On-column formation of arsenic-glutathione species detected by size-exclusion chromatography in conjunction with arsenic-specific detectors.
The 'retention analysis method', which is based on size-exclusion chromatography (SEC) in conjunction with an arsenic-specific detector (graphite furnace atomic absorption spectrometer, GFAAS), was used to study the effect of pH (range 2.0-10.0), temperature (4, 25 and 37 degrees C), and the concentration of glutathione in the mobile phase (0.5-7.5 mM) on the formation of arsenic-glutathione species after injection of sodium arsenite using phosphate-buffered saline solutions as mobile phases. The formation of arsenic-GSH species was facilitated by low temperatures (4 degrees C), pH 6.0-8.0 and high concentrations of glutathione (7.5 mM) in the mobile phase. Simulating the physicochemical parameters found inside human red blood cells (approximately 3.0 mM glutathione, 37 degrees C, pH 7.4) and hepatocytes (approximately 7.5 mM glutathione, 37 degrees C, pH 7.4), SEC-GFAAS provided evidence for the formation of arsenic-glutathione species under these conditions. In addition, the 'chelating agent', sodium DL-2,3-dimercapto- -propanesulfonate (1.0 and 2.0 mM) was demonstrated to bind arsenous acid stronger in the presence of glutathione (7.5 mM) under these conditions (PBS buffer, pH 7.4, 37 degrees C).
Gailer
J
J
University Department of Molecular and Cellular Biology, The University of Arizona, Tucson 85721, USA.
Lindner
W
W
D001151:Arsenic / Q000032:analysis / Q000737:chemistry*; D002021:Buffers; D002614:Chelating Agents; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002850:Chromatography, Gel; D004912:Erythrocytes / Q000737:chemistry; D005978:Glutathione / Q000008:administration & dosage / Q000032:analysis / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000737:chemistry; D008954:Models, Biological; D013054:Spectrophotometry, Atomic; D013696:Temperature; D014494:Unithiol / Q000494:pharmacology
D002021:Buffers; D002614:Chelating Agents; D014494:Unithiol; D005978:Glutathione; D001151:Arsenic