Optimised method for determination hypoglycine A in maple plant material by multidimensional gas chromatography/mass spectrometry.

In 2018, more than 50 cases of horse death by equine atypical myopathy (AM) were reported in the Czech Republic. This disease is often associated with the toxin hypoglycine A (HGA), which is found in several maple plant materials. To monitor this toxin in products of these trees that grow in or around horse pastures, a rapid and inexpensive analytical method that can provide the required accuracy is needed. Until now, maple samples have been prepared for gas chromatography using time-consuming methods, with preparation processes taking longer than 1 h. In this work, a shorter method (25 min) with an accuracy of 90-94 %, reproducibility of 2-5%, precision of 3-9%, and linearity, with an R of 0.999, is presented. This sample preparation consists of a procedure without an SPE extraction step and consumes a lower volume of solvent during the extraction. The limit of quantitation for HGA in plant material was improved from 0.5 μg/g of plant material in previous studies to 0.2 μg/g. The method was validated according to the guideline CD 2002/657/EC and ISO 17025, and was found to have good performance characteristics. This simple and rapid method was tested for the monitoring of hypoglycine A level in maple sycamore plant material (seeds, seedlings, and leaves) during the entire growth of the trees.