Yeo Chia-Rou, Yong Jin-Jie, Popovich David G
National University Hospital, Singapore; National University of Singapore, Singapore.
Department of Chemistry, National University of Singapore, Singapore.
J Pharm Biomed Anal. 2017 May 30;139:148-155. doi: 10.1016/j.jpba.2017.02.054. Epub 2017 Mar 1.
Panax ginseng has been studied for its chemo-preventive properties and pharmaceutical potential. Polyacetylenic compounds isolated from Panax ginseng root typically comprised of non-polar C compound have been reported to exhibit bioactive properties. The objective of this project is to extract, isolate, and characterize bioactive polyacetylenes from Panax ginseng root using various extraction and separation methods Ginseng was extracted by reflux using methanol, ethanol, hexane, ethyl acetate, methanolic ultrasonication. The extracts were partitioned with hexane to obtain water-soluble portion and hexane-soluble portion. Hexane was subsequently removed under vacuum, and formed a crude polyacetylenes extract (crude PA). Silica gel chromatography and semi-preparative HPLC were utilized to prepare 5 fractions and the polyacetylenes were measure by HPLC and molecular weights confirm my APCI-MS and MNR. The bioactive effect was measured by MTT viability assay using murine 3T3-L1 cells. Extraction with methanol under reflux produced significantly larger amount of polyacetylenes (p<0.05). Liquid-liquid extraction and column chromatography were used to separate polyacetylenic compounds into five different fractions. Major polyacetylenes, panaxynol and panaxydol were found in fraction 1 and 2 respectively. Dose-response relationships were observed in 3T3-L1 cells and LC50 were 13.52±3.05μg/mL (fraction 1), 3.69±1.09μg/mL (fraction 2), 52.88±11.16μg/mL (fraction 3), 85.91±27.37μg/mL (fraction 4) and 135.52±32.91μg/mL (fraction 5). Fraction 2 containing panaxydol was found to have exhibited the greatest anti-proliferative effects on 3T3-L1 preadipocytes. Extraction with methanol under reflux produced significantly more polyacetylenes. Fractions that contain panaxydol was the most cytotoxic.
人参已被研究其化学预防特性和药用潜力。据报道，从人参根中分离出的聚乙炔化合物通常由非极性碳化合物组成，具有生物活性。本项目的目的是使用各种提取和分离方法从人参根中提取、分离和表征具有生物活性的聚乙炔。人参通过甲醇、乙醇、己烷、乙酸乙酯回流提取以及甲醇超声提取。提取物用己烷分配以获得水溶性部分和己烷溶性部分。随后在真空下除去己烷，形成粗聚乙炔提取物（粗PA）。利用硅胶柱色谱和半制备HPLC制备了5个馏分，并通过HPLC测定聚乙炔含量，通过大气压化学电离质谱（APCI-MS）和核磁共振（MNR）确定分子量。使用小鼠3T3-L1细胞通过MTT活力测定法测量生物活性效果。回流甲醇提取产生的聚乙炔量显著更多（p<0.05）。液液萃取和柱色谱用于将聚乙炔化合物分离成五个不同的馏分。主要的聚乙炔，人参炔醇和人参二醇分别存在于馏分1和馏分2中。在3T3-L1细胞中观察到剂量反应关系，半数致死浓度（LC50）分别为13.52±3.05μg/mL（馏分1）、3.69±1.09μg/mL（馏分2）、52.88±11.16μg/mL（馏分3）、85.91±27.37μg/mL（馏分4）和135.52±32.91μg/mL（馏分5）。含有人参二醇的馏分2对3T-L细胞显示出最大的抗增殖作用。回流甲醇提取产生的聚乙炔显著更多。含人参二醇的馏分细胞毒性最大。
