Pharmeuropa bio & scientific notes最新文献

The Quality of Medicines & HealthCare (EDQM, Council of Europe) and the European Union (EU) Commission to evaluate the reproducibility of clinical serology results for seasonal influenza vaccines and to assess the impact of technical differences between laboratories on the compliance with the Committee for Human Medicinal Products (CHMP) criteria set by the European Medicines Agency (EMA). The study was run in 2 phases. The present article reports the 1st phase of the study, which aimed at evaluating the variability of the results obtained by 11 laboratories (5 national control laboratories and 6 influenza vaccine manufacturers) using their routine haemagglutination inhibition (HI) assay to test a common panel of clinical trial sera. The results confirmed the limited inter-laboratory reproducibility of the HI testing of influenza vaccine clinical trial samples. In some cases a good agreement was found between laboratories, while a systematic bias or a random scatter of results was observed in other cases. Analysis of estimated systematic bias confirmed that differences between laboratories can be significant (up to 16-fold) in some cases. Correction for this bias resulted in limited improvement. Differences between laboratories were found to result in discrepant decisions on marketing acceptance of vaccines or to decisions based on compliance to different criteria. The study showed that the seroconversion (SC) and mean fold increase (MFI) criteria are more robust against systematic over- or under-estimation of titres whereas the protection rate (PR) is very sensitive to this effect. The fundamental issues with the PR criteria are discussed.

An international collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) to establish the WHO second International Standard (IS) for gramicidin as the stocks of the 1st IS, established in 1964, were close to depletion. The candidate material did not show any sign of potency loss when kept at elevated temperatures of + 4 °C, + 20 °C, + 37 °C and + 45 °C for 3 months. Six laboratories from 5 countries as well as the EDQM laboratory participated in the collaborative study. Potencies of the candidate material were estimated by microbiological assays with sensitive micro-organisms. To ensure continuity between consecutive batches, the 1st IS for gramicidin was used as standard. Based on the results of the study, the 2nd IS for gramicidin was adopted at the meeting of the WHO Expert Committee on Biological Standardization (ECBS) in 2008 with an assigned potency of 1070 International Units per mg (IU/mg). The 2nd IS for gramicidin is available from the EDQM.

The European Pharmacopoeia Biological Reference Preparation (Ph. Eur. BRP) Batch 4 was established as an international common working standard for potency determination of human coagulation factor VIII (FVIII) preparations to replace the dwindling stocks of the BRP Batch 3, the current European standard. Similarly, stocks of the current World Health Organisation 7th International Standard (WHO 7th IS) were also running low. Therefore a project was jointly organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM, Council of Europe) and the National Institute for Biological Standards and Control (NIBSC, UK) in order to replace both standards concomitantly. The potency of the BRP Batch 4 was assigned during an international collaborative study involving 38 laboratories with reference to the WHO 7th IS and the BRP Batch 3. Four candidate materials, 2 plasma-derived (samples A and C) and 2 recombinant (samples B and D) have been evaluated, sample C being the specific candidate for the replacement of the BRP Batch 3. Participants were instructed to perform 8 independent assays following their own routine validated methods, by either the one-stage clotting assay or the chromogenic assay, or both. Laboratories returned 22 data sets for the clotting assay and 30 data sets for the chromogenic assay. This publication reports the results obtained with both assays but only the results of the chromogenic assay are highlighted in the conclusions, as it is the assay prescribed by the European Pharmacopoeia. Data were analysed separately for both assays. The consensus potency value was calculated as the unweighted geometric mean of the unweighted geometric means of each individual laboratory. For sample C, there was a significant difference in potency estimate between the chromogenic and the clotting assay. It was therefore not possible to reconcile both results. The chromogenic potencies however were in very good agreement being 10.4 IU/ampoule (n = 30), when assessed against both standards. The inter-laboratory geometric coefficient of variation (GCV) was 4.8 % and 7.1 % against the WHO 7th IS and the BRP Batch 3 respectively. The Ph. Eur. BRP Batch 4 is a freeze-dried, plasma-derived concentrate. The material was filled in approximately 20,000 ampoules and lyophilised. The final residual water content is 0.33 %. Based on accelerated degradation studies, the stability of the material is suitable for a reference preparation. The candidate Ph. Eur. BRP Batch 4 was adopted at the 136th session of the European Pharmacopoeia Commission in March 2010. The standard will be available from the EDQM with the catalogue number H0920000 upon exhaustion of the current batch.

