Improved buffer system for plasma lipid analysis

Abstract

The invention relates to the field of biological detection, and discloses an improved buffer system for plasma lipid analysis, which comprises a buffer substance, a stable substance, a preservative and a densifier, the concentration of the buffer substance is 0.5 M-2M, the concentration of the stable substance is 1%-30%, the concentration of the preservative is 0.1% o, and the concentration of the densifier is 0.1%-5%; the buffer substance is prepared from one or more of tris (hydroxymethyl) aminomethane, triethanolamine, 4-(2-hydroxyethyl) piperazine ethanesulfonic acid and a barbital salt buffer solution. The buffer substance in the buffer system is added to stabilize the pH of the gel, and the shelf life of the gel preloaded column can be prolonged under the synergistic effect of the stable substance and the densifier, so that the buffer system in the gel column for plasma lipid electrophoresis is improved, and the system achieves the effects of high resolution, good performance and long preservation time; meanwhile, the strip separation after lipid electrophoresis in the plasma is clearer.

Description

Technical Field

[0001] The invention relates to the technical field of biological detection, in particular to an improved buffer system for plasma lipid analysis. Background Art

[0002] Potential pathogenic factors of cardiometabolic diseases (myocardial infarction, stroke) are closely related to lipid molecules such as total triglycerides (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). These biomarkers and molecular pathways will be focused on in clinical research. Analysis of lipids in the blood helps to understand the risk of cardiometabolic diseases and thus regulate fat in the diet.

[0003] The main sources of lipids in the blood are adipose tissue, liver and dietary lipids. Blood lipids are insoluble or slightly soluble in water. Except for fatty acids combined with albumin, the rest of the blood is combined with apolipoproteins to form lipoproteins, which are transported to various tissues through blood circulation to complete their physiological and metabolic functions. Plasma lipoproteins can be divided into many types due to the different types and proportions of lipid components and proteins they contain.

[0004] Clinical laboratories already have a variety of methods for plasma lipid analysis. Due to the diverse composition and complex structure of plasma lipids, it is usually necessary to extract and separate them first, and then select an appropriate method for analysis based on their characteristics, properties and analytical purposes. The most classic of these is ultracentrifugation, which separates various lipoproteins in plasma based on their density differences. Ultracentrifugation generally takes a long time, is cumbersome to operate and requires an ultracentrifugation device, which is not easy to promote and is not widely used. The most commonly used method is electrophoresis, which uses the different mobilities of lipids in plasma for analysis. Depending on the supporting medium, it can be divided into cellulose acetate film lipoprotein electrophoresis, agarose gel lipoprotein electrophoresis and polyacrylamide gel lipoprotein electrophoresis. However, the buffer system of the traditional electrophoresis method has limited buffering capacity, insufficient separation capacity for lipoproteins in plasma, and difficulty in distinguishing between various lipoprotein types, which restricts the realization of people's demand for accurate blood lipid analysis. In addition, the existing buffer system is prone to degradation of the electrophoresis gel, has a short storage time, and is prone to errors in lipid analysis.

[0005] Therefore, there is an urgent need in the art for a buffer system for plasma lipid analysis with high resolution, good performance and long storage time. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention provides an improved buffer system for plasma lipid analysis to solve the problems mentioned in the above background technology.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] The first aspect of the present invention provides an improved buffer system for plasma lipid analysis, comprising a buffer substance, a stabilizing substance, a preservative, and a densifier, wherein the concentration of the buffer substance is 0.5M-2M, the concentration of the stabilizing substance is 1%-30%, the concentration of the preservative is 1‰, and the concentration of the densifier is 0.1%-5%.

[0009] Preferably, the buffer substance is composed of one or more of tris(hydroxymethyl)aminomethane, triethanolamine, 4-hydroxyethylpiperazineethanesulfonic acid, and barbiturate buffer;

[0010] The stabilizing substance is composed of one or more of biological macromolecules and high molecular polymers, and the stabilizing substance includes a stabilizer, and the stabilizer is composed of one or more of sucrose, trehalose, polyethylene glycol, and polypropylene glycol;

[0011] The preservative is composed of one or more of sodium azide, Proclin 300, 5-chloro-2-methyl-4-isothiazoline-3-one, and 2-methyl-4-isothiazoline-3-one;

[0012] The densifier is composed of one or more of glycerol, ethylene glycol and propylene glycol.

