A triglyceride and cholesterol mixed quality control solution, and a preparation method and application thereof

Abstract

The application discloses a triglyceride and cholesterol mixed quality control liquid and a preparation method and application thereof, relates to the technical field of in vitro diagnostic reagents, and solves the problem of poor compatibility of components of the triglyceride and cholesterol mixed quality control liquid. The triglyceride and cholesterol mixed quality control liquid comprises a mother liquor and a diluent, the mother liquor comprises the following components in percentage by mass: glyceryl trioleate 7-10%, Triton X-100 85-90% and cholesterol 3-5%; and the diluent comprises the following components in percentage by mass: isopropyl alcohol 50-51%, sodium phosphate buffer 39-40%, sodium cholate 9.50-9.75% and sodium benzoate 0.25-0.50%. The mixed quality control liquid is high in stability and accuracy, can detect triglyceride and cholesterol at the same time, and the detection results do not interfere with each other.

Description

Technical Field

[0001] This invention relates to the field of in vitro diagnostic reagent technology, and in particular to a mixed quality control solution of triglycerides and cholesterol, its preparation method and application. Background Technology

[0002] Triglycerides (TG), also known as neutral fats, are composed of three fatty acid molecules and one glycerol ester molecule and are a major source of energy in the body. Triglyceride levels are positively correlated not only with total cholesterol (TC), body mass index (BMI), and blood glucose (GLU), but also closely related to obesity. Measuring triglycerides in the blood is crucial because high concentrations (normal range: 40-160 mg / dL for men and 35-135 mg / dL for women) can lead to hyperlipidemia. Hypertriglyceridemia is one of the main manifestations of coronary heart disease. Timely detection helps in the diagnosis and treatment of diabetes, kidney disease, liver obstruction, and various endocrine disorders related to lipid metabolism. Therefore, triglyceride testing is of great significance.

[0003] Cholesterol, also known as cholesterol sterol, is a derivative of cyclopentanoperhydrophenanthrene. Cholesterol is an essential substance for animal tissue cells, participating not only in cell membrane formation but also serving as a raw material for the synthesis of bile acids, vitamin D, and steroid hormones. While cholesterol plays a wide range of physiological roles in the body, excessive amounts can lead to hypercholesterolemia, adversely affecting the body. Modern research has found a close correlation between hypercholesterolemia and atherosclerosis, venous thrombosis, and gallstones; therefore, cholesterol testing is of great significance.

[0004] The detection systems for triglycerides and cholesterol in blood are generally lipid analyzers and lipid analysis systems. These systems require the use of quality control solutions for quality control. The quality control solutions can simulate blood for testing. If the quality control solutions fall within the marked range during testing, it indicates that there is no major problem with the test strip instrument.

[0005] Existing domestic quality control solutions can only measure cholesterol or triglycerides individually, while most current analytical instruments can simultaneously detect both. When assessing instrument performance, additional quality control solutions are required, making operation inconvenient. Patent application number 201811317402.9 discloses a triglyceride quality control solution comprising a mother liquor and a diluent. The mother liquor contains trioleic acid glyceride, Triton X-100, ethylene glycol, fetal bovine serum, and BSA; the diluent contains ethylene glycol, fetal bovine serum, and BSA. This quality control solution can only measure triglycerides individually and cannot detect cholesterol, making it unsuitable for multi-component biochemical analyzers. Imported quality control solutions are expensive, provide unsuitable values, and are prone to interference, limiting their applicability.

[0006] Stability and accuracy are crucial for ensuring the functionality of quality control solutions. Through numerous experiments, the applicant discovered that compared to single-component quality control solutions, the components of mixed triglyceride and cholesterol quality control solutions are more difficult to harmonize, making it harder to maintain stability and accuracy, and their test results are more prone to mutual interference. Therefore, there is an urgent need to develop a mixed quality control solution for triglycerides and cholesterol with high stability and accuracy to complement the use of a multi-component biochemical analyzer, ensuring that the test results do not interfere with each other. Summary of the Invention

[0007] To address the above shortcomings, this invention provides a mixed control solution of triglycerides and cholesterol and its preparation method. This mixed control solution exhibits high stability and accuracy, enabling simultaneous detection of triglycerides and cholesterol without interference between the detection results. The specific technical solution is as follows:

[0008] A mixed control solution of triglycerides and cholesterol includes: a stock solution and a diluent.

