The impact of household processes on fenugreek leaves and seeds was analyzed for total phenolic (TP) and total flavonoid content (TF), and in-vitro biological activities such as antioxidant, antimicrobial, and anti-inflammatory properties. Processes included air-drying of leaves and germinating, soaking, and boiling of seeds. Air-dried fenugreek leaves (ADFL) had high TP (15.27 mg GAE/g D.W.) and TF (7.71 mg QE/g D.W.). The TF of unprocessed, germinated, soaked, and boiled seeds had 6.54, 5.60, 4.59, and 3.84 mg GAE/g D.W., respectively. The TF in UFS, GFS, SFS, and BFS were 4.23, 2.11, 2.10, and 2.33 mg QE/g D.W., respectively. Sixteen phenolic and nineteen flavonoid compounds were identified using the HPLC. Antioxidant activity using DPPH•, ABTS+•, and FRAP• assays indicated that ADFL had high activity. Antimicrobial activity was evaluated against each eight pathogenic bacterial and fungal strains. ADFL showed a strong activity with MIC values ranging from 0.03 to 1.06 and 0.04 to 1.18 mg ml− 1 against bacterial and fungal strains, respectively. Anti-inflammatory activity was evaluated in-vitro against RAW 264.7 macrophage cells using of NO assay. Results revealed that ADFL had the highest cytotoxicity and anti-inflammatory activity according to NO assay. Household processes significantly declined the in-vitro biological properties of processed seeds.
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