HPLC波长转换法同时检测双黄连口服液中4种有效成分的含量

目的建立高效液相色谱波长转换法同时测定双黄连口服液中绿原酸、连翘苷、黄芩苷和汉黄芩素含量。方法采用Phenomenex C18柱(4.6 mm×150 mm,5μm);流动相A为甲醇,流动相B为乙腈,流动相C为0.1%磷酸水溶液;流速为1.0 mL/min;柱温为30℃;进样器温度为5℃;洗脱程序:0～8 min为B∶C=12∶88,9～19 min为A∶B∶C=35∶12∶53,20～27 min为A∶B∶C=35∶40∶25,28～35 min为B∶C=12∶18;检测波长程序:0～8 min为326 nm,9～19 min为226 nm、278 nm,20～27 min为278 nm。结果绿原酸、连翘苷、黄芩苷和汉黄芩素保留时间分别为6.33,15.48,16.69,24.04 min。以峰面积对进样质量线性回归,绿原酸、连翘苷、黄芩苷、汉黄芩素的回归方程分别为Y=41.647X-0.158(r=0.9999),Y=31.391X-0.164(r=0.9999),Y=55.847X+2.290(r=0.9996),Y=74.652X+0.045(r=0.9997),线性范围分别为0.06～0.48μg,0.02～0.45μg,0.375～4.500μg,0.002～0.090μg。绿原酸、连翘苷、黄芩苷和汉黄芩素回收率分别为95.90%、97.02%、95.16%、91.34%。结论本方法简便可行、准确快速,可用于双黄连口服液中4种主要有效成分的含量测定。

Simultaneous Determination of Chlorogenic Acid, Forsythin, Baicalin and Wogonin in Shuanghuanglian Oral Liquid by Wavelength Conversion HPLC

Objective To establish an HPLC quantitative method for simultaneous determination of chlorogenic acid,forsythin,baicalin and wogonin in Shuanghuanglian Oral Liquid by wavelength conversion HPLC.Methods RP-HPLC was performed on a Phenomenex C18column(4.6 mm×150 mm i.d.,5 μm)at 30 ℃.The mobile phase consisted of methanol(A),acetonitrile(B)and 0.1% phosphate solution(C,pH value being 2.1).The gradient elution was manipulated within 0 ～ 8 min with B ∶ C=12 ∶ 88,9 ～ 19min with A ∶ B ∶ C=35 ∶ 12 ∶ 53,20 ~ 27min with A ∶ B ∶ C=35 ∶ 40 ∶ 25,28~35 min with B ∶ C=12 ∶ 18.The flow rate was 1.0 mL/min.The automatic sample injector was set at 5 ℃.The ultraviolet detector was operated at 326nm(0 ～ 8min),226nm and 278nm(9 ～ 19min),and 278nm(20 ～ 27min).Results The retention time of chlorogenic acid,forsythin,baicalin and wogonin was 6.33,15.48,16.69,24.04 min,respectively.Chlorogenic acid showed good linearity in the range of 0.06 ～ 0.48 μg(Y=41.647X-0.158,r=0.9999),forsythin in the range of 0.02 ~ 0.45 μg(Y=31.391X-0.164,r=0.9999),baicalin in the range of 0.375 ~ 4.500 μg(Y=55.847X+2.290,r=0.9996)and wogonin in the range of 0.002 ~ 0.090 μg(Y=74.652X+0.045,r=0.9997).The average recovery of chlorogenic acid,forsythin,baicalin and wogonin was 95.90%,97.02%,95.16%,91.34%,respectively.Conclusion The method is simple,fast and accurate.It can be used for simultaneous analysis of chlorogenic acid,forsythin,baicalin and wogonin in Shuanghuanglian Oral Liquid.