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.
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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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