CN109613282B - High-density lipoprotein cholesterol determination kit and application thereof - Google Patents
High-density lipoprotein cholesterol determination kit and application thereof Download PDFInfo
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Abstract
The invention relates to a high-density lipoprotein cholesterol determination kit, and belongs to the technical field of biological detection. The kit consists of a reagent R1 and a reagent R2; the reagent R1 includes: surfactant, magnesium chloride, 4-aminoantipyrine and buffer; the reagent R2 includes: cholesterol esterase, cholesterol oxidase, peroxidase, sodium deoxycholate, a color developing agent, magnesium chloride and buffer solution; the reagent R1 also includes tert-butyl-4-hydroxyanisole. The kit adopts a direct method for determination, and has the advantages of high accuracy, high specificity, wide linear range and the like.
Description
Technical Field
The invention belongs to the technical field of biological detection, and particularly relates to a high-density lipoprotein cholesterol determination kit and application thereof.
Background
Lipoproteins in human serum are mainly classified into: chylomicrons (CM), very low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL). The HDL is a lipoprotein with minimum particles, promotes the removal of cholesterol from peripheral tissues, transfers the cholesterol to the liver or other tissues for redistribution, can reduce the deposition of the cholesterol on blood vessel walls, and plays a role in resisting atherosclerosis.
Cholesterol, also known as cholesterol, is present in blood lipoproteins either as free cholesterol or as cholesterol esters bound to fatty acids, for example: very low density lipoprotein cholesterol (VLDL-C), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), etc. HDL is synthesized primarily by the liver and small intestine and is the smallest particle lipoprotein with almost half of each of the lipid and protein fractions. HDL is a heterogeneous class of lipoproteins that can be separated into different subfractions using different separation methods due to the varying amounts and qualities of lipids, apolipoproteins, enzymes and lipid transporters contained in the HDL particles. These HDL subfractions differ in shape, density, particle size, charge, and anti-atherosclerotic properties. HDL transports cholesterol from the surrounding tissues (including atherosclerotic plaques) to the liver for recirculation or excretion in the form of bile acids, a process known as reverse cholesterol transport.
Gotto et al found that serum HDL-C levels were inversely related to risk of atherosclerotic cardiovascular disease (ASCVD) (Gotto AM Jr, Brinton EA. insulating low levels of high-Densitiypolipoproteinecholesterol as an insulating word in a cardiovascular disease: an active group report and update. J. AmColllCardiol, 2004,43:717 724.). Therefore, the HDL-C level is clinically used as one of auxiliary diagnostic indicators of coronary heart disease and atherosclerosis.
According to the clinical blood lipid determination suggestion provided in Chinese adult dyslipidemia prevention and treatment guidelines (revised 2016), the conventional determination of serum HDL-C mainly adopts a precipitation method, which can realize high analysis specificity, but has the main disadvantage that a specimen needs to be subjected to treatment such as precipitation and centrifugation in advance, and the result is easily influenced by hyperlipidemia, the ultracentrifugation method is mainly used for determination of a target value and evaluation of various HDL-C detection methods, but has strict requirements on equipment and technical operation, and is difficult to develop in a common laboratory, at present, the homogenization method is commonly used for determination of HDL-C content, wherein a catalase clearing method is the most commonly used method, for example, Chinese patent CN107449748A discloses a high-density lipoprotein cholesterol detection kit and a use method thereof, a plurality of mixed surfactants are adopted, including a series of polyethylene sulfate, sodium dodecyl sulfate and the like, but the addition of a large amount of surfactants is very likely to consume HDL-C, further causes content determination to be inaccurate, a cholesterol-reducing kit is disclosed, and a cyclodextrin-cholesterol-C selective sulfation method is preferably used for determination of a poloxamer-C-cholesterol-C complex, and a poloxamer-cholesterol-C complex is formed by adding of a high-cholesterol-reducing method, and a high-cholesterol-reducing reagent kit is preferably used for a high-cholesterol-.
