Quality control product or calibration product of 25-hydroxyvitamin D and related application of quality control product or calibration product

Abstract

The invention discloses a quality control product or a calibration product of 25-hydroxyvitamin D and related application thereof, and relates to the field of biological detection. In the preparation of the calibration product and / or the quality control product of the 25-hydroxyvitamin D, ascorbic acid and 2, 6-butylated hydroxytoluene are used as protective agents, so that the 25-hydroxyvitamin is more stable, the use and the transportation of the product are convenient, and the production cost is reduced. The freeze-drying loss of effective components of the product is reduced, the cost is saved, the concentration deviation of product preparation can be effectively reduced, the controllable product quality is ensured, and a way is provided for accurate detection of the 25-hydroxyvitamin.

Description

Technical Field

[0001] The present invention relates to the field of biological detection, and more particularly, to a quality control product or calibration product for 25-hydroxyvitamin D and related applications thereof. Background Art

[0002] 25-hydroxyvitamin D usually uses methanol, ethanol, etc. as standard solutions. When preparing its calibration product or quality control product, it is usually mixed with a matrix to obtain a calibration product working solution or a quality control product working solution. During the process of injecting it into an aqueous solution of the matrix, problems such as solute precipitation and precipitation, which are difficult to dissolve, are likely to occur, which will cause deviations in the prepared concentration of the product and affect the product quality.

[0003] In view of this, the present invention is specifically proposed. Summary of the Invention

[0004] The purpose of the present invention is to provide a quality control product or calibration product for 25-hydroxyvitamin D and related applications thereof.

[0005] The present invention is implemented as follows:

[0006] In a first aspect, an embodiment of the present invention provides a composition, which includes: ascorbic acid and 2,6-di-tert-butyl-p-cresol.

[0007] In a second aspect, an embodiment of the present invention provides a matrix containing a protective agent, and the protective agent includes ascorbic acid and 2,6-di-tert-butyl-p-cresol.

[0008] In a third aspect, an embodiment of the present invention provides the application of the composition or the matrix described in the foregoing embodiments in the preparation of a quality control product, calibration product, quality control product working solution or calibration product working solution for 25-hydroxyvitamin D.

[0009] In a fourth aspect, an embodiment of the present invention provides a quality control product working solution or calibration product working solution for 25-hydroxyvitamin D, which includes: a standard solution of 25-hydroxyvitamin D, a matrix and a protective agent; wherein, the protective agent includes: ascorbic acid and 2,6-di-tert-butyl-p-cresol.

[0010] In a fifth aspect, an embodiment of the present invention provides a quality control product working solution or calibration product working solution for 25-hydroxyvitamin D, which includes mixing a standard solution of 25-hydroxyvitamin D, a matrix and a protective agent; the standard solution of 25-hydroxyvitamin D, the matrix and the protective agent are the standard solution of 25-hydroxyvitamin D, the matrix and the protective agent described in the foregoing embodiments.

[0011] In a sixth aspect, an embodiment of the present invention provides a quality control product or calibration product for 25-hydroxyvitamin D, which is obtained by freeze-drying the quality control product working solution or calibration product working solution described in the foregoing embodiments.

[0012] In a seventh aspect, the embodiments of the present invention provide the use of the composition as described in the foregoing embodiments, or the matrix as described in the foregoing embodiments, or the quality control product working solution or calibration product working solution as described in the foregoing embodiments, or the quality control product or calibration product as described in the foregoing embodiments, in the detection of 25-hydroxyvitamin D for non-disease diagnosis or treatment purposes.

[0013] The present invention has the following beneficial effects:

[0014] In the preparation of calibration products and / or quality control products, using ascorbic acid and 2,6-di-tert-butyl-p-cresol as protective agents can make 25-hydroxyvitamin more stable, facilitate the use and transportation of products, reduce the freeze-drying loss of the active ingredients of products, save costs, and effectively reduce the concentration deviation in product preparation, ensuring controllable product quality and providing a way for the accurate detection of 25-hydroxyvitamin D. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as limiting the scope. For those of ordinary skill in the art, other related drawings can be obtained based on these drawings without creative efforts.

