Разработка и валидация методики изолирования ставудина из биологических жидкостей

Abstract

Objective - study of isolating Stavudine from biological fluids (urine, blood plasma). Materials and methods. Objects of research: substance and tablets of 30 mg Stavudine. Reagents: purified water, chloroform, ethyl acetate, dichloromethane, ether, heptane, toluene, acetone, 95% ethyl alcohol, 0.1 M hydrochloric acid solution, 10% ammonium hydroxide solution, 5%solutions of sodium sulfate, 20%sodium sulfate saturated, sodium chloride, sodium chloride saturated, 20%ammonium sulfate, ammonium sulfate saturated. Universal ionomer IT-1101. Stavudine was isolated by liquid-liquid extraction. To detect and quantify Stavudine, thin - layer sorbent chromatography (TLC) and UV spectrophotometry were used. Statistical data processing was performed using the software package for Windows XP (Microsoft Excel), and using the Student's t-test. Results. When using thin-layer chromatography to identify Stavudine, it was found that the Rf value of the spots of Stavudine extracts from biological material corresponded to the Rf value range of 0.67-0.69. The Rf spot value of the standard Stavudine witness sample corresponded to (0.68±0.01). The stability study showed that the Stavudine solution is stable in a hydrochloric acid solution of 0.1 M, in which the absorption spectrum of Stavudine is characterized by absorption maxima at wavelengths of 209±1 and 267±1 nm. The wavelength of 267±1 nm was chosen as the analytical wavelength for the quantitative determination of Stavudine. In the course of the study, various factors affecting the degree of extraction of Stavudine from aqueous solutions were studied. The extractant is ethyl acetate, pH 3, the electrolyte is a saturated solution of ammonium sulfate. The extraction time is 7 minutes, twice. Conclusion. Methods of identification and quantitative determination of Stavudine in extracts from urine and blood plasma by TLC and UV spectrophotometry have been developed. Methods of isolation of Stavudine from urine and blood plasma using the liquid-liquid extraction method have been developed (93.33±2.02%; 89.77±2.02% respectively).