7 (719-59-5)General reagent grade of commerce.A yellow, crystalline powder; melting point, about 97.Complies with the following test.HOMOGENEITY Carry out the test for Related substances described under ChlordiazepoxideHydrochloride applying to the plate 5 l of a 0.05% w/v solution in methanol. The chromatogramshows only one spot with an Rf value of about 0.9.Store protected from light.

6-Aminohexanoic Acid 6-Aminocaproic acid; C6H13NO2 = 131.2 (60-32-2)General reagent grade of commerce.Melting point, about 205.

p-Aminohippuric Acid Aminohippuric acid; N-(4-Aminobenzoyl)glycine; C9H10N2O3 = 194.2(61-78-9)General reagent grade of commerce.A white or almost white powder; melting point, about 200.

Aminohippuric Acid Reagent Dissolve 3 g of phthalic acid and 0.3 g of p-aminohippuric acid insufficient ethanol (96%) to produce 100 ml.

Aminohydroxynaphthalenesulphonic Acid See 4-amino-3-hydroxynaphthalene-1-sulphonic acid.

4-Amino-3-hydroxynaphthalene-1-sulphonic Acid 1-Amino-2-naphthol-4-sulphonic acid;C10H9NO4S = 239.3 (116-63-2)General reagent grade of commerce.A crystalline powder; melting point, about 300, with decomposition.Store protected from light.

Aminohydroxynaphthalenesulphonic Acid Solution Mix 5.0 g of anhydrous sodium sulphite with94.3 g of sodium hydrogensulphite and 0.7 g of 4-amino-3-hydroxynaphthalene-1-sulphonic acid. Dissolve1.5 g of the mixture in water and dilute to 10 ml with the same solvent. Prepare the solution daily.

Aminohydroxynaphthalenesulphonic Acid Solution, Strong Dissolve 0.25 g of 4-amino-3-hydroxynaphthalene-1-sulphonic acid in 75 ml of a 15% w/v solution of sodium metabisulphite, warmingto assist solution if necessary. Add 2.5 ml of a 20% w/v solution of sodium sulphite, mix and addsufficient of the sodium metabisulphite solution to produce 100 ml.

5-Aminoimidazole-4-carboxamide Hydrochloride C4H6N4O,HCl = 162.6 (72-40-2)General reagent grade of commerce.Melting point, about 251, with decomposition.

3-Aminomethylalizarin-N,N-diacetic Acid Aminomethylalizarindiacetic acid; alizarincomplexone; C19H15NO8,2H2O = 421.4 (3952-78-1)A fine, ochre to orange-brown powder; melting point, about 185.Complies with the following test.LOSS ON DRYING Not more than 10.0%, determined on 1 g.

74-8Aminomethylalizarindiacetic Acid Reagent SOLUTION I Dissolve 0.36 g of cerium(III) nitrate in sufficient water to produce 50 ml.SOLUTION II Suspend 0.7 g of 3-aminomethylalizarin-N,N-diacetic acid in 50 ml of water. Dissolvewith the aid of about 0.25 ml of 13.5M ammonia, add 0.25 ml of glacial acetic acid and dilute to100 ml with water.SOLUTION III Dissolve 6 g of sodium acetate in 50 ml of water, add 11.5 ml of glacial acetic acid anddilute to 100 ml with water.To 33 ml of acetone add 6.8 ml of solution III, 1.0 ml of solution II and 1.0 ml of solution I and diluteto 50 ml with water. Use within 5 days.Complies with the following test.SENSITIVITY To 1.0 ml of fluoride standard solution (10 ppm F) add 19.0 ml of water and 5.0 ml of thereagent being examined. After 20 minutes, a distinct blue colour is produced.

Aminomethylalizarindiacetic Acid Solution Dissolve 0.192 g of 3-aminomethylalizarin-N,N-diacetic acid in 6 ml of freshly prepared 1M sodium hydroxide. Add 750 ml of water, 25 ml of succinatebuffer solution pH 4.6 and, dropwise, 0.5M hydrochloric acid until the colour changes from violet-red toyellow (pH 4.5 to 5). Add 100 ml of acetone and dilute to 1000 ml with water.

3-(Aminomethyl)pyridine (3-Pyridylmethyl)amine; 3-picolylamine; C6H8N2 = 108.1 (3731-52-0)General reagent grade of commerce.

8-Aminonaphthalene-2-sulphonic Acid 8-Amino-2-naphthalenesulphonic acid; 1-naphthyl-amine-7-sulphonic acid; C10H9NO3S = 223.2 (119-28-8)General reagent grade of commerce.

Aminonaphthalenesulphonic Acid Solution Mix 0.5 g of 8-aminonaphthalene-2-sulphonic acid,30 ml of glacial acetic acid and 120 ml of water and heat with stirring until dissolved. Allow to cooland filter.Use the solution within 3 weeks.

Aminonitrobenzophenone See 2-amino-5-nitrobenzophenone.

2-Amino-5-nitrobenzophenone Aminonitrobenzophenone; C13H10N2O3 = 242.2 (1775-95-7)General reagent grade of commerce.A yellow, crystalline powder; melting point, about 160; A(1%, 1 cm), 690 to 720, determined at233 nm (0.001% w/v in methanol).

4-Aminophenazone Aminopyrazolone; 4-aminoantipyrine; 4-amino-2,3-dimethyl-1-phenyl-5-pyrazolone; C11H13N3O = 203.3 (83-07-8)General reagent grade of commerce.Light yellow needles or powder; melting point, about 108.

Aminophenazone Solution Aminopyrazolone solutionA 0.1% w/v solution of 4-aminophenazone in borate buffer pH 9.0.

4-Aminophenol C6H7NO = 109.1 (123-30-8)General reagent grade of commerce.A white or slightly coloured, crystalline powder; melting point, about 186, with decomposition.Store protected from light.

Aminopolyether 4,7,13,16,21,24-Hexaoxo-1,10-diazabicyclo[8,8,8]-hexacosane; C18H36N2O6 =376.5 (23978-09-8)General reagent grade of commerce.Melting point, 70 to 73.

