Paroni, R
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
,
De Vecchi, E
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
,
Cighetti, G
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
,
Arcelloni, C
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
,
Fermo, I
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
,
Grossi, A
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
,
Bonini, P
(Department of Laboratory Medicine, IRCCS, H. San Raffaele, Milan, Italy)
AbstractThis HPLC assay with o-phthalaldehyde precolumn derivatization is used to measure the total, oxidized, and protein-bound forms of glutathione in human blood, plasma, and rat tissue. Total glutathione (i.e., sum of reduced, oxidized, and protein-bound fractions) was determined after reduction...
AbstractThis HPLC assay with o-phthalaldehyde precolumn derivatization is used to measure the total, oxidized, and protein-bound forms of glutathione in human blood, plasma, and rat tissue. Total glutathione (i.e., sum of reduced, oxidized, and protein-bound fractions) was determined after reduction with dithiothreitol and protein precipitation with perchloric acid (PCA). A preliminary selective blockage of free sulfhydryl groups with N-ethylmaleimide was necessary to evaluate the different oxidized forms. The assay showed high sensitivity ( 0.999). Samples, after PCA acidification, were stable at room temperature and 4 degrees C for 3 days, and at -20 degrees C and -80 degrees C for > 1 month. The method (involving automated derivatization) not only is very rapid and simple but also allows immediate processing of many different biological samples.
AbstractThis HPLC assay with o-phthalaldehyde precolumn derivatization is used to measure the total, oxidized, and protein-bound forms of glutathione in human blood, plasma, and rat tissue. Total glutathione (i.e., sum of reduced, oxidized, and protein-bound fractions) was determined after reduction with dithiothreitol and protein precipitation with perchloric acid (PCA). A preliminary selective blockage of free sulfhydryl groups with N-ethylmaleimide was necessary to evaluate the different oxidized forms. The assay showed high sensitivity ( 0.999). Samples, after PCA acidification, were stable at room temperature and 4 degrees C for 3 days, and at -20 degrees C and -80 degrees C for > 1 month. The method (involving automated derivatization) not only is very rapid and simple but also allows immediate processing of many different biological samples.
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