Development of Methods of Analysis of Biological Active Substances for Standardization of Dense Extract of Creeping Thyme

Abstract

Pharmaceutical development of mucolytic agent with dense extract of creeping thyme and essential oil of thyme involves obtaining a dense thyme extract of creeping, that is, choosing of optimal conditions that will allow in a short time to obtain a dense extract enriched with the main biological active substances (BAS) - which is a resultant extract and finished drug. Since in the analysis of creeping thyme herb in order to standardize this raw material as a quality index, among others, we chose the composition of flavonoids and hydroxycinnamic acids and the quantitative content of flavonoids, then it is appropriate to study the qualitative and quantitative composition of these bioactive substances and in the obtained extract. The aim of our work was to develop methods of analysis of BAS for the standardization of dense extract of creeping thyme; Identification and determination of the quantitative content of the analyzed biological active substances in herbal medicinal products and their respective extracts were carried out using modern sensitive and selective methods of analysis (thin-layer chromatography, differential spectrophotometry); As a result of TLC, the presence of fructose, glucose, arabinose, xylose, and rhamnose were found in the dense creeping thyme extract. According to the ratio of the size and intensity of staining on the chromatograms, it has been concluded that the predominant monosaccharide is fructose, and from the reducing monosaccharides - glucose, which will allow them to choose markers of the quality of the test extract. We also identified tyrosine, alanine, leucine, glycine, glutamic and aspartic acids at the levels of the respective zones on the chromatogram of the comparison solution. Chromatographic studies of a dense extract of creeping thyme by the thin-layer chromatography method have allowed identifying phenol carboxylic acids – rosmarinic (main representative), caffeic and chlorogenic; flavonoids – luteolin-7-O-glucoside, apigenin-7-O-glucoside, luteolin, apigenin, and rutin. Two acids were not identified, small quantities of which are found in the upper part of a chromatogram and of the unidentified structure of luteolin glucoside, which, by size and intensity of fluorescence, are not the main representative of flavonoids in the extract. Based on the literature, this representative of flavonoids may be Luteolin-7-O-di or triglycoside; Qualitative composition of a dense extract of creeping thyme herb in BAS groups – flavonoids, amino acids, polysaccharides has been studied. In order to standardize the extract, identification markers from phenolic compounds – luteolin-7-O-glucoside, luteolin, rutin, chlorogenic, rosmarinic, and caffeic acids, luteolin glycoside of unknown composition has been proposed. The quantitative content of flavonoids, amino acids, and reconstructive monosaccharides in the dense extract has been investigated. The correlation of content of the main BAS groups for dense extract indicates that the selected conditions for transferring the liquid extract to the dense are correct. When standardizing the dense extract of creeping thyme by the flavonoid content is proposed to be at least 2.5% in terms of apigenin.