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Simultaneous analysis of antihyperglycemic small molecule drugs and peptide drugs by means of dual liquid chromatography high-resolution mass spectrometry

  • Aline C. Vollmer , Lea Wagmann , Armin A. Weber and Markus R. Meyer EMAIL logo
Published/Copyright: April 4, 2023
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 61 Issue 7

Abstract

Objectives

The study aimed to evaluate dual liquid chromatography (LC) coupled to high-resolution mass spectrometry (HRMS) for the simultaneous analysis of small and large molecule drugs by development and application of a validated bioanalytical method.

Methods

The oral antihyperglycemic drugs (OAD) dapagliflozin, empagliflozin, glibenclamide, glimepiride, metformin, pioglitazone, repaglinide, saxagliptin, sitagliptin, and vildagliptin, as well as the antihyperglycemic peptides exenatide, human insulin, insulin aspart, insulin degludec, insulin detemir, insulin glargine, insulin glulisine, insulin lispro, and semaglutide were included in the analytical procedure. Analytes were extracted using a combination of protein precipitation and solid-phase extraction. Two identical reversed-phase columns were used for separation followed by Orbitrap high-resolution mass spectrometry. The whole procedure was validated according to international recommendations.

Results

Different MS parameters had to be used for the two analyte groups, but dual LC separation allowed elution of all analytes within 12 min using the same column type. The analytical procedure was accurate and precise for most of the compounds except for exenatide, semaglutide, and insulin glargine, which were included qualitatively in the method. Analysis of proof-of-concept samples revealed OAD concentrations mostly within their therapeutic range, insulins could be detected in five cases but at concentrations below the lower limit of quantification except for one case.

Conclusions

Dual LC in combination with HRMS was shown to be a suitable platform to analyze small and large molecules in parallel and the current method allowed the determination of a total of 19 antihyperglycemic drugs in blood plasma within 12 min.

Keywords: antihyperglycemic drugs; dual liquid chromatography; LC-HRMS/MS; method development; peptides; small molecules
Corresponding author: Prof. Dr. Markus R. Meyer, Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Center for Molecular Signaling (PZMS), Saarland University, Kirrberger Str., Building 46, 66421 Homburg, Germany, Phone: +49 6841 16 26430, Fax: +49 6841 16 26431, E-mail:

Acknowledgments

The authors like to thank Cathy M. Jacobs, Fabian Frankenfeld, Juel Maalouli Shaar, Sascha K. Manier, Matthias J. Richter, Gabriele Ulrich, Claudio De Nardi, Yvonne Song, PD Dr. Andreas Thomas, Dr. Christoph Sauer, and Synlab Deutschland for their support and/or helpful discussion.

  1. Research funding: None declared.

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

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

  4. Informed consent: Not applicable.

  5. Ethical approval: Not applicable.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2022-1316).

Received: 2022-12-27
Accepted: 2023-03-20
Published Online: 2023-04-04
Published in Print: 2023-06-27

© 2023 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2022-1316
Keywords for this article
antihyperglycemic drugs; dual liquid chromatography; LC-HRMS/MS; method development; peptides; small molecules

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  2. Editorial
  3. ChatGPT: Angel or Demond? Critical thinking is still needed
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  12. Analytical aspects of the antinuclear antibody test by HEp-2 indirect immunofluorescence: EFLM report on an international survey
  13. Guidelines and Recommendations
  14. Variability of cardiac troponin levels in normal subjects and in patients with cardiovascular diseases: analytical considerations and clinical relevance
  15. General Clinical Chemistry and Laboratory Medicine
  16. Pre-analytical long-term stability of neopterin and neurofilament light in stored cerebrospinal fluid samples
  17. Development of a candidate reference measurement procedure by ID-LC-MS/MS for total tau protein measurement in cerebrospinal fluid (CSF)
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