A multi-component composite quality control product for urine biochemical analysis and its preparation method

Abstract

A multi-component composite quality control product for urine biochemical analysis and a preparation method thereof. The quality control product includes human urine matrix, buffer matrix, quality control item addition substances, stabilizing substances, freeze-dried addition substances, and antibacterial agents. The quality control item addition substances at least include ionic substances, enzyme substances, and protein substances. The quality control product obtained by the technical solution of this application has the advantages of good stability, covering many items, and can include ionic substances, enzyme substances, and specific protein urine biochemical detection items, which is beneficial to the daily quality monitoring of testing institutions.

Description

Technical Field

[0001] The present invention relates to the technical field of in vitro diagnostic reagent detection, and particularly relates to a multi-item composite quality control product for urine biochemical analysis and a preparation method thereof. Background Art

[0002] Urine biochemical detection is a method of medical detection, including routine urine analysis, analysis of formed components in urine, quantitative analysis of urinary protein components, determination of urinary enzymes, etc. By detecting urine, doctors can know the types of kidney diseases and judge the severity of the diseases. Urine biochemical detection plays a crucial role in renal function monitoring.

[0003] Currently, most domestic urine biochemical detection reagents are equipped with separate quality control products. The multi-item composite urine quality control products mainly include enzyme-based multi-item composite urine quality control products and special protein-based multi-item composite urine quality control products. However, the stability of this kind of quality control product is not ideal, which is not friendly to the daily quality monitoring of testing institutions; and there are not many existing technologies that combine various types of multi-item urine quality control products, and there is basically no relevant report on combining ion types. Summary of the Invention

[0004] In view of the above deficiencies of the prior art, the present application provides a freeze-dried multi-item composite quality control product for urine biochemical analysis with good stability, covering many items, which can include ion types, enzyme types, and specific protein-based urine biochemical detection items, and is beneficial to the daily quality monitoring of testing institutions.

[0005] To solve the above technical problems, the technical solution adopted by the present application is: a multi-item composite quality control product for urine biochemical analysis, the quality control product includes human urine matrix, buffer matrix, quality control item addition substances, stabilizing substances, freeze-drying addition substances, preservatives (antibacterial agents); the quality control item addition substances at least include ion types, enzyme types, and protein substances.

[0006] Further, the human urine matrix is obtained by the following method: taking a human urine matrix solution that has been overnight at a temperature below 0°C, centrifuging to extract the supernatant, and then filtering it through a filter paper with a pore size of 0.22 μm at a low speed by suction filtration.

[0007] Further, the buffer matrix of the quality control product is HEPES buffer solution, preferably 100 mmol / L HEPES buffer solution.

[0008] Further, the quality control item additive substances are composed of inorganic phosphorus at a concentration of 10 - 40 mmol / L, potassium ions at 30 - 130 mmol / L, sodium ions at 60 - 200 mmol / L, chloride ions at 80 - 250 mmol / L, calcium ions at 1.5 - 4 mmol / L, magnesium ions at 3 - 13 mmol / L; 150 - 500 mmol / L urea (UREA), 0.7 - 1.4 mmol / L uric acid (UA), 7 - 16 mmol / L creatinine (CR), 3 - 15 mmol / L glucose (GLU), 20 - 150 mg / L albumin antigen (human albumin (ALB) antigen), 0.5 - 10 mg / L β2-microglobulin (β2-MG), 1.5 - 10 mg / L urinary transferrin (UTRF), 100 - 150 ng / mL neutrophil gelatinase-associated lipocalin (NGAL), 160 - 900 U / L α-amylase (α-AMY), 8 - 60 U / L N-acetyl-β-D-glucosaminidase (NAG), 20 - 30 mg / L immunoglobulin IgG (IgG), 0.6 - 5 mg / L retinol-binding protein (Retinol-Binding Protein, RBP), 10 - 50 mg / L α1-microglobulin (α1-microglobulin, α1-MG), and 5 - 40 mg / dL human albumin (human serum albumin).

[0009] Further, the stabilizing substances are one or more of sucrose, glycine, PVP30 (polyvinylpyrrolidone K30), and HSA (human serum albumin).

[0010] Furthermore, the stabilizing substances are sucrose and glycine; preferably, they are 5 g / L sucrose and 100 mmol / L glycine.

