Determination of microbiological purity of tablets for thetreatment and prevention of type II Diabetes mellitus

Abstract

Introduction. Diabetes mellitus refers to a group of metabolic diseases characterized by hyperglycaemia, which occurs as a result of a defect in secretion or action of insulin. Chronic hyperglycaemia in diabetes mellitus (DM) leads to a damage, dysfunction and insufficiency of various organs, especially the eyes, kidneys, nerves, heart and blood vessels. Consistent hyperglycaemia activates the formation of free radicals, reduces the activity of antioxidant protection agents: superoxide dismutase, catalase, glutathione peroxidase, vitamins C and E. Therefore, special attention deserve anti-diabetic drugs which, along with the hypoglycaemic effect, have antioxidant properties, the ability to maintain or improve the secretory function of β-cells. The optimal compositions have been established and the technologies of medicinal products under the conventional names "Glycverin" with Voglibose and quercetin and "Thioquerin " on the basis of solid dispersion of thioctic acid were developed by previous studies. As a result of the study of specific pharmacological activity, it has been experimentally proved that the above drugs have a pronounced hypoglycaemic effect and increase the bioavailability of their components, which suggests its relevance in the treatment of type II diabetes. In order to increase biological availability, the method of solid dispersion has been utilized, which has significant advantages, among which: the use of drugs in solution or finely dispersed state provides a high dissolution rate, reduced irritating action on the gastric mucosa, a high therapeutic effect. However, during the technological process of obtaining non-sterile dosage forms, the preparation may be contaminated with microorganisms, which may reduce its stability during storage, which negatively affects its safety profile. Today, SPU clearly regulates the maximum level of contamination by microorganisms for each dosage form. Material & methods. The objects of the study were tablets under the conventional names "Glycverin" and "Thioquerin". Studies on the microbiological purity of the tablets were conducted in accordance with the requirements of the SPU, 2nd ed., p.2.6.12 and 2.6.13, category 3A (5.1.4, N). For the study used: soybean casein broth (HIMedia Laboratories Pvt.Ltd India » expiration date of the medium until XI 2019, Manufactured in India). For Candida albicans used Saburo-Dextrose Agar (Indian Production, «HIMedia Laboratories Pvt.Ltd India » expiration date of the environment until XI 2019). The environments were prepared according to the manufacturer's requirements (amount of powder per litre, pH, conditions of autoclaving, etc.). Each series used in the experiment was tested on growth quality in accordance with the reference documents. For tests on microbiological purity the following media were obtained on the basis of soy-casein agar: Chistovich's medium, blood agar. Results & discussion. Before conducting research on microbiological purity it was mandatory to test the compliance of the growth qualities of the nutrient media used. For this purpose, the nutrient media were inoculated with a small amount of an appropriate test strain of the microorganism (10 - 10 ² colony forming units per ml of medium - CFU / ml). The obtained data indicate that all microorganisms were consistent with the taxonomic designation of the strain, the morphology of the colonies when cultured on the media and cell morphology in microscopy were typical. Nutrient broths (soy-casein and thioglycolic medium) met the requirements for sterility - there was no growth of microorganisms, the environment was transparent. When testing on microbiological purity, the direct sowing method was used for liquid nutrient media. After 14 days of incubation, at cultivation on a Saburo, soy-casein broth and thioglycolic medium the growth of fungi was absent. In a deep sowing study, which was to add 0.1 g of preparation to agar and surface sowing of 0.1 g to agar, the amount of viable cells of microorganisms and fungi has been determined. The study of surface and deep sowing of drugs on Saburo dishes has shown no growth of fungi. The data obtained in Table 3 indicate that the number of microorganisms does not exceed 10 ³ CFU / ml, which meets the requirements of the State Pharmacopoeia of Ukraine. Conclusion. The study of microbiological purity of tablets "Glycverin" and "Thioquerin" has been conducted. The results indicate absence of viable fungal cells. It has been established that the amount of viable cells of microorganisms does not exceed 10 3 CFU / ml in 1 g of the preparation, which meets the requirements of the SPU for internal use preparations. Strains S. aureus, P. aeruginosa and representatives of Enterobacteriaceae sp. family have not been detected. Investigated samples of drugs meet the requirements of the Pharmacopoeia of Ukraine on the indicators of microbiological purity for oral preparations. The obtained results allow asserting the safety and expediency of further development of medicinal products for use in the treatment of type II diabetes mellitus.