Determination of 5-OH-DMT (bufotenine) in the Asian toad Duttyphrynus melanostictus, Hoplobatrachus tigerinus and Haemopis sanguisiga in Madagascar by UHPLC-MS/MS

5-hydroxy-N,N-dimethyltryptamine (5-OH-DMT), known as bufotenine, is proven to have psychoactive effects in high concentrations. Duttyphrynus melanostictus, which produces bufotenine, has migrated to the city of Toamasina in Madagascar, thus, the determination bufotenine's levels in the species' samples is necessary. This study aimed to quantify bufotenine in eggs, tadpoles and toad's skin samples of the Duttyphrynus melanostictus as well as in its predator Hoplobatrachus tigerinus and Haemopis sanguisiga. Two extraction protocols were applied depending on the sample type both based on liquid liquid extraction. Spiked calibration samples were prepared at six concentration levels (10-1000 ng/mL), and performance parameters were evaluated: linearity, accuracy (bias%), precision (CV%) and limit of detection. The proposed protocol is simple and quick, and the UHPLC-MS/MS system used exhibited adequate sensitivity. The target analyte was detected in all samples except for the eggs of Duttyphrynus melanostictus, with concentrations increasing in line with the animal's development.