LC-PDA/APCIitMS identification of algal carotenoid and oxysterol precursors to fatty acid esters in hydrolyzed extracts of the freshwater mussel Dreissena bugensis.

Carotenyl fatty acid esters (carotenyl-FAEs) were extracted in acetone from freeze-dried Dreissena bugensis (Lakes Erie and Ontario) and hydrolyzed to identify the carotenoid precursors. Analysis by liquid chromatography (LC) with photodiode array (PDA) and atmospheric pressure chemical ionization-ion trap mass spectrometry (APCIitMS) revealed the major hydrolysis products: fucoxanthinol (FOH) from fucoxanthin (FX, diatoms); mactraxanthin (MX) from violaxanthin (VX, chlorophytes); 4-fold higher levels of an unknown, tentatively identified as an adduct of two closely eluting CHO and CHO steryl triols. Enzymatic hydrolysis (Candida rugosa) of dreissenid extracts yielded FOH and MX, but residual carotenyl-FAEs remained.

Separation and identification of fatty acid esters of algal carotenoid metabolites in the freshwater mussel Dreissena bugensis, by liquid chromatography with ultraviolet/visible wavelength and mass spectrometric detectors in series.

LC with photodiode array and APCI-ion trap mass spectrometry has made it possible to tentatively identify 76 carotenyl fatty acid esters (cFAEs) in solvent extracts from Dreissena bugensis, collected from Lake Erie: 16 mono- and 33 diFAEs of fucoxanthinol (FOH), and 27 diFAEs of mactraxanthin (MX). FOH and MX, previously identified in cFAE hydrolysates, were confirmed as parent carotenoids of the cFAEs, and as primary metabolites of fucoxanthin and violaxanthin, respectively, derived from diatoms and chlorophytes in the dreissenids' diet. The most abundant fatty acid substituents of cFAEs were 16:0 and 16:1; abundant fatty acid biomarkers were 16:1 and 20:5, from diatoms, and 17:0, from bacteria.