Design of experiments for the optimization of electrospray ionization in the LC‐MS/MS analysis of ciguatoxins

Liquid chromatography (LC) coupled to mass spectrometry (MS) is being widely applied as an analytical tool in the field of marine biotoxins both for regulated and for new and emerging compounds. LC‐MS/MS recently became the reference method for the control of lipophilic toxins in the European Union, and new methods are being developed and optimized to extend the applicability of this technique to other toxin groups. In this work, conditions for the analysis of ciguatoxins (CTXs) by LC‐MS/MS were investigated using standard solutions of CTX1B and CTX3C, which are structurally representative compounds for the rest of the main congeners of Pacific group toxins (P‐CTXs). Preliminary studies were carried out for the selection of precursor and product ions used for multiple reaction monitoring. Two transitions based on the chemical structures of CTXs were set up, and mass spectrometer parameters were adjusted for selected reactions monitored. The electrospray ionization source has been carefully optimized through a design of experiments that consisted of a two‐level fractional factorial design of resolution IV for the screening of adequate source conditions and of response surface designs for optimization of the main interactions between factors. The statistical approach allowed maximizing the sensitivity on the MS analyzer that provides a good specificity in P‐CTX detection, which can be also used for confirmation purposes.