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Enzymatic methods for measuring plasma and tissue sphingomylelin and phosphatidylcholine

A technology of phosphatidylcholine and tissue sheath, which can be used in the test method of plasma SM and PC measurement, and the field of enzymatic measurement method, which can solve the problems of time-consuming and insensitive

Inactive Publication Date: 2009-01-28
THE RES FOUND OF STATE UNIV OF NEW YORK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method is time consuming and insensitive

Method used

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  • Enzymatic methods for measuring plasma and tissue sphingomylelin and phosphatidylcholine
  • Enzymatic methods for measuring plasma and tissue sphingomylelin and phosphatidylcholine
  • Enzymatic methods for measuring plasma and tissue sphingomylelin and phosphatidylcholine

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Embodiment Construction

[0009] Traditionally, SM and PC are measured in 4 steps: 1) lipid extraction; 2) thin-layer chromatography (TLC); 3) extraction of SM and PC from corresponding spots on the TLC plate, and 4) in individual extracts Quantitative phosphate. The whole process is time-consuming and insensitive. Although a simple method exists (Wako Pure Chemical) for measuring total phospholipids including choline (PC+SM), there is no corresponding method for direct measurement of SM and PC. The present invention relates to two rapid, specific and sensitive assays for the measurement of SM and PC in plasma.

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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.

Description

Background technique [0001] In addition to cholesterol and triglycerides, lipoproteins also contain phospholipids, of which phosphatidylcholine (PC) and sphingomyelin (SM) are the two main components, the former accounting for about 70% of the total phospholipids and the latter about 20% of total phospholipids. In human case-control studies, it has been shown that both plasma SM and SM / PC ratio are independent risk factors for coronary heart disease. [0002] It has been known for some time that SM accumulates in atherosclerosis in humans and animal models. Low-density lipoprotein (LDL) extracted from human atherosclerotic lesions is much more abundant in SM than LDL from plasma. Plasma SM levels in apoE knockout (apoE KO) mice were 4-fold higher than in wild-type mice, which may partly explain the increased atherosclerosis in these animals. The SM / PC ratio was 5-fold higher in VLDL from hypercholesterolemic rabbits. [0003] Recently, it has been demonstrated that adminis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/28
CPCC12Q1/44G01N2405/04G01N2333/916C12Q1/28G01N2405/08C12Q2326/96C12Q1/42
Inventor 江贤成穆罕默德·礼萨·霍贾提
Owner THE RES FOUND OF STATE UNIV OF NEW YORK
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