[0009] The mother liquor comprises the following components by weight percentage: 7-10% trioleic acid glycerides, 85-90% Triton X-100, and 3-5% cholesterol;

[0010] The diluent comprises the following components in weight percentage: 50-51% isopropanol, 39-40% sodium phosphate buffer, 9.50-9.75% sodium cholate, and 0.25-0.50% sodium benzoate.

[0011] Preferably, the mother liquor consists of the following components by mass percentage: 7% trioleic acid glyceride, 90% Triton X-100, and 3% cholesterol; the diluent consists of the following components by mass percentage: 50% isopropanol, 40% sodium phosphate buffer, 9.75% sodium cholate, and 0.25% sodium benzoate.

[0012] Preferably, the mother liquor consists of the following components by mass percentage: 10% trioleate, 85% Triton X-100, and 5% cholesterol; the diluent consists of the following components by mass percentage: 51% isopropanol, 39% sodium phosphate buffer, 9.5% sodium cholate, and 0.5% sodium benzoate.

[0013] The present invention also provides a method for preparing the above-mentioned triglyceride and cholesterol mixed control solution, comprising the following steps:

[0014] (1) Weigh the raw materials according to the above mass percentage, add trioleic acid glycerides and cholesterol into Triton X-100, and stir magnetically at 40-50℃ for 30-90 min to obtain the mother liquor;

[0015] (2) Weigh the raw materials according to the above mass percentages, and stir isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate magnetically at 40-50℃ for 30-90 min to obtain a diluted solution;

[0016] (3) Add a certain amount of the diluent to the mother liquor as needed to dilute it, so as to obtain a mixed quality control solution of triglycerides and cholesterol of different concentrations.

[0017] Preferably, the sodium phosphate buffer solution has a concentration of 0.2 mol / L and a pH of 6-7.

[0018] Preferably, the mixed control solution is used for quality control applications in a blood lipid testing system.

[0019] Compared with the prior art, the beneficial effects of the present invention are:

[0020] 1. The theoretical concentration of the mixed quality control solution prepared in this invention has a high degree of agreement with the test value of the biochemical analyzer, a low deviation, and an error of less than 5% from the theoretical value, indicating high accuracy.

[0021] 2. In the mixed quality control solution of the present invention, the combined action of Triton X-100 and isopropanol can fully dissolve trioleic acid glycerides and cholesterol, and sodium cholate can emulsify lipids to further improve stability. The combined action of Triton X-100, isopropanol, sodium cholate, sodium benzoate and sodium phosphate buffer can make the mixed quality control solution form a relatively homogeneous and stable state with good stability.

[0022] 3. This invention can provide test results that simulate clinical samples for laboratory research and development of triglycerides and cholesterol. Attached Figure Description

[0023] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below.

[0024] Figure 1 This is a curve showing the consistency between the actual and detected values ​​of the Triton X-100 content in the raw material of this invention.

[0025] Figure 2 This is a curve showing the consistency between the actual and detected values ​​of the isopropanol content in the raw material of this invention.

[0026] Figure 3 This is a graph showing the relationship between the sodium cholate content of the raw material of this invention and the deviation after aging. Detailed Implementation

[0027] The specific embodiments of the present invention will be described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.

[0028] Example 1

[0029] Triglyceride and cholesterol mixed control solution, including stock solution and diluent;

[0030] The mother liquor contains the following components and quantities: 0.7g trioleic acid glycerides, 9g Triton X-100, and 0.3g cholesterol;

[0031] The diluent contains the following components and quantities: 25g isopropanol, 20g sodium phosphate buffer, 4.875g sodium cholate, and 0.125g sodium benzoate.

[0032] The method for preparing the stock solution of triglyceride and cholesterol quality control products includes the following steps:

[0033] (1) Add trioleic acid glycerides and cholesterol to Triton X-100 stock solution according to the above mass, and dissolve them by magnetic stirring at 45°C for 60 minutes to obtain a homogeneous and stable stock solution.

[0034] (2) Mix isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate at 45°C and stir magnetically for 30 minutes to dissolve them to obtain a diluted solution;

[0035] (3) Dilute the above diluent with the mother liquor to obtain a mixed quality control solution of triglycerides and cholesterol.

