HPLC同时测定小儿肺热咳喘口服液中10个成分的含量

Simultaneous determination of ten ingredients in Xiaoer Feire Kechuan oral solution by HPLC

目的:建立HPLC法同时测定小儿肺热咳喘口服液中10个成分(盐酸麻黄碱、盐酸伪麻黄碱、苦杏仁苷、(R,S)-告依春、芒果苷、黄芩苷、连翘苷、绿原酸、连翘酯苷A、木犀草苷)的含量,为小儿肺热咳喘口服液质量标准完善提供依据。方法:采用ZORBAX SB-C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,体积流量为0.8 mL·min^(-1),柱温为35℃,检测波长为210 nm(盐酸麻黄碱、盐酸伪麻黄碱和苦杏仁苷)、258 nm[(R,S)-告依春、芒果苷、黄芩苷和连翘苷]和330 nm(绿原酸、连翘酯苷A和木犀草苷)。结果:上述10个成分进样量的线性关系分别为0.20~1.20μg(r=0.9990)、0.20~1.20μg(r=0.9996)、0.25~1.50μg(r=0.9994)、0.40~2.40μg(r=0.9995)、0.40~2.40μg(r=0.9992)、0.40~2.40μg(r=0.9991)、0.40~2.40μg(r=0.9993)、0.20~1.20μg(r=0.9993)、0.10~0.60μg(r=0.9994)、0.50~3.00μg(r=0.9996),平均加样回收率为98.5%~99.5%,RSD≤0.89%(n=9)。单因素方差分析结果表明,不同批次间的盐酸麻黄碱、盐酸伪麻黄碱、苦杏仁苷、黄芩苷、连翘苷、连翘酯苷A、芒果苷含量无显著性差异(P>0.01),(R,S)-告依春、绿原酸和木犀草苷的含量有显著差异(P<0.01)。盐酸麻黄碱、盐酸伪麻黄碱、苦杏仁苷、(R,S)-告依春、绿原酸、木犀草苷、黄芩苷、连翘苷、连翘酯苷A、芒果苷的平均含量分别为0.174、0.067、2.326、3.548、2.851、0.958、3.484、3.456、5.389、3.891 mg·mL^(-1)。结论:该方法简便、准确、灵敏,重复性好,可用于同时测定小儿肺热咳喘口服液中10个成分的含量。

Objective:To develop an HPLC method for simultaneous determination of ten constituents[ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalim,(R,S)epigoitrin,mangiferin,baicalin,phillyrin,chlorogenic acid,forsythiaside A and cynaroside]in Xiaoer Feire Kechuan oral solution,so as to offer basis for improvement of the drug’s standard.Methods:The analysis was performed on a ZORBAX SB-C_(18) column(4.6 mm×250 mm,5μm)with acetonitrile-0.1%phosphoric acid aqueous solution as mobile phases in gradientelution,0.8 mL·min^(-1) as flow rate and 35℃as column temperature.The detection wavelengths were at 210 nm for ephedrine hydrochloride,pseudoephedrine hydrochloride and amygdalim,258 nm for(R,S)epigoitrin,mangiferin,baicalin and phillyrin and 330 nm for chlorogenic acid,forsythiaside A and cymaroside,respectively.Results:The linear ranges were 0.20-1.20μg(r=0.9990)for ephedrine hydrochlorid,0.20-1.20μg(r=0.9996)for pseudoephedrine hydrochloride,0.25-1.50μg(r=0.9994)for amygdalim,0.40-2.40μg(r=0.9995)for(R,S)-epigoitrin,0.40-2.40μg(r=0.9992)for chlorogenic acid,0.40-2.40μg(r=0.9991)for cymaroside,0.40-2.40μg(r=0.9993)for baicalin,0.20-1.20μg(r=0.9993)for phillyrin,0.10-0.60μg(r=0.9994)for forsythiaside A and 0.50-3.00μg(r=0.9996)for mangiferin,respectively.The average recovery rates were in the range of 98.5%-99.5%with RSDs≤0.89%(n=9).The results of one-factor analysis of variance showed that there was no significant difference in the contents of ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalim,baicalin,phillyrin,forsythiaside A and mangiferin,but there was significant difference in the content of(R,S)-epigoitrin,chlorogenic acid and cynaroside between different batches.The average contents of ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalim,(R,S)-epigoitrin,chlorogenic acid,cynaroside,baicalin,phillyrin,forsythiaside A and mangiferin were 0.174,0.067,2.326,3.548,2.851,0.958,3.484,3.456,5.389,3.891 mg·mL^(-1),respectively.Conclusion:The developed method is simple,accurate,specific and reproducible,which can be used for determining the contents of ten constituents in Xiaoer Feire Kechuan oral solution.