HPLC-MS/MS method optimisation for matrix metalloproteinase 3 and matrix metalloproteinase 9 determination in human blood serum using target analysis

Petra Kotnik; Metka Koren Krajnc; Artur Pahor; Matjaž Finšgar; Željko Knez

doi:10.1016/j.jpba.2017.11.067

HPLC-MS/MS method optimisation for matrix metalloproteinase 3 and matrix metalloproteinase 9 determination in human blood serum using target analysis

J Pharm Biomed Anal. 2018 Feb 20:150:137-143. doi: 10.1016/j.jpba.2017.11.067. Epub 2017 Nov 29.

Authors

Petra Kotnik¹, Metka Koren Krajnc², Artur Pahor³, Matjaž Finšgar⁴, Željko Knez⁵

Affiliations

¹ Department of Chemistry, Faculty of Medicine, University of Maribor, Taborska 8, SI-2000 Maribor, Slovenia; Laboratory for Separation Processes and Product Design, Faculty of Chemistry and Chemical Engineering, University of Maribor, Smetanova 17, SI-2000 Maribor, Slovenia.
² Clinic of Internal Medicine, Department of Rheumatology, University Medical Centre Maribor, Ljubljanska cesta 5, SI-2000 Maribor, Slovenia.
³ Clinic of Internal Medicine, Department of Rheumatology, University Medical Centre Maribor, Ljubljanska cesta 5, SI-2000 Maribor, Slovenia; Department of Internal Medicine, Faculty of Medicine, University of Maribor, Taborska 8, SI-2000 Maribor, Slovenia.
⁴ Laboratory for Analytical Chemistry and Industrial Analysis, Faculty of Chemistry and Chemical Engineering, University of Maribor, Smetanova 17, SI-2000 Maribor, Slovenia.
⁵ Department of Chemistry, Faculty of Medicine, University of Maribor, Taborska 8, SI-2000 Maribor, Slovenia; Laboratory for Separation Processes and Product Design, Faculty of Chemistry and Chemical Engineering, University of Maribor, Smetanova 17, SI-2000 Maribor, Slovenia. Electronic address: zeljko.knez@um.si.

PMID: 29223062
DOI: 10.1016/j.jpba.2017.11.067

Abstract

A quantitative analysis of zinc endopeptidases matrix metalloproteinase 9 (MMP9) and matrix metalloproteinase 3 (MMP3) from human blood serum are presented. Both matrix metalloproteinases (MMP) are present in human blood serum and can be used as biomarkers for different diseases. The analysis was performed using LC-MS/MS with a triple quadrupole mass spectrometer, based on two specific peptides of each MMP in comparison with an enzyme-linked immunosorbent assay (ELISA). While the conditions for the LC-MS/MS analysis of MMP9 peptides were previously reported for bronchoalveolar lavage fluid, the analysis of MMP3 peptides was newly quantified for human blood serum herein for the first time. For MMP3, the linear behaviour was determined in the concentration range from 1.0-200.0ng/mL (R²=0.997) with an LLOD of 0.5ng/mL. For MMP9, linearity was determined in the concentration range from 6.5-65.0ng/mL (R²=0.995) with an LLOD of 2.0ng/mL.

Keywords: ELISA; Human blood serum; LC–MS/MS; Matrix metalloproteinase 3; Matrix metalloproteinase 9.

Publication types

Comparative Study

MeSH terms

Chromatography, High Pressure Liquid / methods*
Enzyme-Linked Immunosorbent Assay
Humans
Limit of Detection
Matrix Metalloproteinase 3 / blood*
Matrix Metalloproteinase 9 / blood*
Tandem Mass Spectrometry / methods*

Substances

MMP3 protein, human
Matrix Metalloproteinase 3
MMP9 protein, human
Matrix Metalloproteinase 9