Total cholesterol quality control liquid and preparation method thereof

Abstract

The invention relates to a total cholesterol quality control solution, which comprises a total cholesterol mother solution and a base solution, the total cholesterol mother solution comprises total cholesterol and an emulsifier, and the base solution comprises a buffer solution, a preservative and a stabilizer. A preparation method of a total cholesterol quality control solution comprises the following steps: fully dissolving different amounts of total cholesterol in an emulsifier, slowly dropwise adding the dissolved system into a certain amount of a base solution, filtering the system containing precipitates to obtain a uniform, stable and transparent total cholesterol mother solution, adding the base solution containing a stabilizer, a preservative and a buffer solution, and uniformly stirring to obtain the total cholesterol quality control solution. The total cholesterol quality control liquid with the required concentration is prepared. The method can be used for detecting the total cholesterol index of the electrochemical method total cholesterol test paper. The total cholesterol quality control liquid has the advantages of good stability, long storage time, no toxicity and environmental protection, can be well applied to an electrochemical test paper detection system, and ensures the accuracy and stability of a quality control result.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological detection, and particularly relates to a total cholesterol quality control solution and a preparation method thereof. BACKGROUND

[0002] Blood glucose detection, uric acid detection or blood ketone detection are all routine detection items in medical detection. The detection of physiological indexes such as blood glucose, uric acid or blood ketone generally involves electrochemical method. In the electrochemical detection method of indexes such as blood glucose, uric acid or blood ketone, a quality control solution is usually used.

[0003] The quality control solution is a solution containing a to-be-detected substance or a substance with the same property as the to-be-detected substance, which functions to confirm whether the electrochemical detection system is normal. When there is a large deviation or suspicious result in the detection process, the detection result of the detection system on the quality control solution can be used as a basis to confirm whether the detection system is in a normal detection state.

[0004] The quality control solution in the prior art contains a biological source component, which has certain limitations in production and sales. SUMMARY

[0005] The application aims to provide a total cholesterol quality control solution and a preparation method thereof, which do not need biological source substances, have low cost, obvious gradient, can be mass-produced, have available resources, and are safe and environmentally friendly.

[0006] In order to achieve the above-mentioned purpose, the technical scheme of the application is as follows.

[0007] A total cholesterol quality control solution comprises a total cholesterol mother liquor and a base solution, wherein the total cholesterol mother liquor comprises total cholesterol and an emulsifier, and the base solution comprises a buffer, a preservative and a stabilizer.

[0008] Further, the emulsifier is one or more of anhydrous ethanol, isopropyl alcohol and dimethyl sulfoxide.

[0009] Further, the buffer is one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate and sodium dihydrogen phosphate.

[0010] Further, the preservative is one or more of sodium benzoate, potassium sorbate, sodium azide and Proclin 300.

[0011] Further, the stabilizer is one or more of PEG-4000, Triton X-100 and EDTA.

[0012] Further, the mass of the total cholesterol is 0.01 g, and the emulsifier is 5 g.

[0013] Further, the addition amount of the preservative is 3-10 g / L.

[0014] Furthermore, the concentration of the buffer solution is 0.1 mol / L.

[0015] Furthermore, the pH value of the buffer solution is 6-8.

[0016] Furthermore, the amount of stabilizer added is 20-30 g / L.

[0017] A method for preparing a total cholesterol quality control solution comprises first fully dissolving different amounts of total cholesterol in an emulsifier, then slowly adding the dissolved system dropwise to a certain amount of base liquid, filtering the system containing the precipitate to obtain a uniform, stable and transparent total cholesterol mother liquor, and simultaneously adding a base liquid containing a stabilizer, a preservative and a buffer solution to prepare a total cholesterol quality control solution of the required concentration.

