An automated ion-exchange assay for glutathione

✍ Scribed by Celia White Tabor; Herbert Tabor

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 597 KB
• Volume: 78
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90115-4

✦ Synopsis

A method has been developed for the automated assay of glutathione and of related compounds utilizing an anion-exchange resin column in an amino acid analyzer. Reduced glutathione. glutathione disuhide, bis(y-glutamyl)cystine. y-glutamylcysteine, and cysteinylglycine have been assayed either directly or after conversion to various derivatives.

' Satisfactory results have also been obtained with the use of Aminex A-27 resin, which is the same material except for a larger particle size (13.5 pm).

? During the use of this column and buffer system, sediment gradually deposits in the reaction coil and occasionally the coil has to be replaced. The reason for this is unclear, but, to minimize deposits in the coil, a Millipore disc (0.45 pm pore size) is placed above the steel mesh disc at the bottom of columns to prevent any resin particles from entering the coil. In addition, care is taken, during regeneration procedures, to divert the effluent away from the coil. Also, we have recently found that satisfactory results are obtained with the metal-free ascorbate-ninhydrin reagent, prepared as described in the Perkin-Elmer Co. manual for the Hitachi amino acid analyzer.