Universität zu Köln

Thomas, Andreas, Yang, Rouxue, Petring, Simon, Bally, Lia and Thevis, Mario (2020). Simplified quantification of insulin, its synthetic analogs and C-peptide in human plasma by means of LC-HRMS. Drug Test. Anal., 12 (3). S. 382 - 391. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

The quantification of peptide hormones by means of liquid chromatography (LC) coupled to mass spectrometry (MS) or other techniques (e.g. immunoassays) has been a challenging task in modern analytical chemistry. Especially for insulin, its synthetic analogs, and C-peptide, reliable determinations are urgently needed due to their diagnostic value in the management of diabetes and insulin resistance and because of the illicit use of insulin as a performance-enhancing agent in professional sports or as an effective toxin in forensic toxicology. The concomitant measurement of C-peptide and insulin offers an established tool for the diagnostic workup of hypoglycemia (endogenous vs. exogenous hyperinsulinemia), characterizing hepatic insulin clearance, and the assessment of beta-cell function (insulin secretion). Thus, the present approach offers the possibility to determine human insulin and its synthetic analogs (lispro, glulisine, aspart, glargine metabolite, degludec, detemir, porcine, and bovine) and C-peptide simultaneously after sample preparation utilizing protein precipitation and a mixed-mode cation-exchange solid-phase extraction, and subsequent detection by LC-high resolution MS. The method was fully validated regarding the following parameters: specificity, limit of detection (0.2 ng/mL), limit of quantification (0.6 ng/mL), recovery (40-90%), accuracy (78-128%), linearity, precision (< 21%), carry over, robustness, and matrix effects. The proof-of-concept was shown by analyzing authentic plasma samples from adults with class II obesity and prediabetes collected in the course of an oral glucose tolerance test. All sample preparation steps were controlled by two stable isotope-labeled internal standards, namely [[H-2(10)] Leu (B6, B11, B15, B17)]-insulin, and [[C-13(6)] Leu (26, 30)] C-peptide.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Thomas, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Yang, Rouxue UNSPECIFIED UNSPECIFIED UNSPECIFIED Petring, Simon UNSPECIFIED UNSPECIFIED UNSPECIFIED Bally, Lia UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-345426
DOI:	10.1002/dta.2765
Journal or Publication Title:	Drug Test. Anal.
Volume:	12
Number:	3
Page Range:	S. 382 - 391
Date:	2020
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1942-7611
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language TANDEM MASS-SPECTROMETRY; IMMUNOAFFINITY PURIFICATION; SERUM Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/34542