[0018] The beneficial effect of the invention is that the invention can detect the total cholesterol index for the electrochemical total cholesterol test paper. The total cholesterol quality control solution does not require biological substances, has low cost, obvious gradient, can be mass-produced, resources are available, and is safe and environmentally friendly. The total cholesterol quality control solution has good stability, a long storage time, and the solution is non-toxic and environmentally friendly. It can be well applied to the electrochemical test paper detection system to ensure the accuracy and stability of the quality control results. The quality control solution of the invention can test the total cholesterol content and can meet the total cholesterol analyzers currently on the market. At the same time, it can also solve the problem of cumbersome procedures for users when using it. The quality control solution of the present invention can ensure that the total cholesterol analyzer and the test paper can operate correctly as a system. DETAILED DESCRIPTION

[0019] The specific embodiments of the present invention are described below in conjunction with examples to facilitate a better understanding of the present invention.

[0020] Example 1:

[0021] The total cholesterol quality control solution in this embodiment includes a total cholesterol mother solution and a base solution. The total cholesterol mother solution is obtained by fully dissolving 0.01g of total cholesterol in 5g of anhydrous ethanol. The amount of base solution added is: 0.1mol of potassium dihydrogen phosphate buffer at pH 8.0, 3g of sodium benzoate, and 20g of PEG-4000 are added to each L of total cholesterol mother solution. According to the above formula, three portions of base solution are prepared. First, the total cholesterol mother solution (3mmol / L) is added to the first portion as level 1, the total cholesterol mother solution (6mmol / L) is added to the second portion as level 2, and the total cholesterol mother solution (9mmol / L) is added to the third portion as level 3 to prepare three concentration levels of total cholesterol quality control solution. After shaking, the three concentrations of total cholesterol quality control solution are divided into small bottles according to a volume of 2±0.2ml and sealed for storage. Used for performance testing.

[0022] (1) repeatability and accuracy test:

[0023] During the test, 1 bottle of each concentration was taken respectively, and the total cholesterol concentration was tested by Mindray BS-430 biochemical analyzer, and the test kit was the total cholesterol kit produced by Mindray Company. After the test, the CV of each test result of each concentration of the total cholesterol quality control liquid was calculated, and the deviation of the total cholesterol concentration result measured by the Mindray biochemical analyzer from the theoretical concentration was calculated. The test results are shown in Table 1.

[0024] Table 1

[0025] According to the test results of the total cholesterol quality control liquid of the Mindray BS-430 biochemical analyzer in Table 1, it can be seen that the repeatability of the test results of each concentration level of the quality control liquid is good, and the CV is within 2%; at the same time, the test results of the biochemical analyzer BS-430 and the theoretical concentration of the test results are small, and are within ±1%. Therefore, the quality control liquid in the application can effectively ensure the repeatability and accuracy of the test results.

[0026] (2) stability test:

[0027] 1 bottle of each concentration of the total cholesterol quality control liquid after sealing was taken respectively, and was stored at room temperature, and 1 bottle of each concentration of the total cholesterol quality control liquid after sealing was taken respectively, and was stored in a constant temperature box at 65℃ for 12 days (equivalent to storage at room temperature for 2 years). After 12 days, the quality control liquid stored in the constant temperature box at 65℃ was taken out to room temperature for at least 30 min. Then the total cholesterol quality control liquid stored at room temperature and 65℃ was tested by the biochemical analyzer BS-430 respectively, and the deviation of the test results of the total cholesterol quality control liquid stored at 65℃ from the total cholesterol quality control liquid stored at room temperature was calculated. The test results are shown in Table 2.

[0028] Table 2

[0029] According to the test results in Table 2, the deviation of the test results of the total cholesterol quality control liquid stored at 65℃ for 12 days from the total cholesterol quality control liquid stored at room temperature is very small, and the total cholesterol test result is controlled within ±4%, therefore, the total cholesterol quality control liquid of the application has good stability.

