RESUMO
Ethynodiol diacetate (EDA) and ethinyl estradiol (EE) tablets are indicated to prevent pregnancy in women who use oral contraceptives as a contraception method. EDA and EE were separated by reversed-phase HPLC using Agilent ZORBAX SB-Phenyl column, 4.6 mm × 15 cm, 5 µm, using a gradient mixture of acetonitrile and Milli-Q water as mobile phase. The linearity and recovery were found in the range of 0.025-0.25 mg/mL and 0.05-0.18 mg/mL for EDA and 0.001-0.01 mg/mL and 0.002-0.007 mg/mL for EE, respectively. The method is validated according to the regulatory guidelines concerning system suitability, specificity, repeatability, recovery, linearity, robustness, and stability of the sample solution.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cromatografia de Fase Reversa/métodos , Anticoncepcionais Orais/química , Diacetato de Etinodiol/análogos & derivados , Diacetato de Etinodiol/análise , Comprimidos/químicaRESUMO
The impurity profile of ethynodiol diacetate was investigated using the HPLC/UV/MS method. Using the slightly modified HPLC method of USP 24 two impurities, earlier isolated by preparative HPLC and investigated by NMR spectroscopy were separated and characterised. The mass spectra amended by the diode-array UV spectra supported the earlier found structures (E and Z isomers of 17alpha-ethinyl-estr-4-ene-3beta,17-diol-3-acetate-17-(3'-acetoxy-2'-butenoate). Another, hitherto not described impurity, 17alpha-ethinyl-estr-4-ene-3beta,17-diol-3-acetate-17-(3-oxo-butanoate) has also been separated and characterised by means of its mass spectrum, NMR and UV spectra.
Assuntos
Anticoncepcionais Orais Sintéticos/análise , Contaminação de Medicamentos , Diacetato de Etinodiol/análise , Preparações Farmacêuticas/normas , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância Magnética , Espectrofotometria Ultravioleta , EstereoisomerismoRESUMO
The usefulness of the joint application of HPLC and NMR spectroscopy in drug impurity profiling is demonstrated by the following examples: (1) identification of Z and E isomers of 17 alpha-ethynyl-4-oestrene-3 beta, 17-diol-3-acetate-17-(3'-acetoxy-2'-butenoate) in ethynodiol diacetate; (2) identification of the p-tolyl analogue as the impurity of enalapril maleate; (3) identification and quantification of 2'-dehydro-pipecuronium bromide in pipecuronium bromide. The possibilities of utilizing NMR spectroscopy for the identification and quantification of the impurities with and without their isolation are discussed.
Assuntos
Androstano-3,17-diol/análogos & derivados , Enalapril/análise , Diacetato de Etinodiol/análise , Bloqueadores Neuromusculares/análise , Piperazinas/análise , Androstano-3,17-diol/análise , Cromatografia Líquida de Alta Pressão , Enalapril/análogos & derivados , Diacetato de Etinodiol/análogos & derivados , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Pipecurônio , EstereoisomerismoRESUMO
In the presence of ethynodiol diacetate the ethynylestradiol concentration in serum and saliva after oral administration was measured by a rapid radioimmunoassay method developed by the authors. By means of the saliva/serum quotient, and from the concentration of ethynylestradiol in the saliva samples, it was possible to infer the ethynylestradiol content of the serum. Attention is called to the relation between the quotient and time, which should be taken into account when values are calculated. 2, 4 and 12 h after oral administration, the saliva/serum quotients for an ethynylestradiol dose of 0.05 mg were 0.27, 0.76 and 0.40, respectively.
Assuntos
Etinilestradiol/análise , Diacetato de Etinodiol/análise , Saliva/análise , Etinilestradiol/sangue , Diacetato de Etinodiol/sangue , Feminino , Humanos , Radioimunoensaio/métodos , Fatores de TempoRESUMO
A procedure is described for the assay of ethynodiol diacetate and ethinyl estradiol/mestranol by HPLC using two UV detectors at 210 and 280 nm. The system was acetonitrile 38% (v/v) in water as mobile phase on a 250 x 3.2-mm i.d. RP-2 column, with butylated hydroxytoluene as the internal standard. There was greater than 99% recovery from synthetic preparations and the coefficient of variation was greater than 2.0% for formulations.
Assuntos
Anticoncepcionais Orais Combinados/análise , Anticoncepcionais Orais/análise , Etinilestradiol/análise , Diacetato de Etinodiol/análise , Mestranol/análise , Cromatografia Líquida de Alta Pressão/métodos , Espectrofotometria Ultravioleta/métodos , Comprimidos/análiseRESUMO
Mestranol in combination with ethynodiol diacetate, an oral contraceptive formulation, is isolated from the sample on a partition chromatographic column prior to colorimetric determination. The color reaction which is specific for estrogens is formed by shaking an aliquot of the heptane eluate of mestranol with a 30% methanol-sulfuric acid solution. A collaborative study of the method gave results of 99.8% of added mestranol for the simulated mix and 100.7% of labelled mestranol for the commercial tablet. The method has been adopted as official first aciton.
Assuntos
Diacetato de Etinodiol/análise , Mestranol/análise , Fenômenos Químicos , Química , Cromatografia , Colorimetria , Combinação de Medicamentos , Métodos , SolventesRESUMO
The estrogen potencies of 9 oral contraceptive pills, Enovid-E, Enovid-5, Ovulen, Demulen, Norinyl+80, Norinyl+50, Ovral, Norlestrin 1 mg. and Norlestrin 2.5 mg., were determined by bioassay. Relative estrogen potency was determined by analysis of variance. Enovid-5, the most estrogenic compound, had a potency of 4.88 compared to ethinyl estradiol, 50 mcg. equal 1.00; Ovral, the least estrogenic compound, had a potency of 0.81, a sixfold difference. Estrogen potencies at a fractional dose of 0.00155 correlate with reports of the incidence of minor side effects and thromboembolic disease. The effect of progestins on estrogen potency was purely additive (norgestrel and norethynodrel), purely antagonistic, or additive at low concentrations and antagonistic at high concentrations (norethindrone, norethindrone acetate, and ethynodiol diacetate). These results suggest that pills with a greater margin of safety might be developed by utilizing greater ratios of progestin to estrogen. In addition, differences in relative estrogen potency of oral contraceptive pills may be used as a basis for better clinical selection.