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2). GC/MS analysis showed that the most abundant 15C:0 and ��OH-17C:0 fatty acids were iso branched (Figure ?(Figure1A)1A) as previously reported (Pitta et al., 1989; Okuyama and Monde, 1996). Figure 2 High Resolution Accurate Mass/Mass Spectrometer (HRAM/MS) fragmentation of (A) Flavobacterium johnsoniae group I lipid at retention time 27.70 min; (B) proposed MS fragmentation of lysine lipids (LL) observed in F. johnsoniae. The relative abundance of ... Table 1 Elemental composition of amino acid containing lipid fragmentation Selleckchem GSK J4 products observed by high performance liquid chromatography-high resolution accurate mass/mass spectrometry (HPLC-HRAM/MS) in the lipid extracts of Flavobacterium johnsoniae and Pseudopedobacter ... IPLs of P. saltans HPLC-ESI/IT/MS analysis of the lipid extract of P. saltans showed abundant PE, OL, and OLHFA IPLs (Figure ?(Figure1B)1B) making up 8, 10, and 12%, respectively, of HPLC-ESI/IT/MS chromatogram base peak area and four clusters with unknown IPLs, groups I, I', II, and II' making up 23, 28, 1.5, and 7% of HPLC-ESI/IT/MS chromatogram base peak area, respectively. OLHFA were hydroxylated on the ester linked fatty acids at the ��-position. P. saltans group I produced MS3 fragmentation products of m/z 129, 130, and 147, identical to the group I LLs identified in F. johnsoniae. The relative retention time of group I IPLs in P. saltans are also similar to the retention time of the LLs in F. johnsoniae (Table ?(Table1),1), and were therefore also identified as LLs. This was confirmed by HPLC-HRAM/MS analysis (Table ?(Table1).1). P. saltans group I' IPLs exhibited the same MS3 fragmentation products (m/z 129, 130, and 147) as group I LLs, but MS2 fatty acid fragmentation losses were 16 Th larger than observed for group I, indicating the IPLs in group I' are potentially hydroxylated-fatty acid versions of group I LLs (LLHFA). This was also confirmed by HPLC-HRAM/MS analysis (Table ?(Table1).1). As with OLHFA, group I' LLHFA were hydroxylated on the ester linked fatty acid ��-position. The most abundant P. saltans fatty acids were iso branched as previously observed (Steyn et al., 1998; Liolios et al., 2011), and the distribution of fatty acid chain lengths and double bond equivalents were identical between the LLs, LLHFA, OLs, and OLHFA, suggesting a biosynthetic link between these IPL classes (Table ?(Table11). Group I LLs and group I' LLHFA were isolated from the P. saltans extract using preparatory HPLC to confirm the lysine head group structure. Approximately 0.6 mg of both LL and LLHFA were individually isolated for NMR analysis. For comparison, roughly 2.0 mg of known OL was also isolated from the F. johnsoniae extract.