指纹图谱与一测多评法相结合的大黄质量控制方法

 目的 建立指纹图谱及一测多评法同时测定8个成分含量的大黄质量控制方法。方法 高效液相色谱法,采用Symmetry C₁₈(4.6 mm×250 mm,5 μm)色谱柱;流动相为甲醇-0.1%磷酸水梯度洗脱,流速1 mL·min^-1;柱温30 ℃;检测波长254 nm。测定11批大黄样品的指纹图谱,建立指纹图谱共有模式。以大黄酸为内参物,建立没食子酸、儿茶素、大黄素-8-O-葡萄糖苷、芦荟大黄素、大黄素、大黄酚和大黄素甲醚与内参物的相对校正因子,并进行含量计算,实现一测多评。同时采用外标法测定11批大黄中8个成分的含量,比较计算值与实测值的差异,验证一测多评法的准确性。结果 11批样品指纹图谱标定了共有峰35个,指认了其中8个成分分别为没食子酸、儿茶素、大黄素-8-O-葡萄糖苷、芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚,这8个成分分别在0.062 4~1.56 μg(r=0.999 5)、0.18~4.5 μg(r=0.999 8)、0.028 8~0.72 μg(r=0.999 9)、0.019 8~0.495 μg(r=0.999 9)、0.050 5~1.262 5 μg(r=0.999 9)、0.063 7~1.592 5 μg(r=0.999 9)、0.098~2.45 μg(r=0.999 9)和0.163~4.075 μg(r=0.999 9)内线性关系良好。建立的相对校正因子重现性良好,采用校正因子计算的含量值与实测值之间无显著差异。结论 所建立的方法准确、可行,可用于大黄质量控制。

Combinational Quality Control Method of Rhei Radix et Rhizoma Based on Fingerprint and QAMS

OBJECTIVE To establish a quality control method of Rhei Radix et Rhizome(RRR) by fingerprint and simultaneous determining eight components by quantitative analysis of multi-components by single marker (QAMS). METHODS An HPLC method was set up. Symmetry C₁₈ column (4.6 mm×250 mm,5 μm) was used. Methanol-0.1% phosphoric acid was used as gradient mobile phase. The flow rate was 1.0 mL·min^-1. The column temperature was 30 ℃ and detection wavelength set at 254 nm. Eleven batches of RRR were determined and a common mode of fingerprint maintained at established. A method was developed for QAMS to determine gallic acid, catechin, emodin-8-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion in RRR. Rhein was selected as internal reference; the relative correction factors (RCF) of other seven components to rhein were calculated. The contents of the eight components in ten batches of RRR were determined by both external standard method and QAMS. The QAMS method was evaluated by comparison of its assay results with that of external standard method. RESULTS There were 35 common peaks in the fingerprints of ten batches RRR, eight of them were identified as gallic acid, catechin, emodin-8-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion. These eight components showed good linear relationship in the range of 0.062 4-1.56 μg(r=0.999 5), 0.18-4.5 μg(r=0.999 8), 0.028 8-0.72 μg(r=0.999 9), 0.019 8-0.495 μg(r=0.999 9), 0.050 5-1.262 5 μg(r=0.999 9), 0.063 7-1.592 5 μg(r=0.999 9), 0.098-2.45 μg(r=0.999 9), and 0.163-4.075 μg(r=0.999 9), respectively. The established RCF had good reproducibility. No significant differences were found between the quantitative results of external standard method and QAMS. CONCLUSION The developed method is accurate, feasible, and can be used for the overall quality control of RRR.