[0013] The second aspect of the present invention provides an application of the improved buffer system for plasma lipid analysis according to the first aspect, characterized in that the application is selected from any one of the following applications:

[0014] A. Application in the analysis of plasma lipoproteins;

[0015] B. Application in the preparation of prepacked gel columns.

[0016] The third aspect of the present invention provides a method for analyzing plasma lipoprotein using an improved buffer system for plasma lipid analysis, comprising the following steps:

[0017] A1. Sample collection: Collect venous blood after fasting for more than 12 hours, and separate serum or plasma within 3 hours; and conduct timely testing. If the sample cannot be tested in time, store it below -70℃ and slowly thaw it when it is taken out for use;

[0018] A2. Preparation of pre-packed column: prepare the gel pre-packed column by corresponding method;

[0019] A3. Sample staining: Mix the collected sample with the staining solution evenly, incubate at room temperature for 10 minutes, and then centrifuge to obtain the supernatant, which is the lipid-stained plasma / serum;

[0020] A4. Electrophoresis analysis: Under the action of the electric field of the electrophoresis tank, the lipoproteins in the stained plasma / serum pass through the gel pre-packed column in sequence according to the particle diameter, thereby obtaining the lipoprotein separation electrophoresis results;

[0021] A5. Scanning analysis: The lipoprotein separation electrophoresis results are scanned with a scanning analysis system to obtain a lipoprotein electrophoresis spectrum, and the distribution of optical density values ​​in the electrophoresis spectrum is converted into a lipoprotein subtype distribution peak diagram.

[0022] The fourth aspect of the present invention provides a gel prepacked column prepared using an improved buffer system for plasma lipid analysis, comprising the following steps:

[0023] B1. Mixing the buffer substance, the stabilizing substance, the preservative, and the densifier according to a certain proportion to obtain a solution mother solution;

[0024] B2, dissolving 30% acrylamide-methylenebisacrylamide in ultrapure water to prepare an acrylamide solution mother solution;

[0025] B3, the solution mother liquor and the acrylamide solution mother liquor are mixed in a ratio of 5:8, and then ammonium persulfate and tetramethylethylenediamine are added, and then mixed evenly, and then divided into transparent glass tubes with sealed bottoms, and then ultrapure water is added to the upper layer, and the mixture is allowed to stand for 30 minutes, thereby obtaining a first section of a gel column with a length of 4 cm-10 cm;

[0026] B4. After the first section of the gel column is polymerized, remove the ultrapure water on the upper layer of the first section of the gel column, and then mix the solution mother liquor and the acrylamide solution mother liquor in a 1:1 ratio, and then add ammonium persulfate and tetramethylethylenediamine, mix them evenly immediately, and then add them to the upper layer of the first section of the gel column to obtain the second section of the gel column. The length of the second section of the gel column is 0.3cm-3cm, and the distance between the second section of the gel column and the upper edge of the transparent proportional tube is 2-3cm; add ultrapure water to the upper layer of the second section of the gel column and let it stand for 30 minutes to obtain a gel pre-loaded column.

[0027] A fifth aspect of the present invention provides a plasma lipid staining solution, wherein the preparation method of the plasma lipid staining solution comprises the following steps:

[0028] S1, take 2g Sudan Black B dye and dissolve it in the first solvent;

[0029] S2, then add dimethyl sulfoxide, mix well and heat to 50°C, stir for 30 minutes and filter with a 0.22μm filter membrane;

[0030] S3, and then store it away from light to obtain the staining solution.

[0031] Preferably, the first solvent is one or more of ethanol, ethylene glycol, propylene glycol, isopropanol, and acetone solvents, the volume ratio of the first solvent to the total dyeing solution is 90%-95%, and the volume ratio of the dimethyl sulfoxide to the total dyeing solution is 1%-10%.

[0032] The sixth aspect of the present invention provides use of the plasma lipid staining solution described in the fifth aspect in the analysis of plasma lipoproteins.

[0033] A seventh aspect of the present invention provides a method for using a plasma lipid staining solution in the analysis of plasma lipoproteins, comprising the following steps:

[0034] C1. Take 100 μl of sample and mix with 10 μl of staining solution;

[0035] C2. Incubate at room temperature for 10 minutes and then centrifuge to obtain the supernatant to obtain the stained sample.

