SK | 18 December |2 | The International Pharmacopoeia and International Chemical Reference Substances Rabat 29/11/2007 Quality Assurance and Safety: Medicines World Health Organization

SK | 18 December |3 | The International Pharmacopoeia – Int.Ph. 1. Introduction 2. Special features 3. Example of a monograph 4. WHO’s strategy in QC 5. WHO’s related activities

SK | 18 December |4 |

5 |5 | WHO Procedure for the preparation of drug Quality Control specifications (1) …..or why it takes so long…. Preliminary consultation and drafting Draft Quality Control specifications Method development + validation: WHO Collaborating Centres and experts, contracted laboratories Circulation for comments + testing of samples Revision process, additional studies, contacts with manufacturers for queries and additional samples as needed

SK | 18 December |6 | WHO Procedure for the preparation of drug Quality Control specifications (2) Adoption by WHO Expert Committee on Specifications for Pharmaceutical Preparations Presentation to WHO Governing Bodies Recommendation to governments for implementation  publication in Technical Reports and  The International Pharmacopoeia and Basic Tests series

SK | 18 December |7 | WHO does the work … with Partners National and regional authorities International organizations (UNAIDS, UNICEF, World Bank, etc.) International professional and other associations, NGOs (including consumer associations, industry) WHO Expert Panels (official nomination process) Specialists from all areas, regulatory, university, industry……… WHO Collaborating Centres (official nomination process) Pharmacopoeia Commissions and Secretariats, national institutions and institutes.. Regional and interregional groups (ICH…)

SK | 18 December |8 | Type of monographs  Drug substances  Excipients  Finished dosage forms  General methods and requirements:  oral dosage forms, e.g. tablets  dissolution testing…

SK | 18 December |9 | Some Figures - 3 rd edition  Volume 1: 42 General methods and requirements  Volume 2: 88 Active pharmaceutical ingredients  Volume 3: 100 Active pharmaceutical ingredients

SK | 18 December | Some Figures - 3 rd edition  Volume 4: 23 Active pharmaceutical ingredients 65 Excipients 25 Oral dosage forms 14 Injectables 11 General methods and requirements

SK | 18 December | Some Figures - 3 rd edition  Volume 5: 20 Oral dosage forms 39 Active pharmaceutical ingredients 9 General methods and requirements antimalarial agents, artemisinin derivatives: Special section on antimalarial agents, artemisinin derivatives: 5 Active pharmaceutical ingredients 8 Oral dosage forms + General guidance texts on INNs, graphic formulae, establishment of RS...

SK | 18 December | 4 th edition - new  Consolidated in 2 Volumes + CD-ROM:  Vol 1 - pharmaceutical substances (A-O)  Vol 2 - pharmaceutical substances (P-X) + dosage forms + radiopharmaceuticals + methods of analysis + reagents  New:  Monographs on antiretrovials  Revision of existing monographs  Improved presentation  Improved cross-referencing to general methods  CD-ROM: improved search functions

SK | 18 December | 4 th edition - new  New list of impurities shown to be controlled by tests  More to come, first supplement in preparation to cover:  New monographs for ARVs, TB and Malaria medicines,  Revision of others, e.g. to include dissolution tests  For pre-information visit the WHO web site: 

SK | 18 December | New Specifications adopted by 40 th WHO Expert Committee ARVs/APIs: - abacavir sulfate, - efavirenz, - lamivudine, - stavudine, - zidovudine; ARVs/finished products: - nelfinavir mesilate tablets, - nelfinavir mesilate oral powder, and - saquinavir mesilate capsules fixed-dose antituberculosis medicines: - rifampicin tablets, - rifampicin capsules, - rifampicin + isoniazid tablets, - rifampicin + isoniazid + pyrazinamide + ethambutol HCl tablets, - isoniazid + ethambutol HCl tablets, - rifampicin + isoniazid + pyrazinamide tablets

SK | 18 December | New Specifications adopted by 41 st WHO Expert Committee - ARVs –Abacavir oral solution –Abacavir sulfate tablets –Didanosine tablets –Didanosine oral solution (adult formulation) –Lamivudine oral solution –Lamivudine tablets –Stavudine capsules –Zidovudine capsules –Zidovudine IV injection –Zidovudine oral solution –Zidovudine and Lamivudine tablets –Zidovudine, Lamivudine and Abacavir tablets

SK | 18 December | New Specifications adopted by 41 st WHO Expert Committee – antimalarial medicines Doxycycline hyclate capsules (new monograph) Doxycycline hyclate tablets (revision) Doxycycline hyclate (revision) Lumefantrine (new monograph) - subject to further studies

