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Determination of Synacthen in human plasma using immunoaffinity purification and liquid chromatography/tandem mass spectrometry
Author(s) -
Thevis Mario,
Bredehöft Michael,
Geyer Hans,
Kamber Matthias,
Delahaut Philippe,
Schänzer Wilhelm
Publication year - 2006
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2774
Subject(s) - chemistry , chromatography , orbitrap , analyte , mass spectrometry , electrospray ionization , liquid chromatography–mass spectrometry , tandem mass spectrometry , detection limit , extraction (chemistry)
Synacthen is a synthetic analogue to human adrenocorticotropin, which plays an important physiological role by stimulating production of cortisol. In sports, corticosteroids as well as releasing factors (corticotropins) are prohibited according to the regulations of the World Anti‐Doping Agency, and the misuse of Synacthen has been reported several times. Hence, an assay enabling the detection of Synacthen in doping control samples has been developed using immunoaffinity chromatographic isolation of Synacthen from human plasma combined with a concentration of collected fractions using solid‐phase extraction. Unambiguous determination of the target analyte was accomplished using microbore liquid chromatography/electrospray ionization tandem mass spectrometry. Diagnostic product ions such as m/z 223 were characterized using high‐resolution/high‐accuracy Orbitrap mass spectrometry and employed for triple quadrupole MS/MS analysis. The established assay requiring 2 mL of plasma allowed a lower limit of detection (LLOD) at 100 fmol/mL, a recovery of 97% and a precision at the LLOD <20%. Authentic plasma samples obtained from a patient undergoing a standard short Synacthen test were used to prove the applicability of the developed procedure. Copyright © 2006 John Wiley & Sons, Ltd.