Asparagus adscendens Roxb. (Asparagaceae), is native to the Himalayas. The present study, for the first time, was undertaken to explore the antimicrobial potential, to determine the minimum inhibitory concentration (MIC) values of the methanol extract of the roots of A. adscendens and its solid-phase extraction (SPE) fractions using resazurin microtitre assay against Gram-positive and negative bacterial-registered strains and to carry out HPLC-photodiode array analysis of the SPE fractions. The methanol extract and all SPE exhibited considerable level of antibacterial potential against Gram-positive bacteria (MIC: 2.5-0.009 mg/mL) than against Gram-negative bacteria (MIC: 1.25-2.5 mg/mL). The use of microtitre plates has the advantage of lower cost, fast and quantitative results. Like other Asparagus species, the presence of phenolic compounds in all SPE fractions was evident in the HPLC-PDA data.