In-Plate Metabolomics of Single Zebrafish Embryos

Single embryos were exposed to the beta blocker propranolol for 120 hpf in 96 well plates, during which apical endpoints were determined through microscopy. The water was then removed and analyzed for its exposure concentration and the embryos were frozen on dry ice. The embryos were then extracted within plates and injected directly from the plates using three different LC-MS methods: Flow-injection lipidomics, C18-Orbitrap and HILIC-Orbitrap. Reproducibility was evaluated using non-exposed embryos from four different 96 well plates incubated a year later than the first exposure plate.