ABSTRACT
A quick, simple and reliable analysis method has been developed in order to determine berenil, phenamidine, diampron and dibromopropamidine by capillary zone electrophoresis in samples of serum and urine. In order to define the operation parameters in CZE, we have carried out a study on how the apparent electrophoretic mobility (mu(app)) varies when pH, buffer concentration, voltage and temperature are modified. Ohm's law plot has been studied, too. With the data obtained from this study we have determined the optimum work conditions, which are: citrate buffer 25 mM, pH=3.70, 14 kV, 30 degrees C, wavelength of the UV detector: 200 nm, capillary tube: 570 mm x 75 microm. Under these conditions, all the products appear in times between: 7.6 min phenamidine and 8.8 min dibromopropamidine, limits of detection being: berenil: 0.50, phenamidine: 0.25, diampron: 0.40 and dibromopropamidine: 0.80 microg ml(-1). We have carried out a recovery study with three kinds of extraction cartridges: Sep-pak C-18 plus, Sep-pak C-8 plus and Oasis HBL for each one of the products in blood and urine.
Subject(s)
Benzamidines/analysis , Carbanilides/analysis , Diminazene/analogs & derivatives , Electrophoresis, Capillary/methods , Animals , Benzamidines/blood , Benzamidines/urine , Carbanilides/blood , Carbanilides/urine , Diminazene/analysis , Diminazene/blood , Diminazene/urine , Electrolytes , Humans , Hydrogen-Ion Concentration , Male , Rats , Rats, Wistar , Reproducibility of Results , Sensitivity and Specificity , TemperatureABSTRACT
A number of parameters influencing the electrokinetic processing of pentamidine by micellar electrokinetic chromatography (MEKC) were studied in order to develop an analytical method for this compound. The parameters considered were: pH, ionic strength, and SDS concentration of electrolyte, temperature and working voltage. On the basis of the results obtained, the best analytical conditions for the detection of pentamidine in serum and urine by MEKC were determined. Analysis by MEKC permitted determination of the drug in 10 min. Good linearity, reproducibility and accuracy were obtained in the range 0-30 micrograms/ml for both samples, with a correlation coefficient r > or = 0.9998 and a recovery of 87-92% in serum and 90-108.9% in urine. We examined the metabolism of pentamidine using rat liver homogenates in order to exclude any possible interference of metabolites in the analysis of pentamidine.
