High-throughput bioaffinity mass spectrometry for screening and identification of designer anabolic steroids in dietary supplements

P. Aqai, E. Cevik, A. Gerssen, W. Haasnoot, M.W.F. Nielen

Research output: Contribution to journalArticleAcademicpeer-review

27 Citations (Scopus)

Abstract

A generic high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of known and unknown recombinant human sex hormone-binding globulin (rhSHBG)-binding designer steroids in dietary supplements. For screening, a semi-automated competitive inhibition binding assay was combined with fast ultrahigh-performance-LC-electrospray ionization-triple-quadrupole-MS (UPLC-QqQ-MS). 17ß-Testosterone-D3 was used as the stable isotope label of which the binding to rhSHBG-coated paramagnetic microbeads was inhibited by any other binding (designer) steroid. The assay was performed in a 96-well plate and combined with the fast LC-MS, 96 measurements could be performed within 4 h. The concentration-dependent inhibition of the label by steroids in buffer and dietary supplements was demonstrated. Following an adjusted bioaffinity isolation procedure, suspect extracts were injected into a chip-UPLC(NanoTile)-Q-time-of-flight-MS system for full-scan accurate mass identification. Next to known steroids, 1-testosterone was identified in three of the supplements studied and the designer steroid tetrahydrogestrinone was identified in a spiked supplement. The generic steroid-binding assay can be used for high-throughput screening of androgens, estrogens, and gestagens in dietary supplements to fight doping. When combined with chip-UPLC-MS, it is a powerful tool for early warning of unknown emerging rhSHBG bioactive designer steroids in dietary supplements.
Original languageEnglish
Pages (from-to)3255-3262
JournalAnalytical Chemistry
Volume85
Issue number6
DOIs
Publication statusPublished - 2013

Fingerprint

Dietary supplements
Testosterone Congeners
Mass spectrometry
Screening
Steroids
Throughput
Sex Hormone-Binding Globulin
Assays
Labels
Electrospray ionization
Liquid chromatography
Progestins
Isotopes
Androgens
Testosterone
Buffers
Estrogens
Doping (additives)

Keywords

  • hormone-binding globulin
  • multi-residue method
  • liquid-chromatography
  • nutritional supplements
  • androgen bioassay
  • discovery
  • ligands
  • urine
  • contamination
  • immunoassay

Cite this

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abstract = "A generic high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of known and unknown recombinant human sex hormone-binding globulin (rhSHBG)-binding designer steroids in dietary supplements. For screening, a semi-automated competitive inhibition binding assay was combined with fast ultrahigh-performance-LC-electrospray ionization-triple-quadrupole-MS (UPLC-QqQ-MS). 17{\ss}-Testosterone-D3 was used as the stable isotope label of which the binding to rhSHBG-coated paramagnetic microbeads was inhibited by any other binding (designer) steroid. The assay was performed in a 96-well plate and combined with the fast LC-MS, 96 measurements could be performed within 4 h. The concentration-dependent inhibition of the label by steroids in buffer and dietary supplements was demonstrated. Following an adjusted bioaffinity isolation procedure, suspect extracts were injected into a chip-UPLC(NanoTile)-Q-time-of-flight-MS system for full-scan accurate mass identification. Next to known steroids, 1-testosterone was identified in three of the supplements studied and the designer steroid tetrahydrogestrinone was identified in a spiked supplement. The generic steroid-binding assay can be used for high-throughput screening of androgens, estrogens, and gestagens in dietary supplements to fight doping. When combined with chip-UPLC-MS, it is a powerful tool for early warning of unknown emerging rhSHBG bioactive designer steroids in dietary supplements.",
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High-throughput bioaffinity mass spectrometry for screening and identification of designer anabolic steroids in dietary supplements. / Aqai, P.; Cevik, E.; Gerssen, A.; Haasnoot, W.; Nielen, M.W.F.

In: Analytical Chemistry, Vol. 85, No. 6, 2013, p. 3255-3262.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - High-throughput bioaffinity mass spectrometry for screening and identification of designer anabolic steroids in dietary supplements

AU - Aqai, P.

AU - Cevik, E.

AU - Gerssen, A.

AU - Haasnoot, W.

AU - Nielen, M.W.F.

PY - 2013

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AB - A generic high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of known and unknown recombinant human sex hormone-binding globulin (rhSHBG)-binding designer steroids in dietary supplements. For screening, a semi-automated competitive inhibition binding assay was combined with fast ultrahigh-performance-LC-electrospray ionization-triple-quadrupole-MS (UPLC-QqQ-MS). 17ß-Testosterone-D3 was used as the stable isotope label of which the binding to rhSHBG-coated paramagnetic microbeads was inhibited by any other binding (designer) steroid. The assay was performed in a 96-well plate and combined with the fast LC-MS, 96 measurements could be performed within 4 h. The concentration-dependent inhibition of the label by steroids in buffer and dietary supplements was demonstrated. Following an adjusted bioaffinity isolation procedure, suspect extracts were injected into a chip-UPLC(NanoTile)-Q-time-of-flight-MS system for full-scan accurate mass identification. Next to known steroids, 1-testosterone was identified in three of the supplements studied and the designer steroid tetrahydrogestrinone was identified in a spiked supplement. The generic steroid-binding assay can be used for high-throughput screening of androgens, estrogens, and gestagens in dietary supplements to fight doping. When combined with chip-UPLC-MS, it is a powerful tool for early warning of unknown emerging rhSHBG bioactive designer steroids in dietary supplements.

KW - hormone-binding globulin

KW - multi-residue method

KW - liquid-chromatography

KW - nutritional supplements

KW - androgen bioassay

KW - discovery

KW - ligands

KW - urine

KW - contamination

KW - immunoassay

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DO - 10.1021/ac3036052

M3 - Article

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JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

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ER -