An efficient approach for simultaneously obtaining oil and epigoitrin from Orychophragmus violaceus seeds by microwave-mediated immiscible binary solvent extraction.

Microwave-mediated immiscible binary solvent extraction (MIBSE) was applied to simultaneously extract oil and epigoitrin from Orychophragmus violaceus seeds. The upper phase of n-hexane was used to obtain oil, and the lower phase of ethanol solution was used to obtain epigoitrin. Factors potentially affecting the yields of oil and epigoitrin were systematically investigated. The optimum conditions were an ethanol volume fraction of 65%, liquid-solid ratio of lower-phase of 20 mL/g, liquid-solid ratio of upper-phase of 12 mL/g, microwave irradiation power of 393 W, and microwave irradiation time of 29 min. The actual yields of oil and epigoitrin were 34.08% ± 1.38% and 11.86 ± 0.47 mg/g, respectively. GC-MS analysis illustrated that the seed oils obtained by MIBSE and Soxhlet extraction exhibited similar fatty acid compositions. The separated epigoitrin was determined by HPLC analyses, which obtained a purity of 91.25% ± 3.83%, follwed by NMR determinations.

Evaluation of the application of sodium deoxycholate to proteomic analysis of rat hippocampal plasma membrane.

Detergents have been widely used for the solubilization of membrane proteins and the improvement of their digestion. In this paper, we have evaluated the application of sodium deoxycholate (SDC) to the solubilization and digestion of rat hippocampal plasma membrane (PM) proteins. For in-solution digestion, rat hippocampal PM fraction from sucrose-density gradient centrifugation was solubilized by boiling in 1.0% SDC, and directly digested without dilution. During the in-gel digestion of the hippocampal PM proteins separated by SDS-PAGE, 0.1% SDC was added. Before analysis of peptide mixture by liquid chromatography and electrospray mass spectrometry, SDC in the tryptic digests was removed by centrifugation following acidification. Use of 1.0% SDC in solubilization and in-solution digestion of rat PM proteins had led to 77 PM or membrane-associated proteins identified, a more than 2-fold increase over that by use of SDS. The addition of 0.1% SDC to the in-gel digestion of SDS-PAGE-resolved membrane proteins remarkably enhanced the coverage of tryptic peptides and the number of hydrophobic membrane proteins identified. Being a cheaper and more tractable acid-insoluble detergent, SDC could be used at higher concentration in the solubilization and tryptic digestion of proteins including PM proteins with the purpose of enhancing the protein solubility and at the same time making no interference with trypsin activity and subsequent analyses.