SK | 18 December | New Specifications adopted by 41 st WHO Expert Committee – revisions (dissolution ) Metronidazole tablets Doxycycline tablets Isoniazid tablets Chloroquine phosphate tablets Primaquine diphosphate tablets Ethambutol hydrochloride tablets Pyrazinamide tablets Rifampicin capsules Rifampicin tablets

SK | 18 December | Specifications distributed for comments Oral liquids Artemether and lumefantrine tablets Lumefantrine Oseltamivir phosphate Zinc sulfate Zinc sulfate oral solution Zinc sulfate tablets Magnesium sulfate injection

SK | 18 December | WHO’s strategy for quality control  Step-wise approach: Basic tests (identification) Screening tests (TLC) The International Pharmacopoeia International reference materials (ICRS and IR reference spectra)

SK | 18 December | Int. Ph. and links with other programmes and organizations  Establishment of monographs for antiretrovirals collaboration Ph.Eur. USP, JP, IP, Chinese Pharmacopoeia, Brazilian Pharmacopoeia... collaboration with manufacturers links with WHO-UNICEF project on prequalification of suppliers for HIV drugs

SK | 18 December | International Pharmacopoeia Special features: 1. ….when complex, technically demanding methods are described (e.g. HPLC), --> a less technically demanding analytical method (e.g. TLC) proposed as alternative (if possible). 2. …. international validation - impurity profile can vary from country to country!!

SK | 18 December | C 36 H 47 N 5 O 4,H 2 O 4 S Relative molecular mass Chemical name. (2S)-1-[(2S,4R)-4-benzyl-2-hydroxy-5- [[(1S,2R)-2-hydroxy-2,3-dihydro-1H-inden-1-yl] amino]-5-oxopentyl]-N-(1,1-dimethylethyl)-4-(pyridin-3-ylmethyl) piperazine-2-carboxamide sulfate; CAS Reg. No C 36 H 47 N 5 O 4,H 2 O 4 S Relative molecular mass Chemical name. (2S)-1-[(2S,4R)-4-benzyl-2-hydroxy-5- [[(1S,2R)-2-hydroxy-2,3-dihydro-1H-inden-1-yl] amino]-5-oxopentyl]-N-(1,1-dimethylethyl)-4-(pyridin-3-ylmethyl) piperazine-2-carboxamide sulfate; CAS Reg. No Indinaviri sulfas - Indinavir sulfate

SK | 18 December | Description. A white or almost white powder. Solubility. Freely soluble in water, soluble in methanol. Category. Antiretroviral (protease inhibitor). Storage. Indinavir sulfate should be kept in a tightly closed container, protected from light. Additional information. Indinavir sulfate occurs as the monoethanolate which is hygroscopic. It converts to the hydrate upon loss of ethanol and exposure to moist air. Requirements Definition. Indinavir sulfate contains not less than 98.5% and not more than 101.0% of C 36 H 47 N 5 O 4,H 2 O 4 S calculated on anhydrous, ethanol free basis. Description. A white or almost white powder. Solubility. Freely soluble in water, soluble in methanol. Category. Antiretroviral (protease inhibitor). Storage. Indinavir sulfate should be kept in a tightly closed container, protected from light. Additional information. Indinavir sulfate occurs as the monoethanolate which is hygroscopic. It converts to the hydrate upon loss of ethanol and exposure to moist air. Requirements Definition. Indinavir sulfate contains not less than 98.5% and not more than 101.0% of C 36 H 47 N 5 O 4,H 2 O 4 S calculated on anhydrous, ethanol free basis. Indinaviri sulfas - Indinavir sulfate

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Identity tests Either tests A, B and D, or tests C and D may be applied. A. Carry out test A.1. or, where UV detection is not available, test A.2. A.1. Carry out the test as described under Thin-layer chromatography, using silica gel R6 as the coating substance and a mixture of 8 volumes of dichloromethane R and 2 volumes of 2-propanol as the mobile phase. Apply separately to the plate 10 μl of each of 2 solutions in methanol containing (A) 5 mg of the test substance per ml and (B) 5 mg of indinavir RS per ml. After removing the plate from the chromatographic chamber, allow it to dry exhaustively in a current of cool air. Examine the chromatogram in ultraviolet light (254 nm) Thin-layer chromatography The principal spot obtained with solution A corresponds in position, appearance, and intensity with that obtained with solution B.

