三乌胶丸HPLC指纹图谱及多指标成分定量分析研究

Study on HPLC fingerprint and multi-indicator quantitative analysis of Sanwujiaowan

建立三乌胶丸的HPLC指纹图谱及定量研究多种指标性成分的方法,为三乌胶丸的质量评价提供实验根据.色谱分析柱型号为Agilent ZORBAX Eclipse Plus C_(18),进行梯度洗脱,柱温为30℃,流速为1.0 mL/min,测定波长245 nm,流动相为0.2%二乙胺(磷酸调节其pH为4.0)水溶液(A)-乙腈(B).结果表明,建立的15批次三乌胶丸的HPLC指纹图谱一共确定了18个共有峰,辨别了指纹图谱中的6个成分,15批三乌胶丸指纹图谱的相似度均大于0.9.8-去乙酰滇乌碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、大黄素、草乌甲素6个成分的质量分数依次为87.291~1182.018、144.150~2193.629、30.307~2229.562、100.810~646.250、4.474~114.140、4.255~13.591μg/g.通过CA、PCA、PLS-DA等分析,15个批三乌胶丸按照生产年限被分成2类.所建立的三乌胶丸HPLC指纹图谱及6种成分定量分析的方法易操作,重复性好,可为三乌胶丸的质量研究提供参考.

A HPLC fingerprint of Sanwujiaowan and a method for the determination of six components in Sanwujiaowan were built,so as to provide experiment basis for its quality evaluation.The chromatographic conditions were that using Agilent ZORBAX Eclipse Plus C_(18)(φ4.6 mm×250 mm,5μm)liquid chromatographic column,using gradient elution,the temperature of the liquid-analysis column was at 30℃and the wavelength of the detection were set at 245 nm,the flow velocity was set at 1.0 mL/min,the mobile phase is0.2%diethylamine(phosphoric acid adjusted its pH to 4.0)aqueous solution(A)-acetonitrile(B).The fingerprints of 15 batches of Sanwujiaowan were established,18 common peaks and 6 components were identified,and their similarity was more than 0.9.The contents of benzoylmesaconine,benzoylaconitine,benzoylhypacoitine,8-deacetylyunaconitine,emodin and crassicauline A were 144.150~2193.629,30.307~2229.562,100.810~646.250,87.291~1182.018,4.474~114.140,4.255~13.591μg/g respectively.Through the analysis of CA,PCA and PLSDA,15 batches of Sanwujiaowan were divided into 2 types.The HPLC fingerprint and six components quantitative analysis methods established in this paper are easy to operate and reproducible,which can offer reference for the quality research of Sanwujiaowan.