HPLC-FLD法测定清火片中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚

Determination of aloe-emodin, rhein, emodin, chrysophanol, and physcione in Qinghuo Tablets by HPLC-FLD

目的建立高效液相色谱–荧光检测(HPLC-FLD)法测定清火片中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚。方法采用Agilent C18色谱柱(250 mm34.6 mm,5μm);流动相:甲醇–0.1%磷酸溶液(78∶22);体积流量为1 mL/min;柱温30℃;进样量为20μL。对比HPLC-FLD法与HPLC-UV法的测定结果。结果芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚分别在100.6～1 609.6、86.8～1 388.8、359.4～5 750.4、81.2～1 299.2、89.4～1 430.4 ng线性关系良好;游离蒽醌中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的平均回收率分别为93.9%、90.8%、96.2%、92.9%、89.2%,RSD值分别为2.2%、2.4%、1.0%、2.4%、0.6%;总蒽醌中各成分平均回收率分别为100.2%、85.7%、99.2%、104.5%、104.2%,RSD值分别为2.5%、2.2%、3.7%、0.5%、0.7%。结论本方法具有重复性好、灵敏度高、专属性强、内源物干扰小等优点,与HPLC-UV法测定结果一致性较高,可用于中成药清火片的质量控制。

Objective To establish an method HPLC-FLD for the determination of aloe-emodin, rhein, emodin, chrysophanol, and physcione in Qinghuo Tablets. Methods The separation was carried out on Agilent C18 chromatography column(250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-0.1% phosphoric acid solution(78:22). The flow rate was 1 mL/min, temperature of column was set at 30 ℃, and volume of injection was 20 μ L. The results of HPLC-FLD and HPLC-UV were compared. Results The linear ranges of aloe-emodin, rhein, emodin, chrysophanol, and physcione were 100.6 - 1 609.6, 86.8 - 1 388.8, 359.4 -5 750.4, 81.2 - 1 299.2, and 89.4 - 1 430.4 ng, respectively. The average recovery rates of aloe-emodin, rhein, emodin, chrysophanol, and physcione in free anthraquinone were 93.9%, 90.8%, 96.2%, 92.9%, and 89.2%, respectively, with RSD values of 2.2%, 2.4%, 1.0%, 2.4%, and 0.6%, respectively. The average recovery rates of total anthraquinone were 100.2%, 85.7%, 99.2%, 104.5%, and 104.2%, respectively with RSD values of 2.5%, 2.2%, 3.7%, 0.5%, and 0.7%, respectively. Conclusion The method has the advantages of good repeatability, high sensitivity, strong specificity, and little interference from endogenous substances, in good agreement with HPLC-UV method, and can be used for quality control of Qinghuo Tablets.