SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity

P.S. Hjelmborg; Mandana Ghisari; Eva Bonefeld-Jørgensen

doi:10.1007/s00216-006-0463-9

SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity

P.S. Hjelmborg, Mandana Ghisari, Eva Bonefeld-Jørgensen

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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Abstract

Assessment of xenoestrogenic activity in human serum samples requires the removal of endogenous sex hormones to assure that the activity measured originates from xenobiotic compounds only. Serum samples representing high, medium and lower accumulation of persistent organic pollutants (POPs) were extracted using solid-phase extraction (SPE) followed by normal-phase high-performance liquid chromatography (NP-HPLC) for separation of POPs from endogenous hormones. The recovery of polychlorinated biphenyl (PCB) congeners in spiked serum samples was up to 86 %, making the extraction method suitable for the study. MVLN cells, stably transfected with an estrogen receptor (ER) luciferase reporter vector (ERE-CALUX), were exposed to the reconstituted SPE-HPLC extracts for determination of the integrated estrogenic activity. The effects of PCBs were analyzed by direct in vitro exposure of PCBs (138, 153 and 180) and by ex vivo analysis of SPE-HPLC extracts from serum spiked with the PCBs. Similar effects on ER transactivation were observed for the direct in vitro and the ex vivo analysis experiments. The ER transactivation responses determined for actual serum samples were in the linear range of the dose response curve. 17β-Estradiol titrations showed that the xenoestrogenic effects were mediated via ER. Moreover, our SPE-HPLC-ERE-CALUX assay was demonstrated to elicit high interlaboratory correlation. In the present study the combination of SPE-HPLC purification and the ex vivo estrogenic responses measured by ERE-CALUX was validated and considered to be a valuable tool to assess the combined ER effect of lipophilic serum POPs where additive/synergistic and agonistic/antagonistic effects are integrated giving an overall estimate of exposure and bioactivity.

Original language	English
Journal	Analytical and Bioanalytical Chemistry
Volume	385
Issue	5
Pages (from-to)	875 - 887
Number of pages	13
ISSN	1618-2642
DOIs	https://doi.org/10.1007/s00216-006-0463-9
Publication status	Published - 2006

Keywords

SPE-HPLC, ERE-CALUX, MVLN, Endocrine disruption, Xenoestrogens, Environmental chemicals

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10.1007/s00216-006-0463-9

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title = "SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity",

abstract = "Assessment of xenoestrogenic activity in human serum samples requires the removal of endogenous sex hormones to assure that the activity measured originates from xenobiotic compounds only. Serum samples representing high, medium and lower accumulation of persistent organic pollutants (POPs) were extracted using solid-phase extraction (SPE) followed by normal-phase high-performance liquid chromatography (NP-HPLC) for separation of POPs from endogenous hormones. The recovery of polychlorinated biphenyl (PCB) congeners in spiked serum samples was up to 86 %, making the extraction method suitable for the study. MVLN cells, stably transfected with an estrogen receptor (ER) luciferase reporter vector (ERE-CALUX), were exposed to the reconstituted SPE-HPLC extracts for determination of the integrated estrogenic activity. The effects of PCBs were analyzed by direct in vitro exposure of PCBs (138, 153 and 180) and by ex vivo analysis of SPE-HPLC extracts from serum spiked with the PCBs. Similar effects on ER transactivation were observed for the direct in vitro and the ex vivo analysis experiments. The ER transactivation responses determined for actual serum samples were in the linear range of the dose response curve. 17β-Estradiol titrations showed that the xenoestrogenic effects were mediated via ER. Moreover, our SPE-HPLC-ERE-CALUX assay was demonstrated to elicit high interlaboratory correlation. In the present study the combination of SPE-HPLC purification and the ex vivo estrogenic responses measured by ERE-CALUX was validated and considered to be a valuable tool to assess the combined ER effect of lipophilic serum POPs where additive/synergistic and agonistic/antagonistic effects are integrated giving an overall estimate of exposure and bioactivity.",

keywords = "SPE-HPLC, ERE-CALUX, MVLN, Endocrine disruption, Xenoestrogens, Environmental chemicals",

author = "P.S. Hjelmborg and Mandana Ghisari and Eva Bonefeld-J{\o}rgensen",

year = "2006",

doi = "10.1007/s00216-006-0463-9",

language = "English",

volume = "385",

pages = "875 -- 887",

journal = "Analytical and Bioanalytical Chemistry",

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SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity. / Hjelmborg, P.S.; Ghisari, Mandana; Bonefeld-Jørgensen, Eva.
In: Analytical and Bioanalytical Chemistry, Vol. 385, No. 5, 2006, p. 875 - 887.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity

AU - Hjelmborg, P.S.

