UHPLC测定芎菊上清丸中9种成分含量及化学模式分析

Simultaneous Determination of Nine Components in Xiongju Shangqing Pills by UHPLC Combined with Chemical Pattern Recognition

关键词:  芎菊上清丸  超高效液相色谱法  含量测定  模式识别

Key words:  Xiongju Shangqing pills  UHPLC  content determination  pattern recognition

摘要:

目的 建立UHPLC波长切换法同时测定芎菊上清丸中9种成分的含量方法。方法 采用Agilent Ecilipse C₁₈（2.1 mm×100 mm，1.6 μm）色谱柱，流动相：甲醇-0.05%磷酸水溶液，梯度洗脱；流速为0.3 mL·min^-1；检测波长：327，237，320，345，278，254 nm；柱温30℃；进样量2 μL；并采用SPSS 22.0统计软件对含量测定结果进行主成分分析与聚类分析。结果 绿原酸、3，5-二咖啡酰奎宁酸、栀子苷、甘草苷、阿魏酸、盐酸小檗碱、黄芩苷、升麻素苷、5-O-甲基维斯阿米醇苷线性范围分别为4.30~68.80 μg·mL^-1（r=0.999 0）、6.66~106.56 μg·mL^-1（r=0.999 2）、7.67~122.72 μg·mL^-1（r=0.999 4）、4.88~78.08 μg·mL^-1（r=0.999 1）、2.37~37.92 μg·mL^-1（r=0.999 1）、6.50~103.92 μg·mL^-1（r=0.999 2）、8.85~141.60 μg·mL^-1（r=0.999 4）、0.88~14.08 μg·mL^-1（r=0.999 7）、0.74~11.92 μg·mL^-1（r=0.999 3）；平均加样回收率（n=9）均在99.42%~103.10%，RSD均<2.0%。主成分分析与聚类分析均可将不同生产厂家的芎菊上清丸很好地分类，且分类结果一致。结论 所建立的多成分方法快捷、准确、重复性好，可用于芎菊上清丸的质量控制。

Abstract:

OBJECTIVE To develop a UHPLC method for the simultaneous determination of 9 components in Xiongju Shangqing pills. METHODS An Agilent Ecilipse C₁₈(2.1 mm×100 mm, 1.6 μm) chromatographic column was used; the mobile phase was methanol-0.05% phosphoric acid solution with gradient elution at the flow rate of 0.3 mL·min^-1 and the detection wavelengths were 327, 237, 320, 345, 278, 254 nm and the column temperature was 30℃, the injection volume was 2 μL. The principal component analysis(PCA) and cluster analysis(CA) were conducted for the results of content determination by SPSS 22.0 statistical software. RESULTS The linear ranges of chlorogenic acid, 3,5-dicaffeoylquinic acid, geniposide, liquiritin, ferulic acid, berberine hydrochloride, baicalin, prim-O-glucosylcimifugin, 5-O-methylvisammioside were 4.30-68.80 μg·mL^-1 (r=0.999 0), 6.66-106.56 μg·mL^-1(r=0.999 2), 7.67-122.72 μg·mL^-1(r=0.999 4), 4.88-78.08 μg·mL^-1 (r=0.999 1), 2.37-37.92 μg·mL^-1(r=0.999 1), 6.50-103.92 μg·mL^-1(r=0.999 2), 8.85-141.60 μg·mL^-1(r=0.999 4), 0.88-14.08 μg·mL^-1 (r=0.999 7), 0.74-11.92 μg·mL^-1(r=0.999 3), respectively. The recoveries ranged from 99.42%-103.10% and RSDs were all < 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 Xiongju Shangqing pills.