[0016] Figure 1 Schematic diagram of the liquid preparation method using a micro-injection pump in combination with a magnetic stirrer (viewpoint 1);

[0017] Figure 2 Schematic diagram of the liquid preparation method using a micro-injection pump in combination with a magnetic stirrer (viewpoint 2). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0018] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Those not specified in the embodiments are carried out according to conventional conditions or conditions recommended by the manufacturer. Reagents or instruments not indicated by the manufacturer are all conventional products that can be obtained through commercial purchase.

[0019] On the one hand, the embodiments of the present invention provide a composition, which includes: ascorbic acid and 2,6-di-tert-butyl-p-cresol.

[0020] In some embodiments, the mass ratio of ascorbic acid to 2,6-di-tert-butyl-p-cresol is 1 to 10:0.1 to 5. Specifically, it can be any one or the range between any two of 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:3, 1:4, 1:5, 5:0.1, 5:0.5, 5:1, 5:1.5, 5:2, 5:3, 5:4, 5:5, 10:0.1, 10:0.5, 10:1, 10:1.5, 10:2, 10:3, 10:4, 10:5.

[0021] In some embodiments, the composition further comprises: a matrix.

[0022] In some embodiments, the matrix comprises serum and / or serum albumin.

[0023] In some embodiments, the serum comprises bovine serum.

[0024] In some embodiments, the composition further comprises: any one or more of 25-hydroxyvitamin D and a standard solution of 25-hydroxyvitamin D.

[0025] On the other hand, an embodiment of the present invention provides a matrix containing a protective agent, and the protective agent comprises ascorbic acid and 2,6-di-tert-butyl-p-cresol.

[0026] In some embodiments, the mass ratio of ascorbic acid to 2,6-di-tert-butyl-p-cresol is 1 to 10:0.1 to 5. Specifically, it can be any one or the range between any two of 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:3, 1:4, 1:5, 5:0.1, 5:0.5, 5:1, 5:1.5, 5:2, 5:3, 5:4, 5:5, 10:0.1, 10:0.5, 10:1, 10:1.5, 10:2, 10:3, 10:4, 10:5.

[0027] In some embodiments, the matrix comprises serum and / or serum albumin.

[0028] In some embodiments, the serum comprises bovine serum.

[0029] In some embodiments, the final concentration of ascorbic acid in the matrix is 1 to 10 mg / mL, specifically, it can be any one or the range between any two of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 mg / mL.

[0030] In some embodiments, the final concentration of 2,6-di-tert-butyl-p-cresol in the matrix is 0.1 to 5 mg / mL, specifically, it can be any one or the range between any two of 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, and 5 mg / mL. On the other hand, the embodiments of the present invention also provide the use of the composition or the matrix described in any of the foregoing embodiments in the preparation of a quality control product, a calibration product, a quality control working solution, or a calibration working solution of 25-hydroxyvitamin D.

[0031] On the other hand, the embodiments of the present invention also provide a quality control working solution or a calibration working solution of 25-hydroxyvitamin D, which comprises: a standard solution of 25-hydroxyvitamin D, a matrix, and a protective agent;

[0032] Wherein, the protective agent comprises: ascorbic acid and 2,6-di-tert-butyl-p-cresol.

[0033] In some embodiments, the final concentration of ascorbic acid in the quality control working solution or the calibration working solution is 1 to 10 mg / mL, specifically, it can be any one or the range between any two of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 mg / mL.

[0034] In some embodiments, the final concentration of ascorbic acid in the quality control working solution or the calibration working solution is 3 to 7 mg / mL.

[0035] In some embodiments, the final concentration of 2,6-di-tert-butyl-p-cresol in the quality control working solution or the calibration working solution is 0.1 to 5 mg / mL, specifically, it can be any one or the range between any two of 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, and 5 mg / mL.

[0036] In some embodiments, the final concentration of 2,6-di-tert-butyl-p-cresol in the quality control working solution or the calibration working solution is 0.5 to 3 mg / mL.

