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Download fileHigh-Throughput Bioaffinity Mass Spectrometry for Screening and Identification of Designer Anabolic Steroids in Dietary Supplements
journal contribution
posted on 2016-02-19, 16:55 authored by Payam Aqai, Ebru Cevik, Arjen Gerssen, Willem Haasnoot, Michel W. F. NielenA 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). 17β-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.