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

摘要

Graphical abstract Display Omitted Highlights ? LC–MS/MS enabled target analysis of MMP3 and MMP9 from human blood serum. ? Comparison of the results obtained by LC–MS/MS and ELISA gave good agreement in case of MMP3. ? The results obtained by LC–MS/MS in case of MMP9 differ in comparison to ELISA results. ? The LC–MS/MS technique enables determination of MMPs signature peptides. 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 (R 2 =0.997) with an LLOD of 0.5ng/mL. For MMP9, linearity was determined in the concentration range from 6.5–65.0ng/mL (R 2 =0.995) with an LLOD of 2.0ng/mL.