[Objectives] To establish the high performance liquid chromatography( HPLC) determination method for oxysophocarpine and optimize the extraction and purification technology of oxysophocarpine from Sophora alopecuroides by inverse three-phase membrane cycle.[Methods]Based on the single-factor experiment,the effects of aqueous phase and organic phase volume ratio,the concentration of sodium hydroxide,concentration of hydrochloric acid and extraction cycle time on extraction process of oxysophocarpine were investigated using orthogonal design method,to determine the optimal extraction process. [Results]The oxysophocarpine was determined by Shim-pack VP-ODS chromatographic column( 4. 6 mm × 250 mm,5 μm),mobile phase was methanol-0. 2% phosphoric acid aqueous solution( 7∶ 93),gradient elution,flow rate was 1 m L/min,the sample size was 5 μm,column temperature was 30℃,detection wavelength was 221 nm. Aqueous phase and organic phase volume ratio was 1∶ 1,hydrochloric acid concentration was 0. 3 mol/L,sodium hydroxide concentration was 0. 75 mol/L,water pump flow rate was 6 m L/min,60 min cycle time. The extraction rate of oxysophocarpine 98. 21 % within 60 min under the best experimental conditions. Oxysophocarpine has good linearity relationship within the range of 0. 01-0. 7 mg/m L( r^2= 0. 9978,n = 6),the respective average recovery rate was 97. 47%( RSD = 1. 95%). [Conclusions] This extraction process is simple in operation,the organic solvent has low consumption,and can be used for extraction of alkaloid.
目的:建立微量样品羟自由基(·OH)的检测方法,并用于新疆葡萄树伤流液及金银花茶、罗布麻茶、茉莉茶、竹叶茶等样品的检测。方法:采用Mn2+-H2O2-EBT法,检测波长为560 nm,在2 m L的比色皿中,取缓冲溶液(Na B4O7-Na OH)300μL,EBT 120μL,Mn2+300μL,H2O2150μL,将体系体积用水补充至2.0 m L。Mn2+-H2O2-EBT法可作为检测抗氧化物质抗氧化活性的有效可行方法之一,为验证该方法对微量样品抗羟自由基(·OH)活性实时检测的可行性,以抗坏血酸对照。结果:实验结果表明,新疆葡萄树伤流液、金银花茶、罗布麻茶、茉莉茶和竹叶茶均具有较强的清除羟自由基(·OH)的生物活性功能,是一种很好的天然抗氧化及自由基清除剂。结论:微量样品抗羟自由基检测方法可行,实现吸光度测定的同时,对少量样品进行实时、恒温及震荡在线检测,连续观察吸光度的变化。
