坤泰胶囊HPLC特征图谱及多成分含量测定方法的研究

目的：建立HPLC法同时测定坤泰胶囊中没食子酸、芍药苷、黄芩素、黄芩苷、盐酸巴马汀、盐酸小檗碱6个成分的含量，并以盐酸小檗碱为参照峰建立特征图谱。方法：样品用甲醇超声提取，采用Amethyst C₁₈-H色谱柱（4.6 mm×250 mm，5 μm，120 ?）；以乙腈-0.4%磷酸水溶液为流动相，梯度洗脱；检测波长：230 nm；柱温：25℃。结果：没食子酸、芍药苷、黄芩素、黄芩苷、盐酸巴马汀、盐酸小檗碱6个成分在各自的浓度范围内线性关系良好，线性范围分别为0.003 6~0.043 8 mg·mL^-1（r=0.999 6）、0.028 5~0.355 9 mg·mL^-1（r=0.999 5）、0.005 5~0.066 0 mg·mL^-1（r=0.999 5）、0.060 8~0.480 7 mg·mL^-1（r=1.000 0）、0.009 4~0.140 8 mg·mL^-1（r=0.999 9）、0.052 1~0.520 8 mg·mL^-1（r=0.999 9）；平均加样回收率在97.5%~103.0%之间；RSD在0.63%~1.8%之间；17批样品中没食子酸、芍药苷、黄芩素、黄芩苷、盐酸巴马汀、盐酸小檗碱含量范围分别为0.856~1.133 mg·mL^-1、7.030~8.565 mg·mL^-1、1.863~2.510 mg·mL^-1、17.272~21.315 mg·mL^-1、2.870~3.374 mg·mL^-1、11.002~13.245 mg·mL^-1；特征图谱中有21个共有峰，指认了其中6个化学成分，相似度均大于0.990。结论：本研究所建立的坤泰胶囊特征图谱及含量测定方法稳定、可靠，重复性好，可作为坤泰胶囊的质量控制方法。  

Objective:To establish an HPLC method for simultaneous determination of gallic acid,paeoniflorin,baicalein,baicalin,bamatine hydrochloride and berberine hydrochloride in Kuntai capsules,and to establish a characteristic chromatogram with berberine hydrochloride as the reference peak. Methods:The sample was extracted with methanol by ultrasonic extraction. Amethyst C₁₈-H column(4.6 mm×250 mm,5 μm,120?) was used as the solid phase. Acetonitrile-0.4% phosphoric acid aqueous solution was used as mobile phase and gradient elution was performed. Detection wavelength was 230 nm and column temperature was 25℃. Results:Gallic acid,paeoniflorin,baicalin,baicalin,bamatine hydrochloride and berberine hydrochloride had good linear relationship in various concentrations. Linear ranges were 0.003 6-0.043 8 mg·mL^-1(r=0.999 6),0.028 5-0.355 9 mg·mL^-1(r=0.999 5),0.005 5-0.066 0 mg·mL^-1(r=0.999 5),0.060 8-0.480 7 mg·mL^-1(r=1.000 0),0.009 4-0.140 8 mg·mL^-1(r=0.999 9) and 0.052 1-0.520 8 mg·mL^-1(r=0.999 9),respectively. The average recoveries were within 97.5%-103.0% with RSDs ranged from 0.63% to 1.8%. The contents of gallic acid,paeoniflorin,baicalein,baicalin,bamatine hydrochloride and berberine hydrochloride in 17 batches of samples were 0.856~1.133 mg·mL^-1,7.030-8.565 mg·mL^-1,1.863-2.510 mg·mL^-1,17.272-21.315 mg·mL^-1,2.870-3.533 mg·mL^-1 and 11.002-13.245 mg·mL^-1. There were 21 common peaks in the characteristic chromatogram,among which 6 chemical components were identified. Similarities of samples were all above 0.990. Conclusion:The characteristic chromatogram and assay method of Kuntai capsules established in this study are stable,reliable and reproducible,which can be used for quality control of Kuntai capsules. 

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