[0011] Further, the lyophilization additive substances are one or more of mannitol, trehalose, tert-butanol, and Tween 80.

[0012] Furthermore, the lyophilization additive substances are mannitol and Tween 80; preferably, they are 5 g / L mannitol and 1 mL / L Tween 80.

[0013] Further, the preservatives are PC300 and sodium azide; preferably, they are 1 mL / L PC300 and 0.1 g / L sodium azide.

[0014] Further, the pH value of the quality control product is 5.5 - 8.0.

[0015] This application also provides a preparation method for a multi-component composite quality control product for urine biochemical analysis. The preparation steps include:

[0016] (1) Preparation of human urine matrix solution: Take human urine matrix solution that has been stored overnight below 0°C, centrifuge to extract the supernatant, and then filter it through a 0.22 μm pore size filter paper by low-speed suction filtration to obtain it;

[0017] (2) Preparation of HEPES buffer solution: Weigh HEPES into purified water, adjust the pH with NaOH, add purified water to make up to 1 L, and store it at 2 - 8°C for standby;

[0018] (3) Mix and stir evenly the human urine matrix solution and the HEPES buffer solution in a volume ratio of 1:1, and then successively add uric acid, potassium chloride, sodium chloride, phosphoric acid, calcium chloride, α - amylase, creatinine, urea, glucose, stabilizing substances, β2 - microglobulin, α - microglobulin, immunoglobulin IgG, albumin antigen, urinary transferrin, magnesium acetate, N - acetyl - β - D - glucosaminidase, retinol - binding protein, neutrophil gelatinase - associated lipocalin, human serum albumin, freeze - dried additive substances (Tween 80, mannitol), preservatives (PC300, sodium azide), and after mixing evenly, obtain the basic solution of the multi - component composite quality control product for urine biochemical analysis;

[0019] (4) Dispense the prepared multi - component composite quality control product basic solution into freezing bottles with a continuous pipette gun, and perform freeze - drying using a freeze - dryer. After completion, obtain the finished product of the multi - component composite quality control product for urine biochemical analysis.

[0020] Further, in step (1) for preparing the human urine matrix solution, specifically: Take 1 L of human urine matrix solution that has been stored overnight in an - 8°C refrigerator, centrifuge at 3500 revolutions per minute for 15 minutes, extract the supernatant, and filter it through a 0.22 μm pore size filter paper by low - speed suction filtration to obtain it.

[0021] Further, in step (2) for preparing the HEPES buffer solution, specifically: Weigh 23.83 g of HEPES into 980 ml of purified water, adjust the pH with NaOH, add purified water to make up to 1 L, and store it at 2 - 8°C for standby.

[0022] Further, the stabilizing substances in step (3) are sucrose and glycine. The sucrose is added after the addition of glucose, and the glycine is added after the addition of magnesium acetate.

[0023] Further, the freeze - dried additive substances in step (3) are Tween 80 and mannitol, and the Tween 80 and mannitol are successively added after the addition of human serum albumin.

[0024] Further, the operation process of step (3) is specifically as follows: Mix human urine matrix solution and HEPES buffer solution in a ratio of 1:1 and stir evenly. Then, successively add uric acid, potassium chloride, sodium chloride, phosphoric acid, calcium chloride, α-amylase, creatinine, urea, glucose, sucrose, β2-microglobulin, α-microglobulin, immunoglobulin IgG, albumin antigen, urinary transferrin, magnesium acetate, glycine, N-acetyl-β-D-glucosaminidase, retinol-binding protein, neutrophil gelatinase-associated lipocalin, human serum albumin, Tween 80, mannitol, PC300, and sodium azide and mix evenly to obtain a multi-component composite quality control product base solution for urine biochemical analysis.

[0025] Advantages and technical effects of the present application:

[0026] 1. For the first time, the present application combines the quality control substances for urine ion detection items, urine enzyme detection items, and urine specific protein items, thereby obtaining a quality control product with good stability and covering many items.

