Determination of phosphatidylcholine in a flow injection system using immobilized enzyme reactors.
Two alternative procedures are described for the quantitative determination of phosphatidylcholine in a flow-injection system utilizing immobilized enzymes. Phospholipase C from Bacillus cereus and phospholipase D from cabbage were covalently bound to the surface of controlled-pore glass beads and the enzyme-derivatized beads were packed in small columns. In the first procedure, the phospholipase C column was connected with a second column containing coimmobilized alkaline phosphatase and choline oxidase. In the alternative procedure, the column packed with immobilized phospholipase D was connected with a column packed with immobilized choline oxidase. The hydrogen peroxide produced through the action of choline oxidase in both flow-injection systems was detected amperometrically. Both procedures are suitable for an accurate and rapid quantitation of phosphatidylcholine. The sensitivity of the method based on phospholipase C and alkaline phosphatase is higher than that using phospholipase D. Quantitation of phosphatidylcholine at the nanomole level can be easily obtained using the first method.
Masoom
M
M
Institute of Biochemistry, University of Baluchistan, Quetta, Pakistan.
Roberti
R
R
Binaglia
L
L
D000429:Alcohol Oxidoreductases; D000469:Alkaline Phosphatase; D000818:Animals; D001923:Brain Chemistry; D004800:Enzymes, Immobilized; D008297:Male; D010713:Phosphatidylcholines / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012680:Sensitivity and Specificity
D004800:Enzymes, Immobilized; D010713:Phosphatidylcholines; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D000469:Alkaline Phosphatase