Abstract
Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50% B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120%). Little cross-reactivities (< 2%) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 273-279 |
| Number of pages | 7 |
| Journal | Fresenius' Journal of Analytical Chemistry |
| Volume | 369 |
| Issue number | 3-4 |
| State | Published - 2001 |
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ASJC Scopus subject areas
- Biochemistry
- Analytical Chemistry
- Clinical Biochemistry
Cite this
Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine. / Staimer, Norbert; Gee, Shirley J.; Hammock, Bruce D.
In: Fresenius' Journal of Analytical Chemistry, Vol. 369, No. 3-4, 2001, p. 273-279.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine
AU - Staimer, Norbert
AU - Gee, Shirley J.
AU - Hammock, Bruce D.
PY - 2001
Y1 - 2001
N2 - Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50% B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120%). Little cross-reactivities (< 2%) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.
AB - Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50% B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120%). Little cross-reactivities (< 2%) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.
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UR - http://www.scopus.com/inward/citedby.url?scp=0034935568&partnerID=8YFLogxK
M3 - Article
C2 - 11293704
AN - SCOPUS:0034935568
VL - 369
SP - 273
EP - 279
JO - Fresenius Zeitschrift fur Analytische Chemie
JF - Fresenius Zeitschrift fur Analytische Chemie
SN - 0016-1152
IS - 3-4
ER -