title: Evaluation of multiple reaction monitoring cubed for the analysis of tachykinin related peptides in rat spinal cord using a hybrid triple quadrupole-linear ion trap mass spectrometer.
abstract: Targeted peptide methods generally use HPLC-MS/MRM approaches. Although dependent on the instrumental resolution, interferences may occur while performing analysis of complex biological matrices. HPLC-MS/MRM(3) is a technique, which provides a significantly better selectivity, compared with HPLC-MS/MRM assay. HPLC-MS/MRM(3) allows the detection and quantitation by enriching standard MRM with secondary product ions that are generated within the linear ion trap. Substance P (SP) and neurokinin A (NKA) are tachykinin peptides playing a central role in pain transmission. The objective of this study was to verify whether HPLC-MS/MRM(3) could provide significant advantages over a more traditional HPLC-MS/MRM assay for the quantification of SP and NKA in rat spinal cord. The results suggest that reconstructed MRM(3) chromatograms display significant improvements with the nearly complete elimination of interfering peaks but the sensitivity (i.e. signal-to-noise ratio) was severely reduced. The precision (%CV) observed was between 3.5% and 24.1% using HPLC-MS/MRM and in the range of 4.3-13.1% with HPLC-MS/MRM(3), for SP and NKA. The observed accuracy was within 10% of the theoretical concentrations tested. HPLC-MS/MRM(3) may improve the assay sensitivity to detect difference between samples by reducing significantly the potential of interferences and therefore reduce instrumental errors.
authors.affiliation: Groupe de Recherche en Pharmacologie Animal du Québec (GREPAQ), Département de Biomédecine Vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
authors.forename: Floriane
authors.initials: F
authors.lastname: Pailleux
authors.affiliation: Groupe de Recherche en Pharmacologie Animal du Québec (GREPAQ), Département de Biomédecine Vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada. Electronic address: francis.beaudry@umontreal.ca.
authors.forename: Francis
authors.initials: F
authors.lastname: Beaudry
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D051381:Rats; D013116:Spinal Cord / Q000737:chemistry*; D015320:Tachykinins / Q000737:chemistry*
chemical_list: D015320:Tachykinins
keywords: Biomarkers; ESI; HPLC; MRM; MRM(3); MS; Mass spectrometry; Multiple reaction monitoring; NKA; Neuropeptides; SP; TFA; Tachykinins; electrospray ionization; high performance liquid chromatography; mass spectrometry; multiple reaction monitoring; multiple reaction monitoring cubed; neurokinin A; substance P; trifluoroacetic acid