RESUMEN
The chiral separation of norgestrel enantiomers using reversed-phase high-performance liquid chromatography (RP-HPLC) was studied with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as chiral mobile phase additive. The effect of mobile phase composition, concentration of HP-beta-CD and column temperature on enantioselective separation were investigated. The quantification properties of the developed RP-HPLC method were examined. A baseline separation of norgestrel enantiomers was achieved on a Agilent ZORBAX Eclipse XDB-C8 column (150 mm x 4.6 mm i.d., 5 microm). The mobile phase was a mixture of acetonitrile and phosphate buffer (pH 5.0, 20 mM) containing 25 mM HP-beta-CD (30:70, v/v) with a flow rate of 1.0 ml/min. The UV detector was set at 240 nm. Calibration curves were linear (n=8) in the range of 0.2-25 microg/ml, the limit of detection and quantitation were 0.10 and 0.20 microg/ml, respectively, for racemic norgestrel. The values of RSD of repeatability and intermediate precision for spiked sample were less than 4.8%. The method was successfully applied to the enantioselective determination of this drug in stereoselective skin permeation study.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Norgestrel/aislamiento & purificación , 2-Hidroxipropil-beta-Ciclodextrina , Animales , Ratones , Norgestrel/farmacocinética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Absorción Cutánea , Estereoisomerismo , beta-CiclodextrinasRESUMEN
An enantioselective silica rod type chiral stationary phase (CSP) is presented; a novel combination of the well known enantiomer separation properties of beta-cyclodextrin and the unique properties concerning the flow behavior of silica monoliths. Two different synthesis routes are described, and it was found that the in situ modification of a plain silica rod column turned out to be the best. The chromatographic behaviour of the beta-cyclodextrin silica rod was studied and compared with a very similar commercially available beta-cyclodextrin bonded particulate material (ChiraDex). Even if the amount of beta-cyclodextrin bound to the silica rod was only about half of the amount of beta-cyclodextrin bound to ChiraDex) particles, good resolutions were achieved for a set of chiral test components like Chromakalin, Prominal, Oxazepam, Methadone and some other drugs. By taking advantage of the unique features of the silica rods relating to their flat H/u (Van Deemter) curves, fast enantiomer separations could be demonstrated.
Asunto(s)
Cromatografía/métodos , Ciclodextrinas/química , Preparaciones Farmacéuticas/aislamiento & purificación , Dióxido de Silicio/química , Cromatografía/instrumentación , Compuestos Heterocíclicos/química , Compuestos Heterocíclicos/aislamiento & purificación , Mefobarbital/química , Mefobarbital/aislamiento & purificación , Metadona/química , Metadona/aislamiento & purificación , Microscopía Electrónica de Rastreo , Norgestrel/química , Norgestrel/aislamiento & purificación , Compuestos Orgánicos/química , Compuestos Orgánicos/aislamiento & purificación , Oxazepam/química , Oxazepam/aislamiento & purificación , Preparaciones Farmacéuticas/química , Estereoisomerismo , Temperatura , Factores de TiempoRESUMEN
A system is proposed for the classification of related organic impurities in drugs and drug products including among others (separated and non-separated) intermediates, various kinds of by-products, among them products of different side reactions, epimeric/diastereomeric, enantiomeric impurities, impurities in natural products, and finally degradation products. Examples are taken mainly from the author's own experience and from among the named impurities in the European Pharmacopoeia with focus on impurities in hydrocortisone, prednisolone, enalapril maleate, lisinopril, ethynodiol diacetate, pipecuronium bromide, cimetidine, and ethynylsteroids. The methodological aspects of impurity profiling from the detection to the identification/structure elucidation and quantitative determination of impurities are briefly summarized.
Asunto(s)
Contaminación de Medicamentos , Compuestos Orgánicos/análisis , Preparaciones Farmacéuticas/aislamiento & purificación , Cimetidina/química , Cimetidina/aislamiento & purificación , Enalapril/química , Enalapril/aislamiento & purificación , Diacetato de Etinodiol/química , Diacetato de Etinodiol/aislamiento & purificación , Hidrocortisona/química , Hidrocortisona/aislamiento & purificación , Lisinopril/química , Lisinopril/aislamiento & purificación , Noretindrona/química , Noretindrona/aislamiento & purificación , Norgestrel/química , Norgestrel/aislamiento & purificación , Preparaciones Farmacéuticas/química , Pipecuronio/química , Pipecuronio/aislamiento & purificación , Prednisolona/química , Prednisolona/aislamiento & purificaciónRESUMEN
Chiral separation of D,L-Norgestrel was successfully performed by cyclodextrin-modified capillary electrochromatographic (CEC) method. Optimized CEC conditions for purity testing were achieved: Hypersil C-18 packed capillary and 10 mM 2-[N-morpholino]-ethanesulfonic acid buffer (MES) and 40 v/v % methanol as background electrolyte containing 10 mM gamma-cyclodextrin at pH = 5.5. The resolution was found to be strongly dependent on the concentration of chiral selector gamma-cyclodextrin, applied voltage and temperature of the capillary. Our results represent a simple, fast and reproducible method for the separation of D,L-Norgestrel and purity testing of D-Norgestrel. It can be concluded that cyclodextrin modified HPLC, HPCE and CEC offer inexpensive, attractive and reliable chiral separation methods.
