In view of the reputation of genus Salvia in folklore medicine and its abundance in our region, the chemical composition and antimicrobial activity of essential oil from S. santolinifolia Boiss. was analyzed. Chemical analysis, using gas chromatography and gas chromatography mass spectrometry, retention indices and C-13 nuclear magnetic resonance spectroscopy has resulted in identification of 116 constituents, comprising about 97% of the total constituents. Out of these 116, 78 constituents are hitherto unreported from this source. The species belongs to α-pinene chemotype. In antibacterial assay, gram negative gastropathogens (Shigella boydii, S. flexneri, S. dysenteriae, Vibrio cholerae); causative agent of urinary tract infection (Proteus mirabilis and P. vulgaris) and pneumonia (Klebsiella pneumoniae) were found sensitive to this essential oil while Corynebacteria species and Staphylococcus epidermidis were significantly inhibited in antibacterial assay against gram positive bacteria. Clinical and Laboratory Standards Institute protocol was used for determining antimicrobial activity. Thus the essential oil from this species can be utilized as potential chemotherapeutic agent.