基于分析方法质量源于设计理念优化护肝片HPLC指纹图谱的研究

Optimization of the Fingerprint of Hugan Tablets Based on Analytical Quality by Design

目的:应用方法质量源于设计(AQb D)理念,建立护肝片HPLC指纹图谱方法并验证。方法:以峰个数(N)及五味子醇甲含量(SCH)、五味子酯甲量(StA)、五味子甲素量(SchA)和五味子乙素量(SchB)量为评价指标,采用Plackett-Burman设计在风险评估的基础上筛选出关键质量属性(CMAs)及关键质量参数(CMPs),然后对筛选出的显著项采用Box-Behnken设计得到CMAs及CMPs之间的响应模型。通过多变量回归分析得到最佳优化条件,并对最佳优化条件进行验证。结果:在风险评估的6个因子中筛选出了3个CMPs,分别为提取溶剂(A)、料液比(B)及色谱柱柱温(C),而筛选出有统计学意义的CMAs为N、五味子醇甲含量(SCH)及五味子甲素量(SchA)。通过响应面分析各因素与模型的变化趋势,得出其最佳优化条件为提取溶剂为79%甲醇,料液比为23 ml·g^(-1),柱温为32.2℃。在此条件下,验证试验测得N、SCH与SchA分别分别为20、0.628 0mg/片、0.133 0 mg/片,与模型预测值20.4、0.632 0 mg/片、0.138 0 mg/片接近。结论:所建立的方法可靠,为AQb D在中药分析领域的应用提供参考。

Objective: To establish and validate the identification of Hugan tablets by HPLC using the concept of analytical quality by design( AQb D). Methods: The number of peaks and the contents of schisandrin( SCH),schisantherin( St A),deoxyschizandrin( Sch A) and schisandrin B( Sch B) were taken as the analytical target profile( ATP),Plackett-Burman design( PBD) method was used to select critical method parameters( CMPs) and critical method attributes( CMAs) on the basis of risk assessment,and then the response model between CMAs and CMPs was established by Box-Behnken design( BBD). The optimal conditions were obtained via multivariable regression analysis and verified as well. Results: From the six factors of risk assessment,the extraction solvent( A),the column temperature( B) and the liquid to liquid ratio( C) were selected as CMPs. And CMAs were the number of peaks( N),and the contents of schisandrin( SCH) and deoxyschizandrin( SchA). The variation trends of each factor and model were analyzed by response surface analysis. The optimal conditions were as follows: the extraction solvent was 79% methanol,the column temperature was 32. 2℃ and the liquid to liquid ratio was 23 ml·g^-1. Under the optimal conditions,N,SCH and Sch A respectively was 20,0. 628 0 mg/tablet and 0. 133 mg/tablet in the verification test,which was close to the predicted value of 20. 4,0. 632 0 mg/tablet and 0. 138 0 mg/tablet,respectively. Conclusion: The results proved that the method is reliable,which provide reference for the application of AQb D in the analysis of traditional Chinese medicine.