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.
Institute of Biochemistry, University of Baluchistan, Quetta, Pakistan.
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Masoom
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Roberti
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Binaglia
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