一测多评同时测定款冬花中10个成分的含量  

Determination of 10 active constituents in Farfarae Flos by quantitative analysis of multi-components by single marker

作　　者：何兵[1] 刘艳[1] 杨世艳[2] 张燕[3] 

机构地区：[1]泸州医学院药物研究中心,泸州646000 [2]泸州医学院附属中医医院,泸州646000 [3]泸州医学院附属医院,泸州646000

出　　处：《药物分析杂志》2013年第9期1518-1524,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：泸州市2010年重点科技项目(泸市财企[2010]41号);泸州医学院2010年青年基金(泸医院[2010]108号)

摘　　要：目的:建立款冬花中10个活性成分(新绿原酸、绿原酸、隐绿原酸、咖啡酸、芦丁、金丝桃苷、异绿原酸B、异绿原酸A、异绿原酸C及款冬酮)的一测多评含量测定方法。方法:采用Kromasil C18色谱柱(250 mm×4.6 mm,5μm);以乙腈-0.1%磷酸水溶液为流动相;梯度洗脱(0→10→25→30→35→40 min,10%→16%→30%→90%→95%→95%乙腈);分段变波长测定(0～15 min,326 nm,检测新绿原酸、绿原酸、隐绿原酸及咖啡酸;15～20 min,254 nm,检测芦丁、金丝桃苷;20～30 min,326nm,检测异绿原酸B、异绿原酸A及异绿原酸C;30～40 min,220 nm,检测款冬酮);流速1.0 mL·min-1;柱温30℃。以绿原酸为参照物建立与其余成分的相对校正因子,并计算其含量,实现一测多评。同时采用外标法测定该10个成分的含量,并比较二者的差异,以验证一测多评法的准确性和可行性。结果:在一定的线性范围内,绿原酸与新绿原酸、隐绿原酸、咖啡酸、芦丁、金丝桃苷、异绿原酸B、异绿原酸A、异绿原酸C及款冬酮的相对校正因子分别为0.997、0.998、1.762、0.601、0.865、1.229、1.232、1.233、0.743;且在不同实验条件下重现性良好[相对校正因子的RSD(n=6)<2.4%];6批款冬花中10个成分的计算值与实测值间无显著差异。结论:本文建立的一测多评法控制款冬花的质量可行、准确。Objective：To establish a method for simultaneous assay of 10 main active constituents （neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rutin, hyperin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C and tussilagone） in Farfarae Flos by quantitative analysis of multi - components by sin- gle marker （QAMS）. Methods ：The chromatographic separation was achieved on a Kromasil C18column （250 mm × 4. 6 mm,5 μm）, the mobile phase was acetonitrile -0. 1% phosphoric acid aqueous solution with gradient elution（0→10→25→30→35→40 min, 10%→16%→30% →90%→95% →95% acetonitrile）. The detection wavelengths were changeable （0 - 15 rain, 326 nm for detection of neochlorogenic acid, ehlorogenic acid, cryptochlorogenie acid and caffeic acid ; 15 - 20 min, 254 nm for detection of rutin and hyperin ; 20 - 30 rain, 326 nm for detection of isoch- lorogenic acid B, isochlorogenic acid A and isochlorogenic acid C; 30 -40 rain, 220 nm for detection of tussilag- one）. The flow rate was 1.0 mL .min-1 and the column temperature was 30 ℃. Chlorogenic acid was used as the reference, the calibration factor of other 9 constituents to that of chlorogenic acid were calculated respectively to get each relative calibration factor. And the contents of active constituents were calculated by the relative calibration factor to realize QAMS. At the same time,external standard method was used to detect the contents of the 10 constituents,the two methods were compared to validate the accuracy and feasibility of the new method. Results：Relative correction fac- tors of neochlorogenic cryptochlorogenic acid, caffeic acid, rutin, hyperin, isochlorogenic acid B, isochlorogenicacid A, isochlorogenic acid C and tussilagone with reference to chlorogenic acid were 0. 997, 0. 998, 1.762, O. 601, O. 865, 1.229, 1. 232, 1.233, and 0. 743 respectively, and repeatability was good in different experimental conditions [ RSD of elative correction factors less than 2. 4% （n=6）

关 键 词：一测多评 相对校正因子 款冬花 高效液相色谱 新绿原酸 绿原酸 隐绿原酸 咖啡酸 异绿原酸 芦丁 金丝桃苷 款冬酮 

分 类 号：R917[医药卫生—药物分析学]