Simultaneous determination of sulfamethoxazole and trimethoprim in biological fluids for high-throughput analysis: comparison of HPLC with ultraviolet and tandem mass spectrometric detection.

Author(s): Bedor DC, Goncalves TM, Ferreira ML, de Sousa CE, Menezes AL, Oliveira EJ, de Santana DP

Affiliation(s): Nucleo de Desenvolvimento Farmaceutico e Cosmetico, Rua Professor Arthur de Sa s/n, Cidade Universitaria, CEP 50740-520, Recife, Pernambuco, Brazil. danilo_bedor@yahoo.com.br

Publication date & source: 2008-02-15, J Chromatogr B Analyt Technol Biomed Life Sci., 863(1):46-54. Epub 2008 Jan 17.

Publication type: Comparative Study; Randomized Controlled Trial

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.