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

This research article presents a faster, more efficient method for detecting the toxin hypoglycine A (HGA) in maple plants, related to a disease causing horse deaths.

Introduction

• This study addresses a significant issue of concern in animal health care, particularly equine health. In 2018, there were numerous cases of equine deaths in the Czech Republic, associated with the disease equine atypical myopathy (AM). This disease has often been linked with the toxin hypoglycine A (HGA), prevalent in some species of maple plants.

The Need for Improved Detection Methods

• The researchers underline the need for a rapid, inexpensive, and accurate method to monitor HGA in maple trees growing around horse pastures. Until now, the maple sample preparation for detecting this toxin through gas chromatography was laborious and time-consuming, taking over an hour to complete.

Proposed Method

• The researchers present a shorter, more efficient method that reduces the sample preparation time to 25 minutes while maintaining accuracy between 90-94%. Moreover, the new technique also boasts exceptional reproducibility (2-5%), precision (3-9%), and linearity (with a coefficient of determination R of 0.999).
• Unlike previous methods, the new system does not require an SPE extraction step. It also manages to reduce the volume of solvent required during extraction, making it more economically and environmentally friendly.

Improvement in Detection Limit

• An important point of advancement in this new approach is the improved limit of quantitation for HGA in plant material. From the previous detection limit of 0.5 μg/g of plant material, the new method is sensitive enough to detect as low as 0.2 μg/g of HGA, enhancing the effectiveness of monitoring efforts.

Validation and Testing

• The new method was validated according to the guideline CD 2002/657/EC and ISO 17025. The results confirmed its good performance characteristics.
• As a final test, the method was applied to monitor the HGA levels in sycamore maple plant materials – including seeds, seedlings, and leaves – throughout the entire growth of the trees.