基于质量源于设计理念的制川乌UHPLC分析方法研究

Research of the UHPLC Analysis Method for Aconiti Radix Cocta Based on the Concept of Quality by Design

目的 基于质量源于设计理念(quality by design,QbD)建立制川乌的超高效液相色谱(ultra high performance liquid chromatography,UHPLC)分析方法。方法 以制川乌中6个主要成分的定量分析为目标,选取柱温、醋酸铵溶液浓度、流速、pH值、初始乙腈浓度为考察因素进行Plackett-Burman试验设计,根据试验结果确定关键方法参数进行Box-Behnken实验,建立设计空间并选择置信度为95%的置信区间估计法考察设计空间的可靠性,最后选择准确性轮廓方法(accuracy profile,AP)进行方法学验证。以分离度为目标所建最佳模型为二次项式模型,以分析时间为目标所建最佳模型为线性模型。基于以上两个模型建立分析方法的设计空间,并采用置信区间估计对方法的可靠性进行评价。结果 结果显示,从设计空间内选择的30 mmol·L^(-1)的醋酸铵浓度(pH=10)和体积分数28%的初始乙腈比例,在10 min内可完成制川乌中6生物碱的完全分离,该分析方法的准确性、中间精密度、重复性、真实性都在可接受的范围内。结论 优化后的方法可以用于制川乌的UHPLC分析,QbD理念为开发并优化制川乌中6个生物碱的UHPLC分析方法提供了新的思路。

OBJECTIVE To establish a UHPLC analysis method for Aconiti Radix Cocta based on the concept of quality by design(QbD).METHODS Targeting at the quantitative analysis of the six main components of Aconiti Radix Cocta,column temperature,ammonium acetate concentration,flow rate,pH,and initial acetonitrile concentration were selected as investigating factors to carry out the Plackett-Burman design.According to the test results,the key method parameters were determined to carry out the Box-Behnkendesign,the design space was established and the confidence interval estimation method with a confidence interval of 95% was selected to investigate the reliability of the design space.Finally,the accuracy profile(AP) method was selected for methodological verification.The best model with separation as the goal was the quadratic model.with analysis time as the goal,the best model was a linear model.RESULTS At the same time,the design space established based on the two models was reliable.With ammonium acetate concentration of 30 mmol·L^(-1)(pH=10) and initial acetonitrile concentration of 28%,the analytical method could complete elution within 10 min and the accuracy,intermediate precision,repeatability,and authenticity were all acceptable.CONCLUSION The optimized UHPLC method can be used for the analysis of Aconiti Radix Cocta.,so the QbD concept can be used to develop and optimize the UHPLC analysis method of six components in Aconiti Radix Cocta.