HPLC法同时测定双虎清肝颗粒中9种成分的含量

Simultaneous determination of nine constituents in Shuanghu Qinggan Granule by HPLC

目的建立HPLC法同时测定双虎清肝颗粒中鸡矢藤次苷甲酯、车叶草苷酸、车叶草苷、虎杖苷、白藜芦醇、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA含量的方法。方法采用Agilent Zorbax SB C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-体积分数0.1%甲酸溶液,梯度洗脱,流速1.0 mL·min^(-1);柱温30℃,检测波长分别为238 nm(检测鸡矢藤次苷甲酯、车叶草苷酸和车叶草苷)、290 nm(检测虎杖苷和白藜芦醇)和255 nm(检测二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮ⅡA)。结果鸡矢藤次苷甲酯、车叶草苷酸、车叶草苷、虎杖苷、白藜芦醇、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA的质量浓度分别在4.470~89.40 mg·L^(-1)、2.760~55.20 mg·L^(-1)、3.780~75.60 mg·L^(-1)、4.690~93.80 mg·L^(-1)、7.920~158.4 mg·L^(-1)、0.5300~10.60 mg·L^(-1)、1.910~38.20 mg·L^(-1)、1.690~33.80 mg·L^(-1)、2.960~59.20 mg·L^(-1)内与峰面积呈良好的线性关系(r≥0.9991),平均加样回收率分别为98.64%、98.81%、99.15%、100.04%、99.66%、96.92%、97.89%、97.51%和98.40%,RSD分别为1.12%、1.47%、1.08%、0.79%、0.86%、1.06%、1.35%、0.89%和1.22%。结论该方法可作为双虎清肝颗粒的质量控制评价方法。

Objective To establish an HPLC method for simultaneous determination of feretoside,asperulosidic acid,asperuloside,polydatin,resveratrol,15,16-dihydrotanshinone I,cryptotanshinone,tanshinone I and tanshinoneⅡA in Shuanghu Qinggan Granule.Methods The chromatographic separation was carried out on Agilent Zorbax SB C_(18)(250 mm×4.6 mm,5μm)with a mobile phase consisting of acetonitrile-0.1%formic acid solution in gradient elution with a flow rate of 1.0 mL·min^(-1) and the column temperature was set at 30℃.The detection wavelengths were set at 238 nm for feretoside,asperulosidic acid and asperuloside,290 nm for polydatin and resveratrol,and 255 nm for 15,16-dihydrotanshinone I,cryptotanshinone,tanshinone I and tanshinoneⅡA.Results Feretoside,asperulosidic acid,asperuloside,polydatin,resveratrol,15,16-dihydrotanshinone I,cryptotanshinone,tanshinone I and tanshinoneⅡA showed good linear relationships within the ranges of 4.470-89.40 mg·L^(-1),2.760-55.20 mg·L^(-1),3.780-75.60 mg·L^(-1),4.690-93.80 mg·L^(-1),7.920-158.40 mg·L^(-1),0.5300-10.60 mg·L^(-1),1.910-38.20 mg·L^(-1),1.690-33.80 mg·L^(-1),2.960-59.20 mg·L^(-1)(r≥0.9991).The average recoveries were 98.64%,98.81%,99.15%,100.04%,99.66%,96.92%,97.89%,97.51%,98.40%with the RSDs of 1.12%,1.47%,1.08%,0.79%,0.86%,1.06%,1.35%,0.89%and 1.22%,respectively.Conclusion The established method has good specificity,maneuverability,high repeatability and high accuracy,and can be used as a quality control method for Shuanghu Qinggan Granule.