UPLC法同时测定大黄中8个成分的含量

目的：建立UPLC法同时测定大黄中大黄素、大黄酸、大黄酚、大黄素甲醚、芦荟大黄素、番泻苷A、番泻苷B、土大黄苷8个成分的含量。 方法：采用ACQUITY BEH C₁₈色谱柱(2.1 mm×50 mm，1.7 μm)，以乙腈(A)-0.1%甲酸水溶液(B)为流动相，梯度洗脱(0~1 min，10%B→15%B；1~2 min，15%B→20%B；2~3 min，20%B→30%B；3~4 min，30%B→40%B；4~5 min，40%B→60%B；5~6 min，60%B→65%B；6~7 min，65%B→70%B；7~8 min，70%B→80%B；8~9 min，80%B→85%B)，流速0.3 mL·min^-1，柱温35 ℃，254 nm处检测大黄素、大黄酸、大黄酚、大黄素甲醚、芦荟大黄素，320 nm处检测番泻苷A、B和土大黄苷，对测定结果进行主成分分析。 结果：大黄素、大黄酸、大黄酚、大黄素甲醚、芦荟大黄素、番泻苷A、番泻苷B、土大黄苷浓度在一定范围内，与峰面积积分值线性关系良好(r≥0.9996)，方法的精密度RSD为0.51%~0.97%，重复性RSD为0.64%~1.3%，稳定性RSD为0.72%~1.6%，平均加样回收率为96.5%~104.2%。主成分分析结果表明正品大黄与伪品可以明显分开，野生品与栽培品内在质量存在差异。 结论：所建立的方法能同时测定大黄中8个成分的含量，可作为大黄药材与伪品的区分和质量控制的参考方法。

Objective: To develop a ultra-performance liquid chromatography(UPLC)method to determine the content of emodin,rhein,chrysophanol,physcion,aloe-emodin,sennoside A,sennoside B and rhaponticin simultaneously in Radix et Rhizoma Rhei. Methods: The analysis was achieved with an ACQUITY BEH C₁₈ analytical column(2.1 mm×50 mm,1.7 μm)by gradient elution of 0.1%(v/v)formic acid in water and acetonitrile:0-1 min,10%B→15% B;1-2 min,15%B→20%B;2-3 min,20%B→30%B;3-4 min,30%B→40%B;4-5 min,40%B→60%B;5-6 min,60%B→65%B;6-7 min;65%B→70%B;7-8 min,70%B→80%B;8-9 min,80%B→85%B.The flow rate was 0.3 mL·min^-1,the column temperature was maintained at 35 ℃ and the detection wavelength was set at 254 nm and 320 nm.Five free anthraquinones were determined at 254 nm,and sennoside A/B and rhaponticin were determined at 320 nm.Principal component analysis(PCA)was used to analyze the test results. Results: The peak areas and concentrations of emodin,rhein,chrysophanol,physcion,aloe-emodin,sennoside A,sennoside B and rhaponticin showed good linear relationship within a certain concentration range(r≥0.9996);the RSD of precision was 0.51%-0.97%;the RSD of repeatability was 0.64%-1.3%,the RSD of stability was 0.72%-1.6% and the average recoveries were 96.5%-104.2%.Quality and counterfeit of Radix et Rhizoma Rhei could be clearly separated by PCA and there were differences in intrinsic quality between wild variety and cultivation. Conclusion: UPLC method for simultaneous determination of 8 contents of Radix et Rhizoma Rhei could be used to distinguish between qualified products and counterfeits of Radix et Rhizoma Rhei and used as the reference method for quality control.