[0027] 2. For the quality control product of the present application, the method of adding buffer solution using human urine matrix in the preparation can effectively improve the reconstitution and stability of the quality control product; phosphoric acid and calcium chloride can better promote the dissolution of α-amylase; glucose and sucrose can effectively reduce the inactivation of proteins; magnesium acetate and glycine can effectively increase the stability of N-acetyl-β-D-glucosaminidase; mannitol, etc. can effectively increase the collapse temperature during the freeze-drying process, which is beneficial to the stability of the freeze-dried product; the addition of Tween 80, etc. can be beneficial to the improvement of the activity after the quality control product is freeze-dried and reconstituted; the addition of PC300 in combination with sodium azide can greatly increase the antibacterial ability after the quality control substance is reconstituted, and also helps with the reconstitution stability. Therefore, for the quality control product with this specific composition in the present application, by continuously adjusting the reagent additives and the order of adding substances, the quality control substances are well dissolved together and the purpose of stability is achieved. By adding lyoprotectants, the freeze-drying yield of the quality control substances is effectively increased; the obtained freeze-dried quality control product has good stability. The addition of antibacterial substances helps with the reconstitution stability of the quality control product. It is convenient for the daily use of testing institutions.

[0028] 3. The quality control product of the present application is a freeze-dried multi-item composite quality control product for urine biochemistry. This quality control product is more stable, covers a wider range of detection items, and is convenient for the daily quality monitoring of testing institutions.

[0029] 4. Since enzymes and proteins are easily affected by ionic strength, leading to a decrease in stability, in this application, by adding some stabilizing substances and adjusting the addition sequence and concentration range of various substances, ionic substances are made not to affect the stability of enzymes and proteins, truly achieving the stable coexistence of ionic substances, enzyme-based quality control substances, and protein-based quality control substances. Many poorly soluble ionic substances are well dissolved in the solution together with enzyme-based quality control substances and protein-based quality control substances. In addition, specific antibacterial substances are added in this application, further increasing the stability of the composite quality control after reconstitution. Detailed implementation mode

[0030] The purpose of the present invention is to provide a multi-item composite quality control product for urine biochemical analysis. This quality control covers most current urine detection items on the market. The quality control product is stable and can effectively relieve the heavy daily quality monitoring work of testing institutions. The technical solution of the present invention is specifically described through the following embodiments. Relevant personnel can obviously make changes or appropriate modifications and combinations to the methods described herein without departing from the content, spirit, and scope of the present invention to implement and apply the technology of the present invention.

[0031] To further understand the present invention, the present invention will be clearly and completely described below in conjunction with embodiments. It is worth mentioning that the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention. The concentrations of various substances in this application are the concentrations in the final quality control product. Without special instructions, the preparation of the concentrations of various substances in this application is carried out by the conventional preparation methods in the industry.

[0032] Example 1:

[0033] A preparation method of a multi-item composite quality control product for urine biochemical analysis includes the following steps:

[0034] (1) Prepare human urine matrix solution: Take one liter of human urine matrix solution that has been stored in a -8°C refrigerator overnight (the human urine matrix solution here can be the urine of healthy people without virus carriage, and the same applies to the following embodiments), centrifuge at 3500 revolutions per minute for 15 minutes, extract the supernatant, and filter it slowly through a 0.22μm pore size filter paper to obtain it.

[0035] (2) Prepare HEPES buffer solution: Weigh 23.83 g of HEPES into 980 ml of purified water, adjust the pH to 5.5 with NaOH, add purified water to make up to 1 L, and store it at 2 - 8°C for standby.

[0036] (3) Mix human urine matrix solution and HEPES buffer solution in a 1:1 ratio and stir evenly. Then, successively add 0.8 mmol / L uric acid, 50 mmol / L potassium chloride, 80 mmol / L sodium chloride, 20 mmol / L phosphoric acid, 2 mmol / L calcium chloride, 200 U / L α-amylase, 8 mmol / L creatinine, 250 mmol / L urea, 6 mmol / L glucose, 2.5 g / L sucrose, 1.5 mg / L β2-microglobulin, 20 mg / L α-microglobulin, 20 mg / L immunoglobulin IgG, 35 mg / L albumin antigen, 3 mg / L urinary transferrin, 6 mmol / L magnesium acetate, 27 mmol / L glycine, 12 U / L N-acetyl-β-D-glucosaminidase, 1 mg / L retinol-binding protein, 150 ng / mL neutrophil gelatinase-associated lipocalin, 8 mg / dL human serum albumin, 0.5 mL / L Tween 80, 2 g / L mannitol, 0.5 mL / L PC300, and 0.05 g / L sodium azide. After mixing evenly, a multi-component composite quality control product base solution for urine biochemical analysis is obtained;

[0037] (4) Dispense the above-prepared multi-component composite quality control product base solution into freezing bottles using a continuous pipette gun and perform freeze-drying using a freeze dryer. After completion, a finished multi-component composite quality control product for urine biochemical analysis is obtained.