[0037] In some embodiments, the matrix comprises serum and / or serum albumin;

[0038] In some embodiments, the serum comprises bovine serum;

[0039] In some embodiments, the volume ratio of the standard solution of 25-hydroxyvitamin D to the matrix is 1:30 to 60, specifically, it can be any one or the range between any two of 1:30, 1:32, 1:34, 1:36, 1:38, 1:39, 1:40, 1:42, 1:44, 1:46, 1:48, 1:49, 1:50, 1:52, 1:54, 1:56, 1:58, and 1:60.

[0040] In some embodiments, the standard solution of 25-hydroxyvitamin D includes 25-hydroxyvitamin D and a diluent, and the diluent includes methanol and / or ethanol.

[0041] On the other hand, the embodiments of the present invention also provide a quality control product working solution or a calibration product working solution of 25-hydroxyvitamin D, which includes a mixed standard solution of 25-hydroxyvitamin D, a matrix, and a protective agent; the standard solution of 25-hydroxyvitamin D, the matrix, and the protective agent are the standard solution of 25-hydroxyvitamin D, the matrix, and the protective agent described in any of the foregoing embodiments.

[0042] In some embodiments, the mixing includes: first mixing the protective agent and the matrix to obtain a matrix added with the protective agent; adding the standard solution of 25-hydroxyvitamin D to the matrix added with the protective agent to obtain a quality control product working solution or a calibration product working solution.

[0043] In some embodiments, the standard solution of 25-hydroxyvitamin D is added by using a micro-injection pump (micropump).

[0044] The traditional pipette for liquid preparation manually controls the liquid addition amount by the user, which is a huge test for the user's operation skills and proficiency. Therefore, there are unstable factors in itself, affecting its liquid addition accuracy. Specifically, ① Since the traditional pipette uses air displacement (i.e., the equal-mass pipetting principle), when sucking a liquid with a density significantly different from that of water, the liquid volume taken will be too large or too small, resulting in obvious errors and affecting the liquid preparation result; ② The pipette tip and the pipette tip cone at the lower end of the pipette are tightly connected by the user gently pressing the pipette body and turning it left and right. This way of fixing relying on friction is not reliable enough, and the sealing performance is difficult to guarantee, thus affecting the liquid uptake accuracy; ③ The pipetting effect of the traditional pipette is greatly affected by human factors, especially the speed of manually sucking the liquid is not easy to control; ④ The traditional pipette can only perform basic pipetting operations, with a single function and unable to achieve continuous liquid separation. Long-term operation is prone to fatigue, affecting the accuracy and stability of pipetting; ⑤ The traditional pipette can only adjust the volume of the sucked liquid through a single scale adjustment knob, which is relatively troublesome.

[0045] The working principle of the micro-injection pump is based on the pressure difference of the liquid and the absolute control of the fluid. By precisely controlling the movement of the piston or roller, the flow rate and speed of the liquid are controlled. At the same time, the feedback signals from the pressure sensor and the flow sensor ensure the accuracy and stability of the liquid addition. By precisely controlling the drive system, the volume control system and the flow rate adjustment system, the micro-injection pump can meet the accuracy and stability requirements of different experimental needs.

[0046] Using a micro-injection pump eliminates the influence of manual liquid addition operations. Through step-by-step energy conversion, the pipetting rhythm and speed are more uniform and stable, improving the repeatability of the results, enabling more precise control of the liquid flow rate, and ensuring the accuracy of liquid addition. Therefore, selecting a syringe with an appropriate range can further improve the liquid addition accuracy.

[0047] In some embodiments, the flow rate of the micro-injection pump can be 0.1 - 3 mL / min, and the injection volume can be 0.4 mL - 0.8 mL. Optionally, the flow rate can be any one or the range between any two of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, and 3 mL / min, and the injection volume can be any one or the range between any two of 0.4, 0.5, 0.6, 0.7, 0.8 mL.

[0048] In some embodiments, the mixing further includes: stirring when adding the standard solution of 25-hydroxyvitamin D to the matrix. The stirring can be carried out using a magnetic stirrer.

