The embryos
were prepared through
an in-plate extraction method described previously.14 (link) In short, 120 μL of a MeOH:chloroform (80:20) mixture
containing an IS was added to each well along with mixed-size stainless
steel beads. A silicone lid with a polytetrafluorethylene surface
layer was glued onto the plates prior to homogenization, sonication,
and centrifugation. Following extraction, the samples were subjected
to both targeted lipidomics and nontarget metabolomics analysis.13 (link) For the lipidomics analysis, plates were fitted
directly into the autosampler of the same UHPLC–MS/MS system
used for dose characterization (see previous section). After the flow-injection
lipidomics analysis, the plate was moved to the autosampler of another
Ultimate3000 UHPLC fitted with a hydrophilic interaction liquid chromatography
(HILIC) column (BEH amide; Waters, USA) connected to a Q Exactive
Orbitrap HRMS (Thermo, USA) via an electrospray ionization source.
The instrument was operated in positive mode utilizing nontargeted
full scan acquisition with data-dependent MS2 analysis of the three
highest-intensity features.
were prepared through
an in-plate extraction method described previously.14 (link) In short, 120 μL of a MeOH:chloroform (80:20) mixture
containing an IS was added to each well along with mixed-size stainless
steel beads. A silicone lid with a polytetrafluorethylene surface
layer was glued onto the plates prior to homogenization, sonication,
and centrifugation. Following extraction, the samples were subjected
to both targeted lipidomics and nontarget metabolomics analysis.13 (link) For the lipidomics analysis, plates were fitted
directly into the autosampler of the same UHPLC–MS/MS system
used for dose characterization (see previous section). After the flow-injection
lipidomics analysis, the plate was moved to the autosampler of another
Ultimate3000 UHPLC fitted with a hydrophilic interaction liquid chromatography
(HILIC) column (BEH amide; Waters, USA) connected to a Q Exactive
Orbitrap HRMS (Thermo, USA) via an electrospray ionization source.
The instrument was operated in positive mode utilizing nontargeted
full scan acquisition with data-dependent MS2 analysis of the three
highest-intensity features.
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