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Issue 5, 2011
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Determination of IGF-1 and IGF-2, their degradation products and synthetic analogues in urine by LC-MS/MS

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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, R3-IGF-1 and longR3-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 longR3-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-longR3-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.

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

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Publication details

The article was received on 13 Aug 2010, accepted on 19 Nov 2010 and first published on 15 Dec 2010


Article type: Paper
DOI: 10.1039/C0AN00632G
Citation: Analyst, 2011,136, 1003-1012
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    Determination of IGF-1 and IGF-2, their degradation products and synthetic analogues in urine by LC-MS/MS

    A. Thomas, M. Kohler, W. Schänzer, P. Delahaut and M. Thevis, Analyst, 2011, 136, 1003
    DOI: 10.1039/C0AN00632G

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