Spectrofluometry of sulfonamides using fluorescamine

Fluorescamine을 이용한 sulfonamide의 형광 분석법

  • Lim, Jae-young (College of Veterinary Medicine, Seoul National University) ;
  • Han, Su-nam (College of Veterinary Medicine, Seoul National University) ;
  • Lee, Mun-han (College of Veterinary Medicine, Seoul National University) ;
  • Park, Jong-myung (Veterinary Research Institute, Rural Development Administration)
  • 임재영 (서울대학교 수의과대학) ;
  • 한수남 (서울대학교 수의과대학) ;
  • 이문한 (서울대학교 수의과대학) ;
  • 박종명 (농촌진흥청 가축위생연구소)
  • Received : 1989.12.30
  • Published : 1990.01.31


These experiments were carried out to develop a novel, simple, and rapid method to determine urinary sulfonamides using fluorescamine by spectrofluorometry. To get optimal conditions for the sulfonamide-fluorescamine reaction, sulfonamides such as sulfamethazine, sulfamerazine, sulfadimethoxine and sulfamonomethoxine, dissolved in buffers with various pH ranges were reacted with various concentrations of fluorescamine. and then, the fluorescence intensity and stability of the fluorophore were measured. To eliminate the interfering substances in urine, the fluorophore in buffers and urine with a definite pH range was extracted with some organic solvents. After then the fluorescence intensity was measured in organic and aquous phases. The results obtained were summarized as follows: 1. The maximal fluorescence of sulfonamides was presented in acidic state, pH 4.5~5.0, at 30 minutes after reaction. 2. The optimal concentration ratio of sulfamethazine and ffuorescamine was more than 1 : 40 in mole. 3. In pH 4.0, the intensity was maximal but was time-dependent, whereas in pH 8.0, the intensity was time-independent. 4. Sulfamethazine-fluorescamine conjugate could be dissolved in some of organic solvents in acidic state such as chloroform, n-butanol, and ethylacetate. 5. Sulfamethazine-flnorescamine conjugate in swine urine coule be extracted with ethylacetate in acidic state, pH 4.0~5.0.