Comparison of LC-MS methods for the quantitation of ciguatoxins in fish – A collaborative study

Abstract

Ciguatoxins (CTXs) are marine biotoxins that can contaminate seafood and if consumed can result in ciguatera poisoning (CP). The analysis of CTXs is challenging, as they occur in complex tissue matrices, cause CP symptoms at trace amounts (<1 μg kg−1), and certified reference materials are not available. Currently, no standard operating protocols exist for sample preparation or instrumental analysis. Five laboratories worldwide participated in this first-time method comparison study, which used sample extracts containing different CTX groups (CTX4A group, CTX3C group, C-CTX-1) to identify factors impacting CTX quantitation using LC-MS/MS and LC-HRMS. Matrix effects were found to significantly influence CTX quantitation, along with factors such as instrument, eluents, or selected precursor ion. CTXs were quantified using commercially available, non-certified CTX1B and CTX3C standards. Analogues of the CTX groups behaved differently with regard to matrix effects and suitable calibrants with differences between laboratories exceeding a factor of 10 in some cases.