3-Aminopropanol 3-Amino-1-propanol; 3-aminopropan-1-ol; C3H9NO = 75.1 (156-87-6)General reagent grade of commerce.Melting point, about 11; d2020 , about 0.99; nD20 , about 1.461.3-Aminopropionic Acid b-Alanine; C3H7NO2 = 89.1 (107-95-9)General reagent grade of commerce containing not less than 99% of C3H7NO2.Melting point, about 200, with decomposition.

Aminopyrazolone See 4-aminophenazone.Aminopyrazolone Solution See aminophenazone solution.

Ammonia NH3 = 17.03For 18M and 13.5M ammonia use analytical reagent grade solutions of commerce containing 35%and 25% w/w of NH3 and weighing 0.88 g and 0.91 g per ml, respectively.Solutions of molarity xM should be prepared by diluting 75x ml of 13.5M ammonia or 56x ml of 18Mammonia to 1000 ml with water.When ammonia is specified and the strength is not stated, use a reagent prepared by diluting 67 g of13.5M ammonia to 100 ml with water. When used in the limit test for iron, it complies with thefollowing test.IRON Evaporate 5 ml of the reagent being examined to dryness on a water bath, add 10 ml of water,2 ml of a 20% w/v solution of citric acid and 0.1 ml of thioglycollic acid. Add sufficient of the reagent tomake the solution alkaline and dilute to 20 ml with water. No pink colour is produced.

74-9Ammonia, Chloride-free 13.5M ammonia that complies with the following test.CHLORIDE Evaporate 54 ml on a water bath almost to dryness and dilute to 15 ml with water. Thesolution complies with the limit test for chlorides, Appendix VII (1 ppm).

Ammonia, Concentrated Strong Ammonia Solution of the British Pharmacopoeia (about 13.5M).

Ammonia, Methanolic Solutions of the requisite molarity may be obtained by diluting 13.5Mammonia or 18M ammonia with methanol as directed under ammonia.

Ammonia R1, Concentrated Concentrated ammonia containing not less than 32.0% w/w of NH3(about 18M).d2020 , 0.883 to 0.889.Store protected from atmospheric carbon dioxide at a temperature below 20.

Ammonia R1, Dilute Dilute 41 g of concentrated ammonia to 100 ml with water. The solutioncontains not less than 10.0% and not more than 10.4% w/v of NH3 (about 6M).

Ammonia R2, Dilute Dilute 14 g of concentrated ammonia to 100 ml with water. The solutioncontains not less than 3.3% and not more than 3.5% of NH3 (about 2M).

Ammonia R3, Dilute Dilute 0.7 g of concentrated ammonia to 100 ml with water. The solutioncontains not less than 0.16% w/v and not more than 0.18% w/v of NH3.

Ammonium Acetate C2H7NO2 = 77.08 (631-61-8)Analytical reagent grade of commerce.Store in an airtight container.

Ammonium Acetate Solution Dissolve 150 g of ammonium acetate in water, add 3 ml of glacialacetic acid and dilute to 1000 ml with water.Use within 1 week of preparation.

Ammonium and Cerium Nitrate See ammonium cerium(IV) nitrate.

Ammonium and Cerium Sulphate See ammonium cerium(IV) sulphate.

(1R)-()-Ammonium 10-Camphorsulphonate 10-Camphorsulphonic acid, ammonium salt;C10H19O4S = 249.3 (82509-30-6)General reagent grade of commerce containing not less than 97.0% of (1R)-()-ammoniumcamphorsulphonate.[ ]D20 , 18 2 (5% w/v in water).Ammonium Carbonate (507-87-6) Ammonium hydrogen carbonate and ammonium carbamate inapproximately equimolecular proportions containing the equivalent of about 30% w/w of NH3.Analytical reagent grade of commerce.Store in a well-closed container at a temperature not exceeding 20.

Ammonium Carbonate Solution A 15.8% w/v solution of ammonium carbonate.

Ammonium Carbonate Solution, Dilute Dissolve 5 g of ammonium carbonate in a mixture of7.5 ml of 5M ammonia and 50 ml of water, dilute to 100 ml with water and filter, if necessary.

Ammonium Cerium(IV) Nitrate Ceric ammonium nitrate; (NH4)2[Ce(NO3)6] = 548.2(16774-21-3)Analytical reagent grade of commerce.

Ammonium Cerium(IV) Sulphate Ceric ammonium sulphate; 2(NH4)2SO4,Ce(SO4)2,2H2O =632.6 (18923-36-9)General reagent grade of commerce.

Ammonium Chloride NH4Cl = 53.49 (12125-02-9)Analytical reagent grade of commerce.

Ammonium Chloride Solution A 10.7% w/v solution of ammonium chloride.

Ammonium Citrate Diammonium hydrogen citrate; C6H14N2O7 = 226.2 (3012-65-5)General reagent grade of commerce.ACIDITY pH of 2.26% w/v solution, about 4.3.

Ammonium Citrate Solution Dissolve, with cooling, 500 g of citric acid in a mixture of 200 ml ofwater and 200 ml of 13.5M ammonia. Filter and dilute to 1000 ml with water.

Ammonium Cobaltothiocyanate Solution Dissolve 37.5 g of cobalt(II) nitrate and 150 g ofammonium thiocyanate in sufficient water to produce 1000 ml.Use within 1 day of preparation.

Ammonium Dihydrogen Orthophosphate Ammonium dihydrogen phosphate; (NH4)H2PO4 =115.0 (7722-76-1)Analytical reagent grade of commerce.pH of a 2.3% w/v solution, about 4.2, Appendix V L.

74-10Ammonium Formate C2H5NO2 = 63.1 (540-69-2)General reagent grade of commerce.Melting point, 119 to 121.Store in an airtight container.

Ammonium Hydrogen Carbonate Ammonium bicarbonate; NH4HCO3 = 79.06 (1066-33-7)Analytical reagent grade of commerce containing not less than 99% of NH4HCO3.

Ammonium Iron(III) Citrate Ferric ammonium citrate (1185-57-5)General reagent grade (brown) of commerce.

