The use of microfluorometric method for activity-guided isolation of antiplasmodial compound from plant extracts
The use of microfluorometric method for activity-guided isolation of antiplasmodial compound from plant extracts
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DOI：
10.1007/s00436-008-0879-6
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发表时间：
2008-05-01
影响因子：
2
通讯作者：
Kouno, I.
中科院分区：
文献类型：
--
作者：
Shuaibu, M. N.;Wuyep, P. A.;Kouno, I.
用PicoGreen荧光法测定了16种药用植物的甲醇提取物的体外抗疟原虫活性。由寄生虫DNA浓度确定的IC 50，范围为< 11 to >200和< 13 to >200 μ g/ml的恶性疟原虫3D 7和K1，分别和最活跃的提取物是Anogeissus leiocarpus和Terminalia avicennoides（< 11->= 14 μ g/ml）。这两种提取物的水、丁醇、乙酸乙酯和甲醇级分显示丁醇级分具有相对较好的活性（IC 50，10-12 μ g/ml）。活性导向色谱分离的丁醇馏分葡聚糖凝胶LH-20，然后通过核磁共振和相关的高效液相色谱法揭示了已知的水解单宁和一些相关化合物的存在下castalagin，鞣花酸，黄没食子酸，安石榴苷，terchebulin，和其他两个馏分。所有这些化合物对两种菌株的IC（50）在8-21 μ g/ml（8-40 μ M）之间。对小鼠成纤维细胞的毒性试验表明，所有提取物和分离的化合物的IC 50>= 1500 μ g/ml，但苦瓜的IC 50 < 1500 μ g/l。在观察到的IC（50）s下，所有提取物和分离的化合物均不影响人红细胞膜的完整性。然而，副作用以浓度依赖性方式显现（从IC 50>= 500 μ g/ml起）。
In vitro antiplasmodial activity of methanolic extracts of 16 medicinal plants was evaluated by fluorometric assay using PicoGreen. The IC50s, as determined by parasite DNA concentration, ranged from < 11 to > 200 and < 13 to > 200 mu g/ml for Plasmodium falciparum 3D7 and K1, respectively; and the most active extracts were those from Anogeissus leiocarpus and Terminalia avicennoides (< 11->= 14 mu g/ml). Aqueous, butanolic, ethyl acetate, and methanolic fractions of these two extracts revealed butanolic fraction to have a relatively better activity (IC50, 10-12 mu g/ml). Activity-guided chromatographic separation of the butanolic fraction on Sephadex LH-20 followed by nuclear magnetic resonance and correlation high-performance liquid chromatography revealed the presence of known hydrolysable tannins and some related compounds-castalagin, ellagic acid, flavogallonic acid, punicalagin, terchebulin, and two other fractions. The IC(50)s of all these compounds ranged between 8-21 mu g/ml (8-40 mu M) against both the strains. Toxicity assay with mouse fibroblasts showed all the extracts and isolated compounds to have IC50 >= 1500 mu g/ml, except for Momordica balsamina with < 1500 mu g/l. All the extracts and isolated compounds did not affect the integrity of human erythrocyte membrane at the observed IC(50)s. However, adverse effects manifest in a concentration-dependent fashion (from IC50 >= 500 mu g/ml).