A collaborative study was run by the European Directorate for the Quality of Medicines & HealthCare (EDQM) in the context of the Biological Standardisation Programme (BSP), under the aegis of the Council of Europe and the European Commission, to establish replacement batches for the dwindling stocks of the Heparin low-molecular-mass for assay European Pharmacopoeia Biological Reference Preparation (BRP). The replacement batches of BRP are intended to be used in the assays for anti-Xa and anti-IIa activities, as described in the European Pharmacopoeia (Ph. Eur.) monograph Heparins, low-molecular-mass (0828). Three freeze-dried candidate batches were calibrated against the current International Standard (IS) for Heparin, lowmolecular- weight (2nd IS, 01/608). For the purpose of the continuity check between subsequent BRP batches, the current Heparin low-molecular-mass for assay BRP (batch 5) was also included in the test panel. Thirteen official medicines control and manufacturers laboratories from European and non-European countries contributed data. A central statistical analysis of the datasets was performed at the EDQM. On the basis of the results, the 3 candidate materials were assigned a potency of 104 IU/vial for the anti-Xa activity and 31 IU/vial for the anti-IIa activity. Taken into account the preliminary stability data and the results of this collaborative study, the 3 batches of candidate BRP were adopted in June 2010 by the Commission of the Ph. Eur. as Heparin low-molecular-mass for assay BRP batches 6, 7 and 8.

The European Pharmacopoeia (Ph. Eur.) monographs for the water-soluble cellulose ethers require viscosity determination, either in the "Tests" section or in the non-mandatory "Functionality-related characteristics" section. Although the derivatives are chemically closely related and used for similar applications, the viscosity tests strongly differ. Some monographs generically speak of the rotating viscometer method (2.2.10) and a fixed shear rate (e.g. 10 s-1), which would necessitate an absolute measuring system, while others recommend the capillary viscometer method for product grades of less than 600 mPa∙s and the rotating viscometer method and given operating conditions for grades of higher nominal viscosity. Viscometer methods also differ between the United States Pharmacopeia/National Formulary (USP/NF) and the Japanese Pharmacopoeia (JP) monographs. In addition, for some cellulose ethers the tests sometimes diverge from one pharmacopoeia to the other, although the three compendiums are in a harmonisation process. But the main issue is that the viscometer methods originally employed by the product manufacturers are often not those described in the corresponding monographs and generally vary from one manufacturer to the other. The aim of this study was therefore to investigate whether such a situation could invalidate the present pharmacopoeial requirements. 2 per cent solutions of several viscosity grades of hydroxyethylcellulose, hypromellose and methylcellulose were prepared and their (apparent) viscosity determined using both relative and absolute viscometer methods. The viscometer method used not only affects the measured viscosity but experimental values generally do not correspond to the product nominal viscosities. It emerges that, in contrast to Newtonian solutions (i.e. those of grades of up to ca. 50 mPa∙s nominal viscosity), some of the viscometer methods currently specified in the monographs are not able unambiguously to characterise the grades exhibiting non-Newtonian behaviour. It is also concluded that, unless the various manufacturers redefine their product viscosity grades using a single compendial test, two strategies could be adopted, both based on the operating conditions specified in the labeling (i.e those of the manufacturer), the test appearing either in the mandatory section if this is acceptable to the pharmacopoeia (like in some USP/NF monographs) or, for the Ph. Eur., in the "Functionality-related characteristics" section.

This paper presents in-vitro metoprolol release from four different extended-release (ER) formulations, i.e. Metoprolol GEA® Retard, Logimax® forte, Metoprolol Sandoz® and Seloken ZOC® in the presence of 10 to 40% (v/v%) ethanol at pH 1.2 and pH 6.8. The assay of metoprolol in the dissolution media was performed by reversed phase liquid chromatography (RP-LC) using a mixture of methanol and 100 mM phosphate buffer (pH 3.5) in 40:60 ratio as eluent. The dissolution data showed that the metoprolol contents of Metoprolol Sandoz® and Seloken ZOC® were released fast in the presence of 20% ethanol at the investigated conditions, while the other products demonstrated much more stability against ethanol. Unexpectedly it was discovered that the release of metoprolol from Metoprolol GEA® Retard and to some extent also from Logimax® forte decreased in the ethanol containing media.