Therefore, a high-density lipoprotein cholesterol detection kit with high accuracy is urgently needed.
Disclosure of Invention
Aiming at the defect of poor accuracy in the high-density lipoprotein cholesterol content determination process in the prior art, the invention provides the high-density lipoprotein cholesterol determination kit which has high accuracy, high specificity and wider linear range and is suitable for clinical full-white-motion or semi-automatic biochemical analysis.
In order to achieve the technical effects, the invention adopts the following technical scheme:
in one aspect, the present invention provides a high density lipoprotein cholesterol assay kit comprising a reagent R1 and a reagent R2;
the reagent R1 comprises: surfactant, magnesium chloride, 4-aminoantipyrine and buffer;
the reagent R2 comprises: cholesterol esterase, cholesterol oxidase, peroxidase, sodium deoxycholate, a color developing agent, magnesium chloride and buffer solution.
The reagent R1 also comprises tert-butyl-4-hydroxyanisole.
In the kit for measuring high density lipoprotein cholesterol, the reagent R1 includes: 10.0-30.0mmol/L of surfactant, 5.0-15.0mmol/L of magnesium chloride, 0.2-2.0mmol/L of 4-aminoantipyrine, 50.0-150.0mmol/L of buffer solution and 0.1-0.7g/L of tert-butyl-4-hydroxyanisole;
the reagent R2 comprises: 0.2-3.0KU/L cholesterol esterase, 1.0-5.0KU/L cholesterol oxidase, 5.0-15.0KU/L peroxidase, 40.0-70.0mmol/L sodium deoxycholate, 1.0-5.0mmol/L developer, 5.0-15.0mmol/L magnesium chloride and 50.0-150.0mmol/L buffer solution.
Preferably, in the kit for measuring high density lipoprotein cholesterol, the reagent R1 includes: 15.0-25.0mmol/L of surfactant, 6.0-10.0mmol/L of magnesium chloride, 0.3-1.5mmol/L of 4-aminoantipyrine, 70.0-120.0mmol/L of buffer solution and 0.2-0.6g/L of tert-butyl-4-hydroxyanisole;
the reagent R2 comprises: 0.5-2.5KU/L cholesterol esterase, 2.0-4.5KU/L cholesterol oxidase, 7.0-12.0KU/L peroxidase, 45.0-65.0mmol/L sodium deoxycholate, 2.0-4.0mmol/L developer, 6.0-10.0mmol/L magnesium chloride and 70.0-120.0mmol/L buffer solution.
Still preferably, in the kit for measuring high density lipoprotein cholesterol, the reagent R1 includes: 18.0-23.0mmol/L of surfactant, 7.0-9.0mmol/L of magnesium chloride, 0.5-1.0mmol/L of 4-aminoantipyrine, 80.0-100.0mmol/L of buffer solution and 0.3-0.5g/L of tert-butyl-4-hydroxyanisole;
the reagent R2 comprises: 0.8-1.5KU/L cholesterol esterase, 2.5-3.5KU/L cholesterol oxidase, 9.0-11.0KU/L peroxidase, 50.0-62.0mmol/L sodium deoxycholate, 2.5-3.5mmol/L developer, 7.5-9.0mmol/L magnesium chloride and 80.0-100.0mmol/L buffer solution.
Wherein the surfactant is one or more of cetyl polyether-23, tween-60 and Span-60; preferably, the surfactant is ceteth-23.
Wherein the color developing agent is one or more of 2,4, 6-tribromo-3-hydroxybenzoic acid, 2-hydroxy-3-m-toluidine sodium propanesulfonate and phenol; preferably, the color developing agent is 2,4, 6-tribromo-3-hydroxybenzoic acid.
Wherein the buffer solution is one or more of 3-morpholine propanesulfonic acid buffer solution with the pH value of 7.0, phosphate buffer solution with the pH value of 7.0 and borate buffer solution with the pH value of 7.0; preferably, the buffer solution is 3-morpholine propanesulfonic acid buffer solution with the pH value of 7.0.