[0038] Example 2:

[0039] A preparation method of a multi-component composite quality control product for urine biochemical analysis includes the following steps:

[0040] (1) Prepare human urine matrix solution: Take one liter of human urine matrix solution that has been stored in a -8°C refrigerator overnight, centrifuge it at 3500 revolutions per minute for 15 minutes, extract the supernatant, and filter it slowly through a 0.22 μm pore size filter paper to obtain it;

[0041] (2) Prepare HEPES buffer solution: Weigh 23.83 g of HEPES into 980 ml of purified water, adjust the pH to 6.0 with NaOH, add purified water to make up to 1 L, and store it at 2 - 8°C for standby.

[0042] (3) Mix human urine matrix solution and HEPES buffer solution in a ratio of 1:1 and stir evenly. Then, add 1 mmol / L uric acid, 80 mmol / L potassium chloride, 100 mmol / L sodium chloride, 35 mmol / L phosphoric acid, 4 mmol / L calcium chloride, 750 U / L α-amylase, 12 mmol / L creatinine, 300 mmol / L urea, 12 mmol / L glucose, 3 g / L sucrose, 3 mg / L β2-microglobulin, 45 mg / L α-microglobulin, 30 mg / L immunoglobulin IgG, 100 mg / L albumin antigen, 10 mg / L urinary transferrin, 12 mmol / L magnesium acetate, 54 mmol / L glycine, 45 U / L N-acetyl-β-D-glucosaminidase, 5 mg / L retinol-binding protein, 150 ng / mL neutrophil gelatinase-associated lipocalin, 30 mg / dL human serum albumin, 1 ml / l Tween 80, 2.5 g / L mannitol, 0.5 ml / L PC300 and 0.05 g / L sodium azide, and mix evenly to obtain the multi-component composite quality control product base solution for urine biochemical analysis.

[0043] (4) Dispense the above-prepared multi-component composite quality control product base solution into cryovials using a continuous pipette, and perform freeze-drying using a freeze dryer. After completion, obtain the finished multi-component composite quality control product for urine biochemical analysis.

[0044] Example 3:

[0045] A preparation method of a multi-component composite quality control product for urine biochemical analysis includes the following steps:

[0046] (1) Prepare human urine matrix solution: Take 1 liter of human urine matrix solution stored in a -8°C refrigerator overnight, centrifuge at 3500 revolutions per minute for 15 minutes, extract the supernatant, and filter it slowly through a 0.22 μm pore size filter paper to obtain it.

[0047] (2) Prepare HEPES buffer solution: Weigh 23.83 g of HEPES in 980 ml of purified water, adjust the pH to 8.0 with NaOH,

[0048] add purified water to make up to 1 L, and store it at 2-8°C for standby.

[0049] (3) Mix the human urine matrix solution and HEPES buffer in a 1:1 ratio and stir evenly. Then, sequentially add 0.8 mmol / L uric acid, 70 mmol / L potassium chloride, 60 mmol / L sodium chloride, 20 mmol / L phosphoric acid, 1.5 mmol / L calcium chloride, 250 U / L α-amylase, 8 mmol / L creatinine, 200 mmol / L urea, 6.5 mmol / L glucose, 4 g / L sucrose, 1 mg / L β2-microglobulin, 15 mg / L α-microglobulin, 20 mg / L immunoglobulin IgG, 25 mg / L albumin antigen, 2.5 mg / L urinary transferrin, 12 mmol / L magnesium acetate, 67 mmol / L glycine, 10 U / L N-acetyl-β-D-glucosaminidase, 1 mg / L retinol-binding protein, 100 ng / mL neutrophil gelatinase-associated lipocalin, 15 mg / dL human serum albumin, 1 mL / L Tween 80, 3 g / L mannitol, 0.5 mL / L PC300, and 0.05 g / L sodium azide. After mixing evenly, a multi-component composite quality control product base solution for urine biochemical analysis is obtained.

[0050] (4) Dispense the prepared multi-component composite quality control product base solution into freezing bottles using a continuous pipette gun and perform freeze-drying using a freeze dryer. After completion, a finished multi-component composite quality control product for urine biochemical analysis is obtained.

