不同产地扁担藤茎原儿茶酸和香草酸含量测定的研究

Content determination of protocatechuic acid and vanillic acid in stem of Tetrastigma planicaule from different areas

为了测定扁担藤茎原儿茶酸和香草酸的含量,在单因素实验结果基础上,运用正交试验筛选扁担藤茎原儿茶酸和香草酸的提取工艺,采用HPLC同时测定不同产地扁担藤茎中原儿茶酸和香草酸的含量。采用热电Thermo Hypersil ODS C18(5μm,4.6 mm×250 mm)色谱柱,流动相为乙腈-0.1%磷酸(6∶94),流速1.0 mL/min,检测波长370 nm,柱温25℃,进样量为10.0μL。原儿茶酸和香草酸的最佳提取工艺为加入10倍量的甲醇∶0.5%冰醋酸溶液(70∶30),回流提取2 h,蒸干至无醇味后用乙酸乙酯萃取5次;原儿茶酸在进样量为13.76×10-2~213.76×10-2μg呈良好的线性关系(r=0.9997),香草酸在进样量为3.64×10-2~58.24×10-2μg呈良好的线性关系(r=0.9993),原儿茶酸和香草酸的精密度试验、重复性试验和稳定性试验的RSD均小于3%,平均加样回收率分别为100.34%和98.08%,符合含量测定要求。10批扁担藤茎的原儿茶酸含量在24.21~108.31μg/g,香草酸含量在4.26~20.46μg/g。所建立的含量测定方法简便、可靠、准确、重复性好,可用于扁担藤茎药材的质量控制。

To determine the contents of protocatechuic acid and vanillic acid in stem of Tetrastigma planicaule.Orthogonal experiment was used to screen the extraction technology,a HPLC method for determination of protocatechuic acid and vanoxalic acid was used,chromatographic column was Thermo Hypersil ODS C18(5μm,4.6 mm×250 mm),mobile phase was acetonitrile-0.1%phosphoric acid(volume ratio 6∶94),iso-elution,flow rate was 1.0 mL/min,detection wavelength was 370 nm,column temperature was 25℃,the injection volume was 10.0μL.Optimum extraction process of protocatechuic acid and vanillic acid was add 10 times the amount of methanol∶0.5%glacial acetic acid solution(70∶30),reflux extraction for 2 h,steam dry to no alcohol taste,then use ethyl acetate extraction for 5 times;the good linear relationship of Protocatechuic acid was between 13.76×10-2~213.76×10-2μg(r=0.9997),and the good linear relationship of Vanillic acid was 3.64×10-2~58.24×10-2μg(r=0.9993).The RSD of repeatability test,precisionstability tests of protocatechuic acid and vanillic acid were all less than 3%,the average recoveries were 100.34%and 98.08%respectively.The method is simple,accurate,and stable,which can be used to control the quality of Tetrastigma planicaule.