The nutritional and therapeutic benefits of plants are outstanding. Wild mustard (Sinapis arvensis L.) collected from different regions was analysed for its phenolic, flavonoid, protein, and biological activity levels. In this case, the plant ethanol extract was obtained using a soxhlet apparatus. Rel assay kits and the DPPH test were used to assess the plant's antioxidant activity. The agar dilution test was used to determine antimicrobial efficacy. A549 lung cancer cells were used in an MTT assay to measure antiproliferative activity. The Folin−Ciocalteu reagent was used to quantify the total phenolic content of the sample. The amount of flavonoids was determined using an aluminium chloride test. The amount of protein was calculated using the AOAC's standard technique. Based on the research conducted, it was shown that the maximum total antioxidant status (TAS) value for the ethanol extract of wild mustard from various places was 5.232 ± 0.047 mmol L−1. At a concentration of 2 mg mL−1, DPPH activity was measured to be 82.06 ± 1.01. The maximum levels of total phenolic, flavonoid, and protein were 80.57 ± 2.19 mg g−1, 154.07 ± 2.79 mg g−1, and 7.75 ± 0.24%, respectively. Doses of 25–100 μg mL−1 of plant extracts were effective against fungal strains, whereas doses of 50–200 μg mL−1 were beneficial against bacterial strains. The plant extracts were shown to have potent antiproliferative properties. It was found that wild mustard's phenolic, flavonoid, protein, and biological activity levels varied according to the location from which it was gathered. It was also concluded that wild mustard had significant biological activity.
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