[0051] Example 4:

[0052] A preparation method for a multi-component composite quality control product for urine biochemical analysis includes the following steps: (1) Prepare the human urine matrix solution: Take one liter of human urine matrix solution that has been stored in a -8°C refrigerator overnight, centrifuge at 3500 revolutions per minute for 15 minutes, extract the supernatant, and perform low-speed suction filtration using a 0.22 μm pore size filter paper to obtain it.

[0053] (2) Prepare the HEPES buffer: Weigh 23.83 g of HEPES into 980 ml of purified water, adjust the pH to 7.5 with NaOH, add purified water to make up to 1 L, and store it at 2 - 8°C for standby.

[0054] (3) Mix the human urine matrix solution and HEPES buffer in a 1:1 ratio and stir evenly. Then, sequentially add 1.4 mmol / L uric acid, 100 mmol / L potassium chloride, 10 mmol / L sodium chloride, 40 mmol / L phosphoric acid, 4 mmol / L calcium chloride, 900 U / L α-amylase, 16 mmol / L creatinine, 450 mmol / L urea, 15 mmol / L glucose, 4 g / L sucrose, 10 mg / L β2-microglobulin, 50 mg / L α-microglobulin, 30 mg / L immunoglobulin IgG, 135 mg / L albumin antigen, 10 mg / L urinary transferrin, 5 mmol / L magnesium acetate, 80 mmol / L glycine, 55 U / L N-acetyl-β-D-glucosaminidase, 4.5 mg / L retinol-binding protein, 150 ng / mL neutrophil gelatinase-associated lipocalin, 30 mg / dL human serum albumin, 0.5 mL / L Tween 80, 5 g / L mannitol, 1 mL / L PC300, and 0.1 g / L sodium azide. After mixing evenly, a multi-component composite quality control product base solution for urine biochemical analysis is obtained;

[0055] (4) Dispense the prepared multi-component composite quality control product base solution into freezing bottles using a continuous pipette gun, and perform freeze-drying using a freeze dryer. After completion, a finished multi-component composite quality control product for urine biochemical analysis is obtained.

[0056] Example 5

[0057] A preparation method for a multi-component composite quality control product for urine biochemical analysis includes the following steps:

[0058] (1) Prepare the human urine matrix solution: Take one liter of human urine matrix solution stored in an -8°C refrigerator overnight, centrifuge at 3500 revolutions per minute for 15 minutes, extract the supernatant, and perform low-speed suction filtration using a 0.22 μm pore size filter paper to obtain it;

[0059] (2) Prepare HEPES buffer: Weigh 23.83 g of HEPES into 980 ml of purified water, adjust the pH to 7.0 with NaOH, add purified water to make up to 1 L, and store at 2 - 8 °C for later use; (3) Mix the human urine matrix solution and HEPES buffer in a 1:1 ratio and stir well. Then, add 1.2 mmol / L uric acid, 100 mmol / L potassium chloride, 100 mmol / L sodium chloride, 35 mmol / L phosphoric acid, 4 mmol / L calcium chloride, 900 U / L α-amylase, 16 mmol / L creatinine, 400 mmol / L urea, 15 mmol / L glucose, 5 g / L sucrose, 8 mg / L β2-microglobulin, 45 mg / L α1-microglobulin, 30 mg / L immunoglobulin IgG, 120 mg / L albumin antigen, 10 mg / L urinary transferrin, 13 mmol / L magnesium acetate, 100 mmol / L glycine, 60 U / L N-acetyl-β-D-glucosaminidase, 5 mg / L retinol-binding protein, 150 ng / mL neutrophil gelatinase-associated lipocalin, 30 mg / dL human serum albumin, 1 ml / L Tween 80, 5 g / L mannitol, 1 ml / L PC300, and 0.1 g / L sodium azide in sequence. After mixing evenly, a multi-component composite quality control product base solution for urine biochemical analysis is obtained;

[0060] (3) Dispense the prepared multi-component composite quality control product base solution into freezing bottles using a continuous liquid dispenser, and perform freeze-drying using a freeze dryer. After completion, a finished multi-component composite quality control product for urine biochemical analysis is obtained.

