Determination of intracellular choline levels by an enzymatic assay.
A sensitive enzymatic assay for the measurement of intracellular choline is described. The separation of choline from choline-containing phospholipids is accomplished by a minor modification of the Folch technique. The method is based on the specific oxidation of choline by choline oxidase. Phenol and 4-aminoantipyrine in the presence of hydrogen peroxide generated by the oxidation of choline and peroxidase form a red quinone dye which can be detected spectrophotometrically. The assay was useful between 12.5 and 100 nanomoles of choline. The recovery of standard choline in liver homogenates averaged 102 +/- 1.6%. Structurally similar compounds produced minimal interference.
D000429:Alcohol Oxidoreductases; D000818:Animals; D002794:Choline / Q000302:isolation & purification*; D007421:Intestine, Small / Q000032:analysis; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D008168:Lung / Q000032:analysis; D008297:Male; D010084:Oxidation-Reduction; D051381:Rats; D011919:Rats, Inbred Strains
D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D002794:Choline