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.
Department of Biochemistry, Memorial University of Newfoundland, St John's, Newfoundland, A1C 5S7, Canada.
Alejandro M
AM
Cohen
Atef A H
AA
Mansour
Joseph H
JH
Banoub
D000595:Amino Acid Sequence; D000818:Animals; D017756:Aquaculture; D002853:Chromatography, Liquid; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D008297:Male; D017686:Oncorhynchus mykiss / Q000097:blood*; D010455:Peptides / Q000032:analysis / Q000097:blood / Q000737:chemistry; D012098:Reproduction; D012474:Salmon / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization; D014819:Vitellogenins / Q000032:analysis / Q000097:blood* / Q000737:chemistry
D010455:Peptides; D014819:Vitellogenins; D004958:Estradiol