In a preferred embodiment, in the high density lipoprotein cholesterol assay kit, the reagent 1 comprises: 90mmol/L of 3-morpholine propanesulfonic acid buffer solution with 7.0mmol/L value of cetyl polyether-2322.5 mmol/L, 8.0mmol/L of magnesium chloride, 0.6mmol/L, pH of 4-aminoantipyrine and 0.4g/L of tert-butyl-4-hydroxyanisole;
the reagent R2 comprises: 3.0mmol/L of 2,4, 6-tribromo-3-hydroxybenzoic acid, 1.0KU/L of cholesterol esterase, 3.0KU/L of cholesterol oxidase, 10.0KU/L of peroxidase, 56.0mmol/L of sodium deoxycholate, 8.0mmol/L of magnesium chloride and 90mmol/L of 3-morpholine propanesulfonic acid buffer solution with the pH value of 7.0.
The high-density lipoprotein cholesterol determination kit also comprises 2g of high-density lipoprotein cholesterol standard substance freeze-dried powder which is dissolved by normal saline for use. .
On the other hand, the invention also provides a using method of the high-density lipoprotein cholesterol determination kit, which comprises the following steps:
the kit is suitable for full-automatic biochemical analyzers such as DuPont AR, Hitachi 7080/7170/7600/7180, Merrill BS320, Dirrill CS600, Yapeh C8000, Toshiba TBA40FR, Aolin Bass AU2700, Beckman DXC800, Siemens ADVIA2400, Roche P800 and the like.
The determination method of the determination kit is a two-point end point method, and the reaction direction is liter reaction:
(1) uniformly mixing 300 mu L of reagent R1 and 3 mu L of sample to be detected, incubating for 5min at 37 ℃ to obtain reaction liquid 1, and determining absorbance A1Measuring the main wavelength to be 546nm and the sub-wavelength to be 660 nm;
(2) adding 100 μ L reagent R2, mixing, reacting for 5min to obtain reaction solution 2, and measuring absorbance A2Measuring the main wavelength to be 546nm and the sub-wavelength to be 660 nm;
(3) according to the steps (1) to (2), the absorbance of the high-density lipoprotein cholesterol standard substance with different concentrations is measured to obtain the absorbance ABlank spaceAnd AStandard of merit；
(4) The absorbance change value delta A (A) of the standard substance is usedStandard of merit-ABlank space) Taking the concentration c of the corresponding standard substance as a horizontal coordinate, and drawing a standard curve to obtain a standard curve equation;
(5) calculating the absorbance change value delta A (A) of the sample2-A1) And obtaining the concentration of the high-density lipoprotein cholesterol in the sample according to a standard curve equation.
The invention also provides application of the kit in determination of human serum high-density lipoprotein cholesterol content.
Compared with the prior art, the invention has the following beneficial effects:
(1) the high-density lipoprotein cholesterol determination kit provided by the invention has simple components, and the HDL-C content can be accurately determined by proportioning the reagent components according to a better concentration, wherein the linear range can reach 13 mmol/L;
(2) the high-density lipoprotein cholesterol determination kit provided by the invention uses a selective inhibition method;
(3) according to the high-density lipoprotein cholesterol determination kit provided by the invention, the accuracy is obviously improved by adding tert-butyl-4-hydroxyanisole, and the linear correlation coefficient r of theoretical concentration and actual concentration is not less than 0.9950;
(4) the high-density lipoprotein cholesterol determination kit provided by the invention needs a small amount of samples (only 3 mu L of serum), and is basically not influenced by hemoglobin, triglyceride, bilirubin and the like; when hemoglobin, triglyceride, and bilirubin were as high as 8.5g/L, 8.0mmol/L, and 140. mu. mol/L, respectively, there was no significant effect on the HDL-C assay.
Drawings
FIG. 1 is a high density lipoprotein cholesterol standard curve;
FIG. 2 shows the linear range of the high density lipoprotein cholesterol assay kit of the present invention.