[0036] In an eighth aspect of the present invention, there is provided a method for analyzing plasma lipoprotein using a gel prepacked column, comprising the following steps:

[0037] D1. Fix the prepared gel pre-packed column vertically in the electrophoresis tank, so that the second section of the electrophoresis gel column is facing upward and located at the negative end of the electrophoresis tank;

[0038] D2, then slowly add electrophoresis buffer to the positive and negative electrode tanks of the electrophoresis tank, and the electrophoresis buffer in the negative electrode tank is submerged to the upper end of the gel pre-packed column;

[0039] D3, then aspirate the stained sample and slowly add it to the top of the second gel in the gel pre-loaded column, then assemble the electrophoresis tank and turn on the power to perform electrophoresis;

[0040] D4. After the electrophoresis is completed, the gel pre-loaded columns are removed and placed on the scanning analysis system platform for scanning in turn to obtain the plasma lipoprotein electrophoresis map, which is converted into a lipoprotein subtype distribution peak map to complete the analysis.

[0041] The present invention provides an improved buffer system for plasma lipid analysis, which has the following beneficial effects:

[0042] 1. The present invention stabilizes the pH of the gel by adding a buffer substance in the buffer system. The synergistic effect of the stabilizing substance and the densifier can extend the shelf life of the gel pre-packed column, thereby improving the buffer system in the gel column for preparing plasma lipid electrophoresis, so that the system achieves the effects of high resolution, good performance and long storage time, and at the same time makes the band separation of lipids in plasma after electrophoresis clearer.

[0043] 2. The gel prepacked column prepared by the system of the present invention can effectively prevent the rapid degradation of the electrophoresis gel, save the time of repeated preparation of the gel, and reduce the analysis error caused by different batches of gel.

[0044] 3. The improved gel electrophoresis buffer system of the present invention can improve the resolution of blood lipid electrophoresis, efficiently separate the various components of blood lipoproteins, including the various subcomponents of very low density lipoprotein cholesterol, high density lipoprotein cholesterol and low density lipoprotein cholesterol, and achieve accurate separation of blood lipid subcomponents.

[0045] 4. The technical solution of the present invention can be applied to lipoprotein polymorphism analysis and lipoprotein metabolism analysis, provide more accurate cardiovascular and cerebrovascular disease risk assessment indicators, and accurately analyze various subtypes of plasma lipoproteins. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The electrophoresis patterns of 11 different plasma samples in the specific embodiment 5 of the present invention;

[0047] Figure 2 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 1 in the specific embodiment 5 of the present invention;

[0048] Figure 3 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern sample No. 2 in the specific embodiment 5 of the present invention;

[0049] Figure 4 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 3 in the specific embodiment 5 of the present invention;

[0050] Figure 5 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 4 in specific embodiment 5 of the present invention;

[0051] Figure 6 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 5 in the specific embodiment 5 of the present invention;

[0052] Figure 7 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 6 in the specific embodiment 5 of the present invention;

[0053] Figure 8 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 7 in the specific embodiment 5 of the present invention;

[0054] Fig. 9 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 8 in the specific embodiment 5 of the present invention;

[0055] Fig.10 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 9 in the specific embodiment 5 of the present invention;

[0056] Fig.11 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 10 in the specific embodiment 5 of the present invention;

[0057] Fig.12 The lipoprotein subtype distribution peak diagram of the electrophoresis pattern of sample No. 11 in the specific embodiment 5 of the present invention;

[0058] Fig.13 The electrophoresis pattern of the gel prepacked column stored at 4° C. for 18 months in specific embodiment 6 of the present invention;

[0059] Fig.14 The electrophoresis pattern of the gel prepacked column stored at 4° C. for 6 months in specific embodiment 6 of the present invention;

[0060] Fig.15 The electrophoresis patterns of the prepacked gel column in the control example of the present invention and the electrophoresis gel column of the present invention for the same batch of samples are shown. DETAILED DESCRIPTION

[0061] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0062] Embodiment 1:

[0063] An improved method for preparing a buffer system solution stock solution for plasma lipid analysis:

[0064] Weigh 45.53g of tris(hydroxymethyl)aminomethane) and dissolve it in water, add 75g of sucrose and 50g of trehalose, and stir to dissolve; measure 12.5mL of propylene glycol and add to mix evenly; finally, add 500μl of Proclin 300 preservative; adjust the pH, mix thoroughly, make up to 500mL, filter with a 0.45μm filter membrane to obtain the mother solution of the buffer system solution, and refrigerate at 4°C for later use.