Ammonium Iron(II) Sulphate Ferrous ammonium sulphate; (NH4)2Fe(SO4)2,6H2O = 392.1(7783-85-9)Analytical reagent grade of commerce.Store protected from light.

Ammonium Iron(III) Sulphate Ferric ammonium sulphate; NH4Fe(SO4)2,12H2O = 482.2 (7783-83-7)Analytical reagent grade of commerce.

Ammonium Iron(III) Sulphate Solution R1 Dissolve 0.2 g of ammonium iron(III) sulphate in 50 ml of water, add 5 ml of nitric acid and dilute to100 ml with water.

Ammonium Iron(III) Sulphate Solution R2 Ferric ammonium sulphate solution R2A 10% w/v solution of ammonium iron(III) sulphate. If necessary, filter before use.Produces a deep red colour with ammonium thiocyanate in acid solutions.

Ammonium Iron(III) Sulphate Solution R5 Ferric ammonium sulphate solution R5Shake 30.0 g of ammonium iron(III) sulphate with 40 ml of nitric acid and dilute to 100 ml with water.If the solution is turbid, it should be filtered or centrifuged.Store protected from light.

Ammonium Iron(III) Sulphate Solution R6 Ferric ammoniuj sulphate solution R6Dissolve 20 g of ammonium iron(III) sulphate in 75 ml of water, add 10 ml of a 2.8% w/v solution ofsulphuric acid (96% w/w) and dilute to 100 ml with water.

Ammonium Mercaptoacetate Solution Add 300 ml of water to 50 ml of mercaptoacetic acid,neutralise with about 40 ml of 13.5M ammonia and dilute to 500 ml with water.

Ammonium Mercurithiocyanate Reagent Dissolve 30 g of ammonium thiocyanate and 27 g ofmercury(II) chloride in sufficient water to produce 1000 ml.

Ammonium Metavanadate Ammonium vanadate; NH4VO3 = 117.0 (7803-55-6)Analytical reagent grade of commerce.

Ammonium Metavanadate Solution Dissolve, with heating, 5 g of ammonium metavanadate in amixture of 10 ml of 5M sodium hydroxide and 90 ml of water, cool and filter, if necessary, throughglass wool.

Ammonium Molybdate (NH4)6Mo7O24,4H2O = 1236 (12054-85-2)Analytical reagent grade of commerce.

Ammonium Molybdate Reagent Mix in the following order 1 volume of a 2.5% w/v solution ofammonium molybdate, 1 volume of a 10% w/v solution of L-ascorbic acid and 1 volume of 3M sulphuricacid and add 2 volumes of water.Use within 1 day of preparation.

Ammonium Molybdate Reagent R1 Mix 10 ml of a 6.0% w/v solution of disodium arsenate, 50 mlof ammonium molybdate solution, 90 ml of 1M sulphuric acid and add sufficient water to produce200 ml.Condition the mixture at 37 for 24 hours and keep in amber flasks.

Ammonium Molybdate Solution A 10% w/v solution of ammonium molybdate.

Ammonium Molybdate Solution R2 Dissolve 5 g of ammonium molybdate in 30 ml of water withheating, cool, adjust the pH to 7.0 with 2M ammonia and dilute to 50 ml with water.

Ammonium Molybdate Solution R3SOLUTION I Dissolve 5 g of ammonium molybdate in 20 ml of water with the aid of heat.SOLUTION II Mix 150 ml of ethanol (96%) with 150 ml of water and add, with cooling, 100 ml ofsulphuric acid.Add 80 volumes of solution II to 20 volumes of solution I immediately before use.

Ammonium Molybdate Solution R4 Dissolve 1.0 g of ammonium molybdate in sufficient water toproduce 40 ml, add 3 ml of hydrochloric acid and 5 ml of perchloric acid and dilute to 100 ml withacetone.Store protected from light and use within 1 month of preparation.

74-11Ammonium Molybdate Solution R5 Dissolve 1 g of ammonium molybdate in 40 ml of a 15% w/vsolution of sulphuric acid.Prepare the solution daily.

Ammonium MolybdateSulphuric Acid Solution Dissolve 10 g of ammonium molybdate insufficient water to produce 100 ml and add the solution slowly to 250 ml of cold 10M sulphuric acid.Store in a plastic bottle protected from light.

Ammonium Nitrate NH4NO3 = 80.04 (6484-52-2)Analytical reagent grade of commerce.Store in airtight container.

Ammonium Nitrate R1 Ammonium nitrate complying with the following requirements.ACIDITY A solution is faintly acid, Appendix V K.CHLORIDE 0.5 g complies with the limit test for chlorides, Appendix VII (100 ppm).SULPHATE 1.0 g complies with the limit test for sulphates, Appendix VII (150 ppm)SULPHATED ASH Not more than 0.05%, Appendix IX A, Method II. Use 1 g.

Ammonium Oxalate (CO2NH4)2,H2O = 142.1 (6009-70-7)Analytical reagent grade of commerce.

Ammonium Oxalate Solution A 4% w/v solution of ammonium oxalate.

Ammonium Persulphate Ammonium peroxodisulphate; (NH4)2S2O8 = 228.2 (7727-54-0)Analytical reagent grade of commerce.

Ammonium Phosphate See diammonium hydrogen orthophosphate.

Ammonium Polysulphide Solution Dissolve a sufficient quantity of precipitated sulphur to producea deep orange solution in a solution prepared in the following manner. Immediately before usesaturate 120 ml of 6M ammonia with hydrogen sulphide and add 80 ml of 6M ammonia.

Ammonium Pyrrolidinedithiocarbamate Ammonium tetramethylenedithiocarbamate;C5H12N2S2 = 164.3 (5108-96-3)General reagent grade of commerce.Store in a bottle containing a piece of ammonium carbonate in a muslin bag.

Ammonium Pyrrolidinedithiocarbamate Solution A 1.0% w/v solution of ammoniumpyrrolidinedithiocarbamate that has been washed immediately before use with three 25-ml quantities of4-methylpentan-2-one.

