Positional Assignment of C-C Double Bonds in Fatty Acyl Chains of Intact Arsenosugar Phospholipids Occurring in Seaweed Extracts by Epoxidation Reactions.

Water-soluble diacyl arsenosugar phospholipids (As-PL) are natural products widespread in marine animals and algae, including the brown alga , also known as wakame. The systematic recognition of As-PL has been hampered by the lack of standard and of qualitative methods to establish the carbon-carbon double bond positions of unsaturated fatty acyl chains. Here, the epoxidation reaction of fatty acyl substituents of As-PL was carried out with high selectivity by -chloroperoxybenzoic acid and the C-C double bond localization was established by collision-induced dissociation of epoxidized species as deprotonated molecules, [epoM - H]. Reversed-phase liquid chromatography (RPLC) separation and a sequential triple-stage MS (i.e., MS) analysis of unsaturated and epoxidized As-PL were very helpful to characterize the carbon-carbon double bond locations of both -1 and -2 fatty acyl chains, starting from a diagnostic product ion pair with 16.0 Da mass difference. These results indicate that intact As-PL can be annotated in terms of fatty acyl chain composition and in terms of their C-C double bond position(s). Interestingly, hexadecenoic (16:1 Δ) and octadecenoic (18:1 Δ) along with octadecadienoic (18:2 Δ) and octadecatrienoic (18:3 Δ) were found to be the most abundant unsaturated fatty acyl chains of As-PL in the brown alga wakame, thus confirming it as a good source of essential fatty acids with a balanced ω6/ω3 ratio. Although the toxicity of As-including metabolites of algal As-PL is still a matter of debate and needs to be studied in more detail, the described approach can be exploited to assess if As-PL could contribute to the supply of essential fatty acids related to the use of algae as nutritious food.