MAE-RPHPLC-FLD法测定丁公藤及其制剂中总东莨菪内酯含量

目的: 建立测定丁公藤及其制剂中总东莨菪内酯含量的微波辅助萃取(MAE)-RPHPLC-FLD法.方法: 样品经MAE优化液液萃取法的提取、水解、净化前处理过程,再以Welch Ultimate XB-C₁₈色谱柱(4.6 mm×150 mm,5 μm)分离,1.0 mL·min^-1流速的流动相[甲醇-BR缓冲液(pH 9.15)(1:100)]洗脱,FLD检测器(λ_发射=462 nm,λ_激发=388 nm;狭缝:10 nm)检出,用峰面积外标法定量.结果: 标准曲线线性范围为0~1000 ng·mL^-1(r=1.0000);方法检出限为0.1 ng·mL^-1,方法定量限为0.3 ng·mL^-1;样品回收率为99%~101%;中间精密度、重复性试验:RSD均≤1%;稳定性试验:RSD≤0.1%.结论: 本法前处理简捷、高效,检出限、定量限、准确度、精密度试验结果均满足要求,可用于测定丁公藤及其制剂中总东莨菪内酯含量.

Objective: To establish an MAE-RPHPLC-FLD method for measurement of scopoletin content in Dinggongteng and its preparations. Methods: The samples were pre-processed by the liquid-liquid extraction method which was optimized with microwave-assisted extraction (MAE)technology for extraction, hydrolysis, and purification, then the Welch Ultimate XB-C₁₈ column(4.6 mm×150 mm, 5 μm)was used for sample separation, and the samples were eluted with the mobile phase at 1.0 mL·min^-1 flow rate [methanol-BR buffer(pH 9.15)(1:100)], the FLD detector(λ_emission=462 nm, λ_excitation=388 nm;slit:10 nm)was used for detection, and finally the samples were quantitated by peak area with the external standard method. Results: Linear range of the standard curve was 0-1000 ng·mL^-1 (r=1.0000);the method detection limit was 0.1 ng·mL^-1, the method quantitation limit was 0.3 ng·mL^-1;the sample recovery was 99%-101%.RSDs for intermediate precision and repeatability were ≤1%, for stability test were ≤0.1%. Conclusion: This method is simple and efficient, the results of detection limit, quantification limit, accuracy, and precision test meet the requirements for the determination of total scopoletin content in Dinggongteng and its preparations.

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