ABSTRACT
Foi desenvolvido e validado neste estudo um método analítico para quantificação de diclofenaco de dietilamônio em pele humana por cromatografia líquida de alta eficiência, segundo a Resolução 899/2003 da Agência Nacional de Vigilância Sanitária (ANVISA). Empregou-se cromatografia em fase reversa com coluna C18 150 x 4,6 mm, 5 µm Shimpack®, à temperatura de 40ºC e fase móvel, constituída por mistura de acetonitrila e tampão fosfato de sódio 20 mM pH 3,0 (70:30, v/v) com fluxo de 1,2 mL min-1. Os analitos foram detectados por UV a 280 nm e o método foi especifico, preciso, exato, robusto e linear no intervalo de 0,05 a 20 µg mL-1 (R2 = 0,998), mostrando que pode ser utilizado em estudos de penetração cutânea in vitro tendo como modelo de membrana a pele humana.
An analytical method has been developed and validated for the quantitation of diclofenac diethylamine (DDA) in human skin by high performance liquid chromatography (HPLC), in accordance with Regulation 899/2003 of the National Sanitary Surveillance Agency (ANVISA). The HPLC column was a reversed-phase Shimpack® C18, with a 5 µm particle bed, measuring 150 x 4.6 mm i.d., eluted isocratically at 40ºC with 20 mM sodium phosphate buffer (pH 3.0):acetonitrile (30:70, v/v), the mobile phase flowing at 1.2 mL min-1. Analytes were measured by a UV detector set at 280 nm. The results revealed that the method was specific, precise, accurate, robust and linear (R2=0.998) in the range from 0.05 to 20 µg mL-1. Therefore, it can safely be used to assess DDA in vitro penetration of human skin in kinetic studies.
Subject(s)
Humans , Diclofenac/administration & dosage , Skin Absorption , Chromatography, High Pressure LiquidABSTRACT
The comparison of two methods based on online solid phase extraction-liquid chromatography with UV (SPE-LC-UV) or mass spectrometry detection (SPE-LC-MS/MS) for the simultaneous quantification of sulfamethoxazole (SMZ) and trimethoprim (TMP) is presented. The methods were validated and proved to be accurate. The analysis of standard samples for SMZ at concentrations of 0.5, 1.5, 25 and 50microg/mL demonstrated a relative standard deviation of less than 6% for both methods (n=18), while TMP samples at concentrations of 0.05, 0.15, 1.5 and 5.0microg/mL were analyzed with R.S.D. of less than 4% (n=18). The method with mass spectrometric detection was approximately six times more sensitive than the method with ultraviolet detection. The total run time for the SPE-LC-MS/MS was 2.5min per sample as opposed to 18.0min for the SPE-LC-UV method. The method with MS detection in comparison with UV detection proved to be more rugged and was successfully applied to pharmacokinetics studies.