[0030] Example 2:

[0031] The total cholesterol control solution in this embodiment includes a total cholesterol stock solution and a base solution. The total cholesterol stock solution is obtained by fully dissolving 0.01g of total cholesterol in 5g of isopropanol, and then adding the base solution: 0.1mol of pH 8.0 buffer, 3g of sodium benzoate, and 20g / L of PEG-4000 are added to each L of total cholesterol stock solution. According to the above formula, three portions of the base solution are prepared. First, the total cholesterol stock solution (3mmol / L) is added to the first portion as level 1, the total cholesterol stock solution (6mmol / L) is added to the second portion as level 2, and the total cholesterol stock solution (9mmol / L) is added to the third portion as level 3 to prepare three concentration levels of total cholesterol control solution. After shaking, the three concentrations of total cholesterol control solution are divided into vials according to a volume of 2±0.2ml and sealed for storage.

[0032] (1) Repeatability and accuracy test:

[0033] During the test, one vial of each concentration was collected and the total cholesterol concentration was measured using a Mindray BS-430 biochemical analyzer. The test kit was a Mindray total cholesterol kit. After the test, the CV of each test result for each concentration of the total cholesterol control solution was calculated. The deviation between the total cholesterol concentration measured by the Mindray biochemical analyzer and the theoretical concentration was also calculated. The test results are shown in Table 3.

[0034] Table 3

[0035] According to the test results of the Mindray BS-430 biochemical analyzer for the total cholesterol quality control solution in Table 3, it can be seen that the repeatability of each test result of each concentration of the total cholesterol quality control solution is good, and the CV is within 5%; however, the test deviation of the biochemical BS-430 test results from the theoretical concentration is large, both above 5%. Therefore, the total cholesterol quality control solution in the present invention cannot simultaneously ensure good accuracy.

[0036] (2) Stability test:

[0037] Take one bottle of sealed aliquot of total cholesterol control solution at each concentration and store it at room temperature. Then, take one bottle of sealed aliquot of total cholesterol control solution at each concentration and store it in a 65°C incubator for 12 days (equivalent to 2 years at room temperature). After 12 days, bring the control solution from the 65°C incubator to room temperature for at least 30 minutes. Then, use a biochemical BS-430 to measure the concentration of the total cholesterol control solution stored at room temperature and at 65°C. Calculate the deviation between the test results for the total cholesterol control solution stored at 65°C and the total cholesterol control solution stored at room temperature. The test results are shown in Table 4.

[0038] Table 4

[0039] According to the test results in Table 4, the deviation of the total cholesterol quality control liquid stored at 65℃ for 12 days relative to the test results stored at room temperature is 5.5% at low concentration, and thus the stability of the total cholesterol quality control liquid of the present application is poor.

[0040] Example 3:

[0041] The total cholesterol mother liquor: 0.01 g of total cholesterol was fully dissolved in 5 g of anhydrous ethanol. The base liquid formula: buffer pH 6.0 (0.1 mol / L), sodium benzoate (3 g / L), PEG-4000 (20 g / L). Three base liquids were prepared according to the above formula, and the first one was added with the total cholesterol mother liquor (3 mmol / L) as level 1, the second one was added with the total cholesterol mother liquor (6 mmol / L) as level 2, and the third one was added with the total cholesterol mother liquor (9 mmol / L) as level 3, to configure three concentration levels of total cholesterol quality control liquid. After shaking, the three concentrations of total cholesterol quality control liquid were divided into 2±0.2 ml in a small bottle and sealed for storage.

[0042] (1) Repeatability and accuracy test:

[0043] During the test, 1 vial was taken for each concentration, and the concentration of total cholesterol was tested by Mindray BS-430 biochemical analyzer, and the reagent kit was the total cholesterol reagent kit produced by Mindray Company. After the test, the CV of each test result of each concentration of the total cholesterol quality control liquid was calculated, and the deviation of the concentration of total cholesterol measured by the Mindray biochemical analyzer from the theoretical concentration was calculated. The test results are shown in Table 5.

