TLC BIOAUTOGRAPHY AND LCMS-MS ANALYSIS FOR IDENTIFICATION OF COMPOUNDS HAVING INHIBITORY ACTIVITY AGAINST STAPHYLOCOCCUS AUREUS IN ABIES WEBBIANA LEAVES EXTRACTAbstract
This study was performed to identify the antimicrobial compounds in Abies webbiana leaves. The antimicrobial compounds were identified by LCMS-MS. The microbial inhibitory activity was evaluated on Staphylococcus aureus ATCC 6538 using REMA technique and Agar-overlay TLC bioautography. The chloroform extract of Abies webbiana leaves was used for the study. The MIC against Staphylococcus aureus was found to be 195.3 µg/ml- 390.6 µg/ml. A mobile phase comprising of toluene: ethyl acetate: glacial acetic acid (9:1:0.3) was used for identification of antimicrobial compounds present in the extract by TLC bioautography. Totally 16 bands were observed in the HPTLC chromatogram when the plate was observed at 254 nm. The band observed at Rf = 0.58 showed the antimicrobial activity. Preparative TLC technique was used to isolate this active band. The compounds present in the isolated band were then identified by LC-QTRAP. Totally 7 compounds were identified to be present in the isolated bands of chloroform extract. The compounds identified include Betuloside, 2,7-Dihydroxy-4’-methoxyisoflavone, Geinstein 7-O-beta-D-glucoside, β-Sitosterol, Abietane, Coniferol, and 1-(3,4-Dihydroxyphenyl)-1-decene-3,5-dione-Pos. This study was useful in identifying the compounds that show synergistic antimicrobial activity. Structural characterization of plant metabolites in Abies webbiana leaves exhibiting synergistic growth inhibition of S. aureus might serve as ‘lead’ molecule for developing antimicrobial agents.
A. Ambre, M. Bulbule, V. Shirsat *, D. R. Kandula, A. Singh and M. Pillai
Bombay College of Pharmacy, Kolivery Village, Mathuradas Colony, Mumbai, Maharashtra, India.
06 February 2019
28 May 2019
14 June 2019
01 October 2019