[0036] The concentration of the sodium phosphate buffer solution mentioned above is 0.2 mol / L, and the pH is 6-7.

[0037] In Example 1, the theoretical concentration of the quality control solution showed a high degree of agreement with the test values ​​of the biochemical analyzer, indicating good stability.

[0038] Example 2

[0039] Triglyceride and cholesterol mixed control solution, including stock solution and diluent;

[0040] The mother liquor contains the following components and quantities: 1g trioleic acid glycerides, 8.5g Triton X-100, and 0.5g cholesterol;

[0041] The diluent contains the following components and quantities: 25.5g isopropanol, 19.5g sodium phosphate buffer, 4.75g sodium cholate, and 0.25g sodium benzoate.

[0042] The method for preparing the stock solution of triglyceride and cholesterol quality control products includes the following steps:

[0043] (1) Add trioleic acid glycerides and cholesterol to Triton X-100 stock solution according to the above mass, and dissolve them by magnetic stirring at 45°C for 60 minutes to obtain a homogeneous and stable mother solution.

[0044] (2) Mix isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate at 45°C and stir magnetically for 30 minutes to dissolve them to obtain a diluted solution;

[0045] (3) Dilute the above diluent with the mother liquor to obtain a mixed quality control solution of triglycerides and cholesterol.

[0046] The concentration of the sodium phosphate buffer solution mentioned above is 0.2 mol / L, and the pH is 6-7.

[0047] In Example 2, the theoretical concentration of the quality control solution showed a high degree of agreement with the test values ​​of the biochemical analyzer, and the solution exhibited good stability.

[0048] Comparative Example 1

[0049] Triglyceride and cholesterol mixed control solution, including stock solution and diluent;

[0050] The mother liquor contains the following components and quantities: 1g trioleic acid glyceride, 8.5g Triton X-100, and 0.5g cholesterol;

[0051] The diluent contains the following components and quantities: 30g isopropanol, 15g sodium phosphate buffer, 4.75g sodium cholate, and 0.25g sodium benzoate.

[0052] The method for preparing the stock solution of triglyceride and cholesterol quality control products includes the following steps:

[0053] (1) Add trioleic acid glycerides and cholesterol to Triton X-100 stock solution according to the above mass, and dissolve them by magnetic stirring at 45°C for 60 minutes to obtain a homogeneous and stable mother solution.

[0054] (2) Mix isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate at 45°C and stir magnetically for 30 minutes to dissolve them to obtain a diluted solution;

[0055] (3) Dilute the above diluent with the mother liquor to obtain a mixed quality control solution of triglycerides and cholesterol.

[0056] The concentration of the sodium phosphate buffer solution mentioned above is 0.2 mol / L, and the pH is 6-7.

[0057] The theoretical concentration of the control solution in Comparative Example 1 showed a high degree of agreement with the test values ​​of the biochemical analyzer, but its stability was poor.

[0058] Comparative Example 2

[0059] Triglyceride and cholesterol mixed control solution, including stock solution and diluent;

[0060] The mother liquor contains the following components and quantities: 1g trioleic acid glyceride, 8.5g Triton X-100, and 0.5g cholesterol;

[0061] The diluent contains the following components and quantities: 25g isopropanol, 21g sodium phosphate buffer, 3.75g sodium cholate, and 0.25g sodium benzoate.

[0062] The method for preparing the stock solution of triglyceride and cholesterol quality control products includes the following steps:

[0063] (1) Add trioleic acid glycerides and cholesterol to Triton X-100 stock solution according to the above mass, and dissolve them by magnetic stirring at 45°C for 60 minutes to obtain a homogeneous and stable mother solution.

[0064] (2) Mix isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate at 45°C and stir magnetically for 30 minutes to dissolve them to obtain a diluted solution;

[0065] (3) Dilute the above diluent with the mother liquor to obtain a mixed quality control solution of triglycerides and cholesterol.

[0066] The concentration of the sodium phosphate buffer solution mentioned above is 0.2 mol / L, and the pH is 6-7.

[0067] The theoretical concentration of the control solution in Comparative Example 2 showed a high degree of agreement with the test values ​​of the biochemical analyzer, but its stability was poor.

