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Type: Journal article
Title: Determination of steroids in human plasma using temperature-dependent inclusion chromatography for metabolomic investigations
Author: Zarzycki, P.
Kulhanek, K.
Smith, R.
Clifton, V.
Citation: Journal of Chromatography A, 2006; 1104(1-2):203-208
Publisher: Elsevier Science BV
Issue Date: 2006
ISSN: 0021-9673
Statement of
Paweł K. Zarzycki, Kathrin M. Kulhanek, Roger Smith and Vicki L. Clifton
Abstract: Clinical and metabolomic investigations of complex human fluids require cost-effective methodologies that can rapidly assess the steroid hormone milieu of individual samples. The efficiency of quantification of many steroids is limited using immunoassays as these methods can only measure a single component of biological samples and are dependent upon the specificity of the antiserum used in the protocol. In this study, we optimised the solid-phase extraction protocol for the extraction of a range of steroids of varied polarity from estetrol to progesterone from human plasma. The final SPE procedure for efficient extraction of steroids was a washing mixture of 5 ml of 30% methanol and an elution solvent of 2 ml of 100% methanol using 0.5 g C-18 cartridges. This protocol resulted in a high recovery rate, ranging from 85.2 to 99.9% for both the internal standard (7,8-dimethoxyflavone) and steroids of interest. We also improved the separation methodology of our previous work using temperature dependent inclusion chromatography with a mobile phase composition of 35% acetonitrile and 12 mM of β-cyclodextrin at 29 °C. Under these conditions most of the fluid components including estetrol were detected in the first 10 min with progesterone appearing at 43 min. This method is simplistic, inexpensive and reproducible with the capabilities of accurate quantification of steroids. Therefore it could have numerous clinical and metabolomic applications.
Keywords: Solid-phase extraction
Inclusion chromatography
Diode-array detection
Steroid quantification
Cord blood
Human plasma
Description: Copyright © 2005 Elsevier B.V. All rights reserved.
DOI: 10.1016/j.chroma.2005.11.119
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Paediatrics publications

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