High-Throughput Bioaffinity Mass Spectrometry for Screening and Identification of Designer Anabolic Steroids in Dietary Supplements

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• 作者：Payam Aqai ; Ebru Cevik ; Arjen Gerssen ; Willem Haasnoot ; Michel W. F. Nielen
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：March 19, 2013
• 年：2013
• 卷：85
• 期：6
• 页码：3255-3262
• 全文大小：281K
• 年卷期：v.85,no.6(March 19, 2013)
• ISSN：1520-6882

文摘

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尾-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.