RESUMEN
Plackett-Burman (P-B) experimental design has been used to optimize the factors affecting separation of Z and E isomers of clomiphene (zuclomiphene and enclomiphene respectively) using capillary electrophoresis. The P-B design was used to simultaneously investigate the following five factors: buffer ionic strength, buffer pH, heptakis (2,3,6-tri-o-methyl) beta-cyclodextrin (TMCD) concentration, methanol concentration and injection time, each at three levels. In addition to these, a dummy variable was added to estimate the variability of the system. Effects on resolution and analysis time were calculated. Based on the information gained from the P-B design, the following set of conditions was chosen: 100 mM phosphate buffer pH 2.3, 5 mM TMCD, 5% methanol, and 1.7 s hydrodynamic injection time. These conditions gave well-resolved peaks for zuclomiphene and enclomiphene.
Asunto(s)
Clomifeno/análisis , Enclomifeno , Fármacos para la Fertilidad Femenina/análisis , Clomifeno/química , Electroforesis Capilar/métodos , Fármacos para la Fertilidad Femenina/química , Análisis Multivariante , EstereoisomerismoRESUMEN
The degradation products formed when 13-cis retinoic acid (13-cis RA) and all-trans RA were exposed to fluorescent light and air were investigated. These retinoids are known to undergo Z-E isomerization (due to the existence of four unsaturated double bonds) and oxidation when exposed to light and air. Analysis by LC was carried out on a 25 cm x 4.6 mm Zorbax Rx-SIL (5 microns) with a mobile phase (1.4 ml min-1) of heptane-THF-acetic acid (96.5:3.5:0.015) and an in-line UV (365 nm) detector. The LC eluate was coupled through a Vestec universal interface to a Finnigan 4023 mass spectrometer. EI-mass spectra were obtained at 77 eV from m/z 200 to 350 with multiplier voltage of 1200 V. Solid samples of 13-cis RA and all-trans RA exposed to light and air and also solutions of these retinoids in the mobile phase exposed to the same conditions were used for the analysis. Tentative identities of the degradation products from the mass spectra suggest the isomerization of the retinoids (Z-E isomerism) and the formation of the 5,6-epoxides of these isomers. Identities of the 5,6-epoxides were confirmed with chromatographic and mass spectral data from synthetic samples of the epoxides. Isomerization occurred more readily in solution than in the solid form and the 13-cis RA isomer oxidized more readily than the all-trans isomer.
Asunto(s)
Tretinoina/análisis , Cromatografía Liquida , Fluorescencia , Indicadores y Reactivos , Isomerismo , Espectrometría de Masas , Oxidación-Reducción , Espectrofotometría Ultravioleta , Tretinoina/análogos & derivados , Tretinoina/química , Tretinoina/efectos de la radiaciónRESUMEN
Two retinoic acid isomers; 13-cis retinoic acid and all-trans retinoic acid and their photodegradation products were resolved with capillary electrophoresis (CE) (UV detector, 345 nm) using three different mobile phases: method 1--an acetonitrile modified borate buffer (pH 8.5); method 2--borate buffer (pH 8.5) modified with acetonitrile and alpha-cyclodextrin; and method 3--borate buffer (pH 8.5) modified with SDS (MEC). Concentration of acetonitrile in the buffer was varied from 10 to 50% in method 1 and resolutions of 0-1.9 were obtained for the two retinoic acid isomers. Similarly in method 2, concentration of alpha-cyclodextrin in the buffer (with 10% acetonitrile) was varied from 0 to 40 mM, giving resolutions of 0-3.8. In method 3, concentration of SDS in the buffer was varied from 5 to 60 mM resulting in resolutions of 1.3-4.1. Optimum separation conditions for the three methods were applied to the separation of photodegradation products of the two retinoids after exposure to fluorescent light for 36 h. A buffer modified with 45% acetonitrile and the same buffer modified with 10 mM SDS gave incompletely resolved electropherograms with a 72 cm x 50 microns capillary (50 cm to the detector). A buffer containing 20 mM alpha-cyclodextrin 10% acetonitrile gave completely resolved peaks for each isomer. The buffer containing 10 mM SDS gave completely resolved peaks for the photodegradation products when a 122 cm x 50 microns capillary (100 cm to detector) was used.
Asunto(s)
Isotretinoína/química , Tretinoina/química , Cromatografía/métodos , Electroforesis/métodos , Concentración de Iones de Hidrógeno , Isotretinoína/análisis , Luz , Factores de Tiempo , Tretinoina/análisisRESUMEN
The rate of release of caffeine from capsules of guarana was compared with that from capsules containing an equivalent amount of caffeine using the British Pharmacopoeia dissolution test apparatus. Determinations were carried out in media of pH 2 and 6.8 and caffeine concentrations in the dissolution fluid were determined by HPLC. No significant differences in release rates were found between the two preparations at either pH. The rate of absorption of caffeine across rat intestine using the everted gut was also compared for a guarana suspension and a solution containing an equivalent amount of caffeine. Experiments were carried out using fluids of pH 4.0 and 7.4. No significant differences in absorption between the two preparations were observed. These results show that the release and uptake of caffeine from guarana is the same as for preparations containing free caffeine.