[0044] Table 5

[0045] According to the test results of the total cholesterol quality control liquid of the Mindray BS-430 biochemical analyzer in Table 5, the repeatability of each test result of each concentration of the total cholesterol quality control liquid is good, and the CV is within 5%; at the same time, the test deviation of the test results of the biochemical BS-430 from the theoretical concentration is within ±5%, and thus the total cholesterol quality control liquid in the present application can effectively ensure the repeatability and accuracy of the test results.

[0046] (2) Stability test:

[0047] Take 1 bottle of each concentration of the total cholesterol quality control liquid after being sealed and divided, and store it at room temperature. Take another 1 bottle of each concentration of the total cholesterol quality control liquid after being sealed and divided, and store it in a thermostat at 65℃ for 12 days (equivalent to 2 years of storage at room temperature). After 12 days, take the quality control liquid stored in the thermostat at 65℃ to room temperature and keep it constant for at least 30 min. Then use the biochemical BS-430 to test the concentrations of the total cholesterol quality control liquid stored at room temperature and 65℃, respectively, and calculate the deviation of the test results of the total cholesterol quality control liquid stored at 65℃ from that stored at room temperature. The test results are shown in Table 6.

[0048] Table 6

[0049] According to the test results in Table 6, the deviation of the test results of the total cholesterol quality control liquid stored at 65℃ for 12 days from that stored at room temperature is small, all within ±4%, so the total cholesterol quality control liquid of the application has good stability.

[0050] Example 4:

[0051] Total cholesterol mother liquor: 0.01 g of total cholesterol is fully dissolved in 5 g of anhydrous ethanol. Base liquid formula: buffer pH 7.0 (0.1 mol / L); sodium benzoate (3 g / L), PEG-4000 (20 g / L). According to the above formula, three base liquids are configured. The first one is added with the total cholesterol mother liquor (3 mmol / L) as level 1, the second one is added with the total cholesterol mother liquor (6 mmol / L) as level 2, and the third one is added with the total cholesterol mother liquor (9 mmol / L) as level 3, to configure three concentration levels of total cholesterol quality control liquid. After shaking, the above three concentrations of total cholesterol quality control liquid are divided into small bottles with a volume of 2±0.2 ml, and sealed for storage.

[0052] (1) repeatability and accuracy test:

[0053] During the test, 1 small bottle of each concentration is taken, and the concentration of total cholesterol is tested by using the Mindray BS-430 biochemical analyzer, and the test kit is the total cholesterol kit produced by Mindray Company. After the test, the CV of each test result of each concentration of the total cholesterol quality control liquid is calculated, and the deviation of the concentration result of the total cholesterol measured by the Mindray biochemical analyzer from the theoretical concentration is calculated. The test results are shown in Table 7.

[0054] Table 7

[0055] According to the test results in Table 2, the deviation of the total cholesterol quality control liquid stored at 65℃ for 12 days relative to the test results of the total cholesterol quality control liquid stored at room temperature is small, the total cholesterol test results are controlled within ±2%, and therefore the total cholesterol quality control liquid has good stability.

[0056] (2) Stability test:

[0057] One bottle of each concentration of the sealed and sub-packaged total cholesterol quality control liquid was stored at room temperature, and one bottle of each concentration of the sealed and sub-packaged total cholesterol quality control liquid was stored in a 65℃ constant temperature box for 12 days (equivalent to 2 years of storage at room temperature). After 12 days, the quality control liquid stored in the 65℃ constant temperature box was taken out and allowed to stand at room temperature for at least 30 minutes. Then the concentrations of the total cholesterol quality control liquid stored at room temperature and at 65℃ were tested using the biochemical BS-430, and the deviation of the test results of the total cholesterol quality control liquid stored at 65℃ relative to the test results of the total cholesterol quality control liquid stored at room temperature was calculated. The test results are shown in Table 8.

[0058]

[0059] Table 8

[0060] According to the test results in Table 8, the deviation of the test results of the total cholesterol quality control liquid stored at 65℃ for 12 days relative to the test results of the total cholesterol quality control liquid stored at room temperature is large at low concentration levels, and therefore the total cholesterol quality control liquid has poor stability in the total cholesterol item.

