Headspace Gas Chromatography: A Technique to Assess Ethanol Concentration in Zebrafish Embryos
Published: April 30, 2023
Abstract
Source: Lovely, C. B. et al. Quantification of Ethanol Levels in Zebrafish Embryos Using Head Space Gas Chromatography. J. Vis. Exp. (2020).
This video demonstrates the use of headspace gas chromatography to determine the ethanol concentrations in zebrafish embryos. This method provides a useful tool for qualitative and quantitative analysis of volatile organic compounds.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Measuring the embryonic volume using water displacement NOTE: In this protocol, 24 h postfertilization (hpf) embryos (Figure 1) are used. The embryos used in the volume measurements are not used in the ethanol analysis. Place 10 embryos and extraembryonic fluid in 1.5 mL micr…
Representative Results
Figure 1: An image of an embryo at 24 h postfertilization (hpf) inside the chorion. The embryo and the yolk are surrounded by the extraembryonic fluid, all located inside the chorion. Figure 2: Protocol to measure embryonic volume and process the embryos for analysis. (A)</…
Declarações
The authors have nothing to disclose.
Materials
| AutoSampler, CP-8400 | Varian | Gas Chromatograph Autosampler | |
| Ethanol | Decon Labs | 2701 | |
| Gas chromatograph vial with polytetrafluoroethylene/silicone septum and plastic cap 2 mL | Agilent | 8010-0198 | Can reuse the vials after cleaning, but not the caps/septa |
| Gas Chromatograph, CP-3800 | Varian | ||
| Helium | Provided by contract to the university | ||
| HP Innowax capillary column | Agilent | 19095N-123I | 30 m x 0.53 mm x 1.0 μm film thick |
| Hyrdogen | Provided by contract to the university | ||
| Microcentrifuge tube 1.5 mL | Fisher Scientific | 2682002 | |
| Micropipette tips 10 μL | Fisher Scientific | 13611106 | |
| Micropipette tips 1000 μL | Fisher Scientific | 13611127 | |
| Micropipette tips 200 μL | Fisher Scientific | 13611112 | |
| Pipetman L p1000L Micropipette | Gilson | FA10006M | |
| Pipetman L p200L Micropipette | Gilson | FA10005M | |
| Pipetman L p2L Micropipette | Gilson | FA10001M | |
| Polytetrafluoroethylene/silicone septum and plastic cap | Agilent | 5190-7021 | Replacement caps/septa for gas chromatograph vials |
| Solid-phase microextraction fiber assembly Carboxen/Polydimethylsiloxane | Millipore Sigma | 57343-U | Replacement fibers |
| Star Chromatography Workstation | Varian | Chromatography software | |
| Thermogreen Low Bleed (LB-2) Septa | Millipore Sigma | 23154 | Replacement inlet septa |
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Citar este artigo
Headspace Gas Chromatography: A Technique to Assess Ethanol Concentration in Zebrafish Embryos. J. Vis. Exp. (Pending Publication), e21104, doi: (2023).