基于质量源于设计理念的三叶糖脂清汤剂定量指纹图谱研究

Quantitative chromatographic fingerprint analysis of Sanye Tangzhiqing Decoction based on quality by design concept

采用分析质量源于设计(AQb D)过程,以高效液相色谱-紫外检测技术建立了三叶糖脂清汤剂的定量指纹图谱分析方法。首先确定需要优化的方法参数为柱温、流速和流动相梯度条件,再通过确定性筛选设计明确关键方法评价指标(CMAs)为峰个数、共有峰面积占总峰面积的百分比和最后一个峰的保留时间,然后采用逐步回归法建立CMAs与方法参数的定量模型。以模拟实验测定误差法计算基于达标概率的设计空间并成功验证。优化分析条件后,定量测定绿原酸、芍药苷、芦丁、金丝桃苷、槲皮素-3-O-β-D-葡萄糖醛酸苷和丹酚酸B 6种成分。指纹图谱中含共有峰19个,共有峰面积占总峰面积比例为96%。指纹图谱和定量检测分析方法均通过方法学考察,并用于新样品测定。

In this work,a high performance liquid chromatography-ultraviolet(HPLC-UV)detection technology was used to establish fingerprint analysis method for Sanye Tangzhiqing Decoction following an analytical quality by design(AQb D)approach.Firstly,column temperature,flow rate,and gradient elution conditions were determined as the method parameters needing to be optimized.Then according to the results of definitive screening design,three critical method attributes(CMAs)were identified,including peak number,the percentage of common peak area to total peak area,and retention time of the last peak.A stepwise regression method was used then to build quantitative models between CMAs and method parameters.Probability-based design space was calculated and successfully verified using the experimental error simulation method.After the analysis conditions were optimized,the contents of six components,namely chlorogenic acid,paeoniflorin,rutin,hyperoside,quercetin-3-O-β-D-glucuronide,and salvianolic acid B were simultaneously determined.There were 19 common peaks in the fingerprint and their common peak area accounted for 96%of the total peak area.Both fingerprint and quantitative analysis methods were validated applicable in methodology study,and they can be applied to determine new samples.