Quantitative determination of various hopanoids in microorganisms

Abstract

A rapid and sensitive method for determination of various hopanoids in microorganisms is described. Tetrahydroxybacteriohopane (THBH), THBH-ether and -glycoside were acetylated with acetanhydride/pyridine and were separated on a C18 reversed-phase HPLC column with a gradient of acetonitrile in methanol and subsequent monitoring of the eluant at 206 nm. Quantification is based on peak area calculation by using purified bacteriohopanoids as external standards. A linear response is found from 10 to 20 micrograms bacteriohopanetetrol and for up to 200 micrograms ether and glycoside of THBH. Diplopterol and diploptene were determined by GLC after silylation with BSTFA/TMSCl in pyridine; the detector response is linear from 0.1 to 2 micrograms.