[0061] Example 5:

[0062] Total cholesterol stock solution: 0.01g total cholesterol was fully dissolved in 5g anhydrous ethanol. Base liquid formula: buffer pH 8.0 (0.1mol / L) sodium benzoate (10g / L) PEG-4000 (20g / L) Three base liquids were prepared according to the above formula, the first one was added with total cholesterol stock solution (3mmol / L) as level 1, the second one was added with total cholesterol stock solution (6mmol / L) as level 2, and the third one was added with total cholesterol stock solution (9mmol / L) as level 3, to configure three concentration levels of total cholesterol quality control liquid. After shaking, the three concentrations of total cholesterol quality control liquid were sub-packaged into vials with a volume of 2±0.2ml and sealed for storage.

[0063] (1) Repeatability and accuracy test:

[0064] At the time of testing, 1 bottle was taken for each concentration, and the concentration of total cholesterol was tested by using the Mindray BS-430 biochemical analyzer, and the reagent kit was the total cholesterol reagent kit produced by the Mindray Company. After the testing, the CV of each test result of each concentration of the total cholesterol quality control liquid was calculated, and the deviation of the concentration result of the total cholesterol tested by the Mindray biochemical analyzer from the theoretical concentration was calculated. The testing result is shown in Table 9.

[0065]

[0066] Table 9

[0067] According to the testing result of the total cholesterol quality control liquid of the Mindray BS-430 biochemical analyzer in Table 9, the repeatability of each test result of each concentration level of the total cholesterol quality control liquid is good, and the CV is within 5%; meanwhile, the deviation of the testing result of the biochemical BS-430 from the theoretical concentration is within ±4%, so the total cholesterol quality control liquid in the application can effectively ensure the repeatability and accuracy of the testing result.

[0068] (2) Stability testing:

[0069] 1 bottle of each concentration of the total cholesterol quality control liquid after the sealing and the sub-packaging was taken and stored at room temperature, and 1 bottle of each concentration of the total cholesterol quality control liquid after the sealing and the sub-packaging was taken and stored in a constant temperature box at 65℃ for 12 days (equivalent to storage at room temperature for 2 years). After 12 days, the quality control liquid in the constant temperature box at 65℃ was taken out and kept at room temperature for at least 30 min. Then, the concentration of the total cholesterol quality control liquid stored at room temperature and the total cholesterol quality control liquid stored at 65℃ was tested by using the biochemical BS-430, and the deviation of the testing result of the total cholesterol quality control liquid stored at 65℃ from the total cholesterol quality control liquid stored at room temperature was calculated. The testing result is shown in Table 8.

[0070]

[0071] Table 10

[0072] According to the testing result in Table 10, the testing result of the total cholesterol quality control liquid stored at 65℃ for 12 days has a larger deviation at the low concentration level compared with the total cholesterol quality control liquid stored at room temperature, so the total cholesterol quality control liquid in the application has poor stability in the total cholesterol item.

[0073] Example 6:

[0074] Total cholesterol mother liquor: 0.01 g total cholesterol was dissolved in 5 g anhydrous ethanol. Base liquid formula: buffer pH 8.0 (0.1 mol / L); sodium benzoate (3 g / L); PEG-4000 (30 g / L); three base liquids were prepared according to the above formula, the first one was added with total cholesterol mother liquor (3 mmol / L) as level 1, the second one was added with total cholesterol mother liquor (6 mmol / L) as level 2, and the third one was added with total cholesterol mother liquor (9 mmol / L) as level 3, to prepare three concentration levels of total cholesterol control solution. After shaking, the three concentration levels of total cholesterol control solution were divided into 2±0.2 ml in a vial, and sealed for storage.

[0075] (1) Reproducibility and accuracy test:

[0076] During the test, 1 vial was taken for each concentration, and the concentration of total cholesterol was tested by Mindray BS-430 biochemical analyzer, and the reagent kit was total cholesterol reagent kit produced by Mindray Company. After the test, the CV of each test result of each concentration of total cholesterol control solution was calculated, and the deviation of the concentration result of total cholesterol measured by the biochemical analyzer from the theoretical concentration was calculated. The test results are shown in Table 11.

