Method for quantification of remnant-like lipoprotein cholesterol and kit for same
A method for quantifying remnant-like lipoprotein cholesterol in a sample simply and accurately without requiring separation operations, and a kit therefor are disclosed. A method for quantifying cholesterol in a remnant-like lipoprotein in a sample containing different lipoproteins including the remnant-like lipoprotein comprises a step (1) of erasing cholesterol in lipoproteins other than the remnant-like lipoprotein; and a step (2) of quantifying cholesterol in the remaining remnant-like lipoprotein. The step (1) is carried out under an action of a cholesterol esterase having a molecular weight of more than 40 kDa and not having a subunit having a molecular weight of not more than 40 kDa; and the step (2) is carried out under an action of a cholesterol esterase having a molecular weight of not more than 40 kDa or a cholesterol esterase having a subunit having a molecular weight of not more than 40 kDa.
1. A method for quantifying remnant-like lipoprotein cholesterol in a sample containing different lipoproteins including the remnant-like lipoprotein, said method comprising:
step (1) decomposing cholesterol in lipoproteins in the sample except for cholesterol in the remnant-like lipoprotein by decomposing the cholesterol to make it undetectable in step (2); and
step (2) quantifying cholesterol in the remaining remnant-like lipoprotein,
wherein said step (1) is carried out under an action of a cholesterol esterase having a molecular weight of more than 40 kDa and not having a subunit having a molecular weight of 40 kDa or less; and said step (2) is carried out under an action of a cholesterol esterase having a molecular weight of 40 kDa or less or a cholesterol esterase having a subunit having a molecular weight of 40 kDa or less.
2. The method according to claim 1 , wherein said step (1) further comprises treating said sample with said cholesterol esterase and a cholesterol oxidase, and removing generated hydrogen peroxide of step (1); and said step (2) further comprises treating said sample with said cholesterol esterase and cholesterol oxidase, and quantifying generated hydrogen peroxide of step (2).
3. The method according to claim 1 or 2 , wherein said step (1) and said step (2) are carried out in the presence of a nonionic surfactant.
4. The method according to claim 1 , wherein a nonionic surfactant is used in said step (1) and said step (2), and wherein said nonionic surfactant comprises at least one selected from the group consisting of a polyoxyethylene derivative having an HLB value of 11 to 13, polyoxyethylene alkyl ether, polyoxyalkylene alkyl ether and polyoxyethylene alkyl phenyl ether.
5. The method according to claim 1 , wherein said step (1) further comprises treating said sample with a phospholipase.