Universität zu Köln

Thomas, Andreas ORCID: 0000-0003-1199-0743, Walpurgis, Katja, Tretzel, Laura, Brinkkoetter, Paul, Fichant, Eric, Delahaut, Philippe, Schaenzer, Wilhelm and Thevis, Mario (2015). Expanded test method for peptides > 2 kDa employing immunoaffinity purification and LC-HRMS/MS. Drug Test. Anal., 7 (11-12). S. 990 - 999. HOBOKEN: WILEY. ISSN 1942-7611

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Abstract

Bioactive peptides with an approximate molecular mass of 2-12 kDa are of considerable relevance in sports drug testing. Such peptides have been used to manipulate several potential performance-enhancing processes in the athlete's body and include for example growth hormone releasing hormones (sermorelin, CJC-1293, CJC-1295, tesamorelin), synthetic/animal insulins (lispro, aspart, glulisine, glargine, detemir, degludec, bovine and porcine insulin), synthetic ACTH (synacthen), synthetic IGF-I (longR(3)-IGF-I) and mechano growth factors (human MGF, modified human MGF, full-length' MGF). A combined initial test method using one analytical procedure is a desirable tool in doping controls and related disciplines as requests for higher sample throughput with utmost comprehensiveness preferably at reduced costs are constantly issued. An approach modified from an earlier assay proved fit-for-purpose employing pre-concentration of all target analytes by means of ultrafiltration, immunoaffinity purification with coated paramagnetic beads, nano-ultra high performance liquid chromatography (UHPLC) separation, and subsequent detection by means of high resolution tandem mass spectrometry. The method was shown to be applicable to blood and urine samples, which represent the most common doping control specimens. The method was validated considering the parameters specificity, recovery (11-69%), linearity, imprecision (<25%), limit of detection (5-100 pg in urine, 0.1-2 ng in plasma), and ion suppression. The analysis of administration study samples for insulin degludec, detemir, aspart, and synacthen provided the essential data for the proof-of-principle of the method. Copyright (c) 2015 John Wiley & Sons, Ltd.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Walpurgis, Katja UNSPECIFIED UNSPECIFIED UNSPECIFIED Tretzel, Laura UNSPECIFIED UNSPECIFIED UNSPECIFIED Brinkkoetter, Paul UNSPECIFIED UNSPECIFIED UNSPECIFIED Fichant, Eric UNSPECIFIED UNSPECIFIED UNSPECIFIED Delahaut, Philippe UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaenzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-387646
DOI:	10.1002/dta.1868
Journal or Publication Title:	Drug Test. Anal.
Volume:	7
Number:	11-12
Page Range:	S. 990 - 999
Date:	2015
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; DOPING CONTROL PURPOSES; LIQUID-CHROMATOGRAPHY; HUMAN URINE; HUMAN INSULIN; ANALOGS; IDENTIFICATION; HORMONES; PLASMA; POPULATION Multiple languages Biochemical Research Methods; Chemistry, Analytical; Pharmacology & Pharmacy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/38764