High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure
Human placental JEG-3 cells conserve a high P450 aromatase activity and are therefore suitable to evaluate how contaminants may interfere with the routes involved in estrogen synthesis during pregnancy. This has been traditionally assessed by measuring aromatase activity through the amount of tritiated water (3H2O) formed during the aromatization of 1β-3H-androst-4-ene-3,17-dione (3H-AD). This work presents a greener and safer analytical approach for this purpose, which consists of the determination of the trace amounts of the steroids (estradiol, estrone, testosterone, and androstenedione) present in the culture medium. Turbulent flow chromatography coupled to liquid chromatography-tandem mass spectrometry (TFC-HPLC-MS/MS) delivered the high selectivity and sensitivity (limits of detection between 2 and 5 pg/mL) required for these measurements. Moreover, its automation allows high-throughput of samples with minimum sample handling and achieves high precision in the analysis (relative standard deviation values <6%). As a proof of concept, the method was applied to evaluate the effect of monohaloacetic acid exposure on the steroid profile of JEG-3 cells. Iodoacetic acid showed an estrogenic effect (statistically significant increase of estradiol levels compared to unexposed cells) at the highest concentration level tested (0.5 µM) that deserves further evaluation.
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Language: | English |
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Elsevier
2022-02-08
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Subjects: | Turbulent flow chromatography, Disinfection byproducts, Placental JEG-3 cells, Reprotoxicity, Sex hormones, Steroid profiling, |
Online Access: | http://hdl.handle.net/10261/262184 http://dx.doi.org/10.13039/501100004837 https://api.elsevier.com/content/abstract/scopus_id/85124791251 |
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dig-idaea-es-10261-2621842024-05-14T20:35:45Z High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure Mestres, Júlia Pérez-Albaladejo, Elisabet Porte Visa, Cinta Postigo, Cristina Ministerio de Ciencia e Innovación (España) 0000-0002-1319-9552 0000-0002-7344-7044 Turbulent flow chromatography Disinfection byproducts Placental JEG-3 cells Reprotoxicity Sex hormones Steroid profiling Human placental JEG-3 cells conserve a high P450 aromatase activity and are therefore suitable to evaluate how contaminants may interfere with the routes involved in estrogen synthesis during pregnancy. This has been traditionally assessed by measuring aromatase activity through the amount of tritiated water (3H2O) formed during the aromatization of 1β-3H-androst-4-ene-3,17-dione (3H-AD). This work presents a greener and safer analytical approach for this purpose, which consists of the determination of the trace amounts of the steroids (estradiol, estrone, testosterone, and androstenedione) present in the culture medium. Turbulent flow chromatography coupled to liquid chromatography-tandem mass spectrometry (TFC-HPLC-MS/MS) delivered the high selectivity and sensitivity (limits of detection between 2 and 5 pg/mL) required for these measurements. Moreover, its automation allows high-throughput of samples with minimum sample handling and achieves high precision in the analysis (relative standard deviation values <6%). As a proof of concept, the method was applied to evaluate the effect of monohaloacetic acid exposure on the steroid profile of JEG-3 cells. Iodoacetic acid showed an estrogenic effect (statistically significant increase of estradiol levels compared to unexposed cells) at the highest concentration level tested (0.5 µM) that deserves further evaluation. CP acknowledges the support of Fundación General del CSIC through the ComFuturo Programme (2nd edition). This publication is part of the grant CEX2018–000794-S, funded by MCIN/AEI(10.13039/501100011033). Peer reviewed 2022-02-28T10:33:14Z 2022-02-28T10:33:14Z 2022-02-08 artículo http://purl.org/coar/resource_type/c_6501 Journal of Chromatography A 1667: 462886 (2022) 00219673 http://hdl.handle.net/10261/262184 10.1016/j.chroma.2022.462886 http://dx.doi.org/10.13039/501100004837 35193068 2-s2.0-85124791251 https://api.elsevier.com/content/abstract/scopus_id/85124791251 en #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/EC/HE/CEX2018–000794-S Journal of chromatography. A Publisher's version https://doi.org/10.1016/j.chroma.2022.462886 Sí open Elsevier |
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Turbulent flow chromatography Disinfection byproducts Placental JEG-3 cells Reprotoxicity Sex hormones Steroid profiling Turbulent flow chromatography Disinfection byproducts Placental JEG-3 cells Reprotoxicity Sex hormones Steroid profiling |
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Turbulent flow chromatography Disinfection byproducts Placental JEG-3 cells Reprotoxicity Sex hormones Steroid profiling Turbulent flow chromatography Disinfection byproducts Placental JEG-3 cells Reprotoxicity Sex hormones Steroid profiling Mestres, Júlia Pérez-Albaladejo, Elisabet Porte Visa, Cinta Postigo, Cristina High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
description |
Human placental JEG-3 cells conserve a high P450 aromatase activity and are therefore suitable to evaluate how contaminants may interfere with the routes involved in estrogen synthesis during pregnancy. This has been traditionally assessed by measuring aromatase activity through the amount of tritiated water (3H2O) formed during the aromatization of 1β-3H-androst-4-ene-3,17-dione (3H-AD). This work presents a greener and safer analytical approach for this purpose, which consists of the determination of the trace amounts of the steroids (estradiol, estrone, testosterone, and androstenedione) present in the culture medium. Turbulent flow chromatography coupled to liquid chromatography-tandem mass spectrometry (TFC-HPLC-MS/MS) delivered the high selectivity and sensitivity (limits of detection between 2 and 5 pg/mL) required for these measurements. Moreover, its automation allows high-throughput of samples with minimum sample handling and achieves high precision in the analysis (relative standard deviation values <6%). As a proof of concept, the method was applied to evaluate the effect of monohaloacetic acid exposure on the steroid profile of JEG-3 cells. Iodoacetic acid showed an estrogenic effect (statistically significant increase of estradiol levels compared to unexposed cells) at the highest concentration level tested (0.5 µM) that deserves further evaluation. |
author2 |
Ministerio de Ciencia e Innovación (España) |
author_facet |
Ministerio de Ciencia e Innovación (España) Mestres, Júlia Pérez-Albaladejo, Elisabet Porte Visa, Cinta Postigo, Cristina |
format |
artículo |
topic_facet |
Turbulent flow chromatography Disinfection byproducts Placental JEG-3 cells Reprotoxicity Sex hormones Steroid profiling |
author |
Mestres, Júlia Pérez-Albaladejo, Elisabet Porte Visa, Cinta Postigo, Cristina |
author_sort |
Mestres, Júlia |
title |
High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
title_short |
High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
title_full |
High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
title_fullStr |
High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
title_full_unstemmed |
High-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
title_sort |
high-throughput analysis of the steroid profile in placental cell cultures to evaluate endocrine disrupting effects of contaminant exposure |
publisher |
Elsevier |
publishDate |
2022-02-08 |
url |
http://hdl.handle.net/10261/262184 http://dx.doi.org/10.13039/501100004837 https://api.elsevier.com/content/abstract/scopus_id/85124791251 |
work_keys_str_mv |
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