Matrix-assisted laser desorption/ionization time-of-flight vs. fast-atom bombardment and electrospray ionization mass spectrometry in the structural characterization of bacterial poly(3-hydroxyalkanoates)

RATIONALE: Bacterial poly(3-hydroxyalkanoates) (PHAs) are an emergent class of plastic materials available fromrenewable resources. Their properties are strictly correlated with the comonomeric composition and sequence, whichmay be determined by various mass spectrometry approaches. In this paper we compare fast-atom bombardment(FAB) and electrospray ionization (ESI) to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOFMS) of partially pyrolyzed samples.METHODS:We determined the compositions and sequences of the medium-chain-length PHAs (mcl-PHAs) prepared bybacterial fermentation of Pseudomonas aeruginosa ATCC 27853 cultured in media containing fatty acids with 8, 12, 14, 18,and 20 carbon atoms as carbon sources by means of MALDI-TOFMS of pyrolyzates, and compared the results with thoseobtained by FAB- and ESI-MS in previous studies. MALDI matrices used were 9-aminoacridine (9-AA) and indoleacrylicacid (IAA).RESULTS: MALDI-TOFMS was carried out in negative ion mode when using 9-AA as a matrix, giving asemi-quantitative estimation of the 3-hydroxyacids constituting the PHAs, and in positive mode when using IAA,allowing us, through statistical analysis of the relative intensity of the oligomers generated by pyrolysis, to establish thatthe polymers obtained are true random copolyesters and not a mixture of homopolymers or copolymers.CONCLUSIONS: MALDI-TOFMS in 9-AA and IAA of partial pyrolyzates of mcl-PHAs represents a powerful methodfor the structural analysis of these materials. In comparison with FAB and ESI, MALDI provided an extended mass rangewith better sensitivity at higher mass and a faster method of analysis.