SK | 18 December | Indinaviri sulfas - Indinavir sulfate B. The absorption spectrum of a mg/ml solution, when observed between 220 nm and 280 nm, exhibits one maximum at about 260 nm; the specific absorbance is between 56 and 65. C. Dissolve 0.1 g in 10 ml of water, add 2 ml of sodium hydroxide (~80g/l)TS and shake. Filter the resulting precipitate and wash with two 3-ml quantities of water. Dry the washed precipitate for one hour at 105°C. Using the dried precipitate thus obtained, carry out the examination as described under "1. 7 Spectrophotometry in the infrared region". The infrared absorption spectrum is concordant with the spectrum obtained from indinavir RS or with the reference spectrum of indinavir. D. A 20 mg/ml solution yields reaction A described under 2.1 General identification tests as characteristic of sulfates.2.1 General identification tests

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Specific optical rotation. Use a 10.0 mg/ml solution and calculate with reference to the anhydrous and ethanol free substance: +27° to +31°. Heavy metals. Use 1.0 g for the preparation of the test solution as described under Limit test for heavy metals, Procedure 1; determine the heavy metals content according to Method A; not more than 10 μg/g Limit test for heavy metals Sulfated ash. Not more than 1.0 mg/g. Water. Determine as described under 2.8 Determination of water by the Karl Fischer method, Method A, using 0.5 g of the substance; the water content is not more than 15 mg/g.2.8 Determination of water by the Karl Fischer method pH value. pH of a 10 mg/ml solution in carbon-dioxide-free water R:

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Ethanol content. Determine by Gas chromatography, using static head-space injection. Use a fused-silica capillary or wide bore column 30 m long and 0.32 mm or 0.53 mm in internal diameter coated with macrogol 20M R (film thickness: 0.25 μm) Gas chromatography As detector use a flame ionization detector. Use nitrogen for chromatography R or helium R as the carrier gas at an appropriate pressure and a split ratio 1:5 with a linear velocity of about 35 cm/sec. Maintain the temperature of the column at 30°C for 7 min, then raise the temperature at a rate of 35°C per min to 180°C and maintain for 10 min, maintaining the temperature of the injection port at 140°C and that of the flame ionization detector at 250°C.

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Test solution. Dissolve g of the test substance in purified water and dilute to 20.0 ml with the same solvent. Introduce 5.0 ml of this solution and 1.0 ml of purified water into a headspace vial. Prepare two more vials. Reference solutions. Add g of ethanol R to purified water and dilute to ml with the same solvent. Transfer respectively 2.0 ml, 3.0 ml and 4.0 ml in separate headspace injection vials and bring the volume to 6.0 ml with purified water. Blank solution. Introduce 6.0 ml of purified water into a headspace vial. The test is not valid unless the relative standard deviation on the areas of the peaks obtained from the test solutions is not more than 5%. Calculate the ethanol content by using the results obtained with the test solution and with the reference solutions; the ethanol content is not less than 50 mg/g and not more than 80 mg/g.

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Related substances. Carry out the test as described under High performance liquid chromatography, using a stainless steel column (25 cm x 4.6 mm) packed with base-deactivated octadecylsilyl silica gel for chromatography R (5μm) High performance liquid chromatography Use the following conditions for gradient elution: Mobile phase A: 30 volumes of acetonitrile R, 5 volumes of phosphate buffer pH 7.5 and 65 volumes of purified water. Mobile phase B: 70 volumes of acetonitrile R, 5 volumes of phosphate buffer pH 7.5 and 25 volumes of purified water. Prepare the following solutions. For solution (1) use 2.0 mg of the test substance per ml. For solution (2) dilute a suitable volume of solution (1) to obtain a concentration equivalent to 2 μg of Indinavir sulfate per ml. For the system suitability test: prepare solution (3) using 2 ml of solution (1) and 2 ml of sulfuric acid (190 g/l), heat carefully in a boiling water bath

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Operate with a flow rate of 1.0 ml per minute. As a detector use an ultraviolet spectrophotometer set at a wavelength of 220 nm. Maintain the column temperature at 40°C. Inject 20 μl of solution (3). The test is not valid unless the resolution factor between the two major peaks, with a retention time between 15 and 20 min, is not less than 3.5. If necessary adjust the amount of acetonitrile in mobile phase A, or adjust the gradient program. Inject alternatively 20 μl each of solutions (1) and (2). In the chromatograms obtained with solution (1), the area of any peak, other than the principal peak, is not greater than the area of the principal peak obtained with solution (2) (0.1 %). The sum of the areas of all peaks, other than the principal peak, is not greater than five times the area of the principal peak obtained with solution (2) (0.5 %). Disregard any peak with an area less than 0.5 times the area of the principal peak in the chromatogram obtained with solution (2) (0.05%).