Due to the depletion in stocks of the World Health Organization (WHO) 2nd International Standard (IS) for nystatin, an international collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) to establish a replacement batch. Seventeen laboratories participated in the collaborative study, performing the microbiological diffusion assay to estimate the potency of the candidate 3rd International Standard for nystatin. The 2nd International Standard for nystatin was used as a standard to ensure the continuity of the unitage. Follow-up accelerated degradation studies demonstrated that the IS is stable when at the customary storage temperature of - 20 °C. The 3rd IS for nystatin was adopted by the WHO Expert Committee on Biological Standardization (ECBS) in 2006 with an assigned potency of 5710 International Units per mg (IU/mg). The 3rd IS for nystatin is available from the EDQM.

European Pharmacopoeia (Ph. Eur.) monograph 0451 on Rabies vaccine (inactivated) for veterinary use describes an in vivo batch potency test that is based on the NIH test. This assay uses a large number of mice and results in a significant degree of suffering. In the interest of replacement, reduction and refinement of animal tests (3R) a serological potency assay for Rabies vaccine (inactivated) for animal use, developed and validated at the Paul-Ehrlich-Institut, has been assessed in a collaborative study organised by the EDQM (European Directorate for the Quality of Medicines & HealthCare). The goal was to demonstrate the wider transferability of the proposed assay and confirm its suitability. The study involved 13 laboratories and assessed 4 different vaccines from the EU market. Results of the study confirm that a limit test using a relatively small number of animals in a serological assay is possible, reproducible and reliable. The optimal number of animals per vaccine is product specific but may roughly be indicated to be between 8 and 10 for the products included in this study. Non-responders should be included in the analysis because they may reflect sub-potent vaccines. However, there may be a need to impose a maximum on the number of non-responders allowed for the reference vaccine as a monitor for assay validity. This assay provides a significant 3R improvement in terms of both the number of animals used and the amount of suffering entailed and provides a more reliable and reproducible assay format than the vaccination challenge assay. It also reduces the time required as compared to the vaccination challenge assay. It has been recommended to the Ph. Eur. group of experts 15V that this assay be included as an alternative to the batch potency assay in the Ph. Eur. monograph 0451.

A joint project (coded BSP089) was run by the European Directorate for the Quality of Medicines & HealthCare (EDQM) of the Council of Europe, the National Institute for Biological Standards and Control (NIBSC) on behalf of the World Health Organization (WHO) and the Center for Biologics Evaluation and Research (CBER) of the U.S. Food and Drug Administration (FDA) to evaluate, in an international collaborative study, 3 lyophilised intravenous immunoglobulin (IVIG) preparations for their suitability to serve as Reference Preparations to standardise and control the highly variable haemagglutination testing for anti-A and anti-B in IVIG products. 23 laboratories tested candidate IVIG reference preparations consisting of a Positive control, a Negative control and a specifically formulated Limit test reference preparation to define the maximum (e.g., pharmacopoeial) limits of anti-A and anti-B haemagglutinins in IVIG products, where limits are applicable. Laboratories performed direct haemagglutination using papain-treated erythrocytes and/or indirect anti-globulin tests. For both methods, there was up to 16-fold variation in anti-A and anti-B titres, although there was good agreement over a 2-fold titre range for anti-A and anti-B between laboratories using the direct method for both the Positive control and Limit reference preparations. Comparative titration data for the Positive control and Limit reference preparations indicated that the use of a 'Limit' test reference preparation would facilitate identification of higher titre batches when the direct haemagglutination method is used. The Positive control, Negative control and Limit test preparations were adopted in November 2008 by the Commission of the European Pharmacopoeia (Ph. Eur.) as Biological Reference Preparations. The same preparations have been established as reference reagents by the WHO and the U.S FDA, including the maximal specifications defined by the Limit test preparation. This will facilitate global standardisation of haemagglutination tests for anti-A and anti-B, ensure that such tests are sufficiently sensitive and specific, and facilitate identification of batches that exceed maximum recommended levels of anti-A and anti-B antibodies.

An international collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) to establish the WHO second International Standard (IS) for amphotericin B as the stocks of the 1st IS, established in 1963, were close to depletion. The candidate material did not show any sign of potency loss when kept at elevated temperatures of + 4 degrees C, + 20 degrees C, + 37 degrees C and + 45 degrees C for 3 months. Ten laboratories from 8 countries participated in the collaborative study. Potencies of the candidate material were estimated by microbiological assays with sensitive micro-organisms. To ensure continuity between consecutive batches, the 1st IS for amphotericin B was used as the standard. Based on the results of the study, the 2nd IS for amphotericin B was adopted at the meeting of the WHO Expert Committee on Biological Standardisation (ECBS) in 2007 with an assigned potency of 944 International Units per mg (IU/mg). The 2nd IS for Amphotericin B is available from the EDQM.