Determination of IGF-1 and IGF-2, their degradation products and synthetic analogues in urine by LC-MS/MS
- 15 December 2010
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in The Analyst
- Vol. 136 (5), 1003-1012
- https://doi.org/10.1039/c0an00632g
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.Keywords
This publication has 34 references indexed in Scilit:
- Detection of His-tagged Long-R3-IGF-I in a black market productGrowth Hormone & IGF Research, 2010
- Confiscated black market products and nutritional supplements with non‐approved ingredients analyzed in the cologne doping control laboratory 2009Drug Testing and Analysis, 2010
- IGF-I abuse in sport: Current knowledge and future prospects for detectionGrowth Hormone & IGF Research, 2009
- Urinary insulin-like growth factor 2 identifies the presence of urothelial carcinoma of the bladderBJU International, 2009
- Immunoassays for the measurement of IGF-II, IGFBP-2 and -3, and ICTP as indirect biomarkers of recombinant human growth hormone misuse in sport: Values in selected population of athletesJournal of Pharmaceutical and Biomedical Analysis, 2008
- Growth hormone, IGF‐I and insulin and their abuse in sportBritish Journal of Pharmacology, 2008
- The Role of Insulin-Like Growth Factor Binding ProteinsNeuroendocrinology, 2006
- Molecular interactions of the IGF systemCytokine & Growth Factor Reviews, 2005
- Urinary insulin-like growth factor-I measurement in an actual sport competition, an additional approach in laboratory antidoping testsClinica Chimica Acta; International Journal of Clinical Chemistry, 2004
- IGFs and IGF-binding proteins in short children with steroid-dependent nephrotic syndrome on chronic glucocorticoids: changes with 1 year exogenous GHActa Endocrinologica, 2001