[0068] Comparative Example 3

[0069] Triglyceride and cholesterol mixed control solution, including stock solution and diluent;

[0070] The mother liquor contains the following components and quantities: 1.5g trioleic acid glycerides, 7.5g Triton X-100, and 1g cholesterol;

[0071] The diluent contains the following components and quantities: 25g isopropanol, 20g sodium phosphate buffer, 4.75g sodium cholate, and 0.25g sodium benzoate.

[0072] The method for preparing the stock solution of triglyceride and cholesterol quality control products includes the following steps:

[0073] (1) Add trioleic acid glycerides and cholesterol to Triton X-100 stock solution according to the above mass, and dissolve them by magnetic stirring at 45°C for 60 minutes to obtain a homogeneous and stable mother solution.

[0074] (2) Mix isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate at 45°C and stir magnetically for 30 minutes to dissolve them to obtain a diluted solution;

[0075] (3) Dilute the above diluent with the mother liquor to obtain a mixed quality control solution of triglycerides and cholesterol.

[0076] The concentration of the sodium phosphate buffer solution mentioned above is 0.2 mol / L, and the pH is 6-7.

[0077] The theoretical concentration of the control solution in Comparative Example 3 showed a low degree of agreement with the test values ​​of the biochemical analyzer, but also exhibited poor stability.

[0078] Each example and comparative example was prepared into three concentrations (concentration 1:14 of mother liquor to diluent mass ratio, concentration 2:1:7 of mother liquor to diluent mass ratio, and concentration 3:1:3 of mother liquor to diluent mass ratio). Concentration 1 was a low concentration, concentration 2 was a medium concentration, and concentration 3 was a high concentration. One sample of each concentration was placed in a 45°C oven for 7 days and 14 days, respectively. The concentrations (mmol / L) of triglycerides and cholesterol in the mixed control solution were then tested using a Mindray fully automated biochemical analyzer (BS-450), and the deviations from the theoretical concentrations were calculated. See Tables 1 and 2 for details.

[0079] Table 1 Results of triglyceride stability test

[0080]

[0081] Table 2 Results of cholesterol stability test

[0082]

[0083] The data from the examples in Tables 1 and 2 show that the test results of the quality control solution stored at 45°C for 7 days and 14 days, respectively, have smaller deviations compared to those stored at room temperature, while the data from Comparative Examples 1 to 3 show larger deviations. Therefore, the mixed quality control solution prepared by the method of the present invention has good stability in the detection of triglycerides and cholesterol.

[0084] Fourteen concentrations were prepared for each example and comparative example. Concentrations 1-5 were low concentrations, concentrations 6-11 were medium concentrations, and concentrations 12-14 were high concentrations. One sample of each concentration was taken and the concentrations (biochemical values) (mmol / L) of triglycerides and cholesterol in the mixed control solution were tested using a Mindray fully automated biochemical analyzer (BS-450). The results are shown in Tables 3 and 4.

[0085] Table 3. Accuracy Test Results of Triglycerides

[0086] concentration Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Biochemical values 1 1.30 1.32 1.25 1.19 1.71 1.29 2 1.70 1.71 1.67 1.63 2.26 1.69 3 2.78 2.69 2.72 2.65 3.35 2.70 4 3.57 3.50 3.46 3.42 4.01 3.49 5 3.77 3.68 3.79 3.65 4.36 3.70 6 4.21 4.41 4.33 4.18 4.89 4.35 7 4.95 4.73 4.99 4.68 5.35 4.89 8 6.25 6.10 6.31 6.06 6.51 6.23 9 6.51 6.57 6.62 6.46 6.68 6.55 10 6.87 6.93 7.01 6.85 6.86 6.91 11 6.98 7.12 7.11 6.92 7.00 7.03 12 9.17 9.26 9.28 9.13 8.88 9.21 13 9.39 9.21 9.41 9.17 8.92 9.36 14 9.44 9.47 9.55 9.39 8.95 9.50