Detailed Description
The present invention will be further explained with reference to specific embodiments in order to make the technical means, the original characteristics, the achieved objects and the effects of the present invention easy to understand, but the following embodiments are only preferred embodiments of the present invention, and not all embodiments are possible. Based on the embodiments in the implementation, other embodiments obtained by those skilled in the art without any creative efforts belong to the protection scope of the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified, and materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Test conditions and methods:
the instrument comprises the following steps: hitachi 7080 full-automatic biochemical analyzer
Parameters are as follows:
| dominant wavelength | 546nm | Sample (I) | 3μL |
| Sub-wavelength | 660nm | Reagent R1 | 300μL |
| Reaction temperature | 37℃ | Reagent R2 | 100μL |
| Reaction direction | Direction of rising | Type of reaction | End point method |
The method comprises the following operation steps:
example 1
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent R1 includes: tween-6010.0 mmol/L, magnesium chloride 5.0mmol/L, 4-aminoantipyrine 0.2mmol/L, buffer solution 50.0mmol/L and tert-butyl-4-hydroxyanisole 0.1 g/L; the reagent R2 includes: 0.2KU/L cholesterol esterase, 1.0KU/L cholesterol oxidase, 5.0KU/L peroxidase, 40.0mmol/L sodium deoxycholate, 1.0mmol/L sodium 2-hydroxy-3-m-toluidine propanesulfonate, 5.0mmol/L magnesium chloride, and 50.0mmol/L phosphate buffer solution with pH of 7.0.
Example 2
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent R1 includes: tween-6030.0 mmol/L, magnesium chloride 15.0mmol/L, 4-aminoantipyrine 2.0mmol/L, buffer solution 150.0mmol/L and tert-butyl-4-hydroxyanisole 0.7 g/L; the reagent R2 includes: cholesterol esterase 3.0KU/L, cholesterol oxidase 5.0KU/L, peroxidase 15.0KU/L, sodium deoxycholate 70.0mmol/L, sodium 2-hydroxy-3-m-toluidine propanesulfonate 5.0mmol/L, magnesium chloride 15.0mmol/L, and phosphate buffer solution with pH value of 7.0 150.0 mmol/L.
Example 3
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent R1 includes: 70.0mmol/L of borate buffer solution with the value of 7.0 of Span-6015.0 mmol/L, 6.0mmol/L of magnesium chloride, 0.3mmol/L, pH of 4-aminoantipyrine and 0.2g/L of tert-butyl-4-hydroxyanisole; the reagent R2 includes: 0.5KU/L cholesterol esterase, 2.0KU/L cholesterol oxidase, 7.0KU/L peroxidase, 45.0mmol/L sodium deoxycholate, 2.0mmol/L phenol, 6.0mmol/L magnesium chloride, and 70.0mmol/L borate buffer solution with pH of 7.0.
Example 4
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent R1 includes: 120.0mmol/L of borate buffer solution with the value of 7.0 of Span-6025.0 mmol/L, 10.0mmol/L of magnesium chloride, 1.5mmol/L, pH of 4-aminoantipyrine and 0.6g/L of tert-butyl-4-hydroxyanisole; the reagent R2 includes: cholesterol esterase 2.5KU/L, cholesterol oxidase 4.5KU/L, peroxidase 12.0KU/L, sodium deoxycholate 65.0mmol/L, phenol 4.0mmol/L, magnesium chloride 10.0mmol/L, and borate buffer 120.0mmol/L with pH 7.0.
