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Evaluation of CD detection in an HPLC system for analysis of DHEA and related steroids

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Abstract

The biological importance of dehydroepiandrosterone (DHEA) is reflected by the fact that DHEA is a crucial precursor of the biosynthesis of the steroidal sex hormones. Simultaneous separation of DHEA, dehydroepiandrosterone sulfate (DHEA-S), pregnenolone, androstenedione and testosterone has been accomplished by reversed-phase ion-pair high-performance liquid chromatography (RP-IP-HPLC) based on isocratic elution applying circular dichroism (CD) detection at 295 nm. Addition of tetrabutylammonium hydrogensulfate to the mobile phase increases the retention of DHEA-S on the C8-silica column by an apparent ion-pairing mechanism without affecting the retention of the other (non-ionic) steroids. CD spectroscopy provides highly selective detection of compounds possessing optically active absorption bands and the separation is even more selective in the higher wavelength range applied. The linearity of the steroid concentration (c, mg mL−1) versus peak area was tested in the concentration range of 0.5–2 mg mL−1 (injected quantities were 10–40 μg). The relative standard deviation (RSD) values for DHEA and DHEA-S indicated a good intra-assay and inter-assay precision of the method.

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Acknowledgements

This work has been supported by the grant of OTKA, No. T 48554 (Budapest, Hungary). The valuable contributions of Ilona Matalin-Foór and Ilona Kovács-Derzsi are highly appreciated.

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Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, Semmelweis University, Hõgyes E. u. 9, 1092, Budapest, Hungary

    A. Gergely, Gy. Szász, A. Szentesi, K. Gyimesi-Forrás, J. Kökösi & D. Szegvári

  2. State Hospital of Cardiology, 8230, Balatonfüred, Hungary

    G. Veress

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  1. A. Gergely
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  2. Gy. Szász
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  3. A. Szentesi
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  4. K. Gyimesi-Forrás
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  5. J. Kökösi
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  6. D. Szegvári
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  7. G. Veress
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Correspondence to A. Gergely.

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Gergely, A., Szász, G., Szentesi, A. et al. Evaluation of CD detection in an HPLC system for analysis of DHEA and related steroids. Anal Bioanal Chem 384, 1506–1510 (2006). https://doi.org/10.1007/s00216-006-0318-4

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  • DOI: https://doi.org/10.1007/s00216-006-0318-4

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