Optimized spectrophotometric determination of aldehyde dehydrogenase activity in erythrocytes.
We describe a reliable and sensitive semiautomated spectrophotometric assay of aldehyde dehydrogenase (ALDH; EC 1.2.1.3) activity in erythrocytes. The hemolysate can be stabilized with sucrose, and the technique involves only microliters of hemolysate on a centrifugal analyzer. The use of microcolumns to remove interfering hemoglobin is avoided, and reproducibility of the assay has been improved by manipulating the inherent lactate dehydrogenase activity of erythrocytes by adding lactate and oxalate to the reaction mixture. These modifications have decreased the analytical imprecision of the assay, allowing a better appraisal of aldehyde dehydrogenase activity in erythrocytes as a biological marker of excess alcohol consumption. Erythrocytic ALDH activity was significantly less in 40 alcoholics than in 145 teetotallers (median activity 128 vs 219 mU/g of hemoglobin, respectively; P = 0.0001), indicating the potential of this assay as a useful marker of excess alcohol consumption.
Johnson
R D
RD
Department of Medicine, University of Adelaide, South Australia.
Bahnisch
J
J
Stewart
B
B
Shearman
D J
DJ
Edwards
J B
JB
D000079:Acetaldehyde / Q000378:metabolism; D000437:Alcoholism / Q000201:enzymology*; D000444:Aldehyde Dehydrogenase / Q000097:blood*; D000447:Aldehydes / Q000378:metabolism; D004355:Drug Stability; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D007773:Lactates; D019344:Lactic Acid; D010070:Oxalates; D019815:Oxalic Acid; D011786:Quality Control; D012016:Reference Values; D013053:Spectrophotometry
D000447:Aldehydes; D007773:Lactates; D010070:Oxalates; D019344:Lactic Acid; D019815:Oxalic Acid; C005556:propionaldehyde; D000444:Aldehyde Dehydrogenase; D000079:Acetaldehyde