Coupling Stable Isotope Labeling and Liquid Chromatography-Trapped
Ion Mobility Spectrometry-Time-of-Flight-Tandem Mass Spectrometry
for De Novo Mosquito Ovarian Lipid Studies
posted on 2022-04-14, 12:04authored byLilian
V. Tose, Cesar E. Ramirez, Veronika Michalkova, Marcela Nouzova, Fernando G. Noriega, Francisco Fernandez-Lima
There is a need to better understand
lipid metabolism during mosquito
ovarian development. Lipids are the major source of energy supporting
ovarian follicles development in mosquitoes. In this paper, we describe
the complementary use of stable isotope labeling (SIL) and high-resolution
mass spectrometry-based tools for the investigation of de
novo triglycerides (TG) and diglycerides (DG) during the
ovarian previtellogenic (PVG) stage (4–6 days posteclosion)
of female adult Aedes aegypti. Liquid chromatography
coupled to high-resolution trapped ion mobility spectrometry-parallel
accumulation sequential fragmentation-time-of-flight tandem mass spectrometry
(LC-TIMS-PASEF-TOF MS/MS) allowed the separation and quantification
of nonlabeled and 2H/13C-labeled TG and DG species.
Three SIL strategies were evaluated (H2O/2H2O with 50:50 and 95:5 mixtures, 13C-sucrose, and 13C-glucose). Results showed wide applicability with no signs
of lipid ovarian impairment by SIL induced toxicity. The analytical
workflow based on LC-TIMS-TOF MS/MS provided high confidence and high
reproducibility for lipid DG and TG identification and SIL incorporation
based on their separation by retention time (RT), collision cross
section (CCS), and accurate m/z.
In addition, the SIL fatty acid chain incorporation was evaluated
using PASEF MS/MS. The 2H/13C incorporation
into the mosquito diet provided information on how TG lipids are consumed,
stored, and recycled during the PVG stage of ovarian development.