Fast isolation of embelin from Embelia ribes Burm. f. by comparative evaluation of extract, HPLC method validation and evaluation of its antioxidant activity

Embelia ribes Burm. f., is an important medicinal plant used as antioxidant, anticancerous, anti-inflammatory, antidiabetic, anthelmintic, and neuroprotective activities. The compound, embelin, a benzoquinone phenolic compound is responsible for its pharmacological activities. However, a lacuna in the optimization process of its extraction restricts it from wider application in the cosmetic industries. This study aims to develop an optimized extraction process to obtain an embelin-enriched extract along with a swift and economical HPLC quantification method and its antioxidant activity. Various solvent extracts of E. ribes fruits were prepared and screened for total polyphenol content (TPC) and antioxidant activity by DPPH assay. The isolated embelin was subjected to flash chromatography from the optimized extract, offering a rapid purification route. A gradient HPLC method development and validation including all parameters were done using phosphate buffer and acetonitrile as mobile phases for accurate embelin quantification. The cold methanolic extract was eminent among all extracts evaluated, showing the highest embelin content (22.33 ± 0.047%), TPC content (4.92 ± 0.50% w/w), whereas, the strongest antioxidant activity (IC 50 value 1.19 ± 0.12 µg AAE) was in cold aqueous-methanolic extract. The validated HPLC method exhibited excellent linearity over 200–1000 ng ( r = 0.999), with a LOD of 96.43 ng and LOQ of 292.22 ng, and achieved a recovery of 95.3%, confirming high sensitivity, accuracy, and reproducibility. The cold methanol extraction yielded an embelin-enriched extract with demonstrated antioxidizing activity, supported by a sensitive and economical HPLC method. It can be further investigated as a reliable source for development of antioxidant agent by cosmetic industries.