Example 5
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent R1 includes: 80.0mmol/L of 3-morpholine propanesulfonic acid buffer solution with 7.0mmol/L of cetyl polyether-2318.0 mmol/L, 7.0mmol/L of magnesium chloride, 0.5mmol/L, pH of 4-aminoantipyrine and 0.3g/L of tert-butyl-4-hydroxyanisole; the reagent R2 includes: 0.8KU/L cholesterol esterase, 2.5KU/L cholesterol oxidase, 9.0KU/L peroxidase, 50.0mmol/L sodium deoxycholate, 2.5 mmol/L2, 4, 6-tribromo-3-hydroxybenzoic acid, 7.5mmol/L magnesium chloride and 80.0 mmol/L3-morpholinopropanesulfonic acid buffer solution with pH value of 7.0.
Example 6
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent R1 includes: cetyl polyether-2323.0 mmol/L, magnesium chloride 9.0mmol/L, 4-aminoantipyrine 1.0mmol/L, pH value 7.0 3-morpholine propanesulfonic acid buffer solution 100.0mmol/L and tert-butyl-4-hydroxyanisole 0.5 g/L; the reagent R2 includes: cholesterol esterase 1.5KU/L, cholesterol oxidase 3.5KU/L, peroxidase 11.0KU/L, sodium deoxycholate 62.0mmol/L, 2,4, 6-tribromo-3-hydroxybenzoic acid 3.5mmol/L, magnesium chloride 9.0mmol/L and pH 7.0 3-morpholinopropanesulfonic acid buffer solution 100.0 mmol/L.
Example 7
The components of the high density lipoprotein cholesterol assay kit and the concentrations thereof in this example are as follows:
the reagent 1 comprises: 90mmol/L of 3-morpholine propanesulfonic acid buffer solution with 7.0mmol/L value of cetyl polyether-2322.5 mmol/L, 8.0mmol/L of magnesium chloride, 0.6mmol/L, pH of 4-aminoantipyrine and 0.4g/L of tert-butyl-4-hydroxyanisole; the reagent R2 includes: 3.0mmol/L of 2,4, 6-tribromo-3-hydroxybenzoic acid, 1.0KU/L of cholesterol esterase, 3.0KU/L of cholesterol oxidase, 10.0KU/L of peroxidase, 56.0mmol/L of sodium deoxycholate, 8.0mmol/L of magnesium chloride and 90mmol/L of 3-morpholine propanesulfonic acid buffer solution with the pH value of 7.0.
Comparative example 1
Chinese patent CN108627510A discloses a kit.
Comparative example 2
Compared with the example 1, the reagent R1 does not contain tert-butyl-4-hydroxyanisole.
Comparative example 3
Compared with example 1, the concentration of tert-butyl-4-hydroxyanisole in the reagent R1 is 0.05 g/L.
Comparative example 4
Compared with the example 1, the concentration of the tert-butyl-4-hydroxyanisole in the reagent R1 is 0.8 g/L.
Comparative example 5
Compared with the example 3, the reagent R1 does not contain tert-butyl-4-hydroxyanisole.
Experimental example 1 high Density lipoprotein Cholesterol Standard Curve
Using the high density lipoprotein cholesterol standards at concentrations of 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5mmol/L, the high density lipoprotein cholesterol standard curve (shown in FIG. 1) was obtained by the above-described measurement procedure in example 7. Wherein the absorbance change value Delta A (A) of the standard substanceStandard of merit-ABlank space) As ordinate, and corresponding standard concentration as abscissa, standard curve equation: y 0.194x +0.0221, R2＝0.9963。
Experimental example 2 testing of the Linear Range of the high Density lipoprotein Cholesterol kit
High density lipoprotein cholesterol standards were prepared using physiological saline at concentrations of 3.0, 6.0, 9.0, 12.0, 13.0mmol/L, respectively, and HDL-C contents were measured using examples and comparative examples, respectively, with the results shown in Table 1:
TABLE 1 kit Linear Range
| Theoretical value mmol/L | 3.00 | 6.00 | 9.00 | 12.00 | 13.00 |
| Comparative example 1 | 2.90 | 5.80 | 8.40 | 11.40 | 11.90 |
| Comparative example 2 | 2.70 | 5.80 | 8.80 | 11.90 | 12.50 |
| Comparative example 3 | 2.60 | 5.90 | 8.50 | 11.70 | 12.10 |
