Phytochemical characterization and evaluation of antioxidant, antimicrobial, antibiofilm and anticancer activities of ethyl acetate seed extract of Hydnocarpus laurifolia (Dennst) Sleummer

Abstract

Various functional groups were observed in the FTIR analysis of Hydnocarpus laurifolia seeds ethyl acetate extract such as O–H, N–H, C–H, –CH₂, O=C=O, C=O, C=O–NH, and CH_3, etc. Eleven bioactive compounds were detected via GC–MS and the predominant compounds include (1S)-2-cyclopentene-1-tridecanoicacid (chaulmoogric acid) (80.59%); 2-cyclopentene-1-undecanoic acid (hydnocarpic acid) (6.76%); cyclobutylamine (5.28%); methyl thioacetate (ethanethioic acid) (4.84%); lignoceric acid (2.21%). The TPC and TFC values were 0.110 ± 0.04 GAE g⁻¹ and 0.175 ± 0.05 g CE g⁻¹ respectively. Ethyl acetate extract showed strong DDPH free radical scavenging activity with IC₅₀ value 10.64 ± 0.48 µg ml⁻¹ and antioxidant activity index 3.759. The ethyl acetate extract also exhibited potential ABTS radical scavenging efficacy with a very low IC₅₀ value, i.e., 07.81 ± 0.48 µg ml⁻¹. P. aeruginosa was the most sensitive bacteria to the extract with 33.16 ± 0.88 mm inhibition zone and MIC: 3.12 mg ml⁻¹, MBC: 6.25 mg ml⁻¹. P. aeruginosa biofilm was inhibited by ethyl acetate extract 99.22% at MIC concentration. The LM images displayed a decrease in the number of biofilm cells and FE-SEM micrographs showcased the extensive decrease as well as disintegration in biofilm. Additionally, ethyl acetate extract was found selectively cytotoxic to the K562 cancer cells having an IC₅₀ of 25.41 μg ml⁻¹ and barely cytotoxic to normal PBMCs having an IC₅₀ of 482.54 μg ml⁻¹, and the selectivity index value was 18.99. Data validate scientifically the traditional use of H. laurifolia seeds in folk medicines and confirmed that it can be used in modern phytomedicines as an antioxidant, antimicrobial, antibiofilm, and anticancer agent and is toxicologically safe.