Ammonium Reineckate Ammonium tetrathiocyanatodiamminochromate(III) monohydrate;NH4[Cr(NH3)2 (CNS)4],H2O = 354.4 (13573-16-5)General reagent grade of commerce.Red powder or crystals.

Ammonium Reineckate Solution A 1.0% w/v solution of ammonium reineckate.Prepare immediately before use.

Ammonium Sulphamate NH2SO3NH4 = 114.1 (7773-06-0)General reagent grade of commerce.A white, crystalline powder or colourless crystals; melting point, about 130.Store in an airtight container.

Ammonium Sulphate (NH4)2SO4 = 132.1 (7783-20-2)Analytical reagent grade of commerce.

Ammonium Sulphide Solution Saturate 120 ml of dilute ammonia R1 with hydrogen sulphide andadd 80 ml of dilute ammonia R1.Prepare immediately before use.

Ammonium Thiocyanate NH4SCN = 76.12 (1762-95-4)Analytical reagent grade of commerce.Store in an airtight container.

Ammonium Thiocyanate Solution A 7.6% w/v solution of ammonium thiocyanate.

Ammonium Vanadate See ammonium metavanadate

Ammonium Vanadate Solution Dissolve 1.2 g of ammonium metavanadate in 95 ml of water anddilute to 100 ml with sulphuric acid.

Amoxicillin Trihydrate Amoxycillin trihydrate; C16H19N3O5S,3H2O = 419.4 (61336-70-7)General reagent grade of commerce.

Amyl Acetate C7H14O2 = 130.2Consists principally of 3-methylbutyl acetate with a small proportion of 2-methylbutyl acetate.Analytical reagent grade of commerce.A colourless liquid with a sharp, fruity odour; boiling point, about 140; weight per ml, about 0.87 g.

Amyl Alcohol See isoamyl alcohol.

74-12aaaaa-Amylase 1,4-a-D-glucane-glucanohydrolaseA white to light brown powder.

aaaaa-Amylase Solution A solution of a-amylase with an activity of 800 FAU per g.

Anethole p-Prop-1-enylanisole; C10H12O = 148.2 (4180-23-8)General reagent grade of commerce.A white crystalline mass at 20 to 21; liquid above 23; boiling point, about 230; nD

25 , about1.56.Anethole used in gas chromatography complies with the following test.ASSAY Examine by gas chromatography, Appendix III B, under the conditions described in the testfor Chromatographic profile in the monograph for Anise Oil using the reagent being examined assolution (1).

The area of the principal peak, corresponding to trans-anethole, with retention time of about 41minutes, is not less than 99.0% of the total area of the peaks.

cis-Anethole (Z)-1-Methoxy-4-prop-1-enylbenzene; C10H12O = 148.2General reagent grade of commerce.A white crystalline mass at 20; liquid above 23; boiling point, about 230; nD

25 , about 1.56.Cis-anethole used in gas chromatography complies with the following test.ASSAY Examine by gas chromatography, Appendix III B, under the conditions described in the test forChromatographic profile in the monograph for Anise Oil using the reagent being examined assolution (1).

The area of the principal peak is not less than 92.0% of the total area of the peaks.

Aniline C6H7N = 93.13 (62-53-3)Analytical reagent grade of commerce.A colourless to pale yellow oily liquid; boiling point, about 183 to 186; d2020 , about 1.02.Store protected from light.

Aniline Hydrochloride C6H7N,HCl = 129.6 (142-04-1)General reagent grade of commerce.

Aniline Hydrochloride Solution Dissolve 2 g of aniline hydrochloride in a mixture of 65 ml ofethanol (96%) and 35 ml of water and add 2 ml of hydrochloric acid.Use within one day of preparation.

Anion Exchange Resin A resin in chlorinated form containing quaternary ammonium groups.[CH2N+(CH3)3] attached to a polymer lattice consisting of polystyrene cross-linked with 2% ofdivinylbenzene. It is available as beads and the particle size is specified in the monograph. Wash theresin with 1M sodium hydroxide on a sintered-glass filter until the washings are free from chloride, thenwash with water until the washings are neutral. Suspend in freshly prepared ammonia-free water andprotect from atmospheric carbon dioxide.

Anion Exchange Resin, Strongly Basic A gel-type resin in hydroxide form containing quaternaryammonium groups [-CH2N+(CH3)3, type 1] attached to a polymer lattice consisting of polystyrenecross-linked with 8% of divinylbenzene.Brown, transparent beads containing about 50% of water; particle size, 0.2 mm to 1.0 mm; totalexchange capacity, at least 1.2 milliequivalents per ml.

Anion-exchange Resin for Chromatography, Strongly Basic A resin with quaternary aminegroups attached to a lattice of latex cross-linked with divinylbenzene.

Anion Exchange Resin R1 A resin containing quaternary ammonium groups [CH2N+(CH3)3]attached to a lattice consisting of methacrylate.

Anisaldehyde 4-Methoxybenzaldehyde; C8H8O2 = 136.2 (123-11-5)General reagent grade of commerce.A colourless to pale yellow, oily liquid with an aromatic odour; boiling point, about 248; weightper ml, about 1.125 g.Anisaldehyde used in gas chromatography complies with the following test.ASSAY Examine by gas chromatography, Appendix III B, under the conditions described in the test forChromatographic profile in the monograph for Anise Oil using the reagent being examined assolution (1).

The area of the principal peak is not less than 99.0% of the total area of the peaks.

Anisaldehyde Solution Mix in the following order 0.5 ml of anisaldehyde, 10 ml of glacial aceticacid, 85 ml of methanol and 5 ml of sulphuric acid.

Anisaldehyde Solution R1 To 10 ml of anisaldehyde add 90 ml of ethanol (96%), mix, add 10 ml ofsulphuric acid and mix again.

p-Anisidine C7H9NO = 123.2 (104-94-9)General reagent grade of commerce containing not less than 97.0% of C7H9NO.CAUTION p-Anisidine is a skin irritant and sensitiser.

