坤泰胶囊HPLC特征图谱及多成分含量测定方法的研究

Study on HPLC fingerprint and simultaneous determination of multi-components for Kuntai capsules

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,5μm,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·m L-1(r=0.999 6)、0.028 5~0.3559 mg·m L-1(r=0.999 5)、0.005 5~0.066 0 mg·m L-1(r=0.999 5)、0.060 8~0.480 7 mg·m L-1(r=1.000 0)、0.009 4~0.140 8 mg·m L-1(r=0.999 9)、0.052 1~0.520 8 mg·m L-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·m L-1、7.030~8.565 mg·m L-1、1.863~2.510 mg·m L-1、17.272~21.315mg·m L-1、2.870~3.374 mg·m L-1、11.002~13.245 mg·m L-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,5μm,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·m L-1(r=0.999 6),0.028 5-0.355 9 mg·m L-1(r=0.999 5),0.005 5-0.066 0 mg·m L-1(r=0.999 5),0.060 8-0.480 7 mg·m L-1(r=1.000 0),0.009 4-0.140 8 mg·m L-1(r=0.999 9)and 0.052 1-0.520 8 mg·m L-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·m L-1,7.030-8.565 mg·m L-1,1.863-2.510 mg·m L-1,17.272-21.315 mg·m L-1,2.870-3.533 mg·m L-1 and 11.002-13.245 mg·m L-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.