Absolute quantification of Atlantic salmon and rainbow trout vitellogenin by the ‘signature peptide’ approach using electrospray ionization QqToF tandem mass spectrometry

In this work, we present a simple method for absolute quantification of plasma vitellogenin from both rainbow trout and Atlantic salmon. Plasma samples obtained from control and beta-estradiol induced fish were digested with trypsin. A characteristic 'signature peptide' was selected and analyzed by high performance liquid chromatography (HPLC) coupled to an electrospray quadrupole-time-of-flight tandem mass spectrometer, using a deuterated homolog peptide as an internal standard. The hybrid tandem mass spectrometer was operated in a 'pseudo' selected reaction-monitoring mode in which three diagnostic product ions were monitored for identification and quantification purposes. The reproducibility (coefficient of variation approximately 5%) and sensitivity (limit of quantification (LOQ) of 0.009 mg/ml) achieved by this simple assay allow it to be considered as an alternative to immunological assays.