Article: Time-resolved fluoroimmunoassay of plasma and urine O-desmethylangolensin.
The Journal of steroid biochemistry and molecular biology
2002 Volume 81, Issue 4-5, Page(s) 353–361
Abstract: We present a method for the determination of the phytoestrogen metabolite O-desmethylangolensin (O ... DMA) in plasma (serum) and in urine. O-DMA is a metabolite of daidzein, which occurs in soybeans ... using a europium chelate as a label. After the synthesis of 4"-O-carboxymethyl-O-DMA, this compound is ...
Abstract | We present a method for the determination of the phytoestrogen metabolite O-desmethylangolensin (O-DMA) in plasma (serum) and in urine. O-DMA is a metabolite of daidzein, which occurs in soybeans. It has been suggested that isoflavones may afford protection against breast and prostate cancer and therefore, also the metabolites are of interest. The method is based on time-resolved fluoroimmunoassay (TR-FIA) using a europium chelate as a label. After the synthesis of 4"-O-carboxymethyl-O-DMA, this compound is coupled to bovine serum albumin, and then used as antigen in immunization of rabbits. The tracers with the europium chelate are synthesized using the same 4"-O-derivative of the alpha-methyldeoxybenzoin. After enzymatic hydrolysis and ether extraction the immunoassay is carried out by time resolved fluoroimmunoassay (TR-FIA). Cross-reactivity was tested with angolensin, dihydrogenistein, dihydrodaidzein, equol, 6'-OH-angolensin, trans-4-OH-equol, 6'-OH-O-DMA, cis-4-OH-equol and 5-OH-equol. The antiserum cross-reacted only with angolensin. This cross-reactivity seems not to influence the results, which were highly specific. Plasma samples are hydrolyzed and extracted. Urine samples are analyzed directly after hydrolysis without extraction. The correlation coefficient between the plasma TR-FIA results and the GC-MS results was high; r value was 0.985. The correlation coefficient between the urine TR-FIA results and the GC-MS results was high over the entire range of concentrations (0-1500 nmol/l); r value was 0.976, but lower in the low concentration range (0-100 nmol/l), i.e. value was 0.631. The intra-assay coefficients of variation (CVs) for plasma O-DMA concentrations and for urine O-DMA concentrations at three different concentrations varied 2.8-7.7 and 3.0-6.0%, respectively and the inter-assay CVs varied 3.8-8.9 and 4.4-6.6%, respectively. The working range of the plasma and urine O-DMA assays was 0.5-512 nmol/l. |
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MeSH term(s) | Animals ; Antibody Formation ; Cross Reactions ; Evaluation Studies as Topic ; Fluoroimmunoassay/methods ; Gas Chromatography-Mass Spectrometry ; Humans ; Immune Sera ; Isoflavones/blood ; Isoflavones/immunology ; Isoflavones/urine ; Rabbits ; Sensitivity and Specificity ; Serum Albumin, Bovine ; Glycine max |
Chemical Substances | Immune Sera ; Isoflavones ; Serum Albumin, Bovine (27432CM55Q) ; O-desmethylangolensin (SCY1S10OK4) |
Language | English |
Publishing date | 2002-07-02 |
Publishing country | England |
Document type | Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't |
ZDB-ID | 1049188-0 |
ISSN | 1879-1220 ; 0960-0760 |
ISSN (online) | 1879-1220 |
ISSN | 0960-0760 |
DOI | 10.1016/s0960-0760(02)00073-0 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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