A radioimmunoassay for serum dehydroepiandrosterone sulfate |
| |
Authors: | H Sekihara N Osawa H Ibayashi |
| |
Institution: | The Third Department of Internal Medicine, University of Tokyo Faculty of Medicine, Kongo, Tokyo, Japan |
| |
Abstract: | A radioimmunoassay for serum dehydroepiandrosterone sulfate (DHEAS) (1) has been developed using anti-DHEA antiserum obtained by immunizing rabbits with DHEA-17 oxime-bovine serum albumin. Serum volume of 0.01 to 0. 1 ml was used for analysis. After the addition of ammonium salt of DHEA-7 3 H sulfate for recovery and a preliminary removal of DHEA, DHEAS was extracted as pyridinium salt by methylene chloride. The dried extract was subjected to solvolysis (Burstein & Lieberman), followed by paper chromatography. The eluates and DHEA-7 3H which was added to determine the % free of DHEA were evaporated and incubated with the antiserum containing pepsin treated human immune serum globulin and bovine serum albumin at 37°C for 1 hour. Ammonium sulfate was used to separate free from bound DHEA. The accuracy, precision and specificity were satisfactory. The sensitivity was 3 ng per sample. The blank values could not be differentiated from zero. Although the antiserum reacts with the other 3βOHΔ5 steroids as well as DHEA, the complete separation of DHEA from the other 3βOHΔ5 steroids was achieved chromatographically. Serum DHEAS levels in normal subjects and patients with adrenocortical disorders obtained with the radioimmunoassay were comparable to those obtained with gasliquid chromatography. |
| |
Keywords: | |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|