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Background  Medicinal plants are rich source of traditional herbal medicine around the globe. Most of the plant’s therapeutic properties are due to the presence of secondary bioactive compounds.    Methods  The present study analyzed the High Pressure Liquid Chromatography (HPLC) fractions of  Puncia granatum  (peel) extracts (aqueous, chloroform, ethanol and hexane) against multidrug resistant bacterial pathogens ( Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus ). All the fractions having antibacterial activity was processed for bioactive compounds identification using LC MS/MS analysis.    Results  Among total HPLC fractions ( n  = 30), 4 HPLC fractions of  P. granatum  (peel) showed potential activity against MDR pathogens. Fraction 1 (F1) and fraction 4 (F4) collected from aqueous extract showed maximum activity against  P. aeruginosa . Fraction 2 (F2) of hexane showed antibacterial activity against three pathogens, while ethanol F4 exhibited antibacterial activity against  A. baumannii . The active fractions were processed for LC MS/MS analysis to identify bioactive compounds. Valoneic acid dilactone (aqueous F1 and F4), Hexoside (ethanol F4) and Coumaric acid (hexane F2) were identified as bioactive compounds in HPLC fractions.    Conclusion    Puncia granatum  peel extracts HPLC fractions exhibited potential inhibitory activity against MDR bacterial human pathogens. Several bioactive compounds were identified from the HPLC fractions. Further characterization of these compounds may be helpful to conclude it as therapeutic lead molecules against MDR pathogens.

Punica granatum peel extracts: HPLC fractionation and LC MS analysis to quest compounds having activity against multidrug resistant bacteria