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Determination of IGF-1 and IGF-2, their degradation products and synthetic analogues in urine by LC-MS/MS.

Authors :
Thomas A
Kohler M
Schänzer W
Delahaut P
Thevis M
Source :
The Analyst [Analyst] 2011 Mar 07; Vol. 136 (5), pp. 1003-12. Date of Electronic Publication: 2010 Dec 15.
Publication Year :
2011

Abstract

Peptide analysis in doping controls by means of nano-UPLC coupled high resolution/high mass accuracy mass spectrometry provides the state-of-the-art technique in modern sports drug testing. The present study describes a recent application of this technique for the qualitative determination of different urinary insulin-like growth factor (IGF) related peptides. After simultaneous isolation by solid phase extraction and magnetic particle-based immunoaffinity purification, target analytes (IGF-1, IGF-2, Des1-3-IGF-1, R(3)-IGF-1 and longR(3)-IGF-1) were separated by nano-liquid chromatography prior to mass spectrometric detection. Endogenously produced IGF-1 and IGF-2, as well as the degradation product Des1-3-IGF-1, were frequently detected in urine samples from healthy volunteers in a concentration range of 20-400 pg mL(-1). The impact of IGF binding proteins (IGFBPs), being also present in urine, was potentially estimated by an additional ultrafiltration step in the sample preparation procedure. The synthetic analogue longR(3)-IGF-1, which is assumed to be subject to misuse by cheating athletes, was also analysed and detected in fortified urine samples. Besides the intact molecule, an N-terminally truncated degradation product Des1-10-longR(3)-IGF-1 was identified as the more stable target for doping controls using urine samples. The method was validated for qualitative purposes considering the parameters specificity, limit of detection (20-50 pg mL(-1)), recovery (10-35%), precision (<20%), linearity, robustness and stability.

Details

Language :
English
ISSN :
1364-5528
Volume :
136
Issue :
5
Database :
MEDLINE
Journal :
The Analyst
Publication Type :
Academic Journal
Accession number :
21157622
Full Text :
https://doi.org/10.1039/c0an00632g