Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis.
[en] Microfluidic liquid chromatography coupled to a nanoelectrospray source ion trap mass spectrometry was used for the absolute and simultaneous quantitation of C3f and the V65 vitronectin fragment in serum. The method was first carefully optimized and then validated in serum biological matrix. Stable isotopes for the two biomarkers of interest were used as stable isotope labeled peptide standards. A weighted 1/x2 quadratic regression for C3f and a weighted 1/x quadratic regression for the V65 vitronectin peptide were selected for calibration curves. Trueness (with a relative bias <10%), precision (repeatability and intermediate precision <15%) and accuracy (risk <15%) of the method were successfully demonstrated. The linearity of results was validated in the concentration range of 2.5-200ng/mL for C3f and 2.5-100ng/mL for the V65 vitronectin fragment. Serum samples (n=147) classified in 7 groups [(healthy volunteers, OA with 5 grades of severity and rheumatoid arthritis (RA) patients] were analyzed with our new quantitative method. Our data confirm that C3f and the V65 vitronectin fragment are biomarkers of OA severity, but also that C3f fragment is further related to OA severity whereas the V65 vitronectin fragment is more related to early OA detection.
Research Center/Unit :
GIGA-I3 - Giga-Infection, Immunity and Inflammation - ULiège CIRM - Centre Interdisciplinaire de Recherche sur le Médicament - ULiège
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
COBRAIVILLE, Gaël ; Université de Liège > Département des sciences cliniques > Rhumatologie
Fillet, Marianne ; Université de Liège > Département de pharmacie > Analyse des médicaments
Sharif, Mohammed
Ourradi, Khadija
Nys, Gwenaël ; Université de Liège > Département de pharmacie > Analyse des médicaments
Malaise, Michel ; Université de Liège > Département des sciences cliniques > Rhumatologie
Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis.
Publication date :
2017
Journal title :
Talanta
ISSN :
0039-9140
eISSN :
1873-3573
Publisher :
Elsevier, Netherlands
Volume :
169
Pages :
170-180
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright (c) 2017 Elsevier B.V. All rights reserved.
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