SK | 18 December | Indinaviri sulfas - Indinavir sulfate Assay. Dissolve g, accurately weighed, in 50 ml of water and titrate with sodium hydroxide (0.1 mol/l) VS, determine the end point potentiometrically. Each ml of sodium hydroxide (0.1 mol/l) VS is equivalent to mg of C 36 H 47 N 5 O 4,H 2 O 4 S; calculate with reference to the anhydrous and ethanol free substance

SK | 18 December | Int. Ph. and links with other programmes and organizations  Monographs for antimalarials, anti-TB drugs (different clusters in WHO)  General requirements for products derived from plant materials (WHO Traditional Medicines Programme)  Monographs for radiopharmaceuticals (with International Atomic Energy Agency - IAEA)  Monographs for excipients (with Pharmacopoeial Discussion Group - PDG)

SK | 18 December | International Chemical Reference Substances (ICRS)  202 ICRS + 12 melting point reference substances  Established by WHO COLLABORATING CENTRE FOR CHEMICAL REFERENCE SUBSTANCES in Sweden  Primary reference standard  Linked to Ph.Int.  Price for ICRS US$ 70  Includes: - Directions for use and  - Certificate of analysis  Monitoring and on-going stability testing  Can be used for tests and analysis not included in Ph.Int.

SK | 18 December |

SK | 18 December | International Chemical Reference Substances (ICRS) For ordering information, please visit

SK | 18 December | International Infra-red Reference Spectra  WHO COLLABORATING CENTRE FOR CHEMICAL REFERENCE SUBSTANCES in Sweden  69 International Infrared Reference Spectra  Linked to Ph.Int.  Price for ICRS US$ 5  Publication under review.. IR-spectrum of lamivudine

SK | 18 December | International Chemical Reference Substances Establishment of 4-ETC ICRS

SK | 18 December | International Chemical Reference Substances 4- ETC CoA of Manufacturer

SK | 18 December | International Chemical Reference Substances 4-ETC Analysis by WHO Collaborating Centre Sweden Intended use The stock of the current batch of the International Chemical Reference Substance (ICRS) for 4-epitetracycline hydrochloride Control No is depleted and has to be replaced. The monograph for Tetracycline hydrochloride in The International Pharmacopoeia, Fourth Edition, requires a reference substance of 4- epitetracycline hydrochloride to be used in the thin-layer chromatographic test for related substances. Material About 15 g of the sample (manufacturer´s batch no 10) were received at the WHO Centre in September The material is being stored in tightly closed containers at + 5 °C, protected from light.

SK | 18 December | International Chemical Reference Substances 4-ETC Infrared spectrum

SK | 18 December | International Chemical Reference Substances 4-ETC UV-spectrum A UV-spectrum in 10 mM hydrochloric acid was recorded

SK | 18 December | International Chemical Reference Substances 4-ETC Thin-layer chromatography Two secondary spots were detected. They were identified as tetracycline hydrochloride and 4-epianhydro- tetracycline hydrochloride and estimated to about 0.1% and 0.3%, respectively

SK | 18 December | International Chemical Reference Substances 4-ETC High performance liquid chromatography The purity was estimated to 99.6% (w/w). Two impurities above the limit of quantification were detected. They were identified as tetracycline hydrochloride and 4- epianhydrotetracycline hydrochloride and estimated with external standards to 0.1% and 0.3% (w/w), respectively (n=6, RSD=0.01% for the main peak, RSD=9.5% for tetracycline, RSD=3.6% for 4-epianhydrotetracycline.

SK | 18 December | International Chemical Reference Substances 4-ETC

SK | 18 December | International Chemical Reference Substances 4-ETC Stability No special stability studies have been performed. Regular re-examinations of this ICRS when stored in the dry state will be performed. Conclusion 4-Epitetracycline hydrochloride, Control No , can be considered suitable as International Chemical Reference Substance for the intended purpose.

SK | 18 December | What needs to be kept in mind!! 1.Quality cannot be tested into the product!

SK | 18 December | What needs to be kept in mind!! 2. Specifications in national and regional pharmacopoeias are: - based on manufacturers' specifications - specific for the product(s) marketed in its legislative territory

SK | 18 December | What needs to be kept in mind!! 3. Specifications should be used in a comprehensive way, i.e. including all tests listed 4. Specifications should be used intelligently, there is no guarantee that all (possible and impossible) impurities and alterations are covered!

SK | 18 December | The International Pharmacopoeia's advantages 1. Specifications validated internationally, through an independent scientific process 2. Input from WHO Collaborating Centres, national Drug Quality Control laboratories 3. Collaboration with manufacturers around the world, especially for new projects 4. Close collaboration with WHO Member States, Drug Regulatory Authorities

SK | 18 December | Further questions ??????