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Determination of progesterone and some of its neuroactive ring A-reduced metabolites in human serum.
- Source :
-
The Journal of steroid biochemistry and molecular biology [J Steroid Biochem Mol Biol] 2000 Oct; Vol. 74 (3), pp. 137-42. - Publication Year :
- 2000
-
Abstract
- A method for the separation and assay of some ring A-reduced metabolites of progesterone (pregnanediones and pregnanolones) is described. Serum was extracted with an organic solvent, and the extract chromatographed using high performance liquid chromatography (HPLC). A total of 50 fractions was collected for each sample and split using a stream splitter so that 30% was collected in counting vials for recovery while 70% was collected in test tubes which were assayed by radioimmunoassay. An antiserum raised in our laboratory to progesterone-3-CMO-BSA cross-reacted with five of these compounds (5alpha- and 5beta-dihydroprogesterone, 3alpha- and 3beta-5alpha-tetrahydroprogesterone, and 3beta, 5beta-tetrahydroprogesterone). Since pregnenolone eluted with 5alpha, 3beta-tetrahydroprogesterone, pregnenolone was assayed separately and its effect subtracted. Using this method it was shown that picogram to nanogram/ml amounts of these metabolites are present in all human sera. Levels in men were comparable to those of women in the follicular phase of the menstrual cycle. 5alpha-Dihydroprogesterone and 3alpha,5alpha-tetrahydroprogesterone rose substantially in the luteal phase of the menstrual cycle and all rose considerably during pregnancy.
- Subjects :
- Adolescent
Adult
Antibody Specificity
Binding, Competitive
Chromatography, High Pressure Liquid
Cross Reactions immunology
Female
Humans
Immune Sera immunology
Male
Menstrual Cycle blood
Menstrual Cycle metabolism
Middle Aged
Pregnancy
Pregnanediones blood
Pregnanediones chemistry
Pregnanediones immunology
Pregnanediones metabolism
Pregnanolone blood
Pregnanolone chemistry
Pregnanolone immunology
Pregnanolone metabolism
Progesterone chemistry
Progesterone immunology
Radioimmunoassay
Reference Standards
Solvents
Progesterone blood
Progesterone metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0960-0760
- Volume :
- 74
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- The Journal of steroid biochemistry and molecular biology
- Publication Type :
- Academic Journal
- Accession number :
- 11086232
- Full Text :
- https://doi.org/10.1016/s0960-0760(00)00098-4