On-Line Solid Phase Extraction Liquid Chromatography-Mass Spectrometry Method for Multiplexed Proteins Quantitation in an Ecotoxicology Test Specie: Gammarus fossarum - Archive ouverte HAL Access content directly
Journal Articles Journal of Applied Bioanalysis Year : 2018

On-Line Solid Phase Extraction Liquid Chromatography-Mass Spectrometry Method for Multiplexed Proteins Quantitation in an Ecotoxicology Test Specie: Gammarus fossarum

Arnaud Chaumot

Abstract

Methods: After protein extraction and tryptic digestion, peptides were cleaned-up onto an on-line SPE cartridge (Oasis HLB, 2.1 mm x 20 mm, 25 μm particle size) coupled to a LC-MS/MS system. The SRM assay was performed on a quadrupole-trap mass spectrometer. Results: The method targeted 30 proteins in G. fossarum (46 reporter peptides used in the SRM assay) was developed and validated. The method duration was 30 min including the on-line SPE step saving up to 6h per sample. The method’s performance was validated according to FDA guidelines Conclusions: Our method substitutes valuably conventional methods like off-line SPE performed beforehand to the LC-MS/MS system. This assay offers higher sensitivity with no loss and/or degradation of reporter peptides and reached good specifications (linearity, precision and accuracy).
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hal-01860702 , version 1 (23-02-2024)

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Aurore Charnot, Duarte Gouveia, Sophie Ayciriex, Jérôme Lemoine, J. Armengaud, et al.. On-Line Solid Phase Extraction Liquid Chromatography-Mass Spectrometry Method for Multiplexed Proteins Quantitation in an Ecotoxicology Test Specie: Gammarus fossarum. Journal of Applied Bioanalysis, 2018, 4 (3), pp.81-101. ⟨10.17145/jab.18.012⟩. ⟨hal-01860702⟩
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