Chinese Medicine Laboratory, Hong Kong Jockey Club Institute of Chinese Medicine, Shatin, N. T., Hong Kong, China.
J Pharm Biomed Anal. 2010 Aug 1;52(4):468-78. doi: 10.1016/j.jpba.2010.01.032. Epub 2010 Jan 22.
In the present study, an ultra performance liquid chromatography coupled with photo-diode array detector and time-of-flight mass spectrometry (UPLC-PDA-TOFMS) based chemical profiling approach to rapidly evaluate chemical consistency between traditional and dispensing granule decoctions of traditional medicine combinatorial formulae was proposed and validated using San-Huang-Xie-Xin-Tang (SHXXT) as a model combinatorial formula. Two different kinds of decoctions, namely traditional decoction: water extract of mixed three constituent herbs of SHXXT, and dispensing granules decoction: mixed water extract of each individual herbs of SHXXT, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-PDA-TOFMS analysis, the datasets of t(R)-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, further statistics was performed to generate S-plot, in which the variables (t(R)-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during decocting of combinatorial formula. The identities of the changed components can be identified by comparing the mass/UV spectra and retention times with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between traditional and dispensing granule decoctions, and berberine, palmatine, epiberberine, baicalin, wonogoside, 2-O-golloyl-1-O-cinnamoylglucose and emodin were identified as the most changed components during decocting SHXXT. It is suggested that this newly established approach could be used practically for the evaluation of chemical consistency between traditional and dispensing granule decoctions of traditional medicine combinatorial formulae.
在本研究中，提出了一种基于超高效液相色谱-光电二极管阵列检测-飞行时间质谱联用（UPLC-PDA-TOFMS）的化学特征分析方法，用于快速评估传统医学配方的传统汤剂和配方颗粒汤剂之间的化学成分一致性，以三黄泻心汤（SHXXT）为模型配方进行了验证。制备了两种不同的汤剂，即传统汤剂：SHXXT 的三种混合草药的水提取物，以及配方颗粒汤剂：SHXXT 的每种单独草药的混合水提取物。对这两种汤剂样品进行了 UPLC-PDA-TOFMS 分析，处理了 t(R)-m/z 对、离子强度和样品代码的数据集，采用监督正交偏最小二乘判别分析（OPLS-DA）对这两种汤剂样品的差异进行整体比较。一旦在得分图中发现了明显的分类趋势，就会进行进一步的统计，生成 S 图，其中对差异贡献最大的变量（t(R)-m/z 对）清晰地描绘为“S”两端的点，与这些离子相关的成分被认为是配方煎煮过程中变化最大的成分。通过比较质量/紫外光谱和保留时间与参考化合物的光谱和保留时间，并通过将经验分子式与文献中已知化合物的分子式进行匹配，可以鉴定出变化成分的身份。使用所提出的方法，发现传统汤剂和配方颗粒汤剂之间存在全局化学差异，并且鉴定出小檗碱、巴马汀、表小檗碱、黄芩苷、吴茱萸苷、2-O-没食子酰基-1-O-肉桂酰基葡萄糖和大黄素是煎煮 SHXXT 过程中变化最大的成分。建议将这种新建立的方法实际用于评估传统医学配方的传统汤剂和配方颗粒汤剂之间的化学成分一致性。
