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Method and reagent for quantifying cholesterol in triglyceride-rich lipoprotein

A technology for triglycerides and lipoproteins, used in biochemical equipment and methods, microbial determination/inspection, measurement devices, etc.

Active Publication Date: 2019-02-05
DENKA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These are all methods that can be measured by a general-purpose automatic analyzer, but only RLP is measured, which is different from the present invention that measures the entire TRL

Method used

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  • Method and reagent for quantifying cholesterol in triglyceride-rich lipoprotein
  • Method and reagent for quantifying cholesterol in triglyceride-rich lipoprotein
  • Method and reagent for quantifying cholesterol in triglyceride-rich lipoprotein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] The reagent composition A for carrying out the step (1) (the reagent composition for carrying out the step (1) is also referred to as "reagent composition A" hereinafter in Example 2) was prepared as follows, and the reagent composition A for carrying out the step (2) was prepared as follows. (The reagent composition used to carry out step (2) is also referred to as "reagent composition B" after Example 2).

[0071] Reagent composition A

[0072] PIPES buffer, pH6.8 50mmol / L

[0073] Various cholesterol esterases (refer to Table 1) 3U / mL

[0074] Cholesterol oxidase 3U / mL

[0075] Catalase 1200U / mL

[0076]TOOS 2.0mmol / L

[0077] Polyoxyethylene stilbene phenyl ether [HLB:12.8] 0.25%(w / v)

[0078] Reagent composition B

[0079] PIPES buffer, pH6.8 50mmol / L

[0080] 4-Aminoantipyrine 4.0mmol / L

[0081] Peroxidase 20 units / mL

[0082] Sodium azide 0.05%(w / v)

[0083] Polyoxyethylene alkyl ether 0.5%(w / v)

[0084] 150 μL of reagent composition A was added to 3 μ...

Embodiment 2

[0090] Reagent Composition A and Reagent Composition B were formulated as described below.

[0091] Reagent composition A

[0092] PIPES buffer, pH6.8 50mmol / L

[0093] Cholesterol esterase [62kDa] 3U / mL

[0094] Cholesterol oxidase 3U / mL

[0095] Catalase 1200U / mL

[0096] TOOS 2.0mmol / L

[0097] Various surfactants (refer to Table 2)※ 0.25%(w / v)

[0098] ※In the case of combining 2 or more types, the total is 0.25% (w / v)

[0099] Reagent composition B

[0100] PIPES buffer, pH6.8 50mmol / L

[0101] 4-Aminoantipyrine 4.0mmol / L

[0102] Peroxidase 20 units / mL

[0103] Sodium azide 0.05%(w / v)

[0104] Polyoxyethylene alkyl ether 0.5%(w / v)

[0105] 150 μL of reagent composition A was added to 3 μL of serum sample, and after reacting at 37° C. for 5 minutes, 50 μL of reagent composition B was added and allowed to react for 5 minutes, and the absorbance at the main wavelength of 600 nm and the sub-wavelength of 700 nm was measured. Table 2 shows the correlation coeffici...

Embodiment 3

[0110] Reagent Composition A and Reagent Composition B were formulated as described below.

[0111] Reagent composition A

[0112] PIPES buffer, pH6.8 50mmol / L

[0113] Cholesterol esterase [62kDa] 3U / mL

[0114] Cholesterol oxidase 3U / mL

[0115] Catalase 1200U / mL

[0116] TOOS 2.0mmol / L

[0117] Polyoxyethylene polycyclic phenyl ether [HLB: 12.8] Various concentrations (refer to Table 3)

[0118] Reagent composition B

[0119] PIPES buffer, pH6.8 50mmol / L

[0120] 4-Aminoantipyrine 4.0mmol / L

[0121] Peroxidase 20 units / mL

[0122] Sodium azide 0.05w / v%

[0123] Polyoxyethylene alkyl ether 0.5w / v%

[0124] 150 μL of reagent composition A was added to 3 μL of serum sample, and after reacting at 37° C. for 5 minutes, 50 μL of reagent composition B was added and allowed to react for 5 minutes, and the absorbance at the main wavelength of 600 nm and the sub-wavelength of 700 nm was measured. Table 3 shows the correlation coefficient obtained by comparing with the TRL-...

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Abstract

Disclosed are a reagent and a method which are for more specifically quantifying cholesterol (TRL-C) in triglyceride-rich lipoprotein in a test sample, and do not require complex manipulation. The method for quantifying cholesterol in triglyceride-rich lipoprotein comprises: a step (1) for selectively removing cholesterol in lipoproteins other than triglyceride-rich lipoprotein (TRL) by activatinga surfactant and a cholesterol esterase having a molecular weight of more than 50 kDa; and a step (2) for quantifying the remaining cholesterol (TRL-C) in the TRL.

Description

technical field [0001] The present invention relates to cholesterol in triglyceride-rich lipoproteins (hereinafter sometimes referred to as TRLs) (hereinafter referred to as "lipoprotein name" cholesterol or "lipoprotein name" -C means the lipoprotein described in " " Cholesterol in the Quantitative Methods and Quantitative Reagents. Background technique [0002] Lipoproteins contained in blood are classified into chylomicrons, very low density lipoproteins (hereinafter sometimes referred to as VLDL), intermediate density lipoproteins (hereinafter sometimes referred to as IDLs), and low density lipoproteins according to differences in densities separated by ultracentrifugation (hereinafter sometimes referred to as LDL), high density lipoprotein (hereinafter sometimes referred to as HDL). It is known that these lipoproteins have different contents of lipids such as triglycerides and cholesterol, and proteins, and the like, and each of them exhibits different actions in vivo....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/60C12Q1/44G01N33/92
CPCC12Q1/60C12Q1/44G01N33/92C12Q1/26G01N2405/02
Inventor 佐藤谦亨伊海田诚平尾裕子伊藤康树
Owner DENKA CO LTD
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