74-13Store protected from light at 0 to 4.On storage, p-anisidine tends to darken as a result of oxidation. A discoloured reagent can bereduced and decolorised in the following manner. Dissolve 20 g of the reagent in 500 ml of water at75. Add 1 g of sodium sulphite and 10 g of activated charcoal and stir for 5 minutes. Filer, cool thefiltrate to about 0 and allow to stand at this temperature for at least 4 hours. Filter, wash thecrystals with a small quantity of water at about 0 and dry the crystals at a pressure of 2 kPa overphosphorus pentoxide.

Anolyte for Isoelectric Focusing pH 3 to 5 0.1M glutamic acid, 0.5M phosphoric acid Dissolve14.71 g of glutamic acid in water, add 33 ml of orthophosphoric acid and dilute to 1000 ml with water.

Anthracene C14H10 = 178.2 (120-12-7)General reagent grade of commerce.A white, crystalline powder; melting point, about 218.

Anthranilic Acid See 2-aminobenzoic acid.

Anthrone C14H10O = 194.2 (90-44-8)Analytical reagent grade of commerce.A pale yellow, crystalline powder; melting point, about 155.When used in an Assay for glucose or dextrans, complies with the following test.SENSITIVITY TO GLUCOSE Add, carefully, 6 ml of a 0.2% w/v solution in a mixture of 19 ml ofsulphuric acid and 1 ml of water to 3 ml of a solution containing 5 g of D-glucose per ml and heat in awater bath for 5 minutes. The solution is a darker green than a solution prepared in the same mannerbut omitting the glucose.

Anthrone Reagent A 0.2% w/v solution of anthrone in nitrogen-free sulphuric acid. The solutionshould be allowed to stand for 4 hours before use and should be discarded after 7 days.

Antimony Potassium Tartrate Antimony potassium oxide (+)-tartrate; KSbO,C4H4O6,H2O =333.9 (28300-74-5)Analytical reagent grade of commerce.

Antimony Trichloride SbCl3 = 228.1 (10025-91-9)Analytical reagent grade of commerce.Colourless crystals or flakes, fuming in moist air.Store in an airtight container, protected from moisture.

Antimony Trichloride in Dichloroethane Solution Prepare a 22.0% w/v solution of antimonytrichloride in dry 1,2-dichloroethane that has been purified by passing it down a column of silica gel andallow the solution to stand for 24 hours. To 100 ml add 2.5 ml of acetyl chloride and allow to standfor a further 24 hours before use.

Antimony Trichloride Solution Rapidly wash 30 g of antimony trichloride with two 15-mlquantities of ethanol-free chloroform. Drain off the washings and dissolve the washed crystalsimmediately in 100 ml of ethanol-free chloroform, warming slightly.Store over a few grams of anhydrous sodium sulphate.

Antimony Trichloride Solution R1 Prepare two stock solutions in the following manner.SOLUTION A Dissolve 110 g of antimony trichloride in 400 ml of 1,2-dichloroethane. Add 2 g ofanhydrous aluminium oxide, mix and filter through sintered glass into a 500-ml graduated flask. Diluteto 500 ml with 1,2-dichloroethane and mix. The absorbance of the resulting solution measured in a 2-cm cell at 500 nm, Appendix II B, is not more than 0.07 using 1,2-dichloroethane in the reference cell.SOLUTION B In a fume cupboard, mix 100 ml of colourless, distilled acetyl chloride and 400 ml of1,2-dichloroethane and store in a cool place.Mix 90 ml of solution A and 10 ml of solution B. Store in an amber, ground-glass-stoppered bottleand use within 7 days. Discard any reagent in which colour develops.

Antithrombin III ATIII (90170-80-2)Reagent grade of commerce.Antithrombin III is purified from human plasma by heparin agarose chromatography and shouldhave a specific activity of at least 6 IU per mg.

Antithrombin III Solution R1 Reconstitute antithrombin III as directed by the manufacturer anddilute with trischloride buffer pH 7.4 to contain 1 IU per ml.

Antithrombin III Solution R2 Reconstitute antithrombin III as directed by the manufacturer anddilute with tris-chloride buffer pH 7.4 to contain 0.5 IU per ml.

Apigenin 4,5,7-Trihydroxyflavone; C15H10O = 270.2 (520-36-5)General reagent grade of commerce.Light yellowish powder.Melting point, about 310, with decomposition.Complies with the following test.CHROMATOGRAPHY Carry out identification test C described under Chamomile Flowers applying

74-1410 l of a 0.025% w/v solution in methanol. The chromatogram shows a principal zone of yellowish-green fluorescence in the upper third.

Apigenin-7-glucoside C21H20O10 = 432.6General reagent grade of commerce.Light yellowish powder.Melting point, 198 to 201.Complies with the following test.CHROMATOGRAPHY Carry out identification test C described under Chamomile Flowers applying10 l of a 0.025% w/v solution in methanol. The chromatogram shows a principal zone of yellowishfluorescence in the middle third.

Aprotinin (9087-70-1)Reagent grade of commerce containing 10 to 20 trypsin inhibitor units per mg.

Arabinose See L-arabinose.

L-Arabinose Arabinose; C5H10O5 = 150.1 (87-72-9)General reagent grade of commerce.A white, crystalline powder; [ ]D20 , +103 to +105 (5.0% w/v in water containing about 0.05% v/v ofNH3).

Arachidic Alcohol Eicosan-1-ol; Arachidyl alcohol; C20H42O = 298.6 (629-96-9)Purified reagent grade of commerce usually containing not less than 95% of C20H42O.A colourless, waxy or crystalline solid.

Arbutin Arbutoside; 4-hydroxyphenyl-b-D-glucopyranoside; C12H16O7 = 272.3 (497-76-7)General reagent grade of commerce.Melting point, about 200; [ ]D20 , about 64, determined on a 2.0% w/v solution.CHROMATOGRAPHY When examined by test C for Identification in the monograph for BearberryLeaf the chromatogram shows only one principal spot.

Arginine L-Arginine; C6H14N4O2 = 174.2 (74-79-3)General reagent grade of commerce.Melting point, about 235, with decomposition.