[0077]

[0078] Table 11

[0079] According to the test results of the total cholesterol control solution of the biochemical analyzer BS-430 in Table 12, the reproducibility of the test results of each concentration of the total cholesterol control solution is good, and the CV is within 5%, and the test deviation of the high concentration test results of the biochemical BS-430 from the theoretical concentration is within 5%, therefore, the reproducibility and accuracy of the total cholesterol control solution in the application are good.

[0080] (2) Stability test:

[0081] 1 bottle of each concentration of the sealed and divided total cholesterol control solution was taken and stored at room temperature, and 1 bottle of each concentration of the sealed and divided total cholesterol control solution was taken and stored in a constant temperature oven at 65℃ for 12 days (equivalent to 2 years of storage at room temperature). After 12 days, the control solution stored in the constant temperature oven at 65℃ was taken out and kept at room temperature for at least 30 min. Then the concentrations of the total cholesterol control solution stored at room temperature and 65℃ were tested by biochemical BS-430, and the deviation of the test results of the total cholesterol control solution stored at 65℃ from the total cholesterol control solution stored at room temperature was calculated. The test results are shown in Table 12.

[0082]

[0083] Table 12

[0084] The test results of the Mindray BS-430 biochemical analyzer of the total cholesterol quality control liquid according to Table 12 can see that the repeatability of the test results of each concentration of the total cholesterol quality control liquid is good, and the CV is within 5%, but the test result of the high concentration of the biochemical BS-430 is greatly deviated from the test result of the theoretical concentration, which is-6.3%, so the accuracy of the total cholesterol quality control liquid in the application is poor.

[0085] The above is the preferred embodiment of the application, it should be pointed out that, for those skilled in the art, without departing from the principles of the application, can make a number of improvements and refinements, these improvements and refinements also regarded as the protection scope of the application.

Claims

1. A total cholesterol quality control solution, characterized by: It includes total cholesterol mother solution and basic liquid, wherein the total cholesterol mother solution includes total cholesterol and emulsifier, and the basic liquid includes buffer, preservative and stabilizer.

2. The total cholesterol quality control solution according to claim 1, wherein: The emulsifier is one or more of anhydrous ethanol, isopropyl alcohol, and dimethyl sulfoxide.

3. The total cholesterol quality control solution according to claim 1, wherein: The buffer solution is one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate.

4. The total cholesterol quality control solution according to claim 1, wherein: The preservative is one or more of sodium benzoate, potassium sorbate, sodium azide, and Proclin 300.

5. The total cholesterol quality control solution according to claim 1, wherein: The stabilizer is one or more of PEG-4000, Triton X-100, and EDTA.

6. The total cholesterol quality control solution according to claim 1, characterized in that: The mass of the total cholesterol is 0.01 g, and the mass of the emulsifier is 5 g.

7. The total cholesterol quality control solution according to claim 1, wherein: The amount of the preservative added is 3-10 g / L.

8. The total cholesterol quality control solution according to claim 1, wherein: The concentration of the buffer solution is 0.1 mol / L, and the pH value of the buffer solution is 6-8.

9. The total cholesterol quality control solution according to claim 1, characterized in that: The amount of the stabilizer added is 20-30 g / L.

10. A method for preparing the total cholesterol quality control solution according to any one of claims 1 to 9, characterized in that: First, different amounts of total cholesterol were fully dissolved in an emulsifier, and then the dissolved system was slowly added dropwise to a certain amount of base liquid. After magnetic stirring for 30 minutes to fully react, the system containing the precipitate was filtered to obtain a uniform, stable and transparent total cholesterol mother liquor. At the same time, a base liquid containing a stabilizer, preservative and buffer was added to prepare a total cholesterol quality control solution of the required concentration.

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