Headspace Gas Chromatography: A Technique to Assess Ethanol Concentration in Zebrafish Embryos

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.

1. Place 10 embryos and extraembryonic fluid in 1.5 mL microcentrifuge tube marked at a volume of 250 μL (Figure 2A). Add water to the 250 μL fill line (see sample with dyed water in....

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.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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