초록
두충잎에서 인간대장암에세포 HCT-116에 세포증식 억제효과를 나타내는 성분을 silica gel column chromatography TLC로 분리하고 HPLC로 정제한 다음 UV-visual spectro-photometer에 의한 흡수 spectra 특성 HPLC에 의한 retention time과 분리패턴 및 FAB/MS로 분자량을 잠정적으로 동정한 결과 TLC 상에서 분리된 Rf 0.19 band 의 화합물은 chlorophyll a에서 $Mg^{2+}$이 제거된 pheophytin a로 Rf 0.25 band의 화합물은 pyropheophytin a로 확인되었다. 동정된 화합물들의 암세포 억제작용을 확인하기 위해 두충잎의 petroleum ether extract column chromatography로 분리한 분획물 및 TLC에서 분리된 각 band들의 cytotoxic activity을 in vitro에서 HCT-116 cancer cell을 사용하여 측정한 결과 모든 시료에서 암세포 증식억제 현상을 보였으며 특히 Rf 0.19와 Rf 0.25 band에서 분리된 화합물들에서 강하게 나타났다. 이 결과로 볼 때 두충잎에서 분리한 클로로필 유도체는 인간대장암세포인 HCT-116 cell에 대해 강한 항암작용이 있는 것으로 생각된다.
This study was attempted to isolate and identify the anticancer compounds from Eucommia ulmoides leaves using a human colon cancer cell line HCT-116. The petroleum ether extracts with anticancer activity was chromatographed on silica gel TLC and finally anticancer compounds was purified by HPLC. Their chemical structures were roughly elucidate by UV-VIS absorption spectral data HPLC elution pattern and FAM/MS spectroscopy. From this study these compounds were suspected to be pheophytin a formed by the removal of $Mg^{2+}$ from chlorophyll a and pyropheophytina formed by the removal of acetate group from pheophytin a respectively. To confirm the anticancer effects against HCT-116 cancer cell petroleum ether extract fractions of column chromatography and fractions separated on TLC were tested. All samples tested including the extract of petroleum ether fractions of column chromatograph and three bands (0.13,0.19,0.25) of TLC appeared to inhibit the growth of HCT-116 cancer cell however especially 0.19 and 0.25 fractions separated on TLC plate revealed the strongest effect. These results suggest that chlorophyll derivatives in Eucommia ulmoides may be potential anticancer agents against a human colon cancer cell HCT-116.