紫连生肌凝胶剂HPLC指纹图谱建立及6个成分含量测定

Establishment of HPLC fingerprints and determination of six components for Zilian Shengji gel

目的:建立紫连生肌凝胶剂指纹图谱,并测定没食子酸、表小檗碱、黄连碱、巴马汀、小檗碱和左旋紫草素的含量。方法:采用Agilent C_(18)-WR(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.15%磷酸水,梯度洗脱,流速0.8 mL·min-1,柱温25℃,检测波长270、345、516 nm,结合聚类分析、主成分分析和正交偏最小二乘法判别分析对紫连生肌凝胶剂的指纹图谱进行化学模式识别,并对指定的6个成分进行定量分析。结果:建立了紫连生肌凝胶剂指纹图谱,共标定26个共有峰,与对照指纹图谱的相似度为0.828~0.997;指认出没食子酸、表小檗碱、黄连碱、小檗碱、巴马汀和左旋紫草素6个色谱峰。6个成分在各自范围内线性关系良好(r≥0.9995),平均加样回收率在98.7%~99.3%,RSD在1.3%~1.8%。11批样品中6个成分含量分别为827.74~1513.50μg·g^(-1)、137.52~296.05μg·g^(-1)、381.83~884.73μg·g^(-1)、2023.81~4051.66μg·g^(-1)、524.15~986.13μg·g^(-1)和47.65~549.46μg·g^(-1)。结论:该方法简便,稳定可靠,重复性好,可用于紫连生肌凝胶剂的质量控制。

Objective:To establish the HPLC fingerprints for Zilian Shengji gel and to determine the contents of gallic acid,tableberberine,coptisine,palmatine,berberine and shikonin.Methods:The establishment of fingerprints was performed on a 25℃thermostatic Agilent C_(18)-WR(250 mm×4.6 mm,5μm)column with the mobile phase comprising of acetonitrile-0.15%phosphoric acid water at the flowing rate of 0.8mL·min^(-1)in a gradient elution manner,and the detection wavelength were set at 270 nm,345 nm and 516 nm.Cluster analysis,principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were adopted in chemical pattern recognition.The six components identified were quantitatively determined.Results:The HPLC fingerprint of Zilian Shengji gel was constructed with twenty-six common chromatographic peaks,eleven batches of samples with the similarities of 0.828-0.997,six chromatographic peaks of gallic acid,tableberberine,coptisine,palmatine,berberine and shikonin were identified by reference substance comparison.Six constituents showed good linear relationships within their own ranges(r≥0.9995)whose average recoveries were 98.7%-99.3%,with the RSDs of 1.3%-1.8%.The content ranges of the above mentioned six components in eleven batches of samples were 827.74-1513.50μg·g^(-1),137.52-296.05μg·g^(-1),381.83-884.73μg·g^(-1),2023.81-4051.66μg·g^(-1),524.15-986.13μg·g^(-1)and 47.65-549.46μg·g^(-1).Conclusion:This simple,stable,reliable and reproducible method can be used for the quality control of Zilian Shengji gel.