A rottlerin enriched fraction was prepared from extract of Mallotus philippensis Muell. Arg leaves in ethylacetate using HP-20 dianion polymeric resin. The procedure of extraction and fractionation was optimized to get fractions enriched with rottlerin. The different fractions obtained were then evaluated for in-vitro anti-inflammatory activity for COX inhibition. The most active fraction (fraction 6) was subjected to standardization by HPLC and found to contain 70% rottlerin, that could be responsible for the highest activity of the fraction. Further, the chromatographic purification of the active fraction yielded rottlerin (1), isoallorottlerin (2), 8-cinnamoyl-5,7-dihydroxy-2,2,6-trimethyl- 3-chromene (3), and mallotophilippen F (4) as markers. The rottlerinenriched fraction possessed good in-vitro anti-inflammatory activity against both COX-1 and COX-2 enzymes and was more active than the reference standards drugs viz. etoricoxib and ibuprofen. The optimized protocol for extraction and resin-based fractionation was found to be efficient, reproducible, time-saving, and eco-friendly as operated in semi-aqueous conditions.