[关键词]
[摘要]
目的 基于分析方法质量源于设计(AQbD)对复方丹参滴丸(CDDP)皂苷类成分指纹图谱的UPLC-UV方法开发进行初步研究。方法 以峰个数(N)及各指标成分(三七皂苷R1、人参皂苷Rg1、人参皂苷Re和人参皂苷Rb1)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7个因子中筛选出了3个CMPs,分别为流动相体积流量(A)、称样量(B)及C18预柱质量(C),而筛选出统计学显著的CMAs为N、三七皂苷R1量(CR1)及人参皂苷Re量(CRe)。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流动相体积流量为0.28 mL/min,称样量为0.30 g,C18预柱质量为1.10 g。在此条件下,N为12个,CR1为1.676 5mg/g,CRe为0.669 6mg/g。与模型预测值相近,相对误差(RE)分别为1.34%、1.33%、2.94%。最终确立的定量方法为采用Acquity UPLC BEH C18色谱柱(50 mm×2.1 mm,1.7 μm),流动相为乙腈-水梯度洗脱,检测波长203 nm,柱温为30℃,体积流量为0.28 mL/min,进样量为2 μL。结论 所建立的方法可靠,为AQbD在中药分析领域的应用提供参考。
[Key word]
[Abstract]
Objective To study the development of analytical quality by design (AQbD) method for the identification of Compound Danshen Dripping Pills (CDDP) by UPLC-UV. Methods The number of peaks and the target peak content (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1) 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. Results From the seven factors of risk assessment, the flow rate (A), sample weight (B), and C18 column weight (C) were selected as CMPs. And CMAs were the number of peaks (N), notoginsenoside R1 (CR1) and ginsenoside Re (CRe). The variation trends of each factor and model were analyzed by response surface analysis. The optimal conditions were as follows:flow rate of 0.28 mL/min, sample weight of 0.30 g, and C18 column weight of 1.10 g. Under the conditions, N was 12, CR1 was 1.676 5mg/g, and CRe was 0.669 6mg/g, with differences of 1.34%, 1.33%, and 2.94% respectively from the predictive values. The final method was as follows:chromatographic column was Acquity UPLC BEH C18 chromatographic column (50 mm×2.1 mm, 1.7 μm), mobile phase was acetonitrile-water gradient elution, detection wavelength was 203 nm, column temperature was 30℃, flow rate was 0.28 mL/min, and injection volume was 2μL. Conclusion The results prove that the method is reliable.
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[基金项目]
国家中药标准化项目(ZYBZH-C-TJ-55)