HPLC-DAD法同时测定小儿退热合剂中绿原酸、栀子苷、黄芩苷和丹皮酚的含量

目的：建立同时测定小儿退热合剂中绿原酸、栀子苷、黄芩苷和丹皮酚含量的高效液相色谱法。方法：采用C₁₈色谱柱（4.6 mm×250 mm，5 μm），以甲醇（A）-0.2%磷酸水溶液（B）为流动相，梯度洗脱，流速1.0 mL·min^-1，检测波长330 nm（绿原酸）、240 nm（栀子苷）、278 nm（黄芩苷和丹皮酚），柱温为30℃。结果：绿原酸、栀子苷、黄芩苷、丹皮酚进样量分别在0.003~0.171 μg（r=0.999 9）、0.025~1.228 μg（r=0.999 9）、0.034~1.679 μg（r=0.999 9）、0.005~0.254 μg（r=0.999 9）与峰面积呈良好的线性关系，平均回收率（n=9）分别为99.8%、99.1%、100.5%、99.9%，精密度RSD均小于0.5%，重复性RSD均小于2.0%，供试品溶液放置48 h内稳定。其中栀子苷与黄芩苷的含量测定结果与药典方法的测定结果无显著性差异。含量测定的结果表明，不同生产企业之间各成分的含量差异较大，部分生产企业不同生产批次间的差异也较大。结论：经方法学验证，本法可用于小儿退热合剂中绿原酸、栀子苷、黄芩苷和丹皮酚含量的测定。

Objective: To develop an HPLC method for simultaneous determination of chlorogenic acid,geniposide,baicalin,and paeonol in Xiao'er Tuire Heji.Methods: The C₁₈ column(4.6 mm×250 mm, 5μm)was adopted.The mobile phase was methanol(A)-0.2% phosphoric acid aqueous solution(B),with a gradient elution.The flow rate was 1.0 mL·min^-1.The detection wavelength was set at 330 nm for chlorogenic acid,240 nm for geniposide,278 nm for baicalin and paeonol.The column temperature was maintained at 30℃.Results: The calibration curves were linear in the quality range of 0.003-0.171μg(r=0.999 9) for chlorogenic acid,0.025-1.228μg(r=0.999 9) for geniposide,0.034-1.679μg(r=0.999 9) for baicalin,and 0.005-0.254μg(r=0.999 9) for paeonol.The average recoveries(n=9) were 99.8%,99.1%,100.5%,and 99.9%,respectively.The RSDs of precision were less than 0.5% for the four components,and the RSDs of reproducibility were less than 2.0%.The solution was stable within 48 h.There was no significant difference in the content determination of geniposide and baicalin by this method and by Chinese pharmacopeia method.The results of determination showed that the content of each component varies in different production enterprises,and the differences among different production batches of some production enterprises were also great.Conclusion: This method is proved by methodology validation that it can be used for determination of chlorogenic acid,geniposide,baicalin,and paeonol in Xiao'er Tuire Heji.

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