Argon Ar = 39.95 (7440-37-1)Laboratory cylinder grade of commerce containing not less than 99.995% v/v of Ar.When used in the test for Carbon monoxide in medicinal gases (Appendix IX E), after passage of 10litres at a flow rate of 4 litres per hour, not more than 0.05 ml of 0.002M sodium thiosulphate VS isrequired for the titration (0.6 ppm).

Arsenious Trioxide Arsenic trioxide; As2O3 = 197.8 (1327-53-3)Analytical reagent grade of commerce.

Arsenite Solution Dissolve 0.50 g of arsenious trioxide in 5 ml of 2M sodium hydroxide solution, add2.0 g of sodium hydrogen carbonate and dilute to 100 ml with water.

Ascorbic Acid See L-ascorbic acid.

L-Ascorbic Acid C6H8O6 = 176.1 (50-81-7)Analytical reagent grade of commerce.A white, crystalline powder; [ ]D20 , about +22 (2% w/v in water).Ascorbic Acid Solution Dissolve 50 mg of L-ascorbic acid in 0.5 ml of water and dilute to 50 mlwith dimethylformamide.

Asiaticoside O-6-Deoxy-a-L-mannopyranosyl-(1 4)-O-b-D-glucopyranosyl-(1 6)-b-D-glucopyranosyl 2a,3b,23-trihydroxy-4a-urs-12-en-28-oate; C48H78O19 = 959 (16830-15-2)General reagent grade of commerce containing not more than 6.0% of water.Melting point,: about 232, with decomposition.Store protected from humidity.

Asiaticoside used in liquid chromatography complies with the following additional test.

ASSAY. When examined by the method described under Assay in the monograph for Centella, thecontent is not less than 97.0% by normalisation.

L-Aspartyl-L-phenylalanine Aspartame; (S)-3-amino-N-[(S)-1-carboxy-2-phenylethyl]succinamicacid; C13H16N2O5 = 280.3 (13433-09-5)General reagent grade of commerce.Melting point, about 210, with decomposition.

Atenolol C14H22N2O3 = 266.3 (56715-13-0)General reagent grade of commerce.

Atropine Sulphate C34H46N2O6,H2SO4,H2O = 694.8 (5908-99-6)General reagent grade of commerce.Colourless crystals or a white, crystalline powder; melting point, about 195.

74-15Azobenzene C12H10N2 = 182.2 (103-33-3)Use a grade of commerce suitable for chromatographic standardisations.Melting point, about 69.

Azomethine H Azomethine H, monosodium salt hydrate; sodium hydrogen 4-hydroxy-5-(2-hydroxybenzylideneamino)naphthalene-2,7-disulphonate; C17H12NNaO8S2 = 445.4 (5941-07-1)General reagent grade of commerce; the degree of hydration is variable.

Azomethine H Solution Dissolve 0.45 g of azomethine H and 1 g of ascorbic acid in water with theaid of gentle heat and add sufficient water to produce 100 ml.

Barbaloin 1,8-Dihydroxy-3-hydroxymethyl-10-(b-D-glucopyranosyl)anthrone; C21H22O9,H2O =436.4 (1415-73-2)General reagent grade of commerce.Lemon yellow to dark yellow needles or crystalline powder, darkening on exposure to air and light.A(1%, 1 cm) determined in methanol, about 192 at 269 nm, about 226 at 296.5 nm and about 259 at354 nm, calculated with reference to the anhydrous substance.Complies with the following test.HOMOGENEITY Carry out the test for Other species of Rhamnus, anthrones described under Frangulabark applying to the plate a solution containing only the reagent being examined. The chromatogramobtained shows only one principal spot.

Barbital See barbitone.

Barbital Sodium See barbitone sodium.

Barbitone Barbital; 5,5-diethylbarbituric acid; C8H12N2O3 = 184.2 (57-44-3)General reagent grade of commerce.Melting point, about 190.

Barbitone Sodium Barbital sodium; sodium 5,5-diethylbarbiturate; C8H11N2NaO3 = 206.2(144-02-5)General reagent grade of commerce.

Barbituric Acid 2,4,6-Trihydroxypyrimidine; C4H4N2O3 = 128.1 (67-52-7)General reagent grade of commerce.A white or almost white powder; melting point, about 253.

Barium Carbonate BaCO3 = 197.3 (513-77-9)Analytical reagent grade of commerce.

Barium Chloride BaCl2,2H2O = 244.3 (10326-27-9)Analytical reagent grade of commerce.

Barium Chloride Solution A 10.0% w/v solution of barium chloride.

Barium Chloride Solution R1 A 6.1% w/v solution of barium chloride.

Barium Chloride Solution R2 A 3.65% w/v solution of barium chloride.

Barium Hydroxide Ba(OH)2,8H2O = 315.5 (12230-71-6)Analytical reagent grade of commerce.

Barium Hydroxide Solution A 4.73% w/v solution of barium hydroxide.

Barium Sulphate BaSO4 = 233.4 (7727-43-7)Precipitated grade of commerce.

Benzaldehyde C7H6O = 106.1 (100-52-7)Analytical reagent grade of commerce.A colourless to pale yellow, oily liquid with an odour of almonds; d2020 , about 1.05; nD20 , about 1.545.DISTILLATION RANGE Not less than 95% distils between 177 and 180.Store protected from light.

Benzalkonium Chloride Of the British Pharmacopoeia.

Benzalkonium Chloride Solution Of the British Pharmacopoeia.Benzamide C7H7NO = 121.1 (55-21-0)General reagent grade of commerce.Melting point, about 128.

Benzene C6H6 = 78.1 (71-43-2)Analytical reagent grade of commerce.A flammable, colourless liquid; boiling point, about 80.

Benzethonium Chloride C27H42ClNO2,H2O = 466.1 (121-54-0)General reagent grade of commerce.A fine, white powder, or colourless crystals; melting point, about 163.Store protected from light.

Benzil Diphenylethanedione; C14H10O2 = 210.2

74-16(134-81-6)General reagent grade of commerce.Melting point, 95.

