Radioimmunoassay for 4-hydroxyoestrone in human urine

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Abstract
Under the protection of ascorbic acid a 4-hydroxyestrone-bovine serum albumin conjugate was prepared containing intact 4-hydroxyestrone as determined by gas chromatography-mass spectrometry. Using this antigen, antibodies with high affinity and specificity for 4-hydroxyestrone were raised in rabbits. An assay procedure for the determination of 4-hydroxyestrone in human urine and the assessment of its reliability are described. The following urinary excretion rates were found: male children 0.29 .mu.g/24 h, female children 0.35 .mu.g/24 h, men (20-40 yr) 1.6 .mu.g/24 h, men (> 50 yr) 1.8 .mu.g/24 h, women, follicular 2.0 .mu.g/24 h, pre-ovulatory 5.3 .mu.g/24 h, luteal 2.4 .mu.g/24 h, women, pregnant, 1st trimester 30.0 .mu.g/24 h, 2nd trimester 64.0 .mu.g/24 h, 3rd trimester 48.0 .mu.g/24 h, women, post-menopause 1.5 .mu./24 h. The amounts of 4-hydroxyestrone excreted in human urine are about 1/3 to 1/10 of those of 2-hydroxyestrone. During the menstrual cycle the excretion rates of 4-hydroxyestrone are in the same order of magnitude as those of estradiol and show a clear-cut pre ovulatory peak.

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