[0061] Comparative example:

[0062] (1): By mass, add 100 mmol / L of the buffer (HEPES buffer), 0.05 g / L of the surfactant (Triton X-100), 30 g / L of the saccharide substance (dextran), 0.15 g / L of the metal chelating agent (sodium ethylenediaminetetraacetate), and 1.0 g / L of the preservative (sodium azide) to purified water respectively. Wait for dissolution and mix evenly. After adjusting the pH range to 7.0, a multi-component composite quality control product matrix solution is obtained and reserved for later use;

[0063] (2): Add 150 μmol / L of creatinine, 15 mg / L of albumin antigen, 2 mg / L of transferrin, 8 mg / L of immunoglobulin G, 0.8 mg / L of retinol-binding protein, 0.3 mg / L of β2-microglobulin, 5.0 mg / L of α1-microglobulin, and 200 ng / mL of neutrophil gelatinase-associated lipocalin to the multi-component composite quality control product matrix solution respectively. Wait for dissolution and mix evenly to obtain the first multi-component composite quality control product mother liquor and reserve it for later use;

[0064] (3): Add 8.0 g / L of the protein stabilizer (casein), 0.005 g / L of the ionic stabilizer (calcium acetate), 250 U / L of the α-amylase, and 8 U / L of the N-acetyl-β-D-glucosaminidase to purified water respectively. After waiting for dissolution, mix evenly to obtain the mother liquor of the second multi-component quality control product. Place the mother liquor of the second multi-component quality control product in a low-temperature environment at 2°C to 8°C for cooling, and the cooling time is not less than 30 min;

[0065] (4): Add the mother liquor of the second multi-component quality control product to the mother liquor of the first multi-component quality control product, mix evenly, measure the pH value after mixing the mother liquor of the first multi-component quality control product and the mother liquor of the second multi-component quality control product, adjust its pH value range to 7.0, and quantitatively obtain the multi-component quality control product for urine biochemical analysis with strong stability by using the matrix solution of the multi-component quality control product;

[0066] (5): Use a precision flow pump to dispense the multi-component quality control product for urine biochemical analysis with strong stability;

[0067] (6): Use a freeze dryer to perform freeze-drying operation on the dispensed multi-component quality control product for urine biochemical analysis with strong stability, and after completion, press the stopper and seal.

[0068] Perform performance testing on the quality control products obtained in Examples 1-5 and the comparative examples of the present application:

[0069] (1) Freeze-drying efficiency verification scheme, test method: Take out 10 ml of the basic solution of the multi-component quality control product for urine before freeze-drying in the examples and dispense them for storage at -80°C for standby. Reconstitute the freeze-dried multi-component quality control products in the examples with purified water respectively. Use the kits produced by Meikang Biotech Co., Ltd. to detect the corresponding items. Compare the test results of each item of the reconstituted quality control product with the measured values of the basic solution stored at -80°C one by one to obtain the freeze-drying yield. The freeze-drying yield results are shown in Table 1 below:

[0070] Table 1 Freeze-drying yield

[0071]

[0072] Conclusion: It can be seen from Table 1 that the freeze-drying yields of the quality control products obtained in Examples 1-5 are all greater than 75%, and the freeze-drying efficiency is relatively high, especially Example 5 is the best; adding the freeze-drying protectant Tween 80 in the examples can help improve the activity of the enzyme after freeze-drying, and mannitol can effectively increase the collapse temperature during freeze-drying, prevent the effective components from sublimating and escaping with water vapor during freeze-drying, and promote the formation of tangible substances by the effective components, which is beneficial to the stability of the freeze-dried products.

[0073] Stability verification scheme, specifically:

[0074] (1) Stability at 2 - 8°C after opening. Test method: After thawing the freeze - dried multiplex urine quality control products obtained in the examples and comparative examples, place them at the specified storage temperature (2 - 8°C), and take samples on the 0th, 5th, 7th, 14th, and 20th days respectively for testing. Statistical data analysis is carried out to observe the trend changes. The results are shown in Tables 2 to 6.

