Martin Hansen

Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites.

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Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites. / Abdel-Khalik, Jonas; Björklund, Erland; Nielsen, Frederik Knud; Hansen, Martin.

I: Journal of Pharmaceutical and Biomedical Analysis, Bind 145, 10.2017, s. 569-575.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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APA

CBE

Abdel-Khalik J, Björklund E, Nielsen FK, Hansen M. 2017. Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites. Journal of Pharmaceutical and Biomedical Analysis. 145:569-575.

MLA

Vancouver

Abdel-Khalik J, Björklund E, Nielsen FK, Hansen M. Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites. Journal of Pharmaceutical and Biomedical Analysis. 2017 okt;145:569-575.

Author

Abdel-Khalik, Jonas ; Björklund, Erland ; Nielsen, Frederik Knud ; Hansen, Martin. / Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites. I: Journal of Pharmaceutical and Biomedical Analysis. 2017 ; Bind 145. s. 569-575.

Bibtex

@article{c2d2caed04094869b6f5bb882ac35aad,
title = "Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites.",
abstract = "This study demonstrates the addition of (14)C-cholesterol to the human cell line H295R will in-situ form radiolabeled steroid hormones allowing for new mechanistic and metabolic insights. The aim of the present study was to in-situ radiolabel steroid hormones from cell line-incorporated (14)C-cholesterol using the OECD guideline 456, H295R steroidogenesis in-vitro assay. Radiodetection of the steroid metabolites of the steroidogenic pathway allows for an improved understanding of the various enzymatic mechanisms involved without necessarily being dependent on quantification. Generated radiolabeled steroids were analyzed using HPLC hyphenated with a Flow Scintillation Analyzer (FSA). H295R cells were incubated with radiolabeled cholesterol and cell media were collected and prepared by solid phase extraction and analyzed with HPLC-FSA. For successful radiolabeling of the steroids in the steroidogenesis of H295R cells, radioactive cholesterol may potentially only need to be added just before the cells are incubated for 72h in well plates. Based on the obtained HPLC-FSA chromatograms, and confirmation of the observations by studies in the literature, a qualitative time profile for the production of steroid hormones was estimated. Multiple radiolabeled steroid hormones were identified by means of analytical standards and UV (ultraviolet) co-chromatography, though the elucidation of multiple metabolites remains unresolved. Although online radiodetection proved to suffer from suboptimal sensitivity, the concept of radiolabeling the steroidogenesis in H295R cells with (14)C-cholesterol and detecting the radiolabeled steroid hormones online was proved and may assist in further toxicological studies.",
author = "Jonas Abdel-Khalik and Erland Bj{\"o}rklund and Nielsen, {Frederik Knud} and Martin Hansen",
year = "2017",
month = oct,
language = "English",
volume = "145",
pages = "569--575",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
issn = "0731-7085",
publisher = "Elsevier BV",

}

RIS

TY - JOUR

T1 - Incorporation of (14)C-cholesterol in human adrenal corticocarcinoma H295R cell line and online-radiodetection of produced (14)C-steroid hormone metabolites.

AU - Abdel-Khalik, Jonas

AU - Björklund, Erland

AU - Nielsen, Frederik Knud

AU - Hansen, Martin

PY - 2017/10

Y1 - 2017/10

N2 - This study demonstrates the addition of (14)C-cholesterol to the human cell line H295R will in-situ form radiolabeled steroid hormones allowing for new mechanistic and metabolic insights. The aim of the present study was to in-situ radiolabel steroid hormones from cell line-incorporated (14)C-cholesterol using the OECD guideline 456, H295R steroidogenesis in-vitro assay. Radiodetection of the steroid metabolites of the steroidogenic pathway allows for an improved understanding of the various enzymatic mechanisms involved without necessarily being dependent on quantification. Generated radiolabeled steroids were analyzed using HPLC hyphenated with a Flow Scintillation Analyzer (FSA). H295R cells were incubated with radiolabeled cholesterol and cell media were collected and prepared by solid phase extraction and analyzed with HPLC-FSA. For successful radiolabeling of the steroids in the steroidogenesis of H295R cells, radioactive cholesterol may potentially only need to be added just before the cells are incubated for 72h in well plates. Based on the obtained HPLC-FSA chromatograms, and confirmation of the observations by studies in the literature, a qualitative time profile for the production of steroid hormones was estimated. Multiple radiolabeled steroid hormones were identified by means of analytical standards and UV (ultraviolet) co-chromatography, though the elucidation of multiple metabolites remains unresolved. Although online radiodetection proved to suffer from suboptimal sensitivity, the concept of radiolabeling the steroidogenesis in H295R cells with (14)C-cholesterol and detecting the radiolabeled steroid hormones online was proved and may assist in further toxicological studies.

AB - This study demonstrates the addition of (14)C-cholesterol to the human cell line H295R will in-situ form radiolabeled steroid hormones allowing for new mechanistic and metabolic insights. The aim of the present study was to in-situ radiolabel steroid hormones from cell line-incorporated (14)C-cholesterol using the OECD guideline 456, H295R steroidogenesis in-vitro assay. Radiodetection of the steroid metabolites of the steroidogenic pathway allows for an improved understanding of the various enzymatic mechanisms involved without necessarily being dependent on quantification. Generated radiolabeled steroids were analyzed using HPLC hyphenated with a Flow Scintillation Analyzer (FSA). H295R cells were incubated with radiolabeled cholesterol and cell media were collected and prepared by solid phase extraction and analyzed with HPLC-FSA. For successful radiolabeling of the steroids in the steroidogenesis of H295R cells, radioactive cholesterol may potentially only need to be added just before the cells are incubated for 72h in well plates. Based on the obtained HPLC-FSA chromatograms, and confirmation of the observations by studies in the literature, a qualitative time profile for the production of steroid hormones was estimated. Multiple radiolabeled steroid hormones were identified by means of analytical standards and UV (ultraviolet) co-chromatography, though the elucidation of multiple metabolites remains unresolved. Although online radiodetection proved to suffer from suboptimal sensitivity, the concept of radiolabeling the steroidogenesis in H295R cells with (14)C-cholesterol and detecting the radiolabeled steroid hormones online was proved and may assist in further toxicological studies.

M3 - Journal article

VL - 145

SP - 569

EP - 575

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

ER -