Onlom Churanya, Phrompittayarat Watoo, Putalun Waraporn, Waranuch Neti, Ingkaninan Kornkanok
Bioscreening Unit, Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences and Centre of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok, 65000, Thailand.
Faculty of Public Health, Naresuan University, Phitsanulok, 65000, Thailand.
Phytochem Anal. 2017 Jul;28(4):316-323. doi: 10.1002/pca.2678. Epub 2017 Feb 15.
Asparagus racemosus Willd (Asparagaceae family), known as Shatavari, is important in Ayurveda and traditional Thai medicines. The saponin glycosides, shatavarin I and IV are major constituents in its roots and may be responsible for their actions including protection against lipid peroxidation and carcinogenesis.
To develop an immunoaffinity column for isolating compounds with structures related to shatavarin IV from crude extracts of A. racemosus root.
The monoclonal antibody recognising shatavarin IV (mAbShavIV) was coupled to an Affi-Gel Hz gel to isolate compounds with structures related to shatavarin IV from the other components of crude extracts of A. racemosus root. The saponin glycosides in each fraction were analysed by mAbShavIV ELISA and LC-MS/MS.
The pooled wash-through fractions contained 3% of loaded mAbShavIV reactive saponin glycosides, while eluted fractions released ~ 90% of shatavarin saponin glycosides in a single step. Using thiobarbiturate (TBARs) to measure lipid-peroxidation, the extract, and the pooled wash-through fractions showed moderate protection against Cu -induced oxidation of human low density lipoprotein (LDL) (IC 11.3 ± 1.4 and 12.6 ± 0.9 μg/mL, respectively). In contrast, the saponin glycosides eluted from the mAbShavIV-column had weaker protectant (IC 29.7 ± 1.8 μg/mL) suggesting that A. racemosus shatavarins do not inhibit carcinogenesis through preventing lipid peroxidation.
The strategy described here demonstrates its utility for isolating a group of related compounds from the rest of the extract with selectivity and recovery rate. Pharmacological efficacy and synergistic effects of the components obtained can be further investigated. Copyright © 2017 John Wiley & Sons, Ltd.
天门冬(天门冬科),又称沙塔瓦里,在阿育吠陀医学和传统泰国医学中具有重要地位。皂苷糖苷,即沙塔瓦林I和IV是其根部的主要成分,可能是其发挥包括抗脂质过氧化和抗癌作用的原因。
开发一种免疫亲和柱,用于从天门冬根粗提物中分离与沙塔瓦林IV结构相关的化合物。
将识别沙塔瓦林IV的单克隆抗体(mAbShavIV)偶联到Affi-Gel Hz凝胶上,以从天门冬根粗提物的其他成分中分离出与沙塔瓦林IV结构相关的化合物。通过mAbShavIV ELISA和LC-MS/MS分析各馏分中的皂苷糖苷。
合并的洗脱前馏分含有3%加载的mAbShavIV反应性皂苷糖苷,而洗脱馏分在一步中释放了约90%的沙塔瓦林皂苷糖苷。使用硫代巴比妥酸(TBARs)测量脂质过氧化,提取物和合并的洗脱前馏分对铜诱导的人低密度脂蛋白(LDL)氧化显示出适度的保护作用(IC分别为11.3±1.4和12.6±0.9μg/mL)。相比之下,从mAbShavIV柱洗脱的皂苷糖苷具有较弱的保护作用(IC为29.7±1.8μg/mL),这表明天门冬沙塔瓦林不能通过预防脂质过氧化来抑制致癌作用。
本文所述策略证明了其从提取物其余部分中选择性地分离一组相关化合物并具有回收率的实用性。所得成分的药理功效和协同作用可进一步研究。版权所有©2017约翰威立父子有限公司。