Development of an HPTLC method for determination of hypoglycin A in aqueous extracts of seedlings and samaras ofAcerspecies

Abstract

26Hypoglycin A (HGA) is a toxin contained in seeds of the sycamore maple tree (Acer pseudoplatanus). Ingestion of this 27 amino acid causes equine atypical myopathy (AM) in Europe. Another variety, A. negundo, is claimed to be present 28where AM cases were reported in the US. For unknown reasons, occurrence of this disease has increased. It is 29 important to define environmental key factors that may influence toxicity of samaras from Acer species. In addition, the 30 content of HGA in seedlings needs to be determined since AM outbreaks, during autumn period when the seeds fall but 31 also during spring when seeds are germinating. The present study aims to validate a reliable method using high 32 performance thin layer chromatography for determination and comparison of HGA in samaras and seedlings. 33The working range of the method was between 20 µg HGA to 408 µg HGA per ml water, corresponding to 12 -244 34 mg/kg fresh weight or 40 -816 mg/kg dry weight, taking into account of an arbitrary average dry matter content of 35 30%. Instrumental limit of detection and limit of quantification were of 10 µg HGA/ml and 20 µg HGA/ml water, 41They also suggest that detected levels of HGA are highly variable. This confirmed the suitability of the method for 42HGA detection in samaras or seedling. 43

Atypical myopathy in Père David’s deer (Elaphurus davidianus) associated with ingestion of hypoglycin A

“…Quantification of hypoglycin A was performed by HPTLC using a CAMAG TLC scanner 3 at 490 nm with WinCATS 4.3 software (CAMAG, Muttenz, Switzerland). Additional details about the procedure can be found elsewhere (Habyarimana et al, 2017).…”

Potential new sources of hypoglycin A poisoning for equids kept at pasture in spring: a field pilot study

“…Quantification of HGA in plant samples in mg/kg fresh weight was performed by measuring the signal intensity of the dots on the HPTL plates corresponding to HGA using a CAMAG TLC scanner 3 at 490 nm with WinCATSV.4.3 software (CAMAG, Muttenz, Switzerland). Additional details about the procedure can be found elsewhere 16 . No statistical analysis was performed in this field study to compare results among pastures since, on a given date, seedlings were at different stages of growing in each pasture.…”

“…Additional details about the procedure can be found elsewhere. 16 No statistical analysis was performed in this ield study to compare results among pastures since, on a given date, seedlings were at diferent stages of growing in each pasture.…”