[0065] Embodiment 2:

[0066] An improved method for preparing a buffer system solution stock solution for plasma lipid analysis:

[0067] Weigh 119.15g of tris(hydroxymethyl)aminomethane) and dissolve it in water, add 100g of trehalose and 10g of polyethylene glycol 10000, and stir to dissolve; measure 10mL of ethylene glycol and add to mix evenly; finally, add 500μl of Proclin 300 preservative; adjust the pH, mix thoroughly, make up to 500mL, filter with a 0.45μm filter membrane to obtain the mother solution of the buffer system solution, and refrigerate at 4°C for later use.

[0068] Embodiment three:

[0069] A gel prepacked column is prepared using an improved buffer system for plasma lipid analysis. The gel prepacked column is divided into two sections, namely, a first gel column and a second gel column from bottom to top. The length of the first gel column is 7 cm, and the length of the second gel column is 2 cm.

[0070] The preparation steps are as follows:

[0071] S1: preparing a buffer system solution stock solution as in Example 1;

[0072] S2: Prepare acrylamide solution mother liquor: weigh 29g acrylamide powder and 1g methylene bisacrylamide and dissolve them in ultrapure water, stir until completely dissolved, and filter with a 0.45μm filter membrane for later use;

[0073] S3: Prepare ammonium persulfate solution: weigh 10 g of ammonium persulfate and add it to 100 mL of ultrapure sodium chloride, stir until completely dissolved, and filter with a 0.45 μm filter membrane for later use;

[0074] S4: Preparation of the first gel column: Mix the buffer system solution mother liquor prepared in step S1 and the acrylamide solution mother liquor prepared in step S2 in a ratio of 5:8, add ammonium persulfate solution and tetramethylethylenediamine, and mix evenly immediately; add the mixture into a transparent glass tube with a sealed bottom, add 50 μl of ultrapure water to the upper layer of the gel column, and let it stand for 30 minutes; the addition amounts of ammonium persulfate solution and tetramethylethylenediamine are 0.1% and 0.1‰ respectively;

[0075] S5: Preparation of the second gel column: After the first gel column is polymerized, remove the ultrapure water on the upper layer of the first gel column; mix the buffer system solution mother liquor and the acrylamide solution mother liquor in a 1:1 ratio, add ammonium persulfate and tetramethylethylenediamine, and mix them evenly immediately; the amount of ammonium persulfate solution and tetramethylethylenediamine added is 0.1% and 0.1‰ respectively; add them to the upper layer of the first gel column; add 50μl ultrapure water to the upper layer of the second gel column and let it stand for 30 minutes; obtain a gel pre-loaded column.

[0076] Embodiment 4:

[0077] A method for preparing a plasma lipid staining solution comprises the following steps: taking 2g of Sudan Black B dye, dissolving it in 90mL of ethylene glycol, and then adding 10mL of dimethyl sulfoxide; mixing evenly, heating to 50°C, stirring for 30min, filtering with a 0.22μm filter membrane, and storing in the dark to obtain a staining solution.

[0078] Embodiment five:

[0079] A method for analyzing plasma lipoprotein using a gel prepacked column comprises the following steps:

[0080] S1: Sample collection: Collect venous blood after fasting for more than 12 hours, and separate serum or plasma within 3 hours; if the sample cannot be tested in time, the sample needs to be stored below -70℃, and slowly thawed when taken out for use, and no repeated freezing and thawing is allowed;

[0081] S2: Sample staining: Take 10 μL of staining solution in a clean centrifuge tube, add 100 μL of serum or plasma sample, mix well, and incubate at room temperature for 10 minutes to allow the staining solution to fully stain the lipoprotein in the sample;

[0082] S3: Fix one end of the gel preloaded column sample port vertically upward in the rubber hole of the disc electrophoresis tank; add appropriate amount of electrophoresis buffer to the positive and negative electrophoresis tanks respectively. The electrophoresis buffer at the bottom of the electrophoresis tank should cover the electrode column, and the electrophoresis buffer at the top of the electrophoresis tank with the preloaded column should immerse the sample loading end of the preloaded column and the electrode column on the upper cover; ensure that there is no leakage in the upper and lower electrophoresis tanks;

