PMID- 16791568
OWN - NLM
STAT- MEDLINE
DCOM- 20070504
LR  - 20160512
IS  - 1618-2642 (Print)
IS  - 1618-2642 (Linking)
VI  - 385
IP  - 5
DP  - 2006 Jul
TI  - SPE-HPLC purification of endocrine-disrupting compounds from human serum for 
      assessment of xenoestrogenic activity.
PG  - 875-87
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 
      (estrogen response element chemically activated luciferase expression, 
      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 (nos. 138, 153, 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. 17beta-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.
FAU - Hjelmborg, Philip Sebastian
AU  - Hjelmborg PS
AD  - Unit of Cellular and Molecular Toxicology, Institute of Public Health, Department 
      of Environmental and Occupational Medicine, University of Aarhus, Vennelyst 
      Boulevard 6, Building 1260, 8000 Aarhus C, Denmark.
FAU - Ghisari, Mandana
AU  - Ghisari M
FAU - Bonefeld-Jorgensen, Eva Cecilie
AU  - Bonefeld-Jorgensen EC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060603
PL  - Germany
TA  - Anal Bioanal Chem
JT  - Analytical and bioanalytical chemistry
JID - 101134327
RN  - 0 (Endocrine Disruptors)
RN  - 0 (Estrogens)
RN  - 0 (Receptors, Estrogen)
RN  - 0 (Trans-Activators)
RN  - 0 (Xenobiotics)
SB  - IM
MH  - Cell Line
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Endocrine Disruptors/*blood/chemistry/*isolation & purification
MH  - *Estrogens
MH  - Humans
MH  - Male
MH  - Molecular Structure
MH  - Receptors, Estrogen/metabolism
MH  - Solid Phase Extraction/*methods
MH  - Time Factors
MH  - Trans-Activators/metabolism
MH  - Xenobiotics/*blood/chemistry/*isolation & purification
EDAT- 2006/06/23 09:00
MHDA- 2007/05/05 09:00
CRDT- 2006/06/23 09:00
PHST- 2005/12/22 00:00 [received]
PHST- 2006/03/29 00:00 [accepted]
PHST- 2006/03/24 00:00 [revised]
PHST- 2006/06/23 09:00 [pubmed]
PHST- 2007/05/05 09:00 [medline]
PHST- 2006/06/23 09:00 [entrez]
AID - 10.1007/s00216-006-0463-9 [doi]
PST - ppublish
SO  - Anal Bioanal Chem. 2006 Jul;385(5):875-87. doi: 10.1007/s00216-006-0463-9. Epub 
      2006 Jun 3.