[0049] Using a micro-injection pump in combination with a magnetic stirrer to achieve a liquid addition and stirring liquid preparation method, replacing the traditional pipette liquid preparation, can achieve the purpose of accurate liquid addition and effectively avoid the situation of local precipitation or precipitation. In addition, selecting a qualified syringe and removing air bubbles during liquid suction can avoid the influence of its sealing performance on the liquid extraction accuracy. Additionally, through the liquid preparation with a micro-injection pump, it has rich functions and partial automation of operation. The liquid addition volume and liquid addition speed can be quantified, and continuous liquid separation can also be achieved. It can be used and stopped at any time, and the accuracy and stability of liquid transfer can also be guaranteed.

[0050] On the other hand, the embodiments of the present invention also provide a quality control product or calibration product of 25-hydroxyvitamin D, which is obtained by freeze-drying the quality control product working solution or calibration product working solution described in any of the foregoing embodiments.

[0051] In some embodiments, the freeze-drying procedure includes the following steps:

[0052] Cool down to -45°C to -35°C for pre-freezing in 75 - 105 min, achieve vacuum, and maintain for 450 - 510 min;

[0053] Heat up to -15°C to -6°C within 90 to 120 minutes and maintain for 390 to 450 minutes;

[0054] Heat up to -2°C to 2°C again within 40 to 50 minutes and maintain for 390 to 450 minutes;

[0055] Cool down to 25°C to 35°C again within 90 to 110 minutes. After continuing for at least 120 minutes, the freeze-drying ends.

[0056] The pre-freezing temperature must be maintained at -45°C to -35°C, otherwise the product may become viscous or difficult to redissolve. During freeze-drying, the temperature increase should not be too fast. During the sublimation process, it needs to be maintained at -10°C to -6°C, otherwise cracks will appear in the product. During the desorption drying process, the temperature and time must be maintained at -2°C to 2°C and 390 to 450 minutes. If the time is too short, the phenomenon of spraying bottles will occur.

[0057] Specifically, the 75 to 105 minutes can be any one of 75, 80, 85, 90, 100, and 105 minutes or the range between any two of them. The -45°C to -35°C can specifically be any one of -45, -43, -40, -38, and -35°C or the range between any two of them. The 450 to 510 minutes can specifically be any one of 450, 460, 470, 480, 490, 500, and 510 minutes or the range between any two of them. The 90 to 120 minutes can specifically be any one of 90, 100, 105, 110, 115, and 120 minutes or the range between any two of them. The -15°C to -6°C can specifically be any one of -15, -14, -12, -10, -8, and -6°C or the range between any two of them. The 390 to 450 minutes can be any one of 390, 400, 410, 420, 430, 440, and 450 minutes or the range between any two of them. The 40 to 50 minutes can specifically be any one of 40, 45, and 50 minutes or the range between any two of them. The -2°C to 2°C can be any one of -2, 0, and 2°C or the range between any two of them. The 90 to 110 minutes can specifically be any one of 90, 95, 100, 105, and 110 minutes or the range between any two of them. The 25°C to 35°C can be any one of 25, 26, 28, 30, 32, 34, and 35°C or the range between any two of them. The "continuing for at least 120 minutes" means greater than or equal to 120 minutes, and specifically can be greater than or equal to any one of 120, 130, 140, 150, 160, 170, 180, 190, 200 minutes or the range between any two of them.

[0058] In addition, the embodiments of the present invention also provide the use of the composition or the matrix or the quality control product working solution or the calibration product working solution or the quality control product or the calibration product as described in any of the foregoing embodiments in the detection of 25-hydroxyvitamin D for non-disease diagnosis or treatment purposes.

[0059] In some embodiments, the 25-hydroxyvitamin includes 25-hydroxyvitamin D2 and / or 25-hydroxyvitamin D3.

[0060] The detection for non-disease diagnosis or treatment purposes includes the detection of standards, quality control products, calibration products and environmental samples.

[0061] The features and properties of the present invention will be further described in detail below in conjunction with embodiments.

