Chen Meng-Lu, Chang Wen-Qi, Zhou Jian-Liang, Yin Ying-Hao, Xia Wen-Rui, Liu Jian-Qun, Liu Li-Fang, Xin Gui-Zhong
State Key Laboratory of Natural Medicines, Department of Chinese Medicines Analysis, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing, China.
Zhejiang Institute for Food and Drug Control, Hangzhou 310052, China.
J Pharm Biomed Anal. 2017 Oct 25;145:666-674. doi: 10.1016/j.jpba.2017.07.054. Epub 2017 Aug 1.
Traditional Chinese medicine (TCM) materials with closely related species are frequently fungible in clinical use. Therefore, holistic comparison of the composition in bioactive compounds is essential to evaluate whether they are equivalent in efficacy. Taking three officinal species of Callicarpa as a case, we proposed and validated a standardized strategy for the discrimination of closely related TCM materials, which focused on the extraction, profiling and multivariate statistical analysis of their biochemome. Firstly, serial liquid-liquid extractions were utilized to prepare different batches of Callicarpa biochemome, and the preparation yields were utilized for the normalization of sampling quantity prior to UHPLC-IT-MS analysis. Secondly, 34 compounds, including 19 phenylethanoid glycosides, 10 flavonoids and 5 terpenoids, were identified based on an untargeted UHPLC-IT-MS method. Thirdly, method validation of linearity, precision and stability showed that the UHPLC-IT-MS system was qualified (R>0.995, RSD<15%) for subsequent biochemome profiling. After PCA and PLS-DA analysis, 30 marker compounds were screened and demonstrated to be of good predictability using genetic algorithm optimized support vector machines. Finally, a heatmap visualization was employed for clarifying the distribution of marker compounds, which could be helpful to determine whether the three Callicarpa species are, in fact, equivalent substitutes. This study provides a standardized biochemome profiling strategy for systemic comparison analysis of closely related TCM materials, which shows promising perspectives in tracking the supply chain of pharmaceutical suppliers.
临床应用中，来源相近的中药材常可相互替代。因此，对生物活性成分进行整体比较，对于评估其药效是否等同至关重要。以三种紫珠属药用植物为例，我们提出并验证了一种用于鉴别来源相近中药材的标准化策略，该策略聚焦于对其生物化学组进行提取、分析和多变量统计分析。首先，采用连续液液萃取法制备不同批次的紫珠生物化学组，并在超高效液相色谱-离子阱质谱（UHPLC-IT-MS）分析前，利用制备产率对取样量进行归一化处理。其次，基于非靶向UHPLC-IT-MS方法鉴定出34种化合物，包括19种苯乙醇苷、10种黄酮类化合物和5种萜类化合物。第三，线性、精密度和稳定性的方法验证表明，UHPLC-IT-MS系统符合后续生物化学组分析的要求（R>0.995，RSD<15%）。经过主成分分析（PCA）和偏最小二乘判别分析（PLS-DA），筛选出30种标记化合物，并通过遗传算法优化的支持向量机证明其具有良好的预测能力。最后，采用热图可视化方法来阐明标记化合物的分布情况，这有助于确定这三种紫珠属植物是否实际上为等效替代品。本研究为来源相近的中药材的系统比较分析提供了一种标准化的生物化学组分析策略，在追踪药品供应商供应链方面显示出广阔的前景。
