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Method for separating and detecting serum lipoprotein

A technology for serum lipoproteins and lipoproteins, which is applied in the field of detection of serum lipoproteins, can solve problems such as poor accuracy of particle size distribution information, inability to completely separate high-density lipoproteins and low-density lipoproteins, and inaccurate test results. Achieve the effects of reducing cross-linking, accurate quantitative analysis, and short analysis time

Active Publication Date: 2019-07-19
HEBEI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] One of the purposes of the present invention is to provide a method for distinguishing lipoproteins from serum samples to solve the problem of inaccurate detection results caused by interference of other proteins or adsorption on the detection membrane in existing serum lipoprotein detection methods
[0005] The second object of the present invention is to provide a method for detecting serum lipoproteins, to solve the problem that high-density lipoprotein and low-density lipoprotein cannot be completely separated due to the interference of other proteins in the existing serum lipoprotein detection method, and the two particles The accuracy of diameter distribution information is poor, etc.

Method used

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  • Method for separating and detecting serum lipoprotein
  • Method for separating and detecting serum lipoprotein
  • Method for separating and detecting serum lipoprotein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Use ethanol solution of Sudan Black B to stain the lipoproteins in the serum sample dilution: take 30 μL of the serum sample dilution, add 3 μL of Sudan Black B ethanol solution to it, and shake at a constant temperature at 37 °C. In the bed, the vibration frequency is 200 rpm, and the vibration is 3 h.

[0061] The stained serum sample dilutions were tested and analyzed under the following conditions: injection volume 20 μL; detector flow rate 1 mL / min, crossflow flow rate 2 mL / min; gasket 350 μm thick; RC filter Membrane; carrier solution is 50mMNaCl+ 10mM PBS+ 3mM NaN 3 mixed aqueous solution; detector wavelength λ=600 nm.

Embodiment 2

[0066] Use ethanol solution of Sudan Black B to stain the lipoproteins in the serum sample dilution: take 30 μL of the serum sample dilution, add 3 μL of Sudan Black B ethanol solution to it, and shake at a constant temperature at 37 °C. In the bed, the shaking frequency was 200 rpm for 6 h, and the serum sample dilution was stained.

[0067] The stained serum sample dilutions were tested and analyzed under the following conditions: injection volume 20 μL; detector flow rate 1 mL / min, crossflow flow rate 2 mL / min; gasket 350 μm thick; RC filter Membrane; carrier solution is 50mMNaCl+ 10mM PBS+ 3mM NaN 3 mixed aqueous solution; detector wavelength λ=600 nm.

[0068] Embodiment 1 ~ 2, comparative example 2 ~ 3 gained serum sample dilutions are tested and analyzed, and the obtained results are as follows: Figure 4~5 shown. It can be seen from the figure that the combination of Sudan Black B and serum lipoprotein is strengthened after the assisted staining with constant temper...

Embodiment 3

[0070] According to the method of Example 1, the lipoprotein in the serum sample dilution was stained with ethanol solution of Sudan Black B. Detection and analysis of the stained serum sample dilution, the analysis conditions are: the injection volume is 20 μL; the carrier liquid is 50mMNaCl + 10 mM PBS + 3 mM NaN 3 350 μm thick gasket; RC filter membrane; detector wavelength λ=600 nm; detector flow rate 1 mL / min, cross flow flow rate 1 mL / min.

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Abstract

The invention provides a method for separating and detecting serum lipoprotein. The method comprises the following steps of mixing a serum sample with a buffer solution, preparing a serum sample diluent, adding an organic solvent solution of Sudan black B into the serum sample diluent, oscillating in a constant-temperature shaker for not less than 3 hours in order to obtain a dyed serum sample diluent, and separating high-density lipoprotein and low-density lipoprotein in the dyed serum sample diluent by adopting asymmetric field-flow fractionation in combination with an ultraviolet visible light detector and detecting particle size distribution of the high-density lipoprotein and the low-density lipoprotein. The method disclosed by the invention is easy to operate and short in analysis time; the lipoprotein in the serum can be qualitatively detected, and the particle size distribution of the high-density lipoprotein and the low-density lipoprotein can be accurately quantitatively analyzed; the interference of free protein in the serum sample on lipoprotein separation is avoided; the crosslinking of a lipoprotein sample and a filter membrane is reduced; and the method has a wide application prospect.

Description

technical field [0001] The invention relates to a method for detecting lipoprotein, in particular to a preparation method for detecting serum lipoprotein. Background technique [0002] The "2017 China Cardiovascular Disease Report" pointed out that cardiovascular disease accounted for more than 40% of residents' death from diseases, and became the first cause of death among Chinese residents. Cardiovascular disease has become a major public health problem, and it is urgent to prevent and treat cardiovascular disease. Coronary artery disease (CAD) is a very common cardiovascular disease. At present, the lipoprotein-cholesterol content is clinically used as a predictor of CAD. However, recent studies have found that when the serum lipoprotein-cholesterol content is at a normal value, there is still a risk of CAD. Therefore, the evaluation of CAD and event probability from the perspective of serum lipoprotein-cholesterol level has certain limitations. Recent studies have foun...

Claims

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Application Information

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IPC IPC(8): G01N30/00G01N15/02
CPCG01N30/0005G01N15/0205G01N2030/003G01N15/01
Inventor 窦海洋张潇月陈雪张晶申世刚
Owner HEBEI UNIVERSITY
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