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[摘要]
目的 建立基于超高效液相色谱(UPLC)法的大黄中10个蒽醌衍生物同时定量分析方法。方法 采用Waters Acquity超高效液相色谱仪,BEH C18色谱柱(100 mm×2.1 mm,1.7μm),柱温为34℃;甲醇和0.1%磷酸水溶液的混合溶液为流动相,梯度洗脱;体积流量为0.20 mL/min;进样量为2.0 μL;检测波长为410 nm。通过考察柱温和体积流量对保留时间、分离度、理论塔板数和对称因子4个重要的色谱参数的影响,以此获得5个蒽醌糖苷的最佳的色谱分离度和稳健性。结果 优化得到柱温34℃和体积流量0.20 mL/min为最佳分离条件,根据最优参数,建立一种新的基于UPLC的大黄中10个蒽醌衍生物同时定量分析方法。20批次大黄测定结果显示,2种不同基原的大黄药材中芦荟大黄素-8-O-β-D-葡萄糖苷、大黄酸-8-O-β-D-葡萄糖苷的含量差异最大,分别可超过60、77倍,表明2种不同基原的大黄存在较大的质量差异。结论 基于UPLC最佳分离参数建立的10个蒽醌衍生物同时定量分析方法具有较好的稳健性,对更加全面地从化学表征的角度评控不同来源的大黄药材的质量具有参考价值。
[Key word]
[Abstract]
Objective To establish a simultaneous quantitative analytical method of 10 anthraquinone derivatives in Rheum officinale based on ultra-high performance liquid chromatography (UPLC) for the quality evaluation and control. Methods The UPLC method was performed on a BEH C18column (100 mm×2.1 mm, 1.7 μm) with a binary gradient mobile phase consisting of a mixture of methanol and 0.1% aqueous phosphoric acid with column temperature at 34℃. Using a gradient elution method; The flow rate was set at 0.20 mL/min; The injection volume was set at 2.0 μL; The detection wavelength was set at 410 nm. This study focused on the influence of analytical parameters on the separation efficiency of five anthraquinone glucosides in R. officinale. To obtain robust chromatographic resolution of the five anthraquinone glucosides, relatively important chromatographic parameters including the retention time, resolution, theoretical plate number, and symmetry factor were investigated by altering the column temperature and flow rate. Results The optimal column temperature was found to be 34℃, and the optimal flow rate was 0.20 mL/min. A new UPLC analytical method was developed for simultaneous quantification of 10 anthraquinone derivatives in R. officinale based on these optimal parameters. This UPLC-based analytical method was successfully applied to quantitative determination of 10 anthraquinone derivatives in Rheum tanguticum and Rheum palmatum. The analysis showed that there were significant content differences between aloe-emodin-8-O-β-D-glucoside (more than 60-fold) and rhein-8-O-β-D-glucoside (more than 77-fold) in the two species of rhubarbs tested. Conclusion The UPLC method established for the simultaneous determination of 10 anthraquinone derivatives in R. officinale based on the optimal parameters is accurate and sensitive, which will provide a more comprehensive evaluation for the chemical characterization of medicinal materials of R. officinale from different sources.
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[基金项目]
国家自然科学基金项目资助(81403126);国家科技基础性工作专项(2015FY111500-080);成都市科技局项目(2016-HM01-00312-SF);成都中医药大学重点实验室开放基金(2016302))