Benzilic Acid Diphenylglycollic acid; C14H12O3 = 228.3 (76-93-7)General reagent grade of commerce.Melting point, about 151.

Benzocaine Ethyl 4-aminobenzoate; C9H11NO2 = 165.2 (94-09-7)Gneral reagent grade of commerce.Melting point, about 90.

Benzoic Acid C7H6O2 = 122.1 (65-85-0)Analytical reagent grade of commerce.Colourless, feathery crystals or a white powder; melting point, about 121.

Benzoin a-Hydroxy-a-phenylacetophenone; C14H12O2 = 212.3 (579-44-2)General reagent grade of commerce.Melting point, about 137.

Benzophenone C13H10O = 182.2 (119-61-9)General reagent grade of commerce.Melting point, about 48.

1,4-Benzoquinone Cyclohexa-2,5-diene-1,4-dione C6H4O2 = 108.1 (106-51-4)General reagent grade of commerce containing not less than 98.0% of C6H4O2.

Benzoyl Chloride C7H5ClO = 140.6 (98-88-4)Analytical reagent grade of commerce.A colourless, lachrymatory liquid, fuming in moist air; d2020 , about 1.21; boiling point, about 197.Benzoyl Peroxide C14H10O4 = 242.2 (94-36-0)General reagent grade of commerce.White or almost white granules; melting point, after drying, about 104.For safety Benzoyl Peroxide should be kept moistened with about 23% w/w of water.

Benzoylarginine Ethyl Ester Hydrochloride C15H23ClN4O3 = 342.8 (2645-08-1)General reagent grade of commerce.[ ]D20 , 15 to 18, determined in a 1% w/v solution; melting point, about 129; A(1%, 1 cm), 310to 340 determined at 227 nm using a 0.001% w/v solution.

N-Benzoyl-L-prolyl-L-phenylalanyl-L-arginine 4-nitroanilide acetate C35H42N8O8 = 703General reagent grade of commerce.

Benzyl Alcohol C7H8O = 108.1 (100-51-6)General reagent grade of commerce.A colourless liquid; boiling point, about 204; weight per ml, about 1.05 g.

Benzyl Benzoate C14H12O2 = 212.3 (120-51-4)General reagent grade of commerce.Leaflets or an oily liquid; melting point, about 20; weight per ml, about 1.12 g.When used in the test for Identification in the monograph for Peru Balsam, complies with the followingrequirement.CHROMATOGRAPHY Carry out test B for Identification described in the monograph for Peru Balsamapplying to the plate 20 l of a 0.3% w/v solution of the reagent being examined in ethyl acetate. Aftervisualisation, the chromatogram exhibits a principal spot with an Rf value of about 0.8.

Benzyl Butyl Phthalate C19H20O4 = 312 (85-68-7)General reagent grade of commerce.

Benzyl Cinnamate Benzyl 3-phenylprop-2-enoate; C16H14O2 = 238.3 (103-41-3)General reagent grade of commerce.Colourless or yellowish crystals; melting point, about 39.Complies with the following requirement.CHROMATOGRAPHY Carry out test B for Identification described in the monograph for Peru Balsamapplying to the plate 20 l of a 0.3% w/v solution of the reagent being examined in ethyl acetate. Aftervisualisation, the chromatogram exhibits a principal spot with an Rf value of about 0.6.

Benzylamine C7H9N = 107.2General reagent grade of commerce usually containing not less than 98% of C7H9N.Colourless to pale yellow, oily liquid; boiling point, about 185; weight per ml, about 0.98 g.

Benzylpenicillin Sodium Of the British Pharmacopoeia.

2-Benzylpyridine C12H11N = 169.2 (101-82-6)General reagent grade of commerce containing not less than 98.0% of C12H11N.

74-17Melting point, 13 to 16.

Benzyltrimethylammonium Chloride C10H16Cl = 185.7 (56-93-9)General reagent grade of commerce.

Bergapten 5-Methoxypsoralen; C12H8O4 = 216.2 (484-20-8)General reagent grade of commerce.Melting point, about 188.

Betamethasone 9a-Fluoro-11b,17a,21-trihydroxy-16b-methylpregna-1,4-diene-3,20-dione;C22H29FO5 = 392.5 (378-44-9)General reagent grade of commerce.Melting point, about 236.

Betulin Lup-20(39)-ene-3b,28-diol; C30H50O2 = 442.7 (473-98-3)General reagent grade of commerce.Melting point, 248 to 251.

Bibenzyl 1,2-Diphenylethane; C14H14 = 182.3 (103-29-7)General reagent grade of commerce.Colourless or white crystals; melting point, about 50 to 53.

Biphenyl C12H10 = 154.2 (92-52-4)General reagent grade of commerce.Melting point, about 70.

Biphenyl-4-ol See 4-hydroxybiphenyl.

Bisbenzimide 4-{5-[5-(4-Methylpiperazin-1-yl)benzimidazol-2-yl]benzimidazol-2-yl}phenoltrihydrochloride pentahydrate; C25H24N6O,3HCl,5H2O = 624(23491-44-3)General reagent grade of commerce.

Bisbenzimide Stock Solution Dissolve 5 mg of bisbenzimide in sufficient water to produce 100 ml.Store protected from light.

Bisbenzimide Working Solution Immediately before use, dilute 100 l of bisbenzimide stock solutionto 100 ml with phosphate-buffered saline pH 7.4.

Bismuth Oxycarbonate Bismuth subcarbonate (5892-10-4)A basic salt corresponding approximately to the formula (BiO)2CO3,H2O, containing theequivalent of about 91% of Bi2O3.

Bismuth Oxynitrate Bismuth subnitrate 4[BiNO3(OH2),BiO(OH)] = 1462 (1304-85-4)A basic salt containing about 80% of Bi2O3.

Bismuth Oxynitrate R1 Bismuth subnitrate R1Bismuth oxynitrate containing not less than 71.5% and not more than 74.0% of bismuth and not lessthan 14.5% and not more than 16.5% of nitrate, calculated as nitrogen pentoxide, N2O5.

