The measurement of the isoflavone daidzein by time resolved fluorescent immunoassay: a method for assessment of dietary soya exposure.
We report a novel method for the measurement of urinary daidzein that is suitable for assessment of dietary soya exposure. The method incorporates the following features: (i) a highly specific monoclonal antibody to daidzein (clone 4E4) raised through the 7 position of daidzein and (ii) a europium labeled ovalbumin daidzein conjugate. In the present format, dilute urine samples of subjects who ingested soy milk are hydrolyzed with beta-glucuronidase for 30 min on rabbit anti-mouse coated plates. Afterwards, the specific monoclonal antibody to daidzein, clone 4E4, and europium labeled ovalbumin daidzein conjugate are added. After 1 h incubation, the wall bound fluorescence of europium is measured by time resolved fluorescence and is inversely proportional to the concentration of daidzein over the range 0.1-10 ng daidzein/well. The method demonstrates good sensitivity, precision and comparability with the chemical method GC-FID. Unlike the chemical method, the present immunoassay technique for daidzein is applicable for the measurement of large amounts of samples in epidemiological studies for the assessment and monitoring of human exposure to soya food.
Kohen
F
F
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot, Israel. lhkohenf@weizmann.weizmann.ac.il
Lichter
S
S
Gayer
B
B
DeBoever
J
J
Lu
L J
LJ
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D002138:Calibration; D004032:Diet; D004965:Estrogen Antagonists / Q000276:immunology / Q000652:urine*; D004968:Estrogens, Non-Steroidal / Q000032:analysis; D005063:Europium / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D007529:Isoflavones / Q000652:urine*; D010047:Ovalbumin / Q000737:chemistry; D048789:Phytoestrogens; D028321:Plant Preparations; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D004965:Estrogen Antagonists; D004968:Estrogens, Non-Steroidal; D007529:Isoflavones; D048789:Phytoestrogens; D028321:Plant Preparations; D005063:Europium; C004742:daidzein; D010047:Ovalbumin; D005966:Glucuronidase