[0083] S5: Take out the stained sample, centrifuge at 3000 rpm for 3 min, take 50 μL of the stained supernatant after centrifugation and slowly add it to the sample loading end of the pre-loaded column to allow it to settle on the upper layer of the gel in the pre-loaded column;

[0084] S6: Cover the electrophoresis tank, connect the positive and negative electrodes, set the voltage to the maximum value, the current to 3mA / tube, and the electrophoresis time to about 70min;

[0085] S7: After the electrophoresis is completed, the pre-loaded column is taken out and the electrophoresis buffer on the surface is wiped off; the pre-loaded column after the electrophoresis is placed on the scanning platform of the scanner for scanning, and after the scanning is completed, the optical density value and proportion of the lipoprotein subtype bands in the electrophoresis spectrum are analyzed.

[0086] Embodiment six:

[0087] Real-time stability test of the gel prepacked column prepared using this system:

[0088] Gel prepacked columns from three different production batches stored at 4° C. were selected, and the same batch of plasma samples were tested according to the electrophoresis method described in Example 5 to compare whether there were differences in the separation effects of different batches of gel prepacked columns on plasma lipoproteins.

[0089] Comparative Example 1:

[0090] Prepare the gel prepacked column according to the gel electrophoresis system buffer described in CN109374717A:

[0091] Buffer system solution Tris-HCl solution: Dissolve 18.15 g Tris in distilled water, adjust the pH to 6.7 with HCl, make up to 100 mL, filter through a 0.45 μm filter membrane and set aside.

[0092] After gel preparation, the same batch of plasma samples were used for testing with the gel column prepared by the present invention to compare the difference in separation effect of the two on plasma lipoprotein.

[0093] As attached Figure 1 -Attached Fig.15 As shown, the gel prepacked column prepared by using the gel buffer system of the present invention can clearly separate the lipoproteins in plasma with sharp bands. Fig.13 and Fig.14 It is shown that the electrophoresis patterns of the gel prepacked columns in Example 6 that were refrigerated for 18 months and 6 months are still clear, and the analytical performance has not been significantly reduced, which illustrates the positive effect of the buffer system of the present invention on extending the shelf life of the gel prepacked columns and improving the analytical performance. Fig.15 From left to right in the figure are four control example gel pre-packed columns and four embodiment gel pre-packed columns of the present invention. When comparing the separation of four identical plasma samples, the separation performance of the four control example gel pre-packed columns on the left is significantly lower than that on the right, indicating that the performance of the gel pre-packed columns prepared by the buffer system of the present invention is significantly better than that of the prior art.

[0094] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An improved buffer system for plasma lipid analysis, comprising a buffer substance, a stabilizing substance, a preservative, and a densifier, characterized in that: The concentration of the buffer substance is 0.5M-2M, the concentration of the stabilizing substance is 1%-30%, the concentration of the preservative is 1‰, and the concentration of the densifier is 0.1%-5%.

2. The improved buffer system for plasma lipid analysis according to claim 1, characterized in that: The buffer substance is composed of one or more of tris(hydroxymethyl)aminomethane, triethanolamine, 4-hydroxyethylpiperazineethanesulfonic acid, and barbiturate buffer; The stabilizing substance is composed of one or more of biological macromolecules and high molecular polymers, and the stabilizing substance includes a stabilizer, and the stabilizer is composed of one or more of sucrose, trehalose, polyethylene glycol, and polypropylene glycol; The preservative is composed of one or more of sodium azide, Proclin 300, 5-chloro-2-methyl-4-isothiazoline-3-one, and 2-methyl-4-isothiazoline-3-one; The densifier is composed of one or more of glycerol, ethylene glycol and propylene glycol.

3. Application of an improved buffer system for plasma lipid analysis, characterized in that: The application is selected from any one of the following applications: A. Application in the analysis of plasma lipoproteins; B. Application in the preparation of prepacked gel columns.