| Comparative example 4 | 2.80 | 5.80 | 8.60 | 11.80 | 12.20 |
| Comparative example 5 | 2.70 | 6.30 | 8.70 | 11.90 | 12.60 |
| Example 1 | 2.90 | 5.70 | 8.90 | 11.80 | 12.70 |
| Example 2 | 3.30 | 5.80 | 9.30 | 12.30 | 12.80 |
| Example 3 | 3.30 | 6.20 | 9.20 | 12.20 | 13.30 |
| Example 4 | 3.20 | 6.30 | 9.30 | 12.30 | 13.20 |
| Example 5 | 3.10 | 6.20 | 9.20 | 12.20 | 13.10 |
| Example 6 | 3.10 | 6.10 | 9.10 | 12.10 | 13.20 |
| Example 7 | 3.00 | 6.00 | 9.20 | 12.10 | 13.10 |
| Example mean value | 3.13 | 6.04 | 9.17 | 12.14 | 13.06 |
The results show that: in the concentration range of 0-13.00mmol/L, the test value of the kit can reach the theoretical value and the correlation between the test value and the theoretical value is good (figure 2). In contrast, in comparative example 1, the test result was lower than the theoretical value at a sample concentration of 9.0mmol/L, and the test could not satisfy the requirements. Therefore, the kit has wide linear range and high accuracy.
Experimental example 3 testing of accuracy of high Density lipoprotein Cholesterol kit
The HDL-C content of clinical specimens was measured using the kit of the present invention (3 times per case, and the average value M was calculated), and the HDL-C content measured by ultracentrifugation was used as a standard for evaluating the accuracy of the kit, and the relative deviation was calculated. As shown in Table 2, the relative deviation of the kit and the result measured by an ultracentrifugation method is less than 5% by adding tert-butyl-4-hydroxyanisole, so that the HDL-C content test result is more accurate; the results obtained in the comparative examples in which t-butyl-4-hydroxyanisole was not added were all significantly different from those obtained in the ultracentrifugation process.
TABLE 2 kit accuracy comparison
Experimental example 4 interference experiment
In normal human serum, a certain amount of hemoglobin, triglyceride and bilirubin were added, and at the same time, an equal volume of deionized water was added as an interferent serum, and the concentrations of these samples were measured using the high density lipoprotein cholesterol kits of examples 1 to 7. The interference degree is 5 percent as the highest tolerance limit of the determination system to the interference substances, and the high-density lipoprotein cholesterol kit is not influenced by blood hemoglobin below 8g/L, triglyceride below 8.0mmol/L and bilirubin below 140 mu mol/L in serum.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (6)
1. A high density lipoprotein cholesterol assay kit, characterized in that: the kit comprises a reagent R1 and a reagent R2;
the reagent R1 comprises: cetyl polyether-2315.0-25.0 mmol/L, magnesium chloride 6.0-10.0mmol/L, 4-aminoantipyrine 0.3-1.5mmol/L, buffer solution 70.0-120.0mmol/L and tert-butyl-4-hydroxyanisole 0.2-0.6 g/L;
the reagent R2 comprises: 0.5-2.5KU/L cholesterol esterase, 2.0-4.5KU/L cholesterol oxidase, 7.0-12.0KU/L peroxidase, 45.0-65.0mmol/L sodium deoxycholate, 2.0-4.0mmol/L developer, 6.0-10.0mmol/L magnesium chloride and 70.0-120.0mmol/L buffer solution.
2. The high-density lipoprotein cholesterol assay kit according to claim 1, characterized in that: the reagent R1 comprises: cetyl polyether-2318.0-23.0 mmol/L, magnesium chloride 7.0-9.0mmol/L, 4-aminoantipyrine 0.5-1.0mmol/L, buffer solution 80.0-100.0mmol/L and tert-butyl-4-hydroxyanisole 0.3-0.5 g/L;
the reagent R2 comprises: 0.8-1.5KU/L cholesterol esterase, 2.5-3.5KU/L cholesterol oxidase, 9.0-11.0KU/L peroxidase, 50.0-62.0mmol/L sodium deoxycholate, 2.5-3.5mmol/L developer, 7.5-9.0mmol/L magnesium chloride and 80.0-100.0mmol/L buffer solution.
