Substantiation of technological parameters of the oil extract obtained from Urtica dioica roots based on the yield determination of phytosterols using capillary gas chromatography method

Due to the content of phytosterols, extractive preparations of Urtica dioica roots are able to show antiandrogenic effect in the case of external therapy of men and women with androgenic alopecia. Oil extracts (OE) are characterized by several advantages when applied to the skin of the scalp compared to water-alcohol extracts. For the development of OE technology from Urtica dioica roots, it is important to choose the optimal extraction parameters, which are based on the quantitative determination of phytosterols in the extractant and the studied samples of extracts. The aim of the work is to choose the optimal parameters for obtaining OE from Urtica dioica roots based on quantitative determination of phytosterols content in experimental samples of OE by gas capillary chromatography. Materials and methods. Objects of the research – Urtica dioica root, refined corn oil, refined sunflower oil, samples of oil extracts. Determination of phytosterol content in experimental samples was carried out by gas capillary chromatography (chromatograph “Crystal 2000”, manufacturer – research and production company “Analytics”). Results. 5 different compounds of steroid structure (stigmasterol, β-sitosterol, etc.) were identified in sunflower oil by gas liquid chromatography, and 10 (campesterol, 2-α stigmasterol, β-sitosterol, Δ5-avenosterol, etc.) were identified in sunflower oil. The quantitative content of β-sitosterol in the sum of sterols of corn oil was significantly higher compared to the content of this substance in sunflower oil and amounted to 59.33 %. Optimal technological parameters were established considering the peculiarities of extraction with oil extractant and quantitative determination of the amount of phytosterols and β-sitosterol in experimental samples of OE. The total content of plant sterols in OE, including considering their amount in the extractant, was in the range of 7880 mg/kg; the amount of β-sitosterol was 4638 mg/kg. Conclusion. The choice of optimal parameters for obtaining OE from UDR based on determination of phytosterol yield by gas capillary chromatography was experimentally substantiated, namely: extractant – corn oil, raw material-extract ratio – 1: 5, extraction time – 6 h, extraction method – maceration

A Combined Method of Protein Extraction from Unorthodox Plant Samples for Proteomics

Aim:: This study aimed to generate an improved method of protein extraction and purification from plant tissues containing very high amounts of phenolic compounds and other interfering biomolecules. Background:: Protein extraction at proteomic studies on some plant species including conifers is challenging and the yield and quality is unpredictable. Objective:: Two popular protocols were combined with each other to construct a novel one with enhanced abilities to produce higher purity of samples compatible for high precision molecular systems and analysis. Method:: The new method was compared with the other two for their efficiencies in classical SDS-PAGE, 2-DE and capillary chromatography applications. Results:: All three methods were comparable in SDS-PAGE procedure; however, only the new method created acceptable gel images in 2-DE. Bioanalyzer results, also, demonstrated that the new method provided protein samples pure enough to be used in capillary chromatography with 2 times more peaks in electropherograms with lower noise and higher total relative protein concentrations closest to the applied amount. Conclusion:: The new combined method is a successful alternative for plant proteomicists with higher yield and quality of proteins from recalcitrant tissues. Other:: The new method could be preferred, especially, for high-tech, sensitive proteomic analysis.