Analytical method to evaluate in vitro intestinal absorption of the Acetazolamide by HPLC.

MARIA JULIA MORA; ANA LUIZA RIBEIRO DE SOUZA; MARCO VINICIUS CHAUD; MARIA PALMIRA D. GREMIÃO; MARCELA RAQUEL LONGHI; GLADYS ESTER GRANERO

Ribeirão Preto, San Pablo, Brasil

Congreso; 7º Congreso Internacional de Ciências Farmacéuticas (CIFARP), Ribeirão Preto; 2009

ANALYTICAL METHOD TO EVALUATE, IN VITRO, THE INTESTINAL ABSORPTION OF THE ACETAZOLAMIDE BY HPLC

ANA LUIZA R. SOUZA¹; JULIA²; GLADYS²; MARCELA²; MARCO VINICIUS CHAUD³; MARIA PALMIRA D. GREMIÃO¹

1 Faculdade de ciências Farmacêuticas; 2Faculdad de Ciências Químicas – Universidad Nacional de Córdoba; 3UNISO

INTRODUCTION: The acetazolamide (ACZ) is a potent and reversible carbonic anhydrase inhibitor, effective in the control of fluid secretion, e. g., glaucomas, in the treatment of certain convulsive disorders, e. g., epilepsy, and in the promotion of diuresis in instances of abnormal fluid retention. It is poorly water- and lipid-soluble drug. Despite this, it is rapidly and almost completely absorbed from the gastrointestinal tract. OBJECTIVE: In this study, a simple HPLC method was developed, in order to monitor the concentration of ACZ in TC-199 buffer in vitro, in the rat everted gut sac absorption model. METHODOLOGY: Varian HPLC system was used for purpose of analysis in the study and it consisted of a ProStar/Dynamax 210/215 Solvent Delivery Module, ProStar 330 UV-VIS PDA Spectrophotometric Detector, and a Rheodine VS 7125, injection valve with a 100μL loop. The drug was analyzed using 250 x 4.6mm C₁₈ (Varian - Chromsep) column. The isocratic mobile phase was a mixture of methanol: water pH 4.0 at 10:90 (v/v). The flow-rate was 1 mL/min and the quantitative determination was carried out at 265 nm, at 25°C. The calibration curve of ACZ was constructed in TC 199 solution. It was used as solvent medium because this method will be used to monitor concentrations of this drug in in vitro rat intestinal sac absorption experiments. RESULTS: The total running time was within 30 min, with acceptable separation of the compound of interest. The calibration curve was linear over the concentration range of 4.0 – 40.0 μg mL^-1, with the correlation coefficient equal to 0.9993. CONCLUSION: The proposed method was found to be rapid and selective and that for can be applied in the monitoring of the absorption of ACZ in in vitro everted gut sac absorption studies.