The present study focused on the phytochemical profile, isolation of bioactive components, and determination of the toxicity of Azadirachta indica seed extract. The crude extracts of seeds of A. indica were prepared by cold maceration in hexane, ethyl acetate, methanol, and water. Phytochemical screening followed by column chromatography separated main fractions. Acute toxicity was studied using albino mice, administering doses to calculate LD50 and observing for behavioral and physical changes. A. indica seed extracts routinely produce a "brown gummy mass" that is dominated by non-polar chemicals, with hexane obtaining the highest extraction efficiency (28.55%), indicating phytochemical diversity and solvent selectivity. Ethyl acetate had the maximum ability to extract phenols and flavonoids, while methanol was efficient for tannin recovery; this clearly shows the influence of the degree of polarity on the solvent concerning phytochemical extraction. Hexane extract of A. The highest phytochemical diversity was observed in the seeds of A. indica, followed by methanol and then ethyl acetate. The 1H-NMR of compound Fa1 isolated from A. indica seeds confirmed the molecular structure, pointing out functional groups, methyls-and stereochemistry. The 13C-NMR spectrum of A. indica extract showed aliphatic, aromatic, and methyl carbons, giving important signals with significant chemical shifts due to functional groups. GC-MS of Fa1 from A. The A. indica showed complex structural features that included hydroxyl groups and alkyl fragments, confirming its bioactivity and possible interactions with biological systems. Hexane crude extract of seeds of A. indica showed minimal acute toxicity profile in albino mice. No mortality or symptoms were observed during a 24-hour observation period in doses as high as 5000 mg/kg. Whereas the ethyl acetate extract of A. indica seed showed no mortality at any concentration, the crude methanol extract in its crude form exhibited no signs of toxicity or fatality, even at 5000 mg/kg. This dictates its safety profile. Acute toxicity studies of A. indica Linn-seed extract fraction F1 in albino mice do not provide any evidence of any damage, even at high dosages up to 5000 mg/kg. Finally, A. indica seed extracts demonstrate varied phytochemicals and minimal toxicity, confirming their potential for safe bioactive uses.