Qualitative detection of desmopressin in plasma by liquid chromatography–tandem mass spectrometry

Abstract

This work describes a liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) method for detection of desmopressin in human plasma in the low femtomolar range.Desmopressin is a synthetic analogue of the antidiuretic hormone arginine vasopressin and it might be used by athletes as masking agent in the frame of blood passport controls. Therefore, it was recently added by the World Anti-Doping Agency (WADA) to the list of prohibited substances in sport as a masking agent. Mass spectrometry characterization of desmopressin was performed with a high-resolutionOrbitrap-based mass spectrometer. Detection of the peptide in the biological matrix was achieved using a triple quadrupole instrument with an electrospray ionization (ESI) interface after protein precipitation, weak cation solid phase extraction and HPLC separation with an octadecyl reverse phase column. Identification of desmopressin was performed using three product ions, m/z 328.0, m/z 120.0, and m/z 214.0, from the parent ion, m/z 535.5.The extraction efficiency of the method at the limit of detection was estimated at 40% (n=10), the ion suppression at 5% (n=10), and the limit of detection was 50 pg/ml (S/N> 3).Selectivity of the method was verified against several endogenous and synthetic desmopressin related peptides. The performance and the applicability of the method were tested by analysis of clinical samples after administration of desmopressin via intravenous, oral, and intranasal routes. Only after intravenous administration, desmopressin could be successfully detected.3

Doping control analysis of desmopressin in human urine by LC‐ESI‐MS/MS after urine delipidation

“…b) with good intensities ( m/z 328.0, 120.0, 214.0, 276.1, 430.4 and 526.7). The origin of these fragments has been explained previously (Esposito et al ., ).…”

“…The high elimination rate from blood circulation of desmopressin and the low bioavailability of oral and intranasal formulations (Agerso et al ., ) were probably the main causes of the nondetection of this compound in plasma after oral and intra‐nasal administration, as observed in our previous work (Esposito et al ., ). In fact, desmopressin was only detectable in plasma after intravenous administration.…”

Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

“…Finally, novel, previously unknown macromolecules (i.e., TB500 17-23 fragment), with no current approval for human therapeutic use (i.e., agents under preclinical or clinical development, designer drugs, or compounds approved only for veterinary use), but illegally marketed (e.g., via Internet websites) [5] are included in section S0 ''Non-approved substances''. Different analytical approaches have already been developed and published to detect these compounds in nutritional supplements [6][7][8][9][10] and in doping control samples (either blood [11][12][13][14][15][16][17][18] or urine [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37]), employing immunological [26,[35][36][37], electrophoretic [16][17][18][19]27], or liquid chromatography-mass spectrometry techniques [6-15, 20-25, 28-34] or a combination of different analytical technologies. In general, the most effective analytical approach (combined sample pretreatment and the instrumental method) has to take into ...…”

“…Leu-Arg-Ile-Val-Gln-Cys-Arg-Ser-Val-Glu-Gly-Ser-Cys-Gly-Phe-OH Cys 7 and Cys14 1812.9ARA290 pGlu-Glu-Gln-Leu-Glu-Arg-Ala-Leu-Asn-Ser-Ser-…”

Development of a High-Sensitivity Quantitation Method for Arginine Vasopressin by High-Performance Liquid Chromatography Tandem Mass Spectrometry, and Comparison with Quantitative Values by Radioimmunoassay

“…This fragmentation was favorable because both compounds have a disulfide link between the two cysteine residues, and thus y-type fragmentation occurs, yielding y3 ions as the predominant products. 22 Effect of the mobile-phase solvent on sensitivity Figure 2 shows the effect of the aqueous solvent acid concentration of the mobile phase (using two different organic solvent mobile phases) on the assay sensitivity. We found that a low acid concentration in the aqueous mobile phase produced higher sensitivity for AVP, using methanol or acetonitrile as the organic mobile phase.…”