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Analysis of B-type natriuretic peptide impurities using label-free data-independent acquisition mass spectrometry technology

  • Peng Xiao ORCID logo EMAIL logo , Fan Zhang , Xinxue Wang , Dewei Song and Hongmei Li EMAIL logo
Published/Copyright: July 27, 2020
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 59 Issue 1

Abstract

Objectives

Synthetic B-type natriuretic peptide (BNP) is employed in most clinical testing platforms as a raw material of calibrator. Characterization of impurities with structures similar (BNPstrimp compounds) to that of BNP is a reasonable way to decrease clinical measurement errors and improve drug safety.

Methods

A novel quantitative method targeted towards BNPstrimp compounds was developed. First, the peptide samples were separated and identified using ultra-performance liquid chromatography, coupled with high-resolution mass spectrometry (MS). To evaluate biological activity further, BNPstrimp immunoaffinity was investigated using western blot (WB) assays. Second, a quantitative label-free data-independent acquisition (DIA) MS approach was developed, and the internal standard peptide (ISP) was hydrolyzed. Absolute quantification was performed using an isotope dilution MS (ID-MS) approach. Third, method precision was investigated using the C-peptide reference material.

Results

Seventeen BNPstrimp compounds were identified in synthetic BNP, and 10 of them were successfully sequenced. The immunoassay results indicated that deaminated, oxidized, and isomerized BNPstrimp compounds exhibited weaker immunoaffinity than intact BNP1-32. The mass fraction of the synthetic solid ISP1-16, quantified by ID-MS, was 853.5 (±17.8) mg/g. Validation results indicated that the developed method was effective and accurate for the quantitation of the well-separated BNP impurities.

Conclusions

The developed approach was easy to perform, and it was suitable for the parallel quantification of low-abundance BNPstrimp compounds when they performed a good separation in liquid chromatography. The quantitative results were comparable and traceable. This approach is a promising tool for BNP product quality and safety assessment.

Keywords: B-type natriuretic peptide (BNP); data-independent acquisition; impurity peptides; mass spectrometry; quantitative research
Corresponding authors: Peng Xiao, Division of Chemical Metrology and Analytical Science, National Institute of Metrology, Beijing, PR China. Phone: +86 10 64228896, Fax: +86 10 64271639, E-mail: ; Hongmei Li, Division of Chemical Metrology and Analytical Science, National Institute of Metrology, Beijing, PR China. Phone: +86 10 64228896, Fax: +86 10 64271639, E-mail:

Funding source: National Natural Science Foundation of China

Award Identifier / Grant number: 21705144

Award Identifier / Grant number: 31800842

Funding source: National Institute of Metrology Fundamental Research Project

Award Identifier / Grant number: AKY1709

Award Identifier / Grant number: AKYZD1802-2

Acknowledgments

This work was supported by the National Natural Science Foundation of China (grant numbers 21705144, 31800842), and also by the National Institute of Metrology Fundamental Research Project (grant number AKY1709, AKYZD1802-2).

  1. Research funding: The National Natural Science Foundation of China (grant numbers 21705144, 31800842), and the National Institute of Metrology Fundamental Research Project (grant number AKY1709, AKYZD1802-2).

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Informed consent: Informed consent was obtained from all individuals included in this study.

  4. Competing interests: Authors state no conflict of interest.

  5. Ethical approval: The local Institutional Review Board deemed the study exempt from review.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2020-0012).

Received: 2020-01-05
Accepted: 2020-07-07
Published Online: 2020-07-27
Published in Print: 2021-01-26

© 2020 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2020-0012
Keywords for this article
B-type natriuretic peptide (BNP); data-independent acquisition; impurity peptides; mass spectrometry; quantitative research

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  1. 10.1515/cclm-2020-frontmatter1
  2. Mario Plebani turns 70
  3. Editorial
  4. Measurement uncertainty and the importance of correlation
  5. Reviews
  6. Untangling the association between prostate-specific antigen and diabetes: a systematic review and meta-analysis
  7. The emerging role of cell senescence in atherosclerosis
  8. Mini Review
  9. Minimum retesting intervals in practice: 10 years experience
  10. Opinion Paper
  11. Cardiotoxic effects and myocardial injury: the search for a more precise definition of drug cardiotoxicity
  12. EFLM Papers
  13. The CRESS checklist for reporting stability studies: on behalf of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for the Preanalytical Phase (WG-PRE)
  14. Thoughts and expectations of young professionals about the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)
  15. Guidelines and Recommendations
  16. Evidence on clinical relevance of cardiovascular risk evaluation in the general population using cardio-specific biomarkers
  17. Genetics and Molecular Diagnostics
  18. A systematic evaluation of stool DNA preparation protocols for colorectal cancer screening via analysis of DNA methylation biomarkers
  19. General Clinical Chemistry and Laboratory Medicine
  20. The new IVD Regulation 2017/746: a case study at a large university hospital laboratory in Belgium demonstrates the need for clarification on the degrees of freedom laboratories have to use lab-developed tests to improve patient care
  21. Evaluation of serum electrolytes measurement through the 6-year trueness verification program in China
  22. Feasibility for aggregation of commutable external quality assessment results to evaluate metrological traceability and agreement among results
  23. Sample size and rejection limits for detecting reagent lot variability: analysis of the applicability of the Clinical and Laboratory Standards Institute (CLSI) EP26-A protocol to real-world clinical chemistry data
  24. Effects of calcium dobesilate (CaD) interference on serum creatinine measurements: a national External Quality Assessment (EQA)-based educational survey of drug-laboratory test interactions
  25. Mass spectrometric sample identification with indicator compounds introduced via labeled sample tubes
  26. Automation and validation of a MALDI-TOF MS (Mass-Fix) replacement of immunofixation electrophoresis in the clinical lab
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