HPLC法测定二仙汤提取物中抗骨质疏松有效成分的含量  

Content Determination of Active Ingredients of Antiosteoporosis in Er-xian Decoction Extracts

作　　者：许红涛[1,2] 李媛[1,2] 何勇静[2] 袁婷婷[1,2] 张乃丹[2] 张巧艳[1,2] 

机构地区：[1]福建中医药大学药学院,福州350108 [2]第二军医大学药学院生药学教研室,上海200433

出　　处：《中国药房》2015年第15期2122-2125,共4页China Pharmacy

基　　金：上海市科学技术委员会科研计划项目(No.12401900702)

摘　　要：目的：建立测定二仙汤提取物中抗骨质疏松有效成分淫羊藿黄酮、盐酸小檗碱和知母皂苷含量的方法。方法：淫羊藿苷和盐酸小檗碱采用高效液相色谱法,色谱柱为Diamonsil C18,流动相分别为甲醇-1%醋酸（60∶40,V/V）和甲醇-乙腈-0.3%三乙胺水溶液（20∶10∶70,V/V/V,磷酸调节p H=4.0）,检测波长分别为270 nm和340 nm,柱温分别为30℃和25℃,进样量均为20μl,流速均为1.0 ml/min。知母皂苷采用紫外-可见分光光度法测定,皂苷类成分用盐酸水解后,以菝葜皂苷元为对照品,与香草醛显色后,在（590±2）nm波长下检测。结果：淫羊藿苷、盐酸小檗碱、知母皂苷质量浓度分别在40.00-90.00、5.00-35.00、8.00-40.00μg/ml范围内具有良好的线性关系（r=0.999 8、0.999 9、0.999 5）;精密度、稳定性、重复性试验的RSD均〈2%;平均加样回收率分别为101.43%、99.55%、101.30%,RSD分别为1.26%、1.93%、1.55%（n=6）。结论：本方法简便、重复性和稳定性良好,适用于二仙汤提取物的质量控制。OBJECTIVE：To establish a method for the content determination of active ingredients（icariin flavone,berberin hydrochloride and timosaponin）of antiosteoporosis in Er-xian decoction extracts.METHODS：HPLC method was used to determine the icariin and berberin hydrochloride.The column was Diamonsil C18 with the mobile phase of methanol-1% acetic acid（60∶40,V/V）and methanol-acetonitrile-0.3% triethylamine water（20∶10∶70,V/V/V,adjusted p H to 4.0）at the flow rate of 1.0 ml/min;the detection wavelength was respectively 270 nm and 340 nm,the temperature was respectively 30 ℃ and 25 ℃ and the volume was20 μl.UV-visible spectrophotometry was conducted to determine timosaponin.Saponins were hydrolysed with hydrochloric acid and detected at the wavelength of（590±2）nm with the reference of sarsasapogenine after color-developed with vanillin.RESULTS ：Icariin,berberin hydrochloride and timosaponin had a good linear relationship in the range of 40.00-90.00 μg/ml,5.00-35.00 μg/ml and 8.00-40.00 μg/ml,respectively（r=0.999 8,0.999 9,0.999 5）;RSDs of precision,stability and reproducibility tests were all lower than 2%.The average recovery was respectively 101.43%（RSD=1.26%,n=6）,99.55%（RSD=1.93%,n=6） and 101.30%（RSD=1.55%,n=6）.CONCLUSIONS：The method is simple and with high repeatability and stability and can be used for the quality control of Er-xian decoction extracts.

关 键 词：二仙汤提取物 淫羊藿苷 盐酸小檗碱 知母皂苷 高效液相色谱法 紫外-可见分光光度法 含量测定 

分 类 号：R917[医药卫生—药物分析学]