Analysis of non-derivatized bacteriohopanepolyols using UHPLC-HRMS reveals great structural diversity in environmental lipid assemblages

• Published in: Organic geochemistry 2021-10, Vol.160, p.104285, Article 104285
• Main Authors: Hopmans, Ellen C., Smit, Nadine T., Schwartz-Narbonne, Rachel, Sinninghe Damsté, Jaap S., Rush, Darci
• Format: Article
• Language: English
• Subjects: Adenosylhopanes; Non-derivatized bacteriohopanepolyols; novel composite BHPs; UHPLC-HRMS
• ISSN: 0146-6380; 1873-5290
• DOI: 10.1016/j.orggeochem.2021.104285

•Reversed phase UHPLC-ESI/HRMS2 detects a broad range of (novel) bacteriohopanepolyols.•BHPs are detected simultaneously with intact polar lipids.•The nucleoside of adenosylhopane type-3 is a methylated adensine.•The nucleoside of adenosylhopane type-2 is N1-methylinosine. Bacteriohopanepolyols (BHPs) are lipids with great chemotaxonomic potential for microbial populations and biogeochemical processes in the environment. The most commonly used methods for BHP analysis are chemical degradation followed by gas chromatography-mass spectrometry (MS) or derivatization followed by high performance liquid chromatography (HPLC)-atmospheric pressure chemical ionization/MS. Here we report on significant advances in the analysis of non-derivatized BHPs using U(ltra)HPLC-electrospray ionization-high resolution MS2. Fragmentation mass spectra provided information on the BHP core, functionalized side chain, as well as the conjugated moiety of composite BHPs. We successfully identified the common bacteriohopanepolyols and their (di)methylated and (di)unsaturated homologues, aminoBHPs, and composite BHPs (e.g., cyclitol ethers and methylcarbamate-aminoBHPs) in biomass of several known BHP-producing bacteria. To show how the method can be exploited to reveal the diversity of BHPs in the environment, we investigated a soil from an active methane seep, in which we detected ca. 130 individual BHPs, including a complex distribution of adenosylhopanes. We identified the nucleoside base moiety of both adenosylhopane type-2 and type-3. Adenosyl hopane type-3 contains a methylated adenine as its nucleobase, while type-2 appears to contain a deaminated and methylated adenine, or N1-methylinosine. In addition, we detected novel adenosylhopanes. Furthermore, we identified a series of novel composite BHPs comprising of bacteriohopanepolyols conjugated to an ethenolamine moiety. The novel ethenolamineBHPs as well as aminoBHPs were also detected acylated to fatty acids. The analytical approach described allows for simultaneous analysis of the full suite of IPLs, now including BHPs, and represents a further step towards environmental lipidomics.