葛根芩连片多成分含量测定的研究

目的：建立葛根芩连片中14个特征性成分的HPLC含量测定方法，并测定11家生产企业的葛根芩连片含量。方法：采用Inertsil ODS-3（250 mm×4.6 mm，5μm）反相高效液相色谱柱，以乙腈-0.02 mol·L^-1醋酸铵+0.03%三乙胺溶液（用冰醋酸调pH4.3）为流动相梯度洗脱，流速1.0 mL·min^-1，葛根素、大豆苷、甘草苷、黄芩苷、汉黄芩苷、大豆苷元、甘草酸铵、黄芩素8个成分检测波长250 nm，汉黄芩素检测波长280 nm，表小檗碱、盐酸药根碱、黄连碱、盐酸巴马汀、盐酸小檗碱5个成分检测波长346 nm。结果：葛根素、大豆苷、甘草苷、黄芩苷、汉黄芩苷、表小檗碱、盐酸药根碱、黄连碱、大豆苷元、盐酸巴马汀、甘草酸铵、盐酸小檗碱、黄芩素、汉黄芩素进样量分别在146.26~5850.24、24.13~965.04、18.45~738.00、79.18~3167.32、9.57~382.80、4.76~190.40、2.57~102.80、13.41~536.40、10.60~424.00、11.33~453.22、12.08~483.20、46.73~1869.25、20.28~811.20、12.11~484.50 ng范围内与峰面积呈良好线性关系（r>-10.9990）；平均加样回收率（n=6）分别为2.18%、1.79%、1.81%、1.68%、2.27%、2.13%、1.96%、1.07%、0.93%、0.61%、2.92%、0.77%、2.79%、0.62%；精密度、重复性、稳定性良好，RSD均小于3%。用该方法测定11家生产企业生产的16批葛根芩连片，除一批样品的汉黄芩苷未被检出外，其余企业14种成分均有检出，但含量存在差异。结论：该方法准确易行，可用于该品种的整体质量控制。

Objective: To establish an HPLC method for simultaneous determination of fourteen index components in Gegen Qinlian tablets and perform the quantitative analysis on Gegen Qinlian tablets from eleven pharmaceutical companies. Methods: RP-HPLC separation was carried on an Inertsil ODS-3 column(250 mm×4.6 mm, 5 μm)with the mobile phase consisting of acetonitrile and ammonium acetate+0.03% triethylamine aq.(adjusting pH to 4.3 by acetic acid)with a gradient elution at the flow rate of 1.0 mL·min^-1. The detection wavelength was set at 250 nm for puerarin, daidzin, liquiritin, baicalin, wogonoside, daidzein, ammonium glycyrrhizinate and baicalein, at 280 nm for wogonin, and at 346 nm for epierberine, jatrorrhizine hydrochloride, coptisine, palmatine hydrochloride and berberine hydrochloride. Results: Good linear correlations(r>-10.999 0) were found in the range of 146.26-5 850.24 ng, 24.13-965.04 ng, 18.45-738.00 ng, 79.18-3 167.32 ng, 9.57- 382.80 ng, 4.76-190.40 ng, 2.57-102.80 ng, 13.41-536.40 ng, 10.60-424.00 ng, 11.33-453.22 ng, 12.08- 483.20 ng, 46.73-1 869.25 ng, 20.28-811.20 ng, 12.11-484.50 ng for puerarin, daidzin, liquiritin, baicalin, wogonoside, epierberine, jatrorrhizine hydrochloride, coptisine, daidzein, palmatine hydrochloride, ammonium glycyrrhizinate, berberine hydrochloride, baicalein and wogonin, respectively. The average recoveries(n=6)of the fourteen components were 2.18%、1.79%、1.81%、1.68%、2.27%、2.13%、1.96%、1.07%、0.93%、0.61%、2.92%、0.77%、2.79%、0.62%, respectively. The accuracy, precision and stability were satisfactory with RSD values less than 3%. Sixteen batches of Gegen Qinlian tablets from eleven pharmaceutical companies were investigated. Wogonoside was not founded in one batch and all the fourteen components were detected in the rest samples but their contents varied. Conclusion: The established method was accurate and feasible, which could be applied in comprehensive quality control of Gegen Qinlian tablets.