色谱技术在有毒及无毒物种指纹图谱中的应用。

Applications of Chromatographic Techniques for Fingerprinting of Toxic and Non-toxic Species.

出版信息

Pak J Biol Sci. 2020 Mar;23(4):552-560. doi: 10.3923/pjbs.2020.552.560.

DOI:10.3923/pjbs.2020.552.560

Abstract

BACKGROUND AND OBJECTIVE

Euphorbia species have historically been used as medicinal plants to treat different ailments. However, some species have been reported to exhibit various degrees of toxicity. It becomes critical to distinguish toxic species from those that are non-toxic, for a particular application. The aim of the study was to determine the method for fingerprinting the chemical constituents of the selected toxic and non-toxic Euphorbia species to identify markers of toxicity.

MATERIAL AND METHODS

Hexane, DCM, methanol, ethyl acetate and water plant extracts of Euphorbia ammak, clavarioides, caerulescens, polygona and trigona were investigated for their cytotoxic activities towards the mammalian Vero cell line using MTT cell viability test assay. The presence of secondary metabolites and proteins were assessed in the plant extracts. Moreover, the study used chromatographic methods to fingerprint the plant extracts to identify toxicity markers.

RESULTS

The DCM extract of E. ammak exhibited the highest cell growth inhibition at all concentrations tested. The non-polar extracts of E. clavarioides exhibited the highest cell growth inhibition activity with hexane extract reaching IC50 at 1 μg mL-1. The DCM extract of E. caerulescens reached IC50 at a concentration of 10 μg mL-1, while other extracts didn't show any activity. The hexane and DCM extracts of E. polygona exhibited the highest cell growth inhibition activity, reaching IC50 at a concentration of 10 μg mL-1. All 4 extracts of E. trigona didn't show cell growth inhibition. All Euphorbia species showed the presence of secondary metabolites. The biuret and xanthoprotein methods indicated that there were no proteins detected in all 5 Euphorbia species. TLC profiles of toxic extracts revealed additional bands which were absent in non-toxic species.

CONCLUSION

It is concluded that the TLC method developed in this study can be used as a quick screen method to possibly distinguish toxic from non-toxic species, as well as in identifying the studied species.

摘要

背景与目的

大戟属植物在历史上一直被用作药用植物来治疗各种疾病。然而，一些物种已被报道具有不同程度的毒性。对于特定的应用，区分有毒物种和无毒物种变得至关重要。本研究的目的是确定鉴定所选有毒和无毒大戟属物种化学成分指纹图谱的方法，以鉴定毒性标志物。

材料与方法

使用 MTT 细胞活力试验检测哺乳动物 Vero 细胞系，研究大戟属植物的 Ammak、Clavarioides、Caerulescens、Polygona 和 Trigona 的正己烷、DCM、甲醇、乙酸乙酯和水植物提取物的细胞毒性。评估植物提取物中次生代谢物和蛋白质的存在。此外，该研究使用色谱方法对植物提取物进行指纹图谱分析，以鉴定毒性标志物。

结果

E. ammak 的 DCM 提取物在所有测试浓度下均表现出最高的细胞生长抑制作用。E. clavarioides 的非极性提取物表现出最高的细胞生长抑制活性，其正己烷提取物的 IC50 达到 1μg mL-1。E. caerulescens 的 DCM 提取物在浓度为 10μg mL-1 时达到 IC50，而其他提取物则没有任何活性。E. polygona 的正己烷和 DCM 提取物表现出最高的细胞生长抑制活性，其 IC50 浓度达到 10μg mL-1。E. trigona 的 4 种提取物均未显示出细胞生长抑制作用。所有大戟属植物均表现出次生代谢物的存在。双缩脲和黄蛋白方法表明，在所有 5 种大戟属植物中均未检测到蛋白质。有毒提取物的 TLC 图谱显示，在无毒物种中不存在其他带。