Among the family of heterocyclic compounds, The nitrogen containing six membered heterocyle, the piperidine structural was the most dominant and very prevalent element in nature and often found to be naturally occurring bioactive compounds such as alkaloids. Piperidin-3-one derivatives are used as precursors for the synthesis of antimalarial agents febrifugine and Isofebrifugine. Piperidin-4-ones mostly display varied and potent biological properties such as antiviral, antitumour, analgesic, antimicrobial, fungicidal, herbicidal, insecticidal, antihistaminic, anti-inflammatory and anticancer. CNS stimulant and recent reports suggest that compounds containing piperidin-4-one moiety elicit excellent activity when aromatic substitutions are present at 2- and/or 6-positions. In this work, the compound N-nitroso-2,6- diphenylpiperidin-4-one semicarbazone has been prepared and analyzed. The product showed positive nitrogen test (Lassign test), a single spot in TLC and sharp melting point for the purity of the compound. The structure of the compound was further confirmed from the CHN analysis FT-IR, and 1H NMR Spectral data and the compound have being screened for its antimicrobial activity against gram positive bacteria Bacillus subtilis, Staphylococcus aureus and Gram-negative bacteria Escherichia coli by using ciprofloxacin as standard and fungi Candida albicans by using cetramazole as standard. The compound exhibited significant activities against all the tested bacterial and fungal strains.