Universität zu Köln

Lopes, Filipe, Cowan, David A., Thevis, Mario, Thomas, Andreas ORCID: 0000-0003-1199-0743 and Parkin, Mark C. (2014). Quantification of intact human insulin-like growth factor-I in serum by nano-ultrahigh-performance liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom., 28 (13). S. 1426 - 1433. HOBOKEN: WILEY. ISSN 1097-0231

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Abstract

RATIONALEInsulin-like growth factor-I is one of the biomarkers used to detect growth hormone administration prohibited in human sport. Current testing approaches for IGF-I rely on commercial immunoassays, which may change from time to time requiring complex revalidation. Mass spectrometry (MS)-based approaches often rely on enzymatically digesting the protein and measuring specific peptide concentrations. In order to reinforce the current available methodology for IGF-I testing, a reliable and equally sensitive MS method is required for the analysis of intact protein using small sample volumes (<25 L). METHODSIGF-I was extracted from human serum samples by a simple protein precipitation procedure. Separation was achieved via nano-ultrahigh-performance liquid chromatography and MS analysis was conducted by nano-electrospray ionisation triple-quadrupole mass spectrometry in the selected reaction monitoring mode using a stable-isotope-labelled internal standard. RESULTSA six-point calibration curve ranging from 50 to 1000ng/mL of human IGF-I in rat serum was used to establish instrument response. The method provided a limit of quantification of 50ng/mL, with intra- and inter-day precision 5% and intra- and inter-day accuracy 95%. CONCLUSIONSA quantitative method was developed for the quantification of intact IGF-I in human serum samples. The data generated provided important information for the development of a new reference method for the growth hormone biomarker test and helped create a reliable system for monitoring peptide hormones in individual athletes, a possible extension to the athlete biological passport system. Nano-electrospray has here been shown to be sufficiently robust for routine use in an analytical laboratory, allowing for the analysis of minute sample volumes. Copyright (c) 2014 John Wiley & Sons, Ltd.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Lopes, Filipe UNSPECIFIED UNSPECIFIED UNSPECIFIED Cowan, David A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Parkin, Mark C. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-433907
DOI:	10.1002/rcm.6908
Journal or Publication Title:	Rapid Commun. Mass Spectrom.
Volume:	28
Number:	13
Page Range:	S. 1426 - 1433
Date:	2014
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1097-0231
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PROTEIN BIOMARKER DISCOVERY; PEPTIDE-HORMONES; IGF-I; IONIZATION; QUANTITATION; CHALLENGES; PLASMA Multiple languages Biochemical Research Methods; Chemistry, Analytical; Spectroscopy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/43390