In-depth chromatographic and spectrally defined phytocompounds in super-critical-CO2 extracted Phyllanthus emblica seeds oil show robust anti-microbial/biofilm effects in the non-mutagenic manner

Abstract

Bioactive-rich plant-based oils are gaining attention for their antimicrobial properties, addressing concerns of antibiotic resistance. Phyllanthus emblica L. known for its medicinal benefits, has been explored for its pharmacological potential. However, limited research exists on the supercritical extraction of bioactive compounds from P. emblica seeds. This study investigates the phytochemical constituents and antimicrobial activity of supercritically extracted P. emblica seed oil (PeSSO) using advanced techniques such as GC-MS/MS, GC-FID, HPTLC, FT-IR, and NMR. GC-MS/MS analysis identified eight non-polar and eight unsaponifiable compounds, whereas GC-FID detected five fatty acids, with linolenic acid (33.5 %) being the major polyunsaturated fatty acid. HPTLC confirmed the presence of nine active phytocompounds, which were further validated by FT-IR and NMR analyses. In addition, NMR technique was applied to evaluate the physico-chemical parameters of PeSSO. Functionally, PeSSO demonstrated significant bactericidal activity against Escherichia coli and Klebsiella pneumoniae (MIC₅₀ of 1.3 and 2.4 mg/mL, respectively); and inhibiting the biofilm formations, indicating its potential as a robust antimicrobial agent. Additionally, the Ames assay, a bacterial mutagenicity test, showed that PeSSO is non-mutagenic in nature. Taken together, the chromatographic and spectroscopic methods applied in this study effectively facilitated the rapid identification and characterisation of bioactive phytocompounds in Phyllanthus emblica seed oil, processed through environment friendly supercritical extraction, for the first time. Antimicrobial properties of PeSSO along with the non-mutagenic nature support its application in the pharmaceutical and food industries, offering a natural, and safe ingredient for enhancing food safety and health benefits.