The cucurbitane-type triterpenoids of Momordica charantia Linn. have blood lipid lowering effect and thereby improving non-alcoholic fatty liver. In this study, microwave-assisted extraction efficiently obtained these compounds which were further confirmed by the use of UPLC-MS/MS for target analysis and LC-HRMS for non-target analysis, combining GNPS techniques. The results indicated that the content of cucurbitane-type triterpenoids in bitter melon Hualien No. 5 was the highest among the others. Except for the immature whole fruit, the other plant parts such as fruits, seeds, vines and the whole plant of Hualien No. 5 with different maturity levels of immature, mature and overripe can significantly inhibit lipid accumulation in hepatocytes induced by oleic acid. The vines were proven as the most significant among the others where the active components for improving non-alcoholic fatty liver were rapidly screened and identified as 3β,7β,25-trihydroxycucurbita-5,23 (E)-dien-19-al and Momordicine II through non-target analysis, multivariate analysis and molecular network.