Venkatesh Uday, Javarasetty Chethan, Murari Satish Kumar
Department of Biochemistry, Bharathiar University, Coimbatore-641 046, Tamilnadu, India.
Central Coffee Research Institute, Balehonoor, Chikkamagalur-577 117, Karnataka, India.
Afr J Tradit Complement Altern Med. 2017 Mar 1;14(3):167-174. doi: 10.21010/ajtcam.v14i3.18. eCollection 2017.
(.) (WT), commonly known as yellow dots or creeping daisy, is a shrub possessing potent biological activities, and is traditionally used a medicinal plant in Ayurveda, Siddha and Unani systems of medicines, and it has also been tried against leukemia cell line MEG- 01. In the present study, purification and screening of the plant was done for bioactive compounds in methanolic extract of WT for apoptotic and anti-leukemia activity.
The methanolic extract of WT was initially purified through thin layer chromatography (TLC) and screened for the apoptotic and anti-leukemia activities. The positive band of TLC was subjected to silica gel column chromatography for further purification and the fractions obtained from it were screened again for anti-leukemia activity through thymidine uptake assay and apoptotic activity by DNA fragmentation, nuclear staining and flow cytometry assays. The fraction with positive result was subjected to HPLC for analysis of bioactive components.
Out of many combinations of solvents, the methanol and dichloromethane combination in the ratio 6:4 has revealed two bands in TLC, among which the second band showed positive results for apoptotic and anti-leukemic activities. Further purification of second band through silica gel chromatography gave five fractions in which the 3 fraction gave positive results and it shows single peak during compositional analysis through HPLC.
The single peak revealed through HPLC indicates the presence of pure compound with apoptotic and anti-leukemia activities encouraging for further structural analysis.
(.)(WT),俗称黄点菊或匍匐雏菊,是一种具有强大生物活性的灌木,在阿育吠陀、悉达和尤纳尼医学体系中传统上被用作药用植物,并且也已尝试用于对抗白血病细胞系MEG - 01。在本研究中,对WT的甲醇提取物中的生物活性化合物进行了纯化和筛选,以检测其凋亡和抗白血病活性。
WT的甲醇提取物最初通过薄层色谱(TLC)进行纯化,并筛选其凋亡和抗白血病活性。TLC的阳性条带进行硅胶柱色谱进一步纯化,从其中获得的馏分再次通过胸苷摄取试验筛选抗白血病活性,并通过DNA片段化、核染色和流式细胞术试验检测凋亡活性。对结果呈阳性的馏分进行高效液相色谱(HPLC)分析生物活性成分。
在多种溶剂组合中,甲醇与二氯甲烷比例为6:4的组合在TLC中显示出两条条带,其中第二条条带对凋亡和抗白血病活性呈阳性结果。通过硅胶色谱对第二条带进一步纯化得到五个馏分,其中第3馏分结果呈阳性,并且在通过HPLC进行成分分析时显示出单峰。
HPLC显示的单峰表明存在具有凋亡和抗白血病活性的纯化合物,这为进一步的结构分析提供了鼓励。