Enzymatic methods for measuring plasma and tissue sphingomylelin and phosphatidylcholine
AssigneeThe Research Foundation of State University of New York
InventorsXian-Cheng Jiang, Mohammad Reza Hojjati
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Abstract
A method for measuring sphingomyelin and phosphatidylcholine comprising incubating sphingomyelin and phosphatidylcholine with bacterial sphingomyelinase and bacterial phospholipase D, alkaline phosphatase, choline oxidase, peroxidase, N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, and 4-aminoantipyrine, preferably for about 45 minutes. A blue dye is generated.
Claims3
A method for measuring plasma and tissue sphingomylelin and phosphatidylcholine comprising 1) catalyzing the hydrolysis of sphingomylelin to phosphorylcholine and n-acylsphingosine with bacterial SMase;
- 2) generating choline from phosphorylcholine produced from step 1) with alkaline phosphatase;
- 3) generating hydrogen peroxide by adding choline oxidase;
- and 4) adding hydrogen peroxide and with DAOS (N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, sodium salt), 4-aminoantipyrine and peroxidase, to generate a blue to purple dye.
A method for measuring plasma and tissue sphingomylelin and phosphatidylcholine comprising 1) catalyzing the hydrolysis of phosphatidlycholine to choline and phosphatidic acid with bacterial phospholipase D;
- 2) generating hydrogen peroxide by adding choline oxidase;
- and 3) adding hydrogen peroxide and with DAOS (N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, sodium salt), 4-aminoantipyrine and peroxidase, to generate a blue to purple dye.
Dependencies are inferred from the claim text.