10.22376/ijpbs.2019.10.1.p1-12 Volume 9 Issue 4 2018 (October - December) lessThan i greaterThan Trimad lessThan /i greaterThan is a polyherbal formulation mentioned in Ayurvedic literature. The formulation is a composite mixture of three plant parts including rhizomes of Nagarmotha ( lessThan i greaterThan Cyperus rotandus lessThan /i greaterThan L.), fruits of Vidang ( lessThan i greaterThan Embelia ribes lessThan /i greaterThan Burm.) and root of Chitrak ( lessThan i greaterThan Plumbago zylenica lessThan /i greaterThan L.). The present study was undertaken to evaluate phytochemical constituents of lessThan i greaterThan Trimad lessThan /i greaterThan polyherbal formulation through Liquid Chromatography-Mass Spectrometry (LC-MS) analysis. Chromatographic conditions for separation are as follows: The mobile phase was a combination of solvents: water (0.1% formic acid) (A) and Acetonitrile (B). The flow rate was 0.35mL/min. The gradient begins at 2% B and reached 98% B by 12 min. Consequently, the mixture returned to 2% B for a 3 min re-equilibration period. The components were subjected to the mass analyzer through electrospray ionization probe operating in positive and negative mode with the following conditions: capillary voltage for negative and positive, 2500, 4000V; fragmentor voltage, 150V; source temperature, 325°C; sheath gas temperature, 295°C; nebulizer pressure, 45 PSI; and gas flow, 8 L/min. Total twenty putative compounds were identified from methanolic extract of lessThan i greaterThan Trimad lessThan /i greaterThan . Negative and Positive ionization mode detected around 17 and 12 compounds respectively. Out of which, nine compounds are detected in both ionization mode. In negative ionization mode, β-glucopyranosyl plumbagic acid; xanthyletin; embelin; sugeonyl acetate; 1-tetradecene; cyclopentadecane; tetracosanoic acid methyl ester and tricosanoic acid methyl ester compounds were detected. In positive ionization mode, rotundines; γ –muurolene and ethyl p methoxycinnamate were identified. Nine compounds such as cyperol; β- asarone; oleic acid; campholaldehyde; thymol; 1-epineo-isoshinanolone; α-cyperone; phenol 2,4- bis(1,1-dimethylethyl) and n-hexadecanoic acid were detected in both ionization modes. Probably, this is the first report of high-throughput screening of lessThan i greaterThan Trimad lessThan /i greaterThan phytoconstituents through LC-MS analysis. Further studies can examine the mechanisms of action of this formulation. SURESH P KHADKE AND ANIKET A KUVALEKAR Trimad polyherbal formulation, LC-MS, α-Cyperone, β-Glucopyranosyl plumbagic acid and Embelin 136-147