The present study was conducted with the aim to extract phytochemical constituents of nutmeg (Myristica fragrans) seeds using methanol and acetone solvents, respectively by soxhlet apparatus with a yield of (1.47gm/100 gm) for methanol solvent and (2.11gm/100gm) for acetone solvent. The two extracts were tested for their antimicrobial, antioxidant and antiinflammatory activities. Agar well diffusion method was adopted against four bacterial species including two Gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis) and two Gram-negative bacteria (Escherichia coli, and Klebsiella sp.) as well as one yeast (Candida albicans) to determine the antimicrobial activity. The present study showed that the tested microorganisms were inhibited by nutmeg acetone extract with an inhibition zone ranging between (11-15 mm), especially toward Candida albicans with 15 mm diameter while no inhibition zone was observed using methanol extract. Chemical constituents of nutmeg acetone extract were estimated using Atomic Absorption Spectrometry (AAS) and Gas Chromatography-Mass Spectroscopy (GC-MS) measurements revealed the presence of 150 µg/dl Cu, 280 µg/dl Zn, 6.1 µg/dl Pb, 0.08 µg/dl Cd, 0.09 µg/dl Cr, 0.0012 µg/dl Ni and 0.027 µg/dl Mn and 32 extra compounds. Other phytochemical detections including alkaloids, tannins, terpenoid, saponins, glycosides, resins, and flavonoids were investigated using standard procedures. Total phenol content and radical scavenging activity were measured quantitatively using Folin-Ciocalteu and 2,2-diphenylpicrylhydrazyl (DPPH) methods, respectively and were compared qualitatively with the microfluidic paperbased analytical device (µPADs) as a new platform. The extract contains a high concentration of phenolic compounds (0.6217 mg/ml) and the DPPH assay for acetone extract indicated a high amount of antioxidant compounds. The effect of nutmeg acetone extract on cyclooxygenase (COX-2) enzyme activity has been done and showed that the nutmeg extract can inhibit COX-2 activity better than the ability of anti-inflammatory drug (Aspirin). Our findings indicate the potential use of nutmeg acetone extract as a source of antimicrobial, antioxidant, and anti-inflammatory agent.