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Development of a solid phase extraction method for the simultaneous determination of steroid hormones in H295R cell line using liquid chromatography–tandem mass spectrometry
Storbritannien.
Kristianstad University, School of Education and Environment, Avdelningen för Naturvetenskap. Kristianstad University, Plattformen för molekylär analys.ORCID iD: 0000-0003-2283-2927
USA.
2013 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 935, no September, 61-69 p.Article in journal (Refereed) Published
Abstract [en]

The H295R in vitro cell line produces the majority of the steroidogenesis, for which reason it is commonly used as a screening tool for endocrine disrupting chemicals. Simultaneous determination of the precursor cholesterol and key steroid hormones could give a broad insight into the mechanistic disruption of the steroidogenesis. Steroid hormones have primarily been extracted from H295R incubation medium by means of liquid-liquid extraction (LLE) and the obtained recoveries and matrix effects have typically not been stated or assessed. In the present study a solid-phase extraction (SPE) method was developed and validated for the simultaneous extraction of cholesterol and five key steroid hormones pregnenolone, 17-hydroxyprogesterone, testosterone, cortisol and aldosterone from H295R incubation medium, and finally detected by LC-MS/MS. Cholesterol was recovered at a level of 55.7%, while steroid hormone recoveries ranged from 98.2 to 109.4%. Matrix effects varied between -0.6% and 62.8%. Intra-day precision was deemed acceptable, but the inter-day precision for pregnenolone and aldosterone exceeded the precision limit of 15% RSD. Although LLE has been the most frequently used extraction method in H295R studies, however, our investigation has shown that SPE may relatively easily extract and recover steroid hormones, potentially replacing LLE.

Place, publisher, year, edition, pages
2013. Vol. 935, no September, 61-69 p.
Keyword [en]
Solid Phase Extraction, LC-MS/MS, Cholesterol, Steroid hormones, H295R incubation medium, Steroidogenesis.
National Category
Medical Biotechnology
Identifiers
URN: urn:nbn:se:hkr:diva-10816DOI: 10.1016/j.jchromb.2013.07.013ISI: 000324084200010PubMedID: 23948237OAI: oai:DiVA.org:hkr-10816DiVA: diva2:639152
Available from: 2013-08-06 Created: 2013-08-06 Last updated: 2017-04-20Bibliographically approved

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CiteExportLink to record
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Citation style
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