[0062] Example 1

[0063] A method for preparing a calibration product / quality control product, which comprises the following steps:

[0064] I. Solution preparation:

[0065] 1. Solution preparation process:

[0066] ① Select a 1 mL syringe of appropriate specification according to the production plan (set the injection pump flow rate to 0.5 mL / min, injection volume 0.4 mL - 0.8 mL), container, magnetic stir bar, and build the solution preparation device and set the syringe manufacturer, specification, working mode, transmission liquid volume, and injection flow rate in the injection pump.

[0067] ② Measure the required matrix in a clean beaker that has been pre-loaded with a stir bar (in this example, the matrix is bovine serum albumin, and a protective agent has been added. The protective agent is ascorbic acid and 2,6-di-tert-butyl-p-cresol. The final concentration of ascorbic acid in the working solution is 5 mg / mL, and the final concentration of 2,6-di-tert-butyl-p-cresol in the working solution is 1 mg / mL), and start magnetic stirring.

[0068] ③ The syringe aspirates the standard solution of 25-hydroxyvitamin D (obtained by mixing 25-hydroxyvitamin D2, D3 and methanol), and the actual liquid aspiration volume needs to be greater than the theoretical liquid addition volume.

[0069] ④ Install the syringe that has aspirated 1 mL of the standard solution of 25-hydroxyvitamin D on the micro-injection pump, set the injection pump flow rate to 0.5 mL / min, injection volume 0.8 mL. After checking the injection pump flow rate and transmission liquid volume, start the injection pump and slowly drip the standard solution into the 39.2 mL matrix that is being stirred. Refer to Figures 1-2 .

[0070] ⑤ After the injection pump stops adding liquid, continue stirring for about 10 minutes. After the liquid preparation is completed, a calibrator working solution or a quality control product working solution is obtained.

[0071] II. Freeze-drying

[0072] Put the semi-finished products after the above-mentioned sub-packaging into a freeze-dryer and start freeze-drying. In the first 90 minutes, cool from the initial temperature to -40°C and reach vacuum to complete pre-freezing. Keep the above temperature for 480 minutes, then, it takes 110 minutes to heat up to -10°C and keep it for 420 minutes; then it takes 45 minutes to heat up to 0°C and keep it for 420 minutes; finally, it takes 100 minutes to cool down to 30°C and continue for 120 minutes, then perform vacuum plugging to end the freeze-drying. That is, the preparation of 25-hydroxyvitamin D calibrator and quality control product is completed.

[0073] Example 2

[0074] Based on the preparation method provided in Example 1, set up a control group. The control group is roughly the same as Example 1, except that a pipette is used for liquid preparation.

[0075] The effects of the two liquid preparation methods are compared as follows.

[0076] Table 1 Comparison of VD2 results of liquid preparation by pipette and micro-injection pump

[0077]

[0078] Table 2 Comparison of VD3 results of liquid preparation by pipette and micro-injection pump

[0079]

[0080]

[0081] Comparing the concentration results of the undried matrix calibrator and quality control product obtained by the two liquid preparation methods of pipette and micro-injection pump, the actual concentration accuracy of the preparation by micro-injection pump is higher, with a deviation within 5%, and the effect is better.

[0082] Example 3

[0083] Verify the influence of different protectants.

[0084] Based on the preparation method of Example 1, set up several experimental groups. Different experimental groups are roughly the same as Example 1, except that different protectants are used (the concentration of the protectant in the matrix is 1 mg / mL), measure the accuracy of its concentration and the theoretical concentration, and measure the deviation of its concentration from the measured value concentration on day 0 after accelerating the sample at 37°C for 5 days.

[0085] Table 3 Results of VD2 protectant screening

[0086]

[0087]

[0088] Table 4 Screening Results of VD3 Protective Agents

[0089]

[0090] From the above results, it shows that ascorbic acid, 2,6-di-tert-butyl-p-cresol and citric acid monohydrate have a certain protective effect on 25-hydroxyvitamin D.

