LnAs. The highest intensity exhibited an isomer with tR 11.two min, which
LnAs. The highest intensity exhibited an isomer with tR 11.two min, which was interpreted as FAME 18:3n-5,7,9. Its MS/MS spectrum (Figure 8A) showed abundant diagnostic peaks m/z 190.two and m/z 290.1, corresponding to an MBR value of 133. The spectrum closely matched the punicic acid methyl ester shown in Figure 2. Interestingly, two less abundant isomers with the identical diagnostic fragments have been detected at tR 12.0 min and tR 14.eight min (Supplementary Materials Figure S2). These species were isomers with the same double bond positions but different double bond geometries. The geometrical isomers of punicic acid, namely FAME 18:3n-5t,7t,9c (-eleostearic acid); FAME 18:3n-5c,7t,9t (-eleostearic acid), and FAME 18:3n-5t,7t,9t (Combretastatin A-1 Epigenetic Reader Domain catalpic acid) have been detected in pomegranate seed oil previously [51,869]. Because the elution of the FAs in reversed-phase systems proceeds from cis to trans isomers [20,90], the later eluting isomers most likely contained a larger variety of trans double bonds. The MS/MS spectra of FAMEs with three PF-05105679 supplier conjugated double bonds in different positions are shown in Figure 8B . In all of them, the MBR value was 133, plus the diagnostic fragments allowed us to interpret them asMolecules 2021, 26,ten ofMolecules 2021, 26,FAME 18:3n-4,six,eight (Figure 8B), FAME 18:3n-3,5,7 (Figure 8C), FAME 18:3n-2,four,6 (Figure 8D), FAME 18:3n-8,10,12 (Figure 8E), and FAME 18:3n-9,11,13 (Figure 8F). The retention instances from the latter two CLnAs had been close to each other, which resulted in mixed spectra. Overall, ten CLnAs, one particular methylene-interrupted (18:2n-6,9), and two monounsaturated (18:1n-9 and 20:1n-9) FAMEs have been identified within the PSO (Table three). The outcomes had been in fantastic agreement with prior analyses of PSO by silver-ion HPLC [51]. HPLC-based approaches to CLnAs evaluation offer a greater number of isomers detected than GC [55,88,91,92]. We found 4 new CLnAs in the PSO, which, to the finest of our understanding, haven’t but been described within the literature: two geometric isomers of 18:3n-2,four,6 (Figures 8D and S2), FAME 18:3n-8,10,12 ten of 22 (Figure 8E), and FAME 18:3n-9,11,13 (Figure 8F). They’re characterized by the double bonds closer for the terminal carbon end (FAME 18:3n-2,four,6) or methyl ester group (FAME 18:3n-8,ten,12 and FAME 18:3n-9,11,13). in Figure 2. Interestingly, two much less abundant isomers together with the same diagnostic fragments have been detected at tR 12.0 min and tR 14.8 min (Supplementary Supplies Figure S2). These Table 3. FAMEs identified in TG fraction of pomegranate seed oil. species had been isomers using the same double bond positions but different double bond getR (min) References ometries.FAME The geometrical isomers of punicicRel. Peak Area FAME 18:3n-5t,7t,9c (-eleosacid, namely tearic acid); FAME 18:3n-5c,7t,9t (-eleostearic acid), and FAME 18:3n-5t,7t,9t (catalpic 18:3n-5,7,9 11.two 36.0 [51,86,87,89,935] acid) were detected in pomegranate seed oil previously [51,869]. Because the elution from the 18:3n-5,7,9 12.0 24.2 [51,86,87,89] 18:3n-4,6,8 two.5 FAs in reversed-phase systems 12.three proceeds from cis to trans isomers [20,90], the [51] eluting later 18:3n-3,five,7 12.9 1.7 [51] isomers most likely contained a higher quantity of trans double bonds. The MS/MS spectra of 13.6 1.5 [86,87,89,935] FAMEs 18:2n-6,9 conjugated double bonds in diverse positions are shown in Figure 8Bwith 3 18:3n-2,4,6 13.7 1.0 F. In all of them, the MBR value was 133, plus the diagnostic fragments allowed us to 18:3n-5,7,9 14.eight 9.4 [51,86,87,89] interpret them as FAME 18:3n-4,6,8 (Figure 8B), FAM.