AU - Ghisari, Mandana

AU - Bonefeld-Jørgensen, Eva

PY - 2006

Y1 - 2006

N2 - Assessment of xenoestrogenic activity in human serum samples requires the removal of endogenous sex hormones to assure that the activity measured originates from xenobiotic compounds only. Serum samples representing high, medium and lower accumulation of persistent organic pollutants (POPs) were extracted using solid-phase extraction (SPE) followed by normal-phase high-performance liquid chromatography (NP-HPLC) for separation of POPs from endogenous hormones. The recovery of polychlorinated biphenyl (PCB) congeners in spiked serum samples was up to 86 %, making the extraction method suitable for the study. MVLN cells, stably transfected with an estrogen receptor (ER) luciferase reporter vector (ERE-CALUX), were exposed to the reconstituted SPE-HPLC extracts for determination of the integrated estrogenic activity. The effects of PCBs were analyzed by direct in vitro exposure of PCBs (138, 153 and 180) and by ex vivo analysis of SPE-HPLC extracts from serum spiked with the PCBs. Similar effects on ER transactivation were observed for the direct in vitro and the ex vivo analysis experiments. The ER transactivation responses determined for actual serum samples were in the linear range of the dose response curve. 17β-Estradiol titrations showed that the xenoestrogenic effects were mediated via ER. Moreover, our SPE-HPLC-ERE-CALUX assay was demonstrated to elicit high interlaboratory correlation. In the present study the combination of SPE-HPLC purification and the ex vivo estrogenic responses measured by ERE-CALUX was validated and considered to be a valuable tool to assess the combined ER effect of lipophilic serum POPs where additive/synergistic and agonistic/antagonistic effects are integrated giving an overall estimate of exposure and bioactivity.

AB - Assessment of xenoestrogenic activity in human serum samples requires the removal of endogenous sex hormones to assure that the activity measured originates from xenobiotic compounds only. Serum samples representing high, medium and lower accumulation of persistent organic pollutants (POPs) were extracted using solid-phase extraction (SPE) followed by normal-phase high-performance liquid chromatography (NP-HPLC) for separation of POPs from endogenous hormones. The recovery of polychlorinated biphenyl (PCB) congeners in spiked serum samples was up to 86 %, making the extraction method suitable for the study. MVLN cells, stably transfected with an estrogen receptor (ER) luciferase reporter vector (ERE-CALUX), were exposed to the reconstituted SPE-HPLC extracts for determination of the integrated estrogenic activity. The effects of PCBs were analyzed by direct in vitro exposure of PCBs (138, 153 and 180) and by ex vivo analysis of SPE-HPLC extracts from serum spiked with the PCBs. Similar effects on ER transactivation were observed for the direct in vitro and the ex vivo analysis experiments. The ER transactivation responses determined for actual serum samples were in the linear range of the dose response curve. 17β-Estradiol titrations showed that the xenoestrogenic effects were mediated via ER. Moreover, our SPE-HPLC-ERE-CALUX assay was demonstrated to elicit high interlaboratory correlation. In the present study the combination of SPE-HPLC purification and the ex vivo estrogenic responses measured by ERE-CALUX was validated and considered to be a valuable tool to assess the combined ER effect of lipophilic serum POPs where additive/synergistic and agonistic/antagonistic effects are integrated giving an overall estimate of exposure and bioactivity.

KW - SPE-HPLC, ERE-CALUX, MVLN, Endocrine disruption, Xenoestrogens, Environmental chemicals

U2 - 10.1007/s00216-006-0463-9

DO - 10.1007/s00216-006-0463-9

M3 - Journal article

C2 - 16791568

SN - 1618-2642

VL - 385

SP - 875

EP - 887

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

IS - 5

ER -

SPE-HPLC purification of endocrine disrupting compounds from human serum for assessment of xenoestrogenic activity

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