Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 164, Issue -, Pages 590-597Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jpba.2018.11.008
Keywords
Superoxide dismutase 3; LC-MS/MS; Pharmacokinetics; Tryptic digestion
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Funding
- National Research Fund of Korea [NRF-2013M3A9A9050567, NRF-2016M3A9E4947695]
- National Research Foundation of Korea [2013M3A9A9050567] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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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-18 column (2.1 x 50 mm, 3.5 mu m) 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 mu g/mL with a lower limit of quantification of 5 mu g/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. (C) 2018 Elsevier B.V. All rights reserved.
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