4. A method for analyzing plasma lipoprotein using an improved buffer system for plasma lipid analysis, characterized in that: An improved buffer system for plasma lipid analysis according to any one of claims 1 to 3, comprising the following steps: A1. Sample collection: Collect venous blood after fasting for more than 12 hours, and separate serum or plasma within 3 hours; and conduct timely testing. If the sample cannot be tested in time, store it below -70℃ and slowly thaw it when it is taken out for use; A2. Preparation of pre-packed column: prepare the gel pre-packed column by corresponding method; A3. Sample staining: Mix the collected sample with the staining solution evenly, incubate at room temperature for 10 minutes, and then centrifuge to obtain the supernatant, which is the lipid-stained plasma / serum; A4. Electrophoresis analysis: Under the action of the electric field of the electrophoresis tank, the lipoproteins in the stained plasma / serum pass through the gel pre-packed column in sequence according to the particle diameter, thereby obtaining the lipoprotein separation electrophoresis results; A5. Scanning analysis: The lipoprotein separation electrophoresis results are scanned with a scanning analysis system to obtain a lipoprotein electrophoresis spectrum, and the distribution of optical density values ​​in the electrophoresis spectrum is converted into a lipoprotein subtype distribution peak diagram.

5. A gel prepacked column prepared with an improved buffer system for plasma lipid analysis, characterized in that: The improved plasma lipid analysis buffer system according to any one of claims 1 to 3 comprises the following steps: B1. Mixing the buffer substance, the stabilizing substance, the preservative, and the densifier according to a certain proportion to obtain a solution mother solution; B2, dissolving 30% acrylamide-methylenebisacrylamide in ultrapure water to prepare an acrylamide solution mother solution; B3, the solution mother liquor and the acrylamide solution mother liquor are mixed in a ratio of 5:8, and then ammonium persulfate and tetramethylethylenediamine are added, and then mixed evenly, and then divided into transparent glass tubes with sealed bottoms, and then ultrapure water is added to the upper layer, and the mixture is allowed to stand for 30 minutes, thereby obtaining a first section of a gel column with a length of 4 cm-10 cm; B4. After the first section of the gel column is polymerized, remove the ultrapure water on the upper layer of the first section of the gel column, and then mix the solution mother liquor and the acrylamide solution mother liquor in a 1:1 ratio, and then add ammonium persulfate and tetramethylethylenediamine, mix them evenly immediately, and then add them to the upper layer of the first section of the gel column to obtain the second section of the gel column. The length of the second section of the gel column is 0.3cm-3cm, and the distance between the second section of the gel column and the upper edge of the transparent proportional tube is 2-3cm; add ultrapure water to the upper layer of the second section of the gel column and let it stand for 30 minutes to obtain a gel pre-loaded column.

6. A staining solution for plasma lipids, characterized in that: The method for preparing the staining solution of plasma lipids comprises the following steps: S1, take 2g Sudan Black B dye and dissolve it in the first solvent; S2, then add dimethyl sulfoxide, mix well and heat to 50°C, stir for 30 minutes and filter with a 0.22μm filter membrane; S3, and then store it away from light to obtain the staining solution.

7. A plasma lipid staining solution according to claim 7, characterized in that: The first solvent is one or more of ethanol, ethylene glycol, propylene glycol, isopropanol and acetone solvents, the volume ratio of the first solvent to the total dyeing solution is 90%-95%, and the volume ratio of the dimethyl sulfoxide to the total dyeing solution is 1%-10%.

8. Use of a plasma lipid staining solution as claimed in claim 6 or 7 in the analysis of plasma lipoproteins.

9. The use according to claim 8, comprising a method for using a plasma lipid staining solution in the analysis of plasma lipoproteins, characterized in that: The following steps are involved: C1. Take 100 μl of sample and mix with 10 μl of staining solution; C2. Incubate at room temperature for 10 minutes and then centrifuge to obtain the supernatant to obtain the stained sample.

10. A method for analyzing plasma lipoprotein using a gel prepacked column, characterized in that: The following steps are involved: D1. Fix the prepared gel pre-packed column vertically in the electrophoresis tank, so that the second section of the electrophoresis gel column is facing upward and located at the negative end of the electrophoresis tank; D2, then slowly add electrophoresis buffer to the positive and negative electrode tanks of the electrophoresis tank, and the electrophoresis buffer in the negative electrode tank is submerged to the upper end of the gel pre-packed column; D3, then aspirate the stained sample and slowly add it to the top of the second gel in the gel pre-loaded column, then assemble the electrophoresis tank and turn on the power to perform electrophoresis; D4. After the electrophoresis is completed, the gel pre-loaded columns are removed and placed on the scanning analysis system platform for scanning in turn to obtain the plasma lipoprotein electrophoresis map, which is converted into a lipoprotein subtype distribution peak map to complete the analysis.

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