Determination of ptaquiloside and pterosin B derived from bracken (Pteridium aquilinum) in cattle plasma, urine and milk

Abstract

Ptaquiloside (PTA) is a toxin from bracken fern (Pteridium sp.) with genotoxic effects. Hydrolysis of PTA leads to the non-toxic and aromatised indanone, pterosin B (PTB). Here we present a sensitive, fast, simple and direct method, using SPE cartridges to clean and pre-concentrate PTA and PTB in plasma, urine and milk followed by LC-MS quantification. The average recovery of PTA in plasma, urine, and milk was 71, 88 and 77%, respectively, whereas recovery of PTB was 75, 82 and 63%. The method LOQ for PTA and PTB in plasma was 1.2 and 3.7ngmL(-1), 52 and 33ngmL(-1) for undiluted urine and 5.8 and 5.3ngmL(-1) for milk. The method is repeatable within and between days, with RSD values lower than 15% (PTA) and 20% (PTB). When PTA and PTB spiked samples were stored at -18°C for 14 days both compounds remained stable. In contrast, the PTA concentration was reduced by 15% when PTA spiked plasma was left for 5h at room temperature before SPE clean-up, whereas PTB remained stable. The method is the first to allow simultaneous quantification of PTA and PTB in biological fluids in a relevant concentration range. After intravenous administration of 0.092mg PTA per kgbw in a heifer, the plasma concentration was more than 300ngmL(-1) PTA and declined to 9.8ngmL(-1) after 6h, PTB was determined after 10min at 50ngmL(-1.)

Keywords: Bracken fern; Cattle; LC–MS; Ptaquiloside; Pterosin B.