HPLC测定复方鱼腥草合剂中8个成分含量及化学模式分析

Simultaneous Determination of 8 Components in Compound Yuxingcao Mixture by HPLC Combined with Chemical Pattern Recognition

目的:建立高效液相色谱法(HPLC)同时测定复方鱼腥草合剂中8个成分含量的方法。方法:采用迪马Diamonsil C_(18)(250 mm×4.6 mm,5μm)色谱柱,流动相:乙腈(A)-0.1%磷酸水溶液(B),梯度洗脱;流速:1.0 ml·min^(-1);检测波长:250 nm(R,S-告依春、连翘酯苷A、连翘苷、黄芩苷、汉黄芩苷和槲皮素)、330 nm(绿原酸和3,5-二-O-咖啡酰奎宁酸);柱温:30℃;进样量:10μl。采用统计分析软件对含量测定结果进行聚类分析与主成分分析。结果:R,S-告依春、绿原酸、连翘酯苷A、3,5-二-O-咖啡酰奎宁酸、连翘苷、黄芩苷、汉黄芩苷和槲皮素检测质量浓度线性范围分别为1.787~16.085μg·ml^(-1)、0.940~8.462μg·ml^(-1)、0.606~5.453μg·ml^(-1)、0.433~3.893μg·ml^(-1)、0.358~3.226μg·ml^(-1)、7.013~63.120μg·ml^(-1)、3.259~29.328μg·ml^(-1)、0.309~2.784μg·ml^(-1)(r≥0.9992);定量限分别为1.352,0.896,0.578,0.416,0.336,6.954,2.963,0.296μg·ml^(-1);精密度、稳定性(48 h)、重复性试验的RSD<2.0%(n=6);平均加样回收率分别为99.17%,98.76%,98.87%,98.48%,98.55%,99.43%,99.19%,99.32%(n=6),RSD均≤2.0%;主成分分析与聚类分析均可将不同生产厂家的复方鱼腥草合剂很好地分类,两种分类结果一致。结论:所建立的多成分方法快捷、准确、重复性好,可用于复方鱼腥草合剂的质量控制。

Objective:To develop an HPLC method for the simultaneous determination of 8 components in compound Yuxingcao mixture.Methods:A Diamonsil C18(250 mm×4.6 mm,5μm)chromatographic column was used;the mobile phase was methanol-0.1%phosphoric acid solution with gradient elution at the flow rate of 1.0 ml·min^(-1),the detection wavelengths were 250 nm and 330 nm,the column temperature was 30℃,and the injection volume was 10μl.The principal component analysis(PCA)and cluster analysis(CA)were conducted for the results of content determination.Results:The linear ranges of(R,S)-goitrin,chlorogenic acid,geniposide,forsythoside A,3,5-O-dicaffeoylquinic acid,forsythin,baicalin,wogonoside and quercetin were 1.787-16.085μg·ml^(-1),0.940-8.462μg·ml^(-1),0.606-5.453μg·ml^(-1),0.433-3.893μg·ml^(-1),0.358-3.226μg·ml^(-1),7.013-63.120μg·ml^(-1),3.259-29.328μg·ml^(-1)and 0.303-2.784μg·ml^(-1),respectively(r≥0.9992).The average recoveries were 99.17%,98.76%,98.87%,98.48%,98.55%,99.43%,99.19%and 99.32%,respectively with the RSDs lower than 2.0%.The samples could be clearly classified by using CA and PCA,and the results of classification were consistent.Conclusion:The established multi-component method is fast,accurate and reproducible.It can be used for the quality control of compound Yuxingcao mixture.