A novel method to measure the 13C composition of intact bacteriohopanepolyols

We present a novel method to measure the 13C/12C isotope ratio (reported as δ13C) of individual, intact bacteriohopanepolyols (BHPs) using semi-preparative ultrahigh pressure liquid chromatography (UPLC) followed by high temperature gas chromatography-isotope ratio mass spectrometry (HT-GC-IRMS). The method is reproducible, as indicated by the precision of δ13C values for bacteriohopanetetrol (BHT) extracted from R. palustris biomass and analyzed across an order-of-magnitude range of IRMS peak areas (δ13C = −33.4 ± 0.6‰ VPDB, n = 94, ±1σ). To show that this method successfully separates individual BHPs from complex environmental matrices, we report δ13C values for BHT and the BHT stereoisomer (BHT-II) from a ca. 2.9 Ma, organic-rich Mediterranean Sea sediment sample. These analyses suggest that intact BHP δ13C measurements can greatly improve the interpretation of environmental signals by alleviating the need for side-chain cleavage which reduces BHP source-specificity.