[0091] Example 4

[0092] Based on the preparation method of Example 1, several experimental groups were set up. Different experimental groups were roughly the same as Example 1, except that different protective agents were used. The accuracy of their concentration and the theoretical concentration was measured, and the deviation of the concentration after accelerating the sample at 37°C for 5 days and the concentration measured at day 0 was measured.

[0093] Table 5 Screening Results of Different Combinations of VD2 Protective Agents

[0094]

[0095]

[0096] Table 6 Screening Results of Different Combinations of VD3 Protective Agents

[0097]

[0098] Note: The concentration of the protective agent in Tables 5-6 is its final concentration in the matrix.

[0099] From the above results, it shows that the 25-hydroxyvitamin D matrix solution with ascorbic acid + citric acid monohydrate protective agent added significantly degrades after accelerating at 37°C for 5 days, while the protective effect of the 25-hydroxyvitamin D matrix solution with ascorbic acid + 2,6-di-tert-butyl-p-cresol combination protective agent is the best.

[0100] Example 5

[0101] Based on the preparation method of Example 1, several experimental groups were set up. Different experimental groups were roughly the same as Example 1, except that different protective agents were used. The accuracy of their concentration and the theoretical concentration was measured, and the deviation of the concentration after accelerating the sample at 37°C for 5 days and the concentration measured at day 0 was measured.

[0102] Table 7 Screening Results of VD2 Protective Agent Concentrations

[0103]

[0104]

[0105] Table 8 Results of screening for concentration of VD3 protectant

[0106]

[0107]

[0108] Note: The concentrations of the protectants in Tables 7 - 8 are their final concentrations in the matrix.

[0109] It can be seen from the above results that the effect is better when the matrix contains 5 mg / mL ascorbic acid + 1 mg / mL 2,6 - di - tert - butyl - p - cresol as the protectant.

[0110] Example 6

[0111] Verification of freeze - drying loss rate

[0112] Based on the preparation method of Example 1, a control group without adding protectant for freeze - drying was set up, and the results were detected as follows.

[0113] Table 9 Freeze - drying loss rate of VD2

[0114]

[0115]

[0116] Table 10 Freeze - drying loss rate of VD3

[0117]

[0118] It can be seen that after adding the protectant, the loss rate of the active ingredient of the freeze - dried product is low, with a deviation within ±10%, and the product quality is better.

[0119] Example 7

[0120] Stability verification

[0121] Based on the preparation method of Example 1, a control group without adding protectant for freeze - drying was set up, and the results were detected. The product was measured after being placed at 2 - 8°C for 12 months, and the deviation was compared with the measured value at 0 day.

[0122] Table 11 VD2 stability results

[0123]

[0124]

[0125] Table 12 VD3 stability results

[0126]

[0127] The results in the above table show that for the products freeze-dried with a protective agent added to the matrix, the deviation of 25-hydroxyvitamin D is less than 10% after being stored for 12 months under refrigerated conditions, while for the freeze-dried products without the protective agent added, the deviation of 25-hydroxyvitamin D exceeds 15% after being stored for 12 months under refrigerated conditions, indicating that the protective agent has the effect of alleviating the degradation of 25-hydroxyvitamin D.

[0128] The above description is only the preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A composition, characterized in that It includes: Ascorbic acid and 2,6-di-tert-butyl-p-cresol.

2. The composition according to claim 1, characterized in that The mass ratio of the ascorbic acid to the 2,6-di-tert-butyl-p-cresol is 1-10:0.1-5; Optionally, the composition further comprises: a matrix; Optionally, the matrix comprises serum and / or serum albumin; Optionally, the serum comprises bovine serum; Optionally, the composition further comprises: any one or more of 25-hydroxyvitamin D and a standard solution of 25-hydroxyvitamin D.

3. A matrix containing a protective agent, characterized in that The protective agent includes ascorbic acid and 2,6-di-tert-butyl-p-cresol.

