Lipidex chromatography in the radioimmunoassay of serum and urinary cortisol. |
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| Authors: | D. Apter O. Jänne R. Vihko |
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| Affiliation: | Steroid Research Laboratory, Department of Medical Chemistry, University of Helsinki, SF-00170 Helsinki 17, and Department of Clinical Chemistry, University of Oulu, SF-90220 Oulu 22 Finland |
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| Abstract: | A highly specific method for the determination of cortisol in human serum and urine is described. The sample is first extracted with diethyl ether/ethyl acetate (1 : 1, by vol.), then chromatographed on a highly lipophilic derivative of Sephadex (hydroxyalkoxypropyl Sephadex, Lipidex) in light petroleum/chloroform (1 : 1, by vol.), and finally cortisol is measured by radioimmunoassay using a cortisol-21-BSA antiserum. Bound and unbound radioactivities are separated using dextran-coated charcoal technique. The 8 a.m. values (mean +/- S.D.) of cortisol among 11 young females and 16 young males were 152 +/- 32 ng/ml (range 111-235) and 185 +/- 21 ng/ml (103-232), respectively. The respective values at 4 p.m. were 84 +/- 29 ng/ml (26-117) and 84 +/- 36 ng/ml (32-172). The importance of Lipidex chromatography was demonstrated with assays of serum samples from children with congenital adrenal hyperplasia; without chromatography cortisol values were 6 times those with chromatography. Specific cortisol assays from pregnancy serum also necessitated Lipidex chromatography. Among 11 young men and 9 young women the mean daily urinary cortisol excretion was 56 +/- 26 mug (32-109) and 60 +/- 24 mug (39-109), respectively. Specific urinary cortisol determination could not be achieved without Lipidex chromatography. |
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| Keywords: | pregnenolone, 3β-hydroxy-5-pregnen-20-one progesterone, 4-pregnene-3,20-dione 11α-hydroxyprogesterone, 11α-hydroxy-4-pregnene-3,20-dione 17-hydroxyprogesterone, 17α-hydroxy-4-pregnene-3,20-dione 21-deoxycortisol, 11β,17α-dihydroxy -4-pregnene-3, 20-dione 11-deoxycortisol, 17α,21-dihydroxy-4-pregnene-3,20-dione deoxycorticosterone, 21-hydroxy-4-pregnene-3,20-dione corticosterone, 11β,21-dihydroxy-4-pregnene3, 20-dione cortisol, 11β,17α,21-trihydroxy-4-pregnene-3,20-dione cortisone, 17α,21-dihydroxy-4-pregnene -3, 11,20-trione aldosterone, 11β,21-dihydroxy-4-pregnene-3,20-dion-18-al tetrahydro cortisol, 3α,11β,17α,21 -tetrahydroxy-5β-pregnan-20-one cortol, 5β-pregnane-3α, 11β,17α,20α,21-pentol prednisone, 17α,21-dihydroxy-l,4-pregnadiene-3,11,20-trione prednisolone, 11β,17α,21-trihydroxy -1, 4-pregnadiene-3,20-dione dexamethasone, 9α-fluoro-16α-methyl-11β,17α,21-trihy droxy-1, 4-pregnadiene-3,20-dione triamcinolone, 9α-fluoro-11β,16α,17α,21-tetrahydroxy-1,4-pregnadiene -3, 20-dione dehydroepiandrosterone, 3β-hydroxy-5-androsten-17-one testosterone, 17β-hydroxy -4-androsten-3-one estradiol-17β, 1,3,5(10)-estratriene-3,17β-diol |
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