代谢组通过定性和定量分析给定生物体或细胞中所有低分子量代谢物的定性分析来监测活细胞中的化学变化。
样品准备和预处理
样品制备和预处理需要考虑到各种因素,例如时间,区域,现场和药物收集的发育阶段,以减少样品变异对代谢组学分析结果的影响。在采集样品后,需要对药用植物的生物灭活,例如在液氮中进行快速冷冻,以减少氧化还原反应和通过各种水解酶测试的样品的降解。
制备样品的过程基于研究对象的性质和选择样品预处理的研究方法。例如,将不同极性的溶剂应用于分离脂质可溶成分的药用植物的水溶性成分,这些植物的脂质可溶成分较小,这些成分的极性较小。此外,大多数初级二级代谢产物都不挥发,因此在样品提取后,样品的衍生化,例如酯化,酰化,电离,烷基化,等等, is required to obtain more samples available for analysis.
分离和鉴定代谢物
这separation techniques of metabolites usually include gas chromatogram (GC), liquid chromatogram (LC) and capillary electrophoresis (CE). The detection and identification techniques include IR spectrum, UV spectrum, nuclear magnetic resonance (NMR) and mass spectrometry (MS).
Data conversion and analysis
After the metabolites are separated and identified, the raw chromatographic data are obtained. These data are voluminous and need to be converted into digital matrix data in order to clarify the statistical relationships between compounds and their biological significance. The methods of converting these numbers include eliminating noise interference, correcting the baseline, and segmenting the data to exclude the interference of environmental factors during the measurement.
After processing the raw data, the data needs to be analyzed. Commonly used data analysis can be divided into two types of identification models, supervised classification and unsupervised classification. The difference between the two models is that unsupervised classification uses known mathematical models to predict samples, while supervised classification classifies and filtersmetabolite assays来自原始测定概况。监督分类包括主要组件分析(PCA),分层聚类分析(HCA)和自组织图(SOM)。无监督的分类主要包括部分最小二乘 - 歧视分析(PLS-DA)和正交部分最小二乘 - 歧视分析(OPLS-DA)。
Figure 1. Various phytochemical techniques used in the field of medicinal plant research (Mukherjeeet al。,2016)。
Fingerprinting technology can effectively provide comprehensive quality control of medicinal plants and ensure safe and stable effects of their use.
变化药用植物的代谢物在不同的治疗条件下,例如外部物理,化学或生物刺激。将比较刺激前后的药用植物代谢产物的差异,并全面分析,以寻找和确定植物代谢的关键步骤和定律奠定基础。
研究药用植物的基因功能。代谢物的分析揭示了相关基因的表达水平的变化,并且可以推断基因对代谢流的影响。
协助培养和收获药用植物。这药用植物的化学成分根据生长期和环境的变化以及在不同时期和环境中代谢物的积累可以有效地确保药草的质量和产量,并为合理有效利用药用药草提供科学基础。
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