A Simple Method for Determination of Urinary Pregnanediol by Sulfuric Acid Fluorescence Reaction

概要

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A simple fluorometric method for the estimation of urinary pregnanednol (P2) was devised.The urine sample (non-pregnancy or early pregnancy urine: 1.0ml, middle or late pregnancy urine: 0.2-0.5ml) was hydrolyzed with β-glucuronidase and extracted with methylene chloride.Urinary ketosteroids were eliminated by the Girard reaction and estrogens were eliminated by washing the methylene chloride extract with 5N-NaOH.The extract was evaporated, treated with sulfuric acid at 100° for 10 minutes, and measured by fluorometry at 485nm excitation wavelength and 515nm emission wavelength. The precision (c.v.) of the method was 6.7% (range: 1.49-172μg/ml, mean: 1.64μg/ml) for the urine specimen of a female in follicular phase (n=10) or 6.31% for 5μg/ml P2 standard solution (n=10).The recovery of P2 added to urine was 96.0% (mean of ten repeated experiments.range: 91.6-102.0%).The detectable limit was 0.2μg per tube or 1μg/ml urie.Some examples of the P2 assay in normal menstrual cycle and in pregnancy have been reported.
Japan Society of Clinical Chemistryの論文