Determination of Glutathione in Biological Samples by Ion-pairing HPLC/FLD

이온쌍 HPLC/FLD를 이용한 생체 시료중의 Glutathione 농도 분석

  • Yoo, Jeong-Yeon (Seoul Medical Science Institute, Seoul Clinical Laboratories(SCL)) ;
  • Lee, Kyoung-Ok (Seoul Medical Science Institute, Seoul Clinical Laboratories(SCL)) ;
  • Shin, Ho-Sang (Department of Environmental Education, Kongju University)
  • 유정연 (서울의과학연구소 서울임상병리검사센터(SCL)) ;
  • 이경옥 (서울의과학연구소 서울임상병리검사센터(SCL)) ;
  • 신호상 (공주대학교 환경교육학과)
  • Received : 1998.10.14
  • Published : 1999.02.25

Abstract

Glutathione(GSH) in biological samples was determined by high performance liquid chromatographic(HPLC) method with fluorescence detector(FLD) after monobromobimane(MBB) or 4-fluoro-7-sulfobenzofurazan(SBD-F) derivatization. The detection limit of $0.03{\mu}g/mL$ was obtained after MBB derivatization, derivative of MBB was about 200 times more sensitive than that of SBD-F. N-acetylcysteine was used as internal standard and tetrabutylammonium ion as counter ion for better separation. The determination by ion-pairing chromatography after MBB derivatization was characterized by linearity in the range between $0.08{\sim}8.33{\mu}g/mL$ with a good correlation coefficient of 0.998. By precision test appeared relative standard deviation at less than 5% at three different concentrations. This method can be used for the analysis of GSH in plasma and tissue.

Keywords

glutathione;HPLC/FLD;monobromobimane;plasma;rat liver tissue

Acknowledgement

Supported by : 보건복지부

References

  1. J. Chromatogr. v.345 M. K. Halbert;R. P. Baldwin
  2. Oncol. biol. Phys. v.10 A. I. Minchinton;I. J. Radiat
  3. J. Chromatogr. v.338 C. Komuro;K. Ono;Y. Shibmoto;T. Nishidal;M. Takahashi;M. Abe
  4. Methods Enzymol. v.143 M. W. Fariss;D. J. Reed
  5. J. Chromatogr. v.282 T. Toyo'oka;K. Imai
  6. J. Chromatogr. v.414 K. Nakashima;C. Umekawa;H. Yoshido;S. Nakatsuji;S. Akiyama
  7. Anal. Biochem. v.140 M. Shea;S. Howell
  8. Cancer Res. v.46 G. C. Rice;E. A. Bump;D. C. Schrieve;W. Lee;M. Kovacs
  9. J. Chromatogr. v.424 S. Velury;S. B. Howell
  10. Anal. Biochem. v.184 A. M. Svardal;M. A. Mansoor;P. M. Ueland
  11. J. Chromatogr. v.381 A. F. Stein;R. L. Dills;C. D. Klaassen
  12. J. Chromatogr. v.420 Y. Imai;S. Ito;K. Fujita
  13. Clin. Chem. v.41 R. Paroni;E. Devecchi;G. Cighetti;C. Arcelloni;I. Fermo;A. Grossi;P. Bonni
  14. Anal. Biochem. v.160 H. Sunaga;E. Kobayashi;N. Shimojo;K. T. Suzuki
  15. Methods Enzymol. v.143 E. G. DeMaster;B. Redfern
  16. J. Chromatogr. v.419 K. Shimada;T. O. Nambara
  17. J. Chromatogr. v.422 A. Araki;Y. Sako
  18. Clin. Chem. v.24 D. L. Radbestein;R. Saetre
  19. J. Chromatogr. v.382 N. K. Burton;G. W. Aherne
  20. Ann. Rev. Biochem. v.52 A. Meister;M. E. Anderson
  21. Life Sci. v.23 I. Mefford;R. N. Adams
  22. Methods Enzymol. v.251 G. L. Newton;R. C. Fahey