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  • Title: A radioimmunoassay for norethindrone (NET): measurement of serum NET concentrations following ingestion of NET-containing oral contraceptive steroids.
    Author: Stanczyk FZ, Brenner PF, Mishell DR, Ortiz A, Gentzschein EK, Goebelsmann U.
    Journal: Contraception; 1978 Dec; 18(6):615-33. PubMed ID: 750188.
    Abstract:
    A sensitive and reliable radioimmunoassay (RIA) for the measurement of norethindrone (NET) in serum has been established employing anti-11 alpha-hydroxynorethindrone 11-hemisuccinyl-bovine serum albumin serum in conjunction with norethindrone-3-(0-carboxymethyl) oximino-[125I]-iodohistamine. Of a number of ring A reduced NET metabolites, only 17 beta-hydroxy-17 alpha-ethinyl-5 beta-estran-3-one (43%) and 17 alpha-ethinyl-5 alpha-estrane-3 beta, 17 beta-diol (15.7%) cross-reacted appreciably in this RIA. Ethinyl estradiol (EE2) and mestranol (MEE2) exhibited cross-reactions of only 1.1 and 0.4%, respectively. Serum NET levels were measured in four groups of 3 women, each ingesting either 1 mg NET plus 0.05 mg MEE2 (Norinyl 1 + 50 or Ortho Novum 1/50), 0.5 mg NET plus 0.035 mg EE2 (Brevicon) or only 0.35 mg NET (Micronor) daily for 5 consecutive days. Peak serum NET levels were observed within 1/2 to 4 hours after oral intake and fell precipitously thereafter. After reaching a maximum, serum NET concentrations declined in a manner consistent with at least two disposition phases. The average half-life for the first disposition phase was 2.3, 3.4, 3.9 and 4.4 hours in subjects ingesting Norinyl 1 + 50, Ortho Novum 1/50, Brevicon and Micronor, respectively. Peak and 3-hour post-ingestion serum NET concentrations were dose-related but showed considerable subject-to-subject variations. Following discontinuation of tablet intake, serum NET levels remained detectable (greater than 0.05 ng/ml) for at least 5 days in all 3 women who had taken Ortho Novum 1/50, but in none of the other 9 volunteers. These results suggest that different preparations of identical doses and combinations of oral contraceptive steroids may yield different serum NET profiles. However, due to considerable subject-to-subject variations, larger numbers of subjects are required for a conclusive investigation. A sensitive and reliable radioimmunoassay (RIA) for measuring norethindrone (NET) in serum is described. Of a number of ring-A-reduced NET metabolites, only 2 cross-reacted appreciably in this RIA. Ethinyl estradiol and mestranol exhibited cross-reactions of only 1.1 and .4%, respectively. Serum concentrations of NET before and after ingestion of 4 NET-containing oral contraceptives (OCs) by 3 women each were measured by this RIA. In all 12 volunteers, serum NET levels rose rapidly after oral intake, reaching peak levels within .5-4 hours (median, 1.5) and fell precipitiously thereafter. Peak serum NET averaged 10.2 ng/ml after Ortho Novum 1/50 ingestion, 10.4 after Norinyl 1+50, 7.5 after Brevicon, and 4.3 after Micronor ingestion. Individual peak NET levels varied considerably in each group. Average half-lives varied from 2.3-4.4 hours, depending on OC compound ingested. After discontinuation of OCs, serum NET levels remained detectable ( .05 ng/ml) for at least 5 days in all 3 women who took Ortho Novum 1/50 but in none of the other subjects, suggesting that different preparations of identical doses and combinations of OCs may yield different serum NET profiles.
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