[0087] Table 4. Results of Cholesterol Accuracy Test

[0088] concentration Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Biochemical values 1 3.09 3.12 3.07 3.13 3.71 3.10 2 3.31 3.35 3.28 3.38 4.01 3.33 3 3.56 3.53 3.49 3.50 4.33 3.52 4 3.95 3.97 3.93 3.92 4.46 3.96 5 4.19 4.21 4.15 4.17 4.57 4.18 6 4.27 4.31 4.23 4.24 4.66 4.28 7 4.51 4.52 4.46 4.48 4.71 4.50 8 4.72 4.75 4.69 4.68 4.80 4.71 9 4.97 5.03 4.95 4.93 4.98 4.98 10 5.21 5.23 5.15 5.17 5.22 5.20 11 5.38 5.42 5.33 5.36 5.37 5.39 12 6.16 6.18 6.12 6.13 6.09 6.15 13 7.13 7.15 7.06 7.10 6.25 7.12 14 8.39 8.44 8.21 8.30 6.60 8.36

[0089] Tables 3 and 4 show that the theoretical concentration of the quality control solution prepared in this invention has a high degree of agreement with the test values ​​obtained by the biochemical analyzer, with a low deviation, and the error is within 3.5% of the theoretical value (obtained by dividing the difference between the example value and the biochemical value by the biochemical value). When added to the blood lipid test strip, the concentration value shown by the result has a high degree of agreement with the theoretical concentration of the quality control solution, with a low deviation, and the error is within 7%, indicating high accuracy. At the same time, the theoretical concentration values ​​of the high and low levels in Comparative Example 1 also have a high degree of agreement with the test values ​​obtained by the biochemical analyzer, but their stability is poor (as shown in Tables 2 and 3). Therefore, the quality control solution prepared in this invention has the advantages of both high accuracy and good stability.

[0090] The applicant, through multiple experimental data analysis, discovered that the mass ratio of the raw materials Triton X-100, isopropanol, and sodium cholate has a significant impact on the accuracy and stability of the mixed quality control solution. Figure 1 , Figure 2 , Figure 3 As shown, the actual values ​​and test values ​​showed the highest agreement when Triton X-100 accounted for 85-90% of the mother liquor and isopropanol accounted for 50-51% of the diluent. The deviation was smallest after aging of the mixed quality control solution when sodium cholate accounted for about 10% of the diluent.

[0091] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A mixed control solution of triglycerides and cholesterol, characterized in that, include: Mother liquor and diluent, The mother liquor comprises the following components by weight percentage: 7-10% trioleic acid glycerides, 85-90% Triton X-100, and 3-5% cholesterol; The diluent comprises the following components in weight percentage: 50-51% isopropanol, 39-40% sodium phosphate buffer, 9.50-9.75% sodium cholate, and 0.25-0.50% sodium benzoate.

2. The triglyceride and cholesterol mixed control solution according to claim 1, characterized in that, The mother liquor consists of the following components by weight percentage: 7% trioleic acid glyceride, 90% Triton X-100, and 3% cholesterol; the diluent consists of the following components by weight percentage: 50% isopropanol, 40% sodium phosphate buffer, 9.75% sodium cholate, and 0.25% sodium benzoate.

3. The triglyceride and cholesterol mixed control solution according to claim 1, characterized in that, The mother liquor consists of the following components by weight percentage: 10% trioleic acid glyceride, 85% Triton X-100, and 5% cholesterol; the diluent consists of the following components by weight percentage: 51% isopropanol, 39% sodium phosphate buffer, 9.5% sodium cholate, and 0.5% sodium benzoate.

4. A triglyceride and cholesterol mixed control solution according to any one of claims 1 to 3, characterized in that, The sodium phosphate buffer solution has a concentration of 0.2 mol / L and a pH of 6-7.

5. A method for preparing a mixed control solution of triglycerides and cholesterol as described in any one of claims 1 to 4, characterized in that, Includes the following steps: (1) Add trioleic acid glycerides and cholesterol to Triton X-100 and stir magnetically at 40-50℃ for 30-90 min to obtain the mother liquor; (2) Mix isopropanol, sodium phosphate buffer, sodium cholate and sodium benzoate at 40-50℃ and stir magnetically for 30-90 minutes to obtain a diluted solution; (3) Add a certain amount of the diluent to the mother liquor as needed to dilute it, so as to obtain a mixed quality control solution of triglycerides and cholesterol of different concentrations.

6. The application of the triglyceride and cholesterol mixed control solution as described in any one of claims 1 to 4 in the quality control of a blood lipid detection system.

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