4. The matrix according to claim 3, characterized in that The mass ratio of the ascorbic acid to the 2,6-di-tert-butyl-p-cresol is 1-10:0.1-5; Optionally, the final concentration of ascorbic acid in the matrix is ​​1 to 10 mg / mL; Optionally, the final concentration of ascorbic acid in the matrix is ​​3-7 mg / mL; Optionally, the final concentration of the 2,6-di-tert-butyl-p-cresol in the matrix is ​​0.1 to 5 mg / mL; Optionally, the final concentration of the 2,6-di-tert-butyl-p-cresol in the matrix is ​​0.5 to 3 mg / mL; Optionally, the matrix comprises serum and / or serum albumin; Optionally, the serum comprises bovine serum.

5. Use of the composition according to claim 1 or 2 or the matrix according to claim 3 or 4 in the preparation of a quality control product, a calibrator, a quality control product working solution or a calibrator working solution of 25-hydroxyvitamin D.

6. A quality control working solution or calibration working solution of 25-hydroxyvitamin D, characterized in that: It includes: 25-Hydroxyvitamin D standard solution, matrix and protective agent; Wherein, the protective agent includes: ascorbic acid and 2,6-di-tert-butyl-p-cresol.

7. The quality control working solution or calibration working solution according to claim 6, characterized in that: The final concentration of ascorbic acid in the quality control working solution or the calibrator working solution is 1 to 10 mg / mL; Optionally, the final concentration of the ascorbic acid in the quality control working solution or the calibrator working solution is 3 to 7 mg / mL; Optionally, the final concentration of the 2,6-di-tert-butyl-p-cresol in the quality control working solution or the calibrator working solution is 0.1 to 5 mg / mL; Optionally, the final concentration of the 2,6-di-tert-butyl-p-cresol in the quality control working solution or the calibrator working solution is 0.5 to 3 mg / mL; Optionally, the matrix comprises serum and / or serum albumin; Optionally, the serum comprises bovine serum; Optionally, the volume ratio of the standard solution of 25-hydroxyvitamin D to the matrix is ​​1:30-60; Optionally, the standard solution of 25-hydroxyvitamin D comprises 25-hydroxyvitamin D and a diluent, and the diluent comprises methanol and / or ethanol.

8. A quality control working solution or calibration working solution of 25-hydroxyvitamin D, characterized in that: The method comprises mixing a standard solution of 25-hydroxyvitamin D, a matrix and a protective agent; The standard solution, matrix and protective agent of 25-hydroxyvitamin D are the standard solution, matrix and protective agent of 25-hydroxyvitamin D described in claim 6 or 7; Optionally, the mixing comprises: first mixing the protective agent and the matrix to obtain a matrix with the protective agent added; adding the standard solution of 25-hydroxyvitamin D to the matrix with the protective agent added to obtain a quality control working solution or a calibration working solution; Optionally, the adding of the standard solution of 25-hydroxyvitamin D is performed using a microinjection pump; Optionally, the mixing further comprises: stirring when adding the standard solution of 25-hydroxyvitamin D to the matrix.

9. A quality control or calibration product of 25-hydroxyvitamin D, characterized in that: It is obtained by freeze-drying the quality control working solution or the calibration working solution according to any one of claims 6 to 8; Optionally, the freeze-drying procedure comprises the following steps: Cool down to -45℃~-35℃ for pre-freezing at 75min~105min, achieve vacuum, and maintain for 450~510min; Raise the temperature to -15°C to -6°C in 90 to 120 minutes and maintain for 390 to 450 minutes; After 40 to 50 minutes, the temperature is raised to -2°C to 2°C and maintained for 390 to 450 minutes; The temperature is then lowered to 25°C to 35°C at 90 to 110 minutes and continued for at least 120 minutes before the freeze-drying is completed.

10. Use of the composition according to claim 1 or 2, or the matrix according to claim 3 or 4, or the quality control working solution or calibration working solution according to any one of claims 6 to 8, or the quality control or calibration product according to claim 9 in detecting 25-hydroxyvitamin D for purposes other than disease diagnosis or treatment; Optionally, the 25-hydroxyvitamin includes 25-hydroxyvitamin D2 and / or 25-hydroxyvitamin D3.

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