高速液体クロマトグラフィーとradioimmunoassayを用いた妊婦血中16α-hydroxyandrost-4-ene-3, 17-dioneの測定に関する研究
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16α-hydroxyandrost-4-ene-3, 17-dione (16αOH-A) was measured in pregnant women by radioimmunoassay (RIA) with specific antiserum. After extraction of the serum the 16αOH-A fraction was separated by the use of Sephadex LH-20 mini column.<BR>Lately, high performance liquid chromatograph (HPLC) was combined with RIA (HPLC-RIA) for the measurement of various kinds of steroids from an extract of a serum or urine sample.<BR>The purpose of this study was to establish the HPLC-RIA method for the measurement of 16αOH-A in pregnant women.<BR>Pooled serum was extracted twice with 5 volumes of ethylether, and the combined extract was separated either by LH-20 mini column with the solvent system n-hexane : benzene : methanol = 8 : 1 : 1 or by HPLC ODS column with MeOH : CH<SUB>3</SUB>CN : H<SUB>2</SUB>O = 60 : 10 : 30. The 16αOH-A fraction was collected and an aliquod was measured by RIA.<BR>The coefficients of variance for intra and inter-assay (n=10) were computed to be 8.6 and 12.1% with LH-20, and 7.2 and 11.7% with HPLC, respectively. Recovery rate was 95.7± 7.4% with LH-20 and 92.5 ± 8.7% with HPLC.<BR>The values of 16αOH-A as measured following two kinds of chromatographies were closely related to each other. The mean serum hormone concentrations±standard errors were as follows. In maternal peripheral blood up to 11 weeks 1.04 ± 0.19 (LH-20) and 0.55 ± 0.10 (HPLC), 12 to 23 weeks 1.22 ± 0.23 (LH-20) and 0.67 ± 0.77 (HPLC), 24 to 36 weeks 2.24 ± 0.42 (LH-20) and 2.27 ± 0.28 (HPLC), above 37 weeks 1.64 ± 0.52 (LH-20) and 1.85± 0.34 (HPLC), second stage of labor 5.07± 1.28 (LH-20) and 4.73 ± 0.54 (HPLC) ng/ml of steroid, respectively, were measured. Steroid concentration in umbilical arterial and venous blood were 7.87 ± 1.41 (LH-20), 3.75 ± 0.83 (HPLC) and 8.69± 1.66 (LH-20), 5.38±0.87 (HPLC) ng/ml, respectively.<BR>The values of 16αOH-A levels tended to increase as gestation proceeded. These data suggest production of 16αOH-A by the feto-placental unit. An elevated level of 16αOH-A is found in the umbilical venous blood as compared to the arterial samples. This indicates an extensive conversion. After onset of labor, the 16αOH-A levels were significantly increased because of precursor steroid 16α-hydroxydehydroepiandrosterone-sulfate is mobilized by the stress of labor.<BR>This study shows that the HPLC-RIA method is applicable to the measurement of related steroids produced in the feto-placental unit from a small amount of samples.
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- 高速液体クロマトグラフィーとradioimmunoassayを用いた妊婦血中16α-hydroxyandrost-4-ene-3, 17-dioneの測定に関する研究