3. The high-density lipoprotein cholesterol assay kit according to claim 2, characterized in that: the color developing agent is one or more of 2,4, 6-tribromo-3-hydroxybenzoic acid, 2-hydroxy-3-m-toluidine sodium propyl sulfonate, phenol and 4-chlorophenol.
4. The high-density lipoprotein cholesterol assay kit according to claim 2, characterized in that: the buffer solution is one or more of 3-morpholine propanesulfonic acid buffer solution with the pH value of 7.0, phosphate buffer solution with the pH value of 7.0 and borate buffer solution with the pH value of 7.0.
5. The high-density lipoprotein cholesterol assay kit according to any one of claims 2 to 4, characterized in that: the reagent 1 comprises: 90mmol/L of 3-morpholine propanesulfonic acid buffer solution with 7.0mmol/L value of cetyl polyether-2322.5 mmol/L, 8.0mmol/L of magnesium chloride, 0.6mmol/L, pH of 4-aminoantipyrine and 0.4g/L of tert-butyl-4-hydroxyanisole;
the reagent R2 comprises: 3.0mmol/L of 2,4, 6-tribromo-3-hydroxybenzoic acid, 1.0KU/L of cholesterol esterase, 3.0KU/L of cholesterol oxidase, 10.0KU/L of peroxidase, 56.0mmol/L of sodium deoxycholate, 8.0mmol/L of magnesium chloride and 90mmol/L of 3-morpholine propanesulfonic acid buffer solution with the pH value of 7.0.
6. The use of the kit of claim 5 in the non-disease detection of serum high density lipoprotein cholesterol levels in humans.
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| CN111534568B (en) * | 2020-05-11 | 2021-08-31 | 中拓生物有限公司 | Serum high density lipoprotein cholesterol determination kit |
| CN111926057A (en) * | 2020-08-14 | 2020-11-13 | 江西乐成生物医疗有限公司 | Method for quantifying high-density lipoprotein 3 cholesterol, determination reagent and preparation method |
| CN112695071B (en) * | 2020-12-16 | 2022-12-30 | 浙江伊利康生物技术有限公司 | High-density lipoprotein 3 determination reagent, method and kit |
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| JP2653755B2 (en) * | 1994-03-08 | 1997-09-17 | 協和メデックス株式会社 | Determination of cholesterol in high density lipoprotein |
| CN1379235A (en) * | 2002-05-10 | 2002-11-13 | 肖洪武 | Process and reagent for measuring high-density lipoprotein and cholesterol |
| US7858384B2 (en) * | 2005-04-29 | 2010-12-28 | Kimberly-Clark Worldwide, Inc. | Flow control technique for assay devices |
| DE102005020384A1 (en) * | 2005-05-02 | 2006-11-09 | Therainvention Gmbh | Spectroscopic method for the detection of analytes |
| GB0623838D0 (en) * | 2006-11-29 | 2007-01-10 | Malvern Cosmeceutics Ltd | Novel compositions |
| WO2009031506A1 (en) * | 2007-09-05 | 2009-03-12 | Arkray, Inc. | Method of measuring low density lipoprotein (ldl) cholesterol and test piece for measuring ldl cholesterol |
| CN105652017A (en) * | 2016-02-02 | 2016-06-08 | 潍坊三维生物工程集团有限公司 | Reagent kit for detecting LAMBDA light chain content, method and application |
| CN108690869A (en) * | 2017-04-05 | 2018-10-23 | 广州市伊川生物科技有限公司 | A kind of high-density lipoprotein cholesterol detection kit and its detection method |
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