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目的:建立同时测定妇科养坤丸中芍药苷、甘草苷、黄芩苷和木香烃内酯含量的HPLC法并通过响应面优化法分别对4种成分的提取工艺进行优化。方法:采用Kromasil C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)-0.2%磷酸溶液(B),梯度洗脱,流速为1.0 ml·min~(-1),检测波长为230,276,280,225 nm,柱温为28℃。以4种成分含量为指标,选取甲醇浓度、料液比和提取时间为考察因素,采用响应面优选妇科养坤丸中4种有效成分的提取工艺。结果:芍药苷、甘草苷、黄芩苷和木香烃内酯分别在1.616~161.600,0.432~43.180,2.045~204.500,0.518~51.840μg·ml~(-1)呈良好的线性关系,平均回收率(n=9)为98.3%,99.6%,97.9%,98.1%。获得的最佳工艺条件为甲醇浓度64%,料液比1∶51,提取时间为25 min。结论:采用响应面优化妇科养坤丸中4种有效成分的提取工艺,方法简便,预测性良好。所建立的含量测定方法简便、准确,可用于妇科养坤丸的质量控制。
OBJECTIVE: To establish a HPLC method for the simultaneous determination of paeoniflorin, liquiritin, baicalin and fondylolide in Gynecological Yangkun pill and optimize the extraction process of the four components by response surface methodology. Methods: A Kromasil C 18 column (250 mm × 4.6 mm, 5 μm) was used with a mobile phase of acetonitrile (A) and 0.2% phosphoric acid (B) ), The detection wavelength was 230,276,280,225 nm, the column temperature was 28 ℃. The contents of four components were taken as indexes to select the concentration of methanol, the ratio of solid to liquid and the extraction time as the investigation factors. The response surface was used to optimize the extraction process of the four effective components in Gynecological Yangkun pill. Results: Paeoniflorin, glycyrrhizin, baicalin and lignocellulose lactone showed a good linear relationship between 1.616 ~ 161.600, 0.432 ~ 43.180, 2.045 ~ 204.500 and 0.518 ~ 51.840μg · ml ~ (-1) (n = 9) were 98.3%, 99.6%, 97.9%, 98.1%. The optimum conditions were 64% methanol concentration, solid-liquid ratio 1:51 and extraction time 25 min. Conclusion: The response surface optimization of gynecological Yangkun pill four kinds of active ingredients extraction process, the method is simple, good predictability. The established determination of content is simple and accurate, can be used for gynecological Yangkun pill quality control.