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Liquid chromatography-tandem mass spectrometry of recombinant human extracellular superoxide dismutase (rhSOD3) in mouse plasma and its application to pharmacokinetic study.
- Source :
-
Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 2019 Feb 05; Vol. 164, pp. 590-597. Date of Electronic Publication: 2018 Nov 15. - Publication Year :
- 2019
-
Abstract
- The antioxidant enzyme human extracellular superoxide dismutase (SOD3) is a promising biopharmaceutical candidate for the treatment of various diseases. To support the early development of SOD3 as a biopharmaceutical, a simple, sensitive, and rapid liquid chromatography tandem mass spectrometry procedure was developed and validated for the determination of SOD3 levels in the plasma of ICR mice. After purification with Ni-NTA magnetic beads and digestion with trypsin, SOD3 signature peptides and internal standard signature peptide (ISP) were separated via high performance liquid chromatography using a Zorbax C <subscript>18</subscript> column (2.1 × 50 mm, 3.5 μm) and a mobile phase consisting of 10 mM ammonium formate, 0.1% formic acid, and acetonitrile. The analyte and ISP were detected via a tandem mass spectrometer in electrospray ionization and multiple reaction monitoring modes to select both the signature peptide for SOD3 at m/z 669 to 969 and the ISP at m/z 655 to 941 in the positive ion mode. The calibration curves were linear (r > 0.99) between 5 and 1000 μg/mL with a lower limit of quantification of 5 μg/mL. The relative standard deviation ranged from 3.08 to 8.84% while the relative error ranged from -0.13 to -9.56%. This method was successfully applied to a preclinical pharmacokinetic study of SOD3 in male ICR mice.<br /> (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Subjects :
- Animals
Biological Products blood
Chemical Fractionation instrumentation
Chromatography, High Pressure Liquid instrumentation
Chromatography, High Pressure Liquid methods
HEK293 Cells
Humans
Injections, Intravenous
Male
Mice
Mice, Inbred ICR
Models, Animal
Peptides blood
Peptides isolation & purification
Recombinant Proteins administration & dosage
Recombinant Proteins blood
Recombinant Proteins isolation & purification
Recombinant Proteins pharmacokinetics
Reference Standards
Reproducibility of Results
Specific Pathogen-Free Organisms
Spectrometry, Mass, Electrospray Ionization instrumentation
Spectrometry, Mass, Electrospray Ionization methods
Superoxide Dismutase administration & dosage
Superoxide Dismutase blood
Superoxide Dismutase isolation & purification
Tandem Mass Spectrometry instrumentation
Tandem Mass Spectrometry methods
Biological Products pharmacokinetics
Chemical Fractionation methods
Superoxide Dismutase pharmacokinetics
Subjects
Details
- Language :
- English
- ISSN :
- 1873-264X
- Volume :
- 164
- Database :
- MEDLINE
- Journal :
- Journal of pharmaceutical and biomedical analysis
- Publication Type :
- Academic Journal
- Accession number :
- 30469108
- Full Text :
- https://doi.org/10.1016/j.jpba.2018.11.008