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Simultaneous quantification and qualification of synacthen in plasma

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Abstract

Tetracosactide (Synacthen), a synthetic analogue of adrenocorticotropic hormone (ACTH), can be used as a doping agent to increase the secretion of glucocorticoids by adrenal glands. The only published method for anti-doping control of this drug in plasma relies on purification by immunoaffinity chromatography and LC/MS/MS analysis. Its limit of detection is 300 pg/mL, which corresponds to the peak value observed 12 h after 1 mg Synacthen IM administration. We report here a more sensitive method based on preparation of plasma by cation exchange chromatography and solid-phase extraction and analysis by LC/MS/MS with positive-mode electrospray ionization using 7–38 ACTH as internal standard. Identification of Synacthen was performed using two product ions, m/z 671.5 and m/z 223.0, from the parent [M + 5H]5+ ion, m/z 587.4. The recovery was estimated at 70%. A linear calibration curve was obtained from 25 to 600 pg/mL (R 2 > 0.99). The lower limit of detection was 8 pg/mL (S/N > 3). The lower limit of quantification was 15 pg/mL (S/N > 10; CV% < 20%). The performance of the method was illustrated by an 8-h kinetic analysis of plasma samples from nine subjects submitted to IM injections of either Synacthen® (five subjects) or Synacthen® Depot, the slow-release form of the drug (four subjects). Concentrations of Synacthen between 16 and 310 pg/mL were observed. A sensitive method for quantitation of Synacthen in plasma is proposed for anti-doping control analyses.

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Acknowledgments

We thank WADA for financing this project. We particularly express our thanks to Professor Katia Collomp who headed the administration study. We are grateful to the CSOB Laboratory for the use of the LTQ Orbitrap and to Chafia Bennaceur for her help in characterizing the peptides by mass spectrometry. This article is dedicated to the memory of Professor Jacques de Ceaurriz.

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Authors and Affiliations

  1. Département des Analyses, Agence Française de Lutte contre le Dopage (afld), 143, avenue Roger Salengro, Châtenay-Malabry, 92290, Paris, France

    Ayman Chaabo, Jacques de Ceaurriz, Corinne Buisson & Françoise Lasne

  2. Université Pierre et Marie Curie—Institut Parisien de Chimie Moléculaire, Chimie Structurale Organique et Biologique (CSOB), (UMR 7201), 4, place Jussieu, 75252, Paris cedex 05, France

    Ayman Chaabo & Jean-Claude Tabet

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  1. Ayman Chaabo
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  2. Jacques de Ceaurriz
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  3. Corinne Buisson
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  4. Jean-Claude Tabet
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  5. Françoise Lasne
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Corresponding author

Correspondence to Françoise Lasne.

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Jacques de Ceaurriz passed away in January 2010.

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Chaabo, A., de Ceaurriz, J., Buisson, C. et al. Simultaneous quantification and qualification of synacthen in plasma. Anal Bioanal Chem 399, 1835–1843 (2011). https://doi.org/10.1007/s00216-010-4565-z

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  • DOI: https://doi.org/10.1007/s00216-010-4565-z

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