复杂天然产物特征组分HPLC筛选方法的研究
摘要
天然产物成分复杂,不同的来源、不同部位、不同的处理方法得到的产物中有效药用组成、含量等皆可能有相当大的差别。天然产物复杂体系研究中的重点为其特征组分的研究。发展系统化的高效液相色谱特征组分筛选方法,对于寻找特征组分、质量控制等具有重要意义。
水飞蓟作为一种重要的中药,具有诸多药理特征,已知水飞蓟素是其最重要的有效组分之一。本文以水飞蓟作为实验样品,系统研究其幼草、种子、花、叶、根、茎等不同部位,采用不同极性的溶剂热回流、超声提取等方法得到的组分的结构特征。说明了不同部位提取物中化学成分的差异,建立了完备的水飞蓟HPLC分析方法。并进一步对水飞蓟幼草和全草的水提物进行抗肿瘤研究,实验发现除种子外,水飞蓟幼草和全草的水提物,对HepG2,MNK45,MCF-7,A431四种不同种类的癌细胞都有抗肿瘤的抑制作用,可以为水飞蓟的全草的植物利用度提供一定参考。
茶叶是中国传统饮品。以茶叶提取物制备的茶饮料为样品,将构建的方法运用于茶饮料中特征组分茶氨酸的快速分析。研究不同处理方法得到的绿茶、红茶、铁观音、乌龙茶、普洱茶等不同品种茶饮料中的茶氨酸水平。通过优化样品预处理方法,结合UPLC技术,发展了相应的快速分析方法,与普通HPLC分析方法时间相比节约了分析时间,且具有灵敏度高,干扰少等特点。所发展的HPLC方法普适、稳健,为复杂天然产物的特征组分的高效液相色谱法筛选提供了系统策略。
The compositons of the traditional Chinese medicines are complex. Moreover, the obvious difference of effect and concentration cause by the different parts, source, parts and processing methods. Identifications of feature compositions of traditional Chinese medicines are crucial. Development of the systematic HPLC method for the analysis of feature compositon and quality control is of great importance.
Milk thistle is an important herb medicine and the silibin is the key acitive component in it. In this study, HPLC method was developed for the analysis of milk thistle. Different parts of the silymarin, including the tender grass, seeds, flowers, leaves, roots, and stems, were extracted by different solvents followed by HPLC analysis. The anti-cancer bioacitivity of the extracts were investigated based on the four cancer cell vitro assay, including theHepG2, MNK45, MCF-7 and A431. The results showed that the extracts of tender grass and the whole plant were able to inhibit the four cancer cell.
The ready-to-drink teas made from tea extracts are popular. Theanine, an amino acid soly found in tea, can be used as a mark to determine the quality of ready-to-drink teas. In this report, a fast method for quantification of the theanine in the ready-to-drink teas was developed using UPLC. Ready-to-drink teas from market included green tea, black tea, Tie Guan Yin, Oolong tea, and Pu'er tea. By optimizing the sample preparation methods combined with UPLC the analysis time was just 6 minutes, much faster than the normal HPLC method. It was proved that the UPLC and SPE method for the analysis of theanine in ready-to-drink teas was reliable by validation.
水飞蓟作为一种重要的中药,具有诸多药理特征,已知水飞蓟素是其最重要的有效组分之一。本文以水飞蓟作为实验样品,系统研究其幼草、种子、花、叶、根、茎等不同部位,采用不同极性的溶剂热回流、超声提取等方法得到的组分的结构特征。说明了不同部位提取物中化学成分的差异,建立了完备的水飞蓟HPLC分析方法。并进一步对水飞蓟幼草和全草的水提物进行抗肿瘤研究,实验发现除种子外,水飞蓟幼草和全草的水提物,对HepG2,MNK45,MCF-7,A431四种不同种类的癌细胞都有抗肿瘤的抑制作用,可以为水飞蓟的全草的植物利用度提供一定参考。
茶叶是中国传统饮品。以茶叶提取物制备的茶饮料为样品,将构建的方法运用于茶饮料中特征组分茶氨酸的快速分析。研究不同处理方法得到的绿茶、红茶、铁观音、乌龙茶、普洱茶等不同品种茶饮料中的茶氨酸水平。通过优化样品预处理方法,结合UPLC技术,发展了相应的快速分析方法,与普通HPLC分析方法时间相比节约了分析时间,且具有灵敏度高,干扰少等特点。所发展的HPLC方法普适、稳健,为复杂天然产物的特征组分的高效液相色谱法筛选提供了系统策略。
The compositons of the traditional Chinese medicines are complex. Moreover, the obvious difference of effect and concentration cause by the different parts, source, parts and processing methods. Identifications of feature compositions of traditional Chinese medicines are crucial. Development of the systematic HPLC method for the analysis of feature compositon and quality control is of great importance.
Milk thistle is an important herb medicine and the silibin is the key acitive component in it. In this study, HPLC method was developed for the analysis of milk thistle. Different parts of the silymarin, including the tender grass, seeds, flowers, leaves, roots, and stems, were extracted by different solvents followed by HPLC analysis. The anti-cancer bioacitivity of the extracts were investigated based on the four cancer cell vitro assay, including theHepG2, MNK45, MCF-7 and A431. The results showed that the extracts of tender grass and the whole plant were able to inhibit the four cancer cell.
The ready-to-drink teas made from tea extracts are popular. Theanine, an amino acid soly found in tea, can be used as a mark to determine the quality of ready-to-drink teas. In this report, a fast method for quantification of the theanine in the ready-to-drink teas was developed using UPLC. Ready-to-drink teas from market included green tea, black tea, Tie Guan Yin, Oolong tea, and Pu'er tea. By optimizing the sample preparation methods combined with UPLC the analysis time was just 6 minutes, much faster than the normal HPLC method. It was proved that the UPLC and SPE method for the analysis of theanine in ready-to-drink teas was reliable by validation.
引文
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[3]王翠艳,赵卫红,白轩,赵亚明.水飞蓟宾提取方法的研究.鞍山师范学院院报.2000：12(4)：91-92
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[5]Ewari, D. S. Jayas, R. A. Holley. Fresher under pressure scientific research high pressure processing of foods:an overview. Science des aliments.1999,19:619-621
[6]吴怀祥.高压食品加工.食品科学.1996,17(11)：3-9
[7]励建容.高压技术在食品工业中的应用研究.食品与发酵业.1997,23(6)：9-14
[8]张守勤,牛新春.常温超高压提取水飞蓟素的研究.农机化研究.2007,(3)：143-145
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