Determination of progesterone and some of its neuroactive ring A-reduced metabolites in human serum.
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.
Pearson Murphy
B E
BE
Departments of Psychiatry, Obstetrics and Gynecology, and Medicine, McGill University, Montreal, Canada. md98@musica.mcgill.ca
Allison
C M
CM
D000293:Adolescent; D000328:Adult; D000918:Antibody Specificity; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions / Q000276:immunology; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008297:Male; D008597:Menstrual Cycle / Q000097:blood / Q000378:metabolism; D008875:Middle Aged; D011247:Pregnancy; D011277:Pregnanediones / Q000097:blood / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D011280:Pregnanolone / Q000097:blood / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D011374:Progesterone / Q000097:blood* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011863:Radioimmunoassay; D012015:Reference Standards; D012997:Solvents
D007106:Immune Sera; D011277:Pregnanediones; D012997:Solvents; D011374:Progesterone; D011280:Pregnanolone