Study summary · research use only
Immunoaffinity purification of peptide hormones prior to liquid chromatography-mass spectrometry in doping controls
Plain-language summary
Paraphrased from the published abstract below — not a verdict on whether anything works.
This method-development study describes an immunoaffinity purification approach combined with nano-scale liquid chromatography and high resolution mass spectrometry for detecting prohibited peptide hormones in doping control, using human plasma/serum or urine samples. The method simultaneously purified 12 prohibited peptides, including porcine insulin, Novolog, Apidra, Lantus DesB30-32 metabolite, Humalog, human insulin, Synacthen, luteinizing hormone-releasing hormone (LH-RH), growth hormone releasing hormone (GH-RH(1-29)), CJC-1295, LongR(3)-IGF-1, and IFG-1. The authors report limits of detection in the low pg/mL range in urine and describe the sample preparation as fast and simple, processing approximately 25 samples per day, with materials that are commercially available and applicable to laboratories in other analytical fields.
Abstract
For most peptide hormones prohibited in elite sports the concentrations in plasma or urine are very low (pg/mL). Accordingly, hyphenated purification and enrichment steps prior to mass spectrometric detection are required to obtain sufficient doping control assays. Immunoaffinity purification in combination with nano-scale liquid chromatography coupled to high resolution/high accuracy mass spectrometry was found to have the potential of providing the necessary sensitivity and unambiguous specificity to produce reliable results. With the presented methodology 12 prohibited peptides (porcine insulin, Novolog, Apidra, Lantus DesB30-32 metabolite, Humalog and human insulin, Synacthen (synthetic ACTH analogue), luteinizing hormone-releasing hormone (LH-RH), growth hormone releasing hormone (GH-RH(1-29)) and CJC-1295 (GH-RH analogue), LongR(3)-IGF-1 and IFG-1) were simultaneously purified from plasma/serum or urine. With limits of detection for each target compound ranging in the low pg/mL level (urine), the method enables the determination of urinary peptides at physiologically relevant concentrations. For each class of peptides an appropriate antibody and a respective internal standard was implemented ensuring robust analysis conditions. Due to the fast and simple sample preparation procedure (∼25 samples per day) and the fact that all materials are commercial available, the implementation of the methodology to laboratories from other analytical fields (forensics, pharmacokinetic sciences, etc.) is enabled.
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