Bismuth Oxynitrate Solution Bismuth subnitrate solutionDissolve 5 g of bismuth oxynitrate R1 in a mixture of 8.4 ml of nitric acid and 50 ml of water and diluteto 250 ml with water; filter if necessary.ACIDITY To 10 ml of the reagent add 0.05 ml of methyl orange solution and titrate with 1M sodiumhydroxide VS. Not less than 5.0 ml and not more than 6.25 ml is required to change the colour of thesolution.

Bismuth Subcarbonate See bismuth oxycarbonate.

Bismuth Subnitrate See bismuth oxynitrate.

Bismuth Subnitrate R1 See bismuth oxynitrate R1.

Bismuth Subnitrate Solution See bismuth oxynitrate solution.

N,O-Bis(trimethylsilyl)acetamide C8H21NOSi2 = 203.4 (10416-59-8)General reagent grade of commerce.A colourless liquid; weight per ml, about 0.83 g.

Bis(trimethylsilyl)trifluoroacetamide C8H18F3NOSi2 = 257.4 (25561-30-2)General reagent grade of commerce.Weight per ml, about 0.97 g; nD

20 , about 1.384.

Biuret C2H5N3O2 = 103.1 (108-19-0)General reagent grade of commerce.White, hygroscopic crystals; melting point, 188 to 190, with decomposition.

Biuret Reagent Dissolve 1.5 g of copper(II) sulphate and 6.0 g of potassium sodium (+)-tartrate in500 ml of water, add 300 ml of a carbonate-free 10% w/v solution of sodium hydroxide and sufficientof the same solution to produce 1000 ml and mix.

Blocking Solution A 10% v/v solution of acetic acid.

74-18Blue Dextran 2000 (9049-32-5) Prepared from dextran having an average molecular weight ofabout 2 106 by introduction of a polycyclic chromophore that colours the substance blue. Thedegree of substitution is 0.017. It is freeze-dried and dissolves rapidly and completely in water andaqueous saline solutions.Complies with the following test.LIGHT ABSORPTION A 0.1% w/v solution in phosphate buffer pH 7.0 exhibits a maximum at 280 nm,Appendix II B.

Boldine C19H21NO4 = 327.3 (476-70-0)General reagent grade of commerce.Melting point, about 163; [ ]D25 , about +27 determined in a 0.1% w/v solution in absolute ethanol.CHROMATOGRAPHY When used in the reference solution in Identification test C in the monographfor Boldo Leaf the chromatogram shows a single principal spot.ASSAY Carry out the test for Alkaloids in the monograph for Boldo Leaf using a 0.012% w/v solutionof the reagent being examined in the mobile phase. The area of the principal peak is not less than99.0% by normalisation.

Borate Solution Dissolve 9.55 g of sodium tetraborate in sulphuric acid (96% w/w), heating on a waterbath, and add sufficient sulphuric acid (96% w/w) to produce 1000 ml.

Boric Acid H3BO3 = 61.83 (10043-35-3)Analytical reagent grade of commerce.

Boric Acid Solution Dissolve 5 g of boric acid in a mixture of 20 ml of water and 20 ml of absoluteethanol and dilute to 250 ml with absolute ethanol.

Bormeol See D-borneol.

D-Borneol Borneol; endo-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ol; C10H18O = 154.3 (507-70-0)General reagent grade of commerce.Melting point, about 208.Complies with the following test.HOMOGENEITY Carry out the method for thin-layer chromatography, Appendix III A, using silica gel Gas the coating substance and chloroform as the mobile phase, but allowing the solvent front to ascend10 cm above the line of application. Apply to the plate 10 l of a 0.1% w/v solution of the substancebeing examined in toluene. After removal of the plate, allow it to dry in air and spray with anisaldehydesolution, using 10 ml for a plate 200 mm 200 mm, and heat at 100 to 105 for 10 minutes. Thechromatogram obtained shows only one principal spot.

Bornyl Acetate See D-bornyl acetate.

D-Bornyl Acetate Bornyl Acetate; C12H20O2 = 196.3 (5655-61-8)General reagent grade of commerce.Melting point, about 28; [ ]D20 , about +44 (10% w/v in ethanol).Complies with the following test.HOMOGENEITY Carry out the method for thin-layer chromatography, Appendix III A, using silica gel Gas the coating substance and chloroform as the mobile phase, but allowing the solvent front to ascend10 cm above the line of application. Apply to the plate 10 l of a 0.2% w/v solution of the reagentbeing examined in toluene. After removal of the plate, allow it to dry in air and spray with anisaldehydesolution, using 10 ml for a plate 200 mm 200 mm, and heat at 100 to 105 for 10 minutes. Thechromatogram obtained shows only one principal spot.

Boron Trichloride BCl3 = 117.2 (10294-34-5)General reagent grade of commerce.Colourless gas which reacts violently with water. Available as solutions in suitable solvents (2-chloro-ethanol, dichloromethane, hexane, heptane, methanol).Boiling point, about 12.6; nD

20 , about 1.420

Boron Trichloride-Methanol Solution A 12% w/v solution of BCl3 in methanol.Store protected from light at 20, preferably in sealed tubes.

Boron Trifluoride BF3 = 67.8 (7637-07-2)General regent grade of commerce.Colourless gas.

Boron TrifluorideMethanol Solution See Boron trifluoride solution.Boron Trifluoride Solution A grade of commerce containing about 14% of BF3 in methanol.

Bovine Coagulation Factor Xa An enzyme which converts prothrombin to thrombin. The semi-purified preparation is obtained from liquid bovine plasma and it may be prepared by activation ofthe zymogen factor X with a suitable activator such as Russells viper venom.Store freeze-dried preparation at 20 and frozen solution at a temperature not higher than 20.

Bovine Factor Xa Solution Reconstitute bovine coagulation factor Xa as directed by themanufacturer and dilute with tris-chloride buffer pH 7.4. Any change in the absorbance of the solution,

74-19Appendix II B, measured at 405 nm using tris-chloride buffer pH 7.4 in the reference cell, is not morethan 0