Одним из основных критериев биологической безопасности иммунобиологических препаратов явля-ется их стерильность. В статье представлена история разработки в нашей стране методов испытания иммунобиологических препаратов по показателю «Стерильность», начиная с 1961 г. и заканчивая со-временными требованиями, регламентируемыми Государственной фармакопеей Российской Федерации XIII издания. Детально проанализированы ключевые подходы по совершенствованию оценки качества по данному показателю, в том числе в отношении выбора оптимальных питательных сред и методик проверки их качества, чувствительных тест-штаммов и условий инкубирования, определения количества отбираемых образцов препарата, необходимого для достоверного подтверждения стерильности всей се-рии (объем выборки), а также по разработке схемы проведения испытания, учитывающей особенности производства и применения иммунобиологических препаратов. Приведена информация о многолетнем опыте использования разработанной в нашей стране схемы испытания стерильности Национальным ор-ганом контроля медицинских иммунобиологических препаратов ГИСК им. Л.А. Тарасевича. Представлен анализ современного состояния проблемы гармонизации требований к проведению испытания стериль-ности иммунобиологических и других лекарственных препаратов, в том числе с ведущими зарубежными фармакопеями, а также перспективы их использования странами -членами Евразийского экономиче-ского союза. БИОпрепараты. Профилактика, диагностика, лечение 2018; 18(1): 5-15.

Culture media with predetermined specific characteristics play a key role in ensuring safety, quality and efficacy of medicinal products, especially biologicals. It is necessary to formulate clear criteria for culture media suitability, because they are used not only in manufacture, but also in quality control of medicinal products. The expansion of the range of complex high-technology medicinal products containing biologically derived substances as active ingredients, whose quality and properties are tested by biological methods, calls for more stringent requirements for the quality of culture media. The aim of the study was to analyse requirements for the culture media used in the production of medicinal products — in order to identify approaches to the development of a national standard. Currently, culture media are not included into the State Register of Medicinal Products, and there are no standards that could be used by manufacturers to confirm that a culture medium complies with specific safety, efficacy and quality requirements. The paper summarises the results of the analysis of the State Pharmacopoeia of the Russian Federation, 14th ed., and nine leading world pharmacopeias’ requirements for culture media used for various purposes (manufacture, auxiliary purposes, and quality control). Most countries have adopted requirements mainly for those culture media that are used in purity tests. The analysis revealed differences both in the lists of requirements (test strains, incubation and storage conditions, preparation for use, suitability assessment), and the approaches to the establishment and presentation of requirements. The paper discusses the main challenges and risks associated with the quality control of culture media in accordance with the requirements of the State Pharmacopoeia of the Russian Federation, 14th ed. It outlines current national and international ways of improving the quality of culture media used in the production of medicinal products, and substantiates the need to develop an appropriate national standard. The paper suggests approaches to the development and standardisation of requirements for the quality of various types of media used in the production and control of medicines. These approaches could be used in the preparation of the general chapter Culture media.

Medicinal products fail sterility testing if visual observation shows the growth of microorganisms that manifests itself as turbidity, sedimentation, flocculation and other changes in the growth medium. A key factor allowing robust determination of changes in the culture that may be suspected of contamination is the quality of growth media used, namely their transparency, and absence of foreign matter detectable by microscopic examination of the growth media smears. The presence of such foreign matter makes it especially difficult to interpret the results of testing of immunobiological products, namely live bacterial vaccines, because they cause turbidity of the media due to their specific composition. The article dwells upon the results of testing (in terms of Transparency and Microbial content) of dehydrated growth media recommended by the State Pharmacopoeia of the Russian Federation, 13th ed., General monograph 1.2.4.0003.15 for sterility testing of immunobiological medicinal products. The study revealed the presence of microorganisms, including pathogenic ones, in the growth media. In view of the fact that certificates of analysis and technical documentation accompanying components of growth media and dehydrated growth media produced by most national and foreign manufacturers do not contain any data on the acceptable levels of microorganisms it is argued that these products have to be tested for microbial content. The study also investigated the ways of improving the quality of commercial dehydrated growth media at the preparation stage.

In accordance with the requirements of the State Pharmacopoeia of the Russian Federation, 14th edition, Clostridium novyi 198 test strain is used to determine growth promoting properties of thioglycollate medium when testing sterility of immunobiological medicinal products. The preservation of C. novyi 198 as spores requires a special liquid medium containing insoluble calcium chloride casein (CChC). At present, there is no reference standard available on the Russian market for the quality control of CChC used as a component of the media for preservation of C. novyi 198.The aim of the study was to develop and certify the first batch of calcium chloride casein industry reference standard (IRS) for the quality control of CChC batches and liquid media containing CChC which is used to preserve C. novyi 198 as spores.Materials and methods: thermally denatured CChC which was chemically synthesized from industrial CChC was used as candidate material. The quality control was performed using the test procedures described in procedural guidelines MUK 4.2.2316-08. C. novyi 198 spores were obtained as described in the «Sterility testing» section of MUK 4.2.2316-08 (preparation, inoculation and incubation in the medium containing calcium chloride casein IRS).Results: a list of quality parameters was developed for calcium chloride casein IRS: «Spore formation» (certifiable parameter), «Appearance», «Loss on drying», «Clarity of medium». The first batch of the candidate calcium chloride casein IRS was certified. The value of the «Spore formation» certifiable parameter was found to be (32.6 ± 7.3) %, the IRS shelf life was 5 years when stored in a dry place protected from light at a temperature of 15 to 25 °C. A documentation package for the IRS 4228-444-2018 was prepared and approved. The liquid medium containing CChC reference material (0.3 g/10 mL) allows for preservation of the biological properties of the C. novyi 198 working culture for at least 12 months.Conclusions: the certification results demonstrate that the calcium chloride casein IRS meets the applicable requirements and can be used as reference material to control the quality of new CChC batches and to assess the suitability of liquid medium containing CChC for preservation purposes. The use of the calcium chloride casein IRS makes it possible to standardise and greatly simplify the procedure for preserving the stability of C. novyi 198, to increase the efficiency and reliability of immunobiologicals quality control in terms of Sterility, and to reduce costs when using a freeze-dried strain.