首乌苷中蒽醌类成分含量测定方法比较研究

Comparative Study on Determination Method of Anthraquinones Content in SWG

目的:研究用两种方法测定首乌苷(SWG)中蒽醌的含量,并比较两者之间差异,以评价用HPLC法和UV-VIS法测定SWG中蒽醌含量的合理性。方法:采用HPLC法,测定大黄素、大黄素甲醚的含量,色谱柱为ZORBAX Eclipse XDB-C18(4.6 mm×150 mm,5μm);流动相为甲醇-水-磷酸(80∶20∶0.1);检测波长为254nm。采用UV-VIS法测定蒽醌含量,用2.5 mol/L硫酸水解前后经氯仿萃取,以0.5%醋酸镁甲醇溶液显色,用紫外-可见风光光度法于509 nm处测定吸光度。结果:用HPLC法测定首乌苷水解前大黄素的含量为0.0050%,大黄素甲醚的含量为0.002 4%;水解后大黄素的含量为0.062 6%,大黄素甲醚的含量为0.003 0%,以大黄素和大黄素甲醚之和含量计算游离蒽醌为0.007 4%,总蒽醌为0.065 6%。用UV-VIS法测定首乌苷药液中游离蒽醌的含量为0.008 2%,总蒽醌含量为0.0958%,结合蒽醌含量为0.087 6%。结论:测定总蒽醌可以用UV-VIS法评价,但能结合HPLC法将更合理;SWG中蒽醌类成分含量极少。

Objective： To study the content of anthraquinones in SWG determined by two methods and the difference between the two and judge the reasonable determination of anthraquinones content with HPLC and UV- VIS method.Methods： Using HPLC method to determine the content of emodin and physcion,chromatographic column was ZORBAX Eclipse XDB- C18（ 4. 6 mm × 150 mm,5 μm）. The mobile phase was methanol- water- phosphoric acid（ 80 ∶ 20 ∶0. 1）. The detection wavelength was 254 nm. Using UV- VIS method,we determined the content of total anthraquinones with 2. 5 mol / L sulfuric acid hydrolyzed and extracted with chloroform. 0. 5% magnesium acetate methanol solution was used for coloration and UV- VIS spectrophotometric absorbance was measured in 509 nm. Results： The content of emodin was 0. 005 0% and physcione was 0. 002 4% before hydrolyzing by HPLC method in SWG. The content of emodin was0. 062 6 % and physcione was 0. 003 0% after hydrolyzing. The content of free anthraquinone was 0. 007 4% and the total anthraquinone was 0. 065 6% with emodin and physcion sum. The content of free anthraquinone was 0. 008 2% and the total anthraquinone was 0. 095 8% and the combined anthraquinone was 0. 087 6% by UV- VIS method in SWG.Conclusions： The UV- VIS method can be used to determine the total anthraquinone and combined with HPLC method will be more reasonable. The contents of anthraquinones were little in SWG.