Method for quantifying cholesterol in high-density lipoprotein 2, and reagent kit for the method

Patent metadata

Field	Value
Publication number	US-10955426-B2
Application number	US-201816019175-A
Country	US
Kind code	B2
Filing date	Jun 26, 2018
Priority date	Feb 28, 2011
Publication date	Mar 23, 2021
Grant date	Mar 23, 2021

Abstract

Disclosed is a method for quantifying HDL2 cholesterol in a test sample without requiring laborious operations. The method for quantifying cholesterol comprises allowing phospholipase to act on HDL to quantify cholesterol. Also disclosed is a method comprising: a first step of transferring cholesterols other than high-density lipoproteins in a test sample to the outside of the reaction system; and a second step of quantifying high-density lipoprotein 2 cholesterol among the high-density lipoproteins remaining in the reaction system; wherein, by performing the second step by the above method, high-density lipoprotein 2 cholesterol in the test sample can be quantified.

First claim

The invention claimed is:

1. A kit for quantifying cholesterol in high-density lipoprotein 2, said kit comprising:
a reagent (i); and
a reagent (ii),
wherein the reagent (i) comprises cholesterol esterase, cholesterol oxidase, N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline (HDAOS), a catalase, a monovalent or divalent cation(s) or a salt(s) thereof and an polyoxyethylene-polyoxypropylene condensate, and the reagent (i) does not include a phospholipase C, a phospholipase D, sphingomyelinase and 4-aminoantipyrine,
wherein the reagent (ii) comprises a phospholipase D, 4-aminoantipyrine, peroxidase and an effective catalase inhibiting amount of sodium azide, and wherein the reagent (ii) does not include cholesterol esterase and cholesterol oxidase.

2. The kit according to claim 1 , wherein the concentration of said phospholipase D in the reagent (ii) is 0.1 to 200 U/mL.

3. The kit according to claim 2 , wherein the concentration of said phospholipase D in the reagent (ii) is 1.0 to 50 U/mL.

4. The kit according to claim 1 , wherein said polyoxyethylene-polyoxypropylene condensate is Pluronic P123.

5. The kit according to claim 1 , wherein the concentration of said polyoxyethylene-polyoxypropylene in the reagent (ii) is 0.002 to 3.0% by weight.

6. The kit according to claim 1 , wherein the monovalent and/or divalent cation(s) and/or the salt(s) is sodium chloride.

7. The kit according to claim 6 , wherein the concentration of said sodium chloride is 5 to 30 g/L.

8. The kit according to claim 1 , wherein the concentration of said catalase is 600 U/mL.

9. The kit according to claim 1 , wherein the concentration of said cholesterol esterase is 0.2 to 2.0 U/mL.

10. The kit according to claim 1 , wherein the concentration of said cholesterol oxidase is 0.1 to 1.0 U/mL.

11. The kit according to claim 1 , wherein the concentration of said N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline is 0.5 to 2.0 mmol/L.

12. The kit according to claim 1 , wherein the concentration of said sodium azide is 0.1 g/L to 1.0 g/L.

13. The kit according to claim 1 , wherein the concentration of said 4-aminoantipyrine is 0.1 to 2.0 mmol/L.

Assignees

• Denka Seiken Kk
• Denka Company Ltd

Inventors

• Higuchi Maiko
• Itoh Yasuki

Classifications

• C12Q1/60

  involving cholesterol · CPC title

• C12Q1/44 Primary

  involving esterase · CPC title

• G01N33/92 Primary

  involving lipids, e.g. cholesterol {, lipoproteins, or their receptors (steroid hormones G01N33/743)} · CPC title

• G01N2333/916

  acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4) · CPC title