Date of Completion

2025

Document Type

Thesis

Degree Name

Bachelor of Science in Life and Health Sciences

Keywords

Armoracia rusticana, Allyl isothiocyanate, Antimicrobial activity, Kirby-Bauer assay, LC-MS, Staphylococcus aureus, P. aeruginosa, S. pneumoniae and E. coli.

Abstract

This study investigates the antimicrobial potential of Armoracia rusticana (horseradish) extract, with a focus on allyl isothiocyanate (AITC)-a bioactive compound derived from glucosinolate hydrolysis. Aqueous enzymatic extraction was employed, followed by phytochemical profiling via liquid chromatography-mass spectrometry (LC-MS), and antimicrobial testing against two Gram-positive (Staphylococcus aureus, Streptococcus pneumoniae) and two Gram-negative strains (Escherichia coli, Pseudomonas aeruginosa). Inhibition zones were measured using the Kirby-Bauer disc diffusion assay across four volumetric extract concentrations (25%, 50%, 75%, 100%). Results revealed dose-dependent inhibition in S. aureus, with 100% extract achieving a mean zone of 9.77 mm and a calculated ECso of 148.79 μg/mL, indicating moderate antimicrobial activity. P. aeruginosa demonstrated weak but statistically significant responsiveness at higher concentrations, while S pneumoniae and E. coli exhibited negligible inhibition, consistent with known resistance mechanisms. Statistical analysis via one-way ANOVA and Tukey's HSD confirmed significant intergroup variation (p < 0.05) in extract-treated groups, supporting the strain-specific efficacy of the extract. Although AITC was not directly detected in LC-MS due to volatility and polarity constraints, unresolved MS/MS peaks suggest the presence of additional bioactive metabolites. Methodological limitations, including the absence of controlled myrosinase hydrolysis and GC-MS quantification, underscore the need for extract standardization and mechanistic validation. Nevertheless, the study affirms the selective in vitro antimicrobial activity of horseradish-derived phytochemicals and identifies methodological avenues for optimizing efficacy and compound detection.

First Advisor

Iluminada A. Ronio

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