HPLC-DAD法同时测定玄参炮制前后8个有效成分的含量
Simultaneous quantitation of eight active components in crude and processed Radix Scrophulariae extracts by high performance liquid chromatography with diode array detector
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摘要: 目的:同时测定不同来源玄参生品与炮制品中8个主要成分的含量;方法:采用Agilent Zorbax Extend C18色谱柱(250 mm 4.6 mm, 5m),二极管阵列检测器;以0.3%磷酸水溶液-乙腈为流动相,梯度洗脱,流速为1.0 mLmin-1,检测波长为280 nm和210 nm,柱温为30℃。结果:8个主要成分均呈现良好的线性关系(r2 0.9992);检测限与定量限分别为0.07-0.32和0.16-0.93gmL-1;日间与日内偏差小于0.45%;平均回收率在99.31%-100.19%, RSD范围为0.29%-1.28%。结论:本方法可用于玄参生品与炮制品内在质量控制。Abstract: AIM: To determine eight major components in crude and processed Radix Scrophulariae, several samples from different arrears were examined. METHODS: The eight components were separated on an Agilent Zorbax Extend C18 column (250 mm 4.6 mm, 5 m) and detected by diode array detector (DAD). Mobile phase was composed of (A) aqueous phosphoric acid (0.03%, V/V) and (B) acetonitrile using a gradient elution. Analytes were performed at 30℃ with a flow rate of 1.0 mLmin-1 and UV detection at 280 nm and 210 nm. RESULTS: All calibration curves showed good linear regression (r2 0.999 2) within tested ranges. The limits of detection and limits of quantification were 0.07-0.32 gmL-1 and 0.16-0.93 gmL-1, respectively. Overall intra-day and inter-day variations were less than 0.45%, and the average recoveries were 99.31%-100.19%, with RSD ranging from 0.29% to 1.28% for the analytes. CONCLUSION: The developed method can be applied to the intrinsic quality control of crude and processed Radix Scrophulariae.