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.