金线风中5种生物碱含量测定及提取工艺研究

Simultaneous determination of five kinds of component and extraction technology of alkaloids in the extract of Cyclea hypoglauca(Schauer)Diels

为了建立同时测定金线风药材中木兰花碱、左旋箭毒碱、非洲防己碱、巴马亭红碱及掌叶防己碱5种生物碱成分的高效液相色谱(HPLC)方法及提取工艺的优化。采用HPLC法,Wetch Xtimate C 18色谱柱(4.6 mm×250 mm,5μm);流动相为乙腈(A)-0.5%磷酸水溶液(B),梯度洗脱,检测波长为282 nm,流速为0.5 mL/min,柱温30℃;采用正交实验以乙醇浓度、料液比、pH和提取次数为考察因素,5种生物碱之和的提取率为考察指标对超声微波提取工艺进行优化并验证。木兰花碱、左旋箭毒碱、非洲防己碱、巴马亭红碱及掌叶防己碱分别在测量范围内线性关系良好。正交优化实验得出金线风生物碱最佳提取条件为乙醇浓度60%,料液比1∶18,用盐酸溶液调节提取液pH为2,提取4次。优化后的提取条件可使金线风生物碱提取率达到2.33%。该HPLC法准确可靠,灵敏快捷,重复性高,可为金线风提取物生物碱的质量评价与控制提供实验依据;优化的超声微波提取工艺可重复、稳定。

To establish an HPLC method for the simultaneous determination of magnoflorine,L-curine,columbamine,palmatrubin and palmatine in Cyclea hypoglauca(Schauer)Diels.Ultrasonic and microwave-assisted extract was used to study the optimal extract process of the alkaloids in Cyclea hypoglauca(Schauer)Diels.HPLC was used with Wetch Xtimate C 18 column(4.6 mm×250 mm,5μm).The mobile phase was acetonitrile(A)and 0.5%phosphoric acid aqueous solution(B),the detection wavelength was 282 nm,the flow rate was 0.5 mL/min,and the column temperature was kept at 30℃.The orthogonal experiment was used to investigate suchfactorsas ethanol concentration,material-to-liquid ratio,pH and extract times.The extraction rate of the sum of the five alkaloids is the inspection index to optimize and verify the ultrasonic microwave extraction process.The linear range of magnoflorine,L-curine,columba-mine,palmatrubin and palmatine were good.Through orthogonal optimization experiments,the optimal extract conditions for alkaloids were as follows:ethanol concentration:60%;material-liquid-ratio:1∶18;pH of the extracting solution was adjusted to 2,and extract was performed four times.The highest extract rate of alkaloids from Cyclea hypoglauca(Schauer)Diels was 2.33%.The method was sensitive,rapid,accurate,reliable and reproducible,and can provide experimental basis for quality evaluation and control of alkaloids extracted from Cyclea hypoglauca(Schauer)Diels.The optimized ultrasonic microwave extraction process is repeatable and stable.