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Method and kit for measuring cholesterol in low density lipoproteins

A low-density lipoprotein and high-density lipoprotein technology, which is applied to the determination/inspection of microorganisms, biological testing, biochemical equipment and methods, etc.

Inactive Publication Date: 2013-06-26
ARKRAY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are complicated to operate and difficult to handle many samples, so it is difficult to use them in clinical examinations, etc.

Method used

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  • Method and kit for measuring cholesterol in low density lipoproteins
  • Method and kit for measuring cholesterol in low density lipoproteins
  • Method and kit for measuring cholesterol in low density lipoproteins

Examples

Experimental program
Comparison scheme
Effect test

reference example 1）

[0133] LDL cholesterol in the samples was measured using reagents 1-1 to 1-4 and reagent 2 shown in Table 1 below. As a sample, the following sample was used: VLDL and LDL recovered from human serum by ultracentrifugation were added to lipoprotein-free human serum, and it was diluted 20 times with reagent 1-3 (LDL concentration: 83mg / dl, VLDL concentration: 61mg / dL).

[0134] 90 μL of reagent 1-1, 30 μL of reagent 1-2, and 15 μL of reagent 1-4 were sequentially added to 20 μL of the sample, and reacted at 37° C. for 240 seconds. Next, 15 μL of reagent 2 containing surfactants shown in Table 2 below was added, and the absorbance was measured using a general-purpose automatic analyzer (trade name: BioMajesty-8, manufactured by JEOL Ltd.) to obtain LDL cholesterol measurement values. The results are shown in Table 2 below.

[0135] [Table 1]

[0136]

[0137] *1 Polyoxyalkylene polycyclic phenyl ether: Newcol 2608F (product name: Nippon Emulsifier Co., Ltd., HLB12.5, conta...

reference example 2）

[0148] Polyoxyethylene distyrenated phenyl ether with an HLB of 12.8 (trade name: EMULGENA60, manufactured by Kao Corporation, concentration 8% by weight (final concentration: 0.71% by weight)) was used as the surfactant of reagent 2, and the following Samples a to f having the compositions shown in Table 3 were measured for absorbance in the same manner as in Reference Example 1. The results are shown in figure 1 . In addition, in figure 1 In the figure, the vertical axis represents the absorbance (Abs.), and the horizontal axis represents the measurement time, and 1Pt. means 9 seconds.

[0149] [table 3]

[0150] (table 3)

LDL (mg / dL)

VLDL (mg / dL)

sample a

51

-

sample b

136

-

sample c

52

19

sample d

52

60

Sample e

51

104

Sample f

50

160

[0151] Such as figure 1 As shown, in samples c and d in which the VLDL concentration was less than 100 mg / dL, it was con...

Embodiment 1）

[0165] Use reagents 1-1 to 1-4 shown in Table 1 above, and surfactant B1 (polyoxyethylene distyrenated phenyl ether with HLB of 12.8, trade name: EMULGEN A60, manufactured by Kao Corporation, concentration 8 % by weight) and reagent 2 (three types) of surfactants (any one) shown in Table 5 below were used to measure LDL cholesterol. The results are shown in Figure 4 A and B.

[0166] LDL cholesterol was measured by the same method as in Reference Example 1. The concentration of polyoxyethylene distyrenated phenyl ether having an HLB of 12.8 was set to 8% by weight (final concentration in the reaction system: 0.71% by weight). The concentration of the surfactant shown in Table 5 below was 0% by weight, 0.1% by weight, 0.2% by weight, or 0.4% by weight. As samples, two types of samples were prepared: a sample containing only LDL (LDL concentration: 50 mg / dL) and a sample containing LDL and VLDL (LDL concentration: 52 mg / dL, VLDL: 156 mg / dL).

[0167] [table 5]

[0168] ...

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Abstract

Provided is a novel measurement method by which LDL cholesterol can be easily and accurately measured. Specifically provided is a method for measuring LDL cholesterol in a sample, which comprises a step wherein a surfactant A1, a surfactant B1 and a surfactant B2 are made to be present in a reaction liquid, which contains the sample having been subjected to an HDL elimination process, and an enzymatic reaction of the cholesterol in LDL is caused. The surfactant A1 is a polyoxyalkylene polycyclic phenyl ether having an HLB of 12.5 or less; the surfactant B1 is selected from the group consisting of a polyoxyethylene polycyclic phenyl ether having an HLB of 12.6 or more, a polyoxyethylene distyrenated phenyl ether having an HLB of 12.7-14.5 (inclusive) and a polyoxyethylene styrenated phenyl ether having an HLB of 12.0-14.5 (inclusive); and the surfactant B2 is selected from the group consisting of a polyoxyethylene lauryl ether having an HLB of 16.9 or more, a polyoxyethylene myristyl ether and a polyoxyethylene (35-40) octyl phenyl ether.

Description

technical field [0001] The present invention relates to a method for measuring cholesterol in low-density lipoprotein (hereinafter referred to as "LDL") and a kit for the measurement. Background technique [0002] LDL is a lipoprotein that functions to transport cholesterol synthesized in the liver to peripheral tissues. Cholesterol in LDL (hereinafter also referred to as "LDL cholesterol") is known to be one of the causative substances of arteriosclerosis. Therefore, from April 2008, the LDL cholesterol level was added as an inspection item in the health diagnosis based on the Japanese Industrial Safety and Health Law. [0003] As a method for measuring LDL cholesterol, for example, there is a method of separating LDL from other lipoproteins (for example, high-density lipoprotein (hereinafter also referred to as "HDL"), very low-density lipoprotein (hereinafter also referred to as "VLDL") by ultracentrifugation ) after separation, using enzymes to measure only LDL cholest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/60C12Q1/32C12Q1/44
CPCC12Q1/32C12Q1/44G01N33/92C12Q1/60
Inventor 今井敏博滨隆志
Owner ARKRAY INC
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