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目的建立岩黄连培养细胞的高效液相色谱法(HPLC)指纹图谱质量控制方法,测定主要生物碱的含量。方法采用高效液相色谱法,色谱柱为Dikma Diamonsil C18柱(4.6 mm×250 mm,5μm),流动相为乙腈和水体积比为40∶60,每升流动相加3.4 g磷酸二氢钾和1.7 g SDS,检测波长345 nm,流速1 mL·min-1,柱温25℃。结果在线性范围内,脱氢卡维丁和小檗碱的浓度与色谱峰面积呈良好的线形关系,回归方程分别为Y=3 645.2X+47.61(r=0.998 9)和Y=4 172.7X+73.52(r=0.999 2),加样回收率分别为97.9%和98.2%,RSD均在3%以内。结论该方法准确、可靠,能有效分离和测定岩黄连培养细胞中脱氢卡维丁和小檗碱含量,所建立的指纹图谱相似度评价和方法学考察结果良好,可用于质量控制和定量分析。
OBJECTIVE To establish a method for the quality control of Rhizoma Coptidis cultured cells by high performance liquid chromatography (HPLC) fingerprinting and to determine the content of major alkaloids. Methods High performance liquid chromatography (HPLC) was performed on a Dikma Diamonsil C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase consisting of acetonitrile and water at a volumetric ratio of 40:60, mobile phase addition of 3.4 g potassium dihydrogen phosphate and 1.7 g SDS, detection wavelength 345 nm, flow rate 1 mL · min-1, column temperature 25 ℃. Results In the linear range, the concentrations of dehydrocavagetin and berberine showed a good linear relationship with the chromatographic peak area. The regression equations were Y = 3 645.2X + 47.61 (r = 0.998 9) and Y = 4 172.7X +73.52 (r = 0.999 2). The recoveries of samples were 97.9% and 98.2%, respectively. The RSDs were within 3%. Conclusion The method is accurate and reliable, and can effectively separate and determine the content of dehydrocavagetin and berberine in the cultured cells of Rhizoma Coptis. The fingerprint similarity evaluation and the methodological study results are good, which can be used for quality control and quantitative analysis .