[0075] Table 2 Stability at 2 - 8°C after opening / Relative deviation of Example 1

[0076]

[0077] Table 3 Stability at 2 - 8°C after opening / Relative deviation of Example 2

[0078]

[0079]

[0080] Table 4 Stability at 2 - 8°C after opening / Relative deviation of Example 3

[0081]

[0082]

[0083] Table 5 Stability at 2 - 8°C after opening / Relative deviation of Example 4

[0084]

[0085] Table 6 Stability at 2 - 8°C after opening / Relative deviation of Example 5

[0086]

[0087]

[0088] Table 7 Stability at 2 - 8°C after opening / Relative deviation of Comparative Example

[0089]

[0090] Conclusion: It can be seen from Tables 1 - 7 that the deviation of the stability at 2 - 8°C after opening of the comparative example within 20 days is within 15%, and the trend is not significant, which is worse than that of Examples 1 - 5. It can be seen from Tables 2 - 6 that the deviation of the stability at 2 - 8°C after opening of the examples is within 10%, and the trend is not significant, indicating that the multiplex urine quality control products in the examples have good stability within 20 days, especially the stability at 2 - 8°C after opening of Example 5 is the best.

[0091] (2) Stability at -20°C during thawing. Test method: After thawing the freeze-dried multi-component urine quality control products obtained in the examples and comparative examples, place them separately at the specified storage temperature (-20°C). Take samples on the 0th, 3rd, 7th, 14th, 21st, and 30th days respectively for testing, and analyze the trend changes in the statistical data. The results are shown in Tables 7 to 11.

[0092] Table 8 Thawing stability / relative deviation of Example 1

[0093]

[0094] Table 9 Thawing stability / relative deviation of Example 2

[0095]

[0096] Table 10 Thawing stability / relative deviation of Example 3

[0097]

[0098]

[0099] Table 11 Thawing stability / relative deviation of Example 4

[0100]

[0101]

[0102] Table 12 Thawing stability / relative deviation of Example 5

[0103]

[0104] Table 13 Thawing stability / relative deviation of the comparative example

[0105]

[0106] Conclusion: It can be obtained from Tables 8 - 13 that the deviation of the thawing stability of the multi-component urine quality control products in the comparative example is within 10%, and the trend is not significant. The deviation of the thawing stability of the multi-component urine quality control products in Examples 1 - 5 is within 8%, and the trend is not significant, indicating that the thawing stability of the quality control products in the examples is good and more stable than that in the comparative example during this period. Among them, Example 5 is the best.

[0107] (3) Antibacterial test, test method: After thawing the urine multi-component composite quality control products in Examples 1-5 and the comparative example, store them in a refrigerator at 2-8°C. Take 20 μl of samples on day 0, day 7, day 15, day 24, and day 35. After spreading on the plate, culture at 28°C for 3 days, and observe the growth of colonies. If there is no growth, it is indicated by " / "; if there are less than 5 colonies, it is indicated by "+"; if there are 5-15 colonies, it is indicated by "++"; if there are 15-30 colonies, it is indicated by "+++"; if there are more than 30 colonies, it is indicated by "++++". The results are shown in Table 14:

[0108] Table 14 Colony degree of the quality control products prepared in the examples and the comparative example after 3-day culture at 28°C

[0109]

[0110] Conclusion: The antibacterial agents PC300 and sodium azide used in this application are broad-spectrum antibacterial agents with excellent antibacterial ability. These two substances have good compatibility and stability and will not interfere with other components in the quality control products. It can be seen from Table 13 that the antibacterial ability of the comparative example is very weak, while Examples 1-5 all have good antibacterial ability, especially Example 5 has excellent antibacterial ability and is suitable for daily quality monitoring in testing institutions.

[0111] From the evaluation and test results of the above several examples and the comparative example, it can be concluded that the freeze-drying yield of the urine multi-component composite quality control product of this application is high, the stability is good, and the antibacterial ability is strong, which is particularly suitable for daily quality control monitoring in testing institutions.

[0112] In summary, the specific examples described in this article are only illustrative of the spirit of the present invention. Simple modifications or equivalent replacements made by the R & D personnel in the technical field of the present invention to the technical solutions of the present invention do not depart from the essence and scope of the technical solutions of the present invention.

