University of Copenhagen, Faculty of Health and Medical Sciences, Department of Veterinary Disease Biology, Ridebanevej 9, DK-1870 Frederiksberg, Denmark; University of Copenhagen, Faculty of Science, Department of Plant and Environmental Sciences, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark.
University of Copenhagen, Faculty of Science, Department of Plant and Environmental Sciences, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Mar 1;951-952:44-51. doi: 10.1016/j.jchromb.2014.01.022. Epub 2014 Jan 23.
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.)
鹿蹄草吡喃酮（PTA）是来自蕨类植物（水龙骨科）的一种具有遗传毒性的毒素。PTA 水解后会生成无毒且芳构化的茚满酮，即鹿蹄草素 B（PTB）。本文介绍了一种灵敏、快速、简单且直接的方法，使用 SPE 小柱对血浆、尿液和牛奶中的 PTA 和 PTB 进行净化和预浓缩，然后采用 LC-MS 进行定量分析。PTA 在血浆、尿液和牛奶中的平均回收率分别为 71%、88%和 77%，而 PTB 的回收率分别为 75%、82%和 63%。PTA 和 PTB 在血浆中的方法定量下限分别为 1.2 和 3.7ngmL(-1)、未稀释尿液中为 52 和 33ngmL(-1)、牛奶中为 5.8 和 5.3ngmL(-1)。该方法日内和日间重复性良好，RSD 值均低于 15%（PTA）和 20%（PTB）。当 PTA 和 PTB 加标样品在-18°C 下储存 14 天时，两种化合物均保持稳定。相比之下，当在 SPE 净化前将 PTA 加标血浆在室温下放置 5 小时时，PTA 的浓度降低了 15%，而 PTB 保持稳定。该方法首次能够在相关浓度范围内同时定量检测生物体液中的 PTA 和 PTB。在一头小母牛中静脉注射 0.092mg PTA/kgbw 后，PTA 的血浆浓度超过 300ngmL(-1)，6 小时后降至 9.8ngmL(-1)，10 分钟后即可检测到 50ngmL(-1)的 PTB。
