Discovery of gymnodimine fatty acid ester metabolites in shellfish using liquid chromatography/mass spectrometry

Abstract

Gymnodimines (GYMs) are fast-acting toxins that belong to the cyclic imine group, a subclass of lipophilic marine toxins. GYMs are considered to be emerging toxins but have not yet been linked to incidents of human poisoning, Limited knowledge on the metabolism of GYMs means that a proper risk assessment has not been possible and caution must be taken when establishing the relevance of GYMs in terms of food safety of marine products. A series of mass spectrometric experiments involving precursor and product ion scans, selected reaction monitoring (SRM), and high-resolution mass spectrometry (MS) were used to detect and confirm 10-O-acyl esters of gymnodimine-A (1). We have detected for the first time the presence of a range of acyl ester derivatives of GYMs in shellfish samples from the Gulf of Gabes, Tunisia. The MS fragmentation pathways of 1 and its esters were also elucidated. Partial synthesis of a palmitic acid ester of 1 facilitated confirmation of identity and calibration of SRM analyses. Evidence of acyl ester metabolites of gymnodimine-B and -C was also obtained. A semi-quantitative analysis indicated that the majority of GYMs present in the sample were in the acylated form (>90%), suggesting that these compounds must not be neglected when trying to understand the risks associated with GYMs. There is a clear need for toxicology studies on these esters and assessment of bio-availability to humans.