Asunto(s)
Preparaciones Farmacéuticas/aislamiento & purificación , gamma-Ciclodextrinas , Ciclodextrinas , Electroquímica , Electroforesis Capilar/métodos , Indicadores y Reactivos , Norgestrel/química , Norgestrel/aislamiento & purificación , Preparaciones Farmacéuticas/química , Preparaciones Farmacéuticas/normas , Estereoisomerismo , TermodinámicaRESUMEN
A new apolar impurity (3,17 alpha-diethinyl-13-ethyl-3,5-gonadiene-17-ol, IIb) was detected and identified in norgestrel with the aid of thin-layer and high-performance chromatography and spectroscopic techniques. IIb is the product of the acid-catalysed dehydration of an overethinylated side product (Ib) of the ethinylation step in the synthesis of norgestrel. IIb can be determined by thin-layer densitometry and high-performance liquid chromatography. Another impurity (17 alpha-ethinyl-13-ethyl-4-gonene-17-ol, IV), originating from a side product of the Birch reduction step in the synthesis of norgestrel was also detected and identified. The spot of IV overlaps with that of IIb in the TLC system of USP XXIII but can be separated and quantification by more selective TLC systems and by gas chromatography.
Asunto(s)
Anticonceptivos Sintéticos Orales/aislamiento & purificación , Norgestrel/aislamiento & purificación , Congéneres de la Progesterona/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Anticonceptivos Sintéticos Orales/síntesis química , Norgestrel/análogos & derivados , Norgestrel/síntesis química , Congéneres de la Progesterona/síntesis química , Análisis EspectralRESUMEN
4-Ene-3-ketosteroids and 17-ketosteroids were quantitatively transformed into the corresponding hydrazones using Girard P and T reagents, respectively. The positively charged derivatives were separated by capillary electrophoresis. The spectrophotometric characteristics of the derivatives permitted their sensitive detection in the 230-280 nm range. The steroids investigated included nortestosterone and its phenylpropionate, norethisterone and its oenanthate, d,l-norgestrel, dehydroepiandrosterone, androstenedione and ethisterone.
Asunto(s)
Cetosteroides/aislamiento & purificación , Androstenodiona/análogos & derivados , Androstenodiona/análisis , Androstenodiona/aislamiento & purificación , Betaína/análogos & derivados , Betaína/química , Deshidroepiandrosterona/análogos & derivados , Deshidroepiandrosterona/análisis , Deshidroepiandrosterona/aislamiento & purificación , Electroforesis Capilar , Etisterona/análogos & derivados , Etisterona/análisis , Etisterona/aislamiento & purificación , Indicadores y Reactivos/química , Cetosteroides/análisis , Nandrolona/análogos & derivados , Nandrolona/análisis , Nandrolona/aislamiento & purificación , Noretindrona/análogos & derivados , Noretindrona/análisis , Noretindrona/aislamiento & purificación , Norgestrel/análogos & derivados , Norgestrel/análisis , Norgestrel/aislamiento & purificación , Espectrofotometría UltravioletaRESUMEN
The metabolism of norgestimate (ORF-10131; 14C-d-13-ethyl-17-acetoxy-18,19-dinor-17 alpha-pregn-4-en-20- yn -3-oxime) was studied in humans. Compound labeled with carbon-14 in the 17 alpha-ethynyl group was administered to four female subjects. An average of 46.8 percent of the administered radioactivity was excreted in the urine and 36.8 percent in the feces over a two-week collection period. About 12 percent of the urinary radioactivity represented freely extractable metabolites and another 57 percent consisted of extractable material released by enzyme hydrolysis. The ethynylated metabolites of norgestimate were separated from endogenous compounds and non- ethynylated metabolites by silver- sulfoethyl cellulose column chromatography. Metabolites were subsequently isolated by high performance liquid chromatography and thin layer chromatography. The identification of five urinary metabolites was accomplished by combined gas-liquid chromatography/mass spectrometry. These metabolites include norgestrel, 16 beta- hydroxynorgestrel , 2 alpha- hydroxynorgestrel , 3 alpha, 5 beta- tetrahydronorgestrel , and a fifth trihydroxylated metabolite of undetermined stereochemical configuration; 3,16-dihydroxy-5- tetrahydronorgestrel .
Asunto(s)
Norgestrel/análogos & derivados , Biotransformación , Radioisótopos de Carbono , Heces/análisis , Femenino , Humanos , Norgestrel/aislamiento & purificación , Norgestrel/metabolismo , Norgestrel/orinaRESUMEN
PIP: A method generally applicable to procedures for extraction and purification of ethynyl steroids in urine and plasma before gas chromatography (GC)/mass spectrometry (MS) analysis is described. The method requires using 20 ml of urine or 10 ml of plasma, which is then extracted on a column of Amberlite (3 gm). The extract is filtered to remove organic cations, and then after neutral steroids and other compounds are separated into groups by passing the efluent through the column, a further filtering on SP-Sephadex yields ethynyl steroids, which are selectively retained by the ion exchanger and are then eluted by displacement with l-hexyne. Using this method, recoveries of ethynyl steroids were from 75-90%. A simplified method is also outlined which can be used when separation of neutral, phenolic, and conjugated steroids is not required. The GC/MS analyses are performed using an OV-1 or SE-30 open tubular capillary column and single ion detection. A combination of external and internal standards is used. Limit of detection of unconjugated ethynylestradiol in urine was 5 pgm/ml. The paper concludes with examples of analyses of urine from a healthy woman taking 250 mcgm of (-)norgestrel and 50 mcgm of ethynylestradiol daily.^ieng