The European Directorate for the Quality of Medicines and HealthCare (EDQM) organised an international collaborative study under the aegis of the Biological Standardisation Programme, a joint programme of the Council of Europe and the European Commission, to establish replacement batches for the European Pharmacopoeia (Ph. Eur.) Heparin sodium Biological Reference Preparation (BRP). The study involved cali brating candidate BRP batches 5 and 6 in International Units with respect to the World Health Organization (WHO) 6th International Standard for Unfractionated Heparin (07/328) using the anti-IIa and anti-Xa chromo genic assays and the sheep plasma clotting assay as prescribed in the respective Ph. Eur. monographs. Their suitability for use as a titration reference in the assay of protamine sulfate according to the Ph. Eur. mono graph on Protamine sulfate (0569) was also confirmed. They have been adopted by the Ph. Eur. Commission as Ph. Eur. BRP batch 5 and batch 6 respectively, and will be available from the EDQM under catalogue number H0200000 once the predecessor batch has been exhausted.
This article reviews the historical use of general animal safety tests in the European Pharmacopoeia, their scientific limitations, and the ethical concerns that have driven their progressive elimination. It highlights the role of the European Pharmacopoeia Commission in implementing the 3Rs principles (Replacement, Reduction, and Refinement) and introducing a fourth 'R', "Removal". Key milestones include the suppression of tests for abnormal toxicity, pyrogens, histamine and depressor substances, culminating in the complete abolition of general animal safety tests by 2026. The article emphasises the scientific superiority of validated in vitro methods, the impact of good manufacturing practices, and the regulatory strategies that ensure product safety without reliance on animal testing. This evolution represents a significant achievement for animal welfare and modern pharmaceutical quality control.
The European Pharmacopoeia (Ph. Eur.) prescribes that low-molecular-mass heparin products be calibrated for their molecular mass distribution using a reference preparation with a defined molecular mass calibration. Heparin low-molecular-mass for calibration Chemical Reference Substance (CRS) batch 5 was established to this end and is used in conjunction with a Broad Standard Table as described in monograph Heparins, low-molecular-mass (0828). Stocks of this reference preparation were running low. Therefore, a replacement batch was established by the European Directorate for the Quality of Medicines & HealthCare (EDQM) under the aegis of the Biological Standardisation Programme. Due to the complexity of preparation of a replacement starting material, a ready-to-use candidate refill material was procured. A confirmatory study of the fitness for use was carried out, which demonstrated that the material was suitable for the intended purpose. The candidate CRS was adopted by the Ph. Eur. Commission in April 2024 as Ph. Eur. Heparin Low-Molecular-Mass for calibration CRS batch 6. It is available from the EDQM under catalogue number H0190000.
A study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM, Council of Europe) to establish a replacement batch for the current European Pharmacopoeia (Ph. Eur.) Human Anti-D immunoglobulin Biological Reference Preparation (BRP) batch 1 whose stocks were dwindling. The BSP176 study was run in the framework of the Biological Standardisation Programme (BSP) of the Council of Europe and the European Union (EU) Commission. In view of the current stocks, the difficulties in sourcing suitable starting materials and the time required to calibrate a replacement batch with the 3 different methods described in Ph. Eur. chapter 2.7.13. Assay of human anti-D immunoglobulin, it had been decided as an interim measure to establish BRP batch 2 (BRP2) using ampoules already qualified for use as the World Health Organization (WHO) 3rd International Standard (IS) for anti-D immunoglobulin (16/332) and obtained from MHRA. For this purpose, a verification study has been performed at the EDQM laboratory on the candidate BRP (cBRP) material, i.e. ampoules of the 3rd IS for anti-D immunoglobulin received at EDQM, to be used as BRP2. The study showed that the anti-D estimated potency of the cBRP, WHO 3rd IS for anti-D Immunoglobulin, relative to BRP1 which is equivalent to the WHO 2nd IS, was consistent with the value of 297 IU/ampoule assigned during the establishment study for the 3rd IS and in continuity with BRP1. Stability was confirmed by checking the potency of ampoules stored at the recommended storage temperature (i.e. -20°C) against ampoules stored at -70°C since the establishment. The molecular-size distribution profile of the cBRP was also similar to the one obtained during the establishment study, again confirming the stability. All these data confirmed the good stability of the WHO 3rd IS for anti-D Immunoglobulin in line with the stability study performed during its establishment and confirmed its suitability for use as BRP2. In October 2024, the Ph. Eur. Commission officially adopted the candidate batch as Ph. Eur. Human Anti-D immunoglobulin BRP batch 2 with an assigned potency of 297 IU/ampoule. The activity of the new batch of Ph. Eur. Anti-D immunoglobulin BRP will be regularly monitored.
Here we report the outcome of a collaborative study aimed at demonstrating the suitability of the candidate Chemical Reference Substance (CRS) as a replacement reference (batch 2) for the physicochemical methods in European Pharmacopoeia monograph 1316 Erythropoietin concentrated solution. Results from the study demonstrated that for the physicochemical methods described in the monograph - capillary zone electrophoresis, polyacrylamide gel electrophoresis and immunoblotting, peptide mapping and glycan mapping - the candidate CRS is essentially identical to CRS batch 1 and is suitable to be established as Erythropoietin for physicochemical tests CRS batch 2. CRS batch 2 is a freeze-dried preparation presented in vials. It was adopted at the 177th session of the European Pharmacopoeia Commission in November 2023 and is available from the European Directorate for the Quality of Medicines & HealthCare (EDQM) under product code Y0001725.
