Doping control analysis of intact rapid-acting insulin analogues in human urine by liquid chromatography-tandem mass spectrometry

Analytical Chemistry
Mario ThevisWilhelm Schänzer

Abstract

Insulin and related synthetic therapeutics have been prohibited by the World Anti-Doping Agency for athletes demonstrably not suffering from diabetes mellitus. The primary specimen for doping controls has been urine, but the renal excretion of intact human insulin as well as synthetic analogues such as the rapid-acting products Humalog LisPro, Novolog Aspart, and Apidra Glulisine has been reported negligible owing to metabolic degradation. Nevertheless, employing solid-phase extraction in combination with immunoaffinity purification followed by a top-down sequencing-based mass spectrometric approach, an assay was established allowing the identification of three intact rapid-acting synthetic insulins in doping control urine samples. A volume of 25 mL of urine was concentrated, insulin analogues were isolated from the concentrate by immunoaffinity chromatography, and the eluate was analyzed using microbore liquid chromatography/tandem mass spectrometry. Characteristic product ion spectra obtained from 5-fold protonated intact analytes as well as isolated insulin B-chains allowed the unambiguous identification of target analytes with detection limits of 0.05 ng/mL (9 fmol/mL). Moreover, assay validation demonstrated recoveries bet...Continue Reading

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Citations

Feb 1, 2007·Analytical and Bioanalytical Chemistry·Mario Thevis, Wilhelm Schänzer
Feb 16, 2008·Analytical and Bioanalytical Chemistry·Marie-Agnes PopotJean-Claude Tabet
Oct 12, 2010·International Journal of Legal Medicine·Frank MusshoffBurkhard Madea
Jul 20, 2010·Annual Review of Analytical Chemistry·Larry D Bowers
Apr 4, 2008·Asian Journal of Andrology·Osquel BarrosoOlivier Rabin
Jul 15, 2011·Forensic Science International : Synergy·Mario ThevisWilhelm Schänzer
Sep 19, 2015·Drug Testing and Analysis·Andreas ThomasMario Thevis
Jun 5, 2007·Endocrinology and Metabolism Clinics of North America·Paul C Carpenter
Apr 1, 2008·British Journal of Pharmacology·R I G Holt, P H Sönksen
Nov 22, 2011·Drug Testing and Analysis·Mario ThevisIlkka Ojanperä
Apr 2, 2009·Organogenesis·Mario Thevis, Wilhelm Schänzer
Dec 10, 2013·Forensic Science International : Synergy·Ilkka OjanperäMario Thevis
Jun 8, 2007·The Analyst·Mario Thevis, Wilhelm Schänzer
Jan 13, 2009·European Journal of Mass Spectrometry·Václav RancKarel Lemr
Nov 14, 2006·Rapid Communications in Mass Spectrometry : RCM·Mario ThevisWilhelm Schänzer
Dec 1, 2006·Journal of Mass Spectrometry : JMS
Jan 4, 2007·Rapid Communications in Mass Spectrometry : RCM·Mario ThevisWilhelm Schänzer
Nov 15, 2007·Mass Spectrometry Reviews·Mario ThevisWilhelm Schänzer
Feb 1, 2009·Drug Testing and Analysis·Andreas ThomasMario Thevis
May 10, 2020·Drug Testing and Analysis·Péter JudákKoen Deventer
Mar 8, 2020·Drug Testing and Analysis·Luca NarduzziCorinne Buisson
Nov 11, 2014·Expert Review of Proteomics·Mario ThevisWilhelm Schänzer
Oct 24, 2013·Clinical Chemistry and Laboratory Medicine : CCLM·Béatrice HeurtaultArnaud Agin
Feb 7, 2007·Rapid Communications in Mass Spectrometry : RCM·Katleen Van UytfangheLinda M Thienpont
Aug 17, 2020·Journal of Chromatography. a·M AndrasiA Gaspar

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