Claims

1. A multi-component composite quality control product for urine biochemical analysis, characterized in that: The quality control product described above includes human urine matrix, buffer matrix, quality control item addition substances, stabilizing substances, freeze-dried addition substances, and antibacterial agents; The added substances in the quality control items are composed of phosphoric acid at a concentration of 10 - 40 mmol / L, potassium chloride at 30 - 130 mmol / L, sodium chloride at 60 - 200 mmol / L, calcium chloride at 1.5 - 4 mmol / L, magnesium acetate at 3 - 13 mmol / L; urea at 150 - 500 mmol / L, uric acid at 0.7 - 1.4 mmol / L, creatinine at 7 - 16 mmol / L, glucose at 3 - 15 mmol / L, albumin antigen at 20 - 150 mg / L, β2-microglobulin at 0.5 - 10 mg / L, urinary transferrin at 1.5 - 10 mg / L, neutrophil gelatinase-associated lipocalin at 100 - 150 ng / mL, α α-amylase at 8 - 60 U / L, N-acetyl-β-D-glucosaminidase at 20 - 30 mg / L, immunoglobulin IgG at 0.6 - 5 mg / L, retinol-binding protein at 10 - 50 mg / L α β2-microglobulin at 5 - 40 mg / dL, and human serum albumin; The buffer matrix of the quality control product is HEPES buffer solution; The stabilizing substances are two kinds: 2.5 - 5 g / L sucrose and 27 - 100 mmol / L glycine; The freeze-dried addition substances are two kinds: 2 - 5 g / L mannitol and 0.5 - 1 ml / L Tween 80.

2. The multiplex composite quality control product for urine biochemical analysis according to claim 1, characterized in that: The human urine matrix described above is obtained by the following method: Take the human urine matrix solution that has been overnight at a temperature below 0 °C, centrifuge to extract the supernatant, and then perform low-speed suction filtration through a filter paper with a pore size of 0.22 μm.

3. The multi-component composite quality control product for urine biochemical analysis according to claim 1, wherein: The stabilizing substances are two kinds: 5 g / L sucrose and 100 mmol / L glycine; the freeze-dried addition substances are two kinds: 5 g / L mannitol and 1 ml / L Tween 80; the antibacterial agents are 1 ml / L PC300 and 0.1 g / L sodium azide.

4. A preparation method of a multi-component composite quality control product for urine biochemical analysis according to any one of claims 1-3, characterized in that: The preparation steps include: (1) Prepare the human urine matrix solution: Take the human urine matrix solution that has been overnight at a temperature below 0 °C, centrifuge to extract the supernatant, and then perform low-speed suction filtration through a filter paper with a pore size of 0.22 μm; (2) Prepare the HEPES buffer solution: Weigh HEPES in purified water, adjust the pH with NaOH, add purified water to make up to 1 L, and store at 2 - 8 °C for standby; (3) Mix and stir evenly the human urine matrix solution and the HEPES buffer solution in a volume ratio of 1:1, and then sequentially add uric acid, potassium chloride, sodium chloride, phosphoric acid, calcium chloride, α-amylase, creatinine, urea, glucose, stabilizing substances, β2-microglobulin, α-microglobulin, immunoglobulin IgG, albumin antigen, urinary transferrin, magnesium acetate, N-acetyl-β-D-glucosaminidase, retinol-binding protein, neutrophil gelatinase-associated lipocalin, human serum albumin, freeze-dried addition substances, antibacterial agents, and after mixing evenly, obtain the multi-component composite quality control product base solution for urine biochemical analysis; (4) Dispense the prepared multi-component composite quality control product base solution into freezing bottles with a continuous pipette gun, and perform freeze-drying using a freeze dryer. After completion, obtain the finished multi-component composite quality control product for urine biochemical analysis.

5. The preparation method of the multiple composite quality control product for urine biochemical analysis according to claim 4, characterized in that: The step (1) of preparing the human urine matrix solution is specifically as follows: Take 1 L of human urine matrix solution that has been overnight in an -8 °C refrigerator, centrifuge at 3500 revolutions per minute for 15 minutes, extract the supernatant, and perform low-speed suction filtration through a filter paper with a pore size of 0.22 μm to obtain it; the step (2) of preparing the HEPES buffer solution is specifically as follows: Weigh 23.83 g of HEPES in 980 ml of purified water, adjust the pH with NaOH, add purified water to make up to 1 L, and store at 2 - 8 °C for standby.

6. The preparation method of the multi-component composite quality control product for urine biochemical analysis according to claim 5, wherein: The stabilizing substances in the step (3) are sucrose and glycine. The sucrose is added after the addition of glucose, and the glycine is added after the addition of magnesium acetate; the freeze-dried addition substances in the step (3) are Tween 80 and mannitol, and the Tween 80 and mannitol are sequentially added after the addition of human serum albumin.

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