Comparative study of the influence of method on the quantity of proteins extracted from the raw material of (Baptisia australis (L.) Willd) roots

Abstract

Baptisia australis (L.) Willd. - perennial plant of the Fabaceae family, which preparations have anti-inflammatory, analgesic, antioxidant effects and can be used for cardiovascular prophylaxis. It has been investigated that Baptisia australis (L.) Willd. herbal raw material, such as grass, roots, accumulates flavonoids, alkaloids, triterpene saponins and other compounds, but proteins were not quantified. In this research, protein concentration was evaluated based on the protein extraction method [1]. The aim of the experiment: To determine the influence of protein extraction from Baptisia australis (L.) Willd. roots methods on the quantitative composition of proteins. Experimental tasks: 1. To determine the amount of proteins extracted from the roots of Baptisia australis (L.) Willd. raw material using PBS (phosphate buffer solution) and precipitating proteins with ammonium sulfate. 2. To determine the amount of proteins extracted from the roots of Baptisia australis (L.) Willd. raw material using PBS and 0.5% thiourea solution and precipitating proteins with ammonium sulfate. 3. To compare the effect of protein extraction methods based on protein content. Material and methods: Material – fresh Baptisia australis (L.) Willd. roots were collected in Vytautas Magnus University botanical garden medical herbs section. For the first method of protein extraction from raw material we used a phosphate buffer solution (PBSx10) prepared from Na2HPO4, NaH2PO4, NaCl and deionized water; For the second method of the protein extraction from raw material we used phosphate buffer solution (PBSx10) adding 0.5% thiourea solution. For both methods we used protein precipitation method by ammonium sulfate, resuspension method by PBSx1 [2], spectrophotometric (at 595 nm) protein determination by Bradford method [3], statistical analysis method based on SPSS 17.0 and Microsoft Excel. Results: Extraction of the raw material by the first method showed that 1,056 mg ± 0,170mg[...