A whole blood composite quality control product for microfluidic biochemical analysis chip

Abstract

The invention relates to the field of quality control products, and specifically discloses a whole blood composite quality control product for a microfluidic biochemical analysis chip; the raw materials of the quality control product include pig whole blood, quality control item additives, Aldrich solution, a buffer, an antioxidant, biocytin hydrochloride, levodopa, xanthan gum, dabigatran, and a freeze-dried additive; the quality control item additives include cholesterol sulfate sodium salt, potassium dihydrogen phosphate, ammonium ferric sulfate dodecahydrate, calcium chloride, magnesium sulfate heptahydrate, urea, sodium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, glycocholic acid, glucose, creatinine, uric acid, triolein, bovine serum albumin, conjugated bilirubin, lactic acid, β-hydroxybutyric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipase, lactate dehydrogenase, γ-glutamyl transferase, amylase, adenosine deaminase, α-fucosidase, and creatine kinase.

Description

Technical Field

[0001] The invention relates to the field of quality control products, and specifically discloses a whole blood composite quality control product for a microfluidic biochemical analysis chip. Background Art

[0002] Microfluidic chip technology has the advantages of less reagent consumption, less sample required, short detection time, low cost, simple operation and easy integration. At present, centrifugal microfluidic disc biochemical analysis chips have gradually been used in the clinical field. The chip integrates sample pretreatment and detection process. During analysis, the sample only needs to be injected into the chip containing freeze-dried reagent through the sample hole for quantitative determination. When this type of chip is tested, the unmeasured blood sample can be converted into multiple aliquots of precisely diluted plasma through capillary action and the centrifugal force when the detection chip rotates, thereby realizing the functions of automated whole blood quantification, blood cell plasma separation, and plasma dilution, and simultaneously analyzing multiple chemical components through optical detection of the reaction hole.

[0003] At present, the items that can be detected by this type of chip have covered multiple fields such as liver function, kidney function, blood sugar, blood lipids, ions, etc. Chinese patent CN201711076990.7 discloses a method for preparing plasma quality control products for centrifugal microfluidic chip quality control. Bovine plasma is used as the material, which is more representative than conventional serum quality control, and avoids the matrix effect caused by the difference between serum samples and plasma samples to a certain extent. Since the ability to directly analyze whole blood samples is one of the important advantages of centrifugal microfluidic disc biochemical analysis chips, if whole blood type quality control products can be used, it can be closest to the matrix of the sample to be tested, and better quality control can be performed on the chip detection. Therefore, providing a whole blood composite quality control product that can be used for centrifugal microfluidic disc biochemical analysis chips has become an urgent problem to be solved in the industry. Summary of the invention

[0004] The object of the present invention is to provide a whole blood composite quality control product for a microfluidic biochemical analysis chip to solve the problems raised in the above background technology.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0006] A whole blood composite quality control product for a microfluidic biochemical analysis chip: the raw materials of the whole blood composite quality control product include porcine whole blood, quality control item additives, Aldrich solution, buffer, antioxidant, biocytin hydrochloride, levodopa, xanthan gum, dabigatran, and freeze-dried additives.

[0007] Preferably, the concentration of biocytin hydrochloride is 1-10 g / L, the concentration of levodopa is 20-50 mmol / L, the concentration of xanthan gum is 5-20 g / L; the concentration of the buffer is 50-200 mmol / L, the concentration of dabigatran is 1-10 g / L, the concentration of the antioxidant is 10-200 μmol / L, the concentration of the freeze-dried additive is 20-100 g / L, and the rest is porcine whole blood and Aldrich solution; the porcine whole blood includes plasma and blood cells, and the ratio of the added amount of Aldrich solution to the blood cell volume is 1:1.

[0008] Preferably, the added substances for the quality control items include cholesterol sulfate sodium salt, potassium dihydrogen phosphate, ammonium ferric sulfate dodecahydrate, calcium chloride, magnesium sulfate heptahydrate, urea, sodium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, glycocholic acid, glucose, creatinine, uric acid, triolein, bovine serum albumin, conjugated bilirubin, lactic acid, β-hydroxybutyric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipase, lactate dehydrogenase, γ-glutamyl transferase, amylase, adenosine deaminase, α-fucosidase, and creatine kinase.

[0009] Preferably, the concentrations of the substances are as follows: cholesterol sodium sulfate 2-7 mmol / L, potassium dihydrogen phosphate 1-3 mmol / L, ammonium ferric sulfate dodecahydrate 10-40 μmol / L, calcium chloride 2-3.5 mmol / L, magnesium sulfate heptahydrate 0.9-1.7 mmol / L, urea 6-22 mmol / L, sodium chloride 80-120 mmol / L, zinc sulfate heptahydrate 20-40 μmol / L, copper sulfate pentahydrate 15-30 μmol / L, glycocholic acid 25-50 μmol / L, glucose 5-16 mmol / L, creatinine 80-400 μmol / L, uric acid 250-700 μmol / L, triolein 1 ~4mmol / L, bovine serum albumin 40~80g / L, conjugated bilirubin 15~90μmol / L, lactic acid 1.5~6mmol / L, β-hydroxybutyric acid 0.2~2mmol / L, alanine aminotransferase 40~130U / L, aspartate aminotransferase 40~150U / L, alkaline phosphatase 150~400U / L, lipase 20~90U / L, lactate dehydrogenase 150~300U / L, γ-glutamyl transferase 40~200U / L, amylase 50~300U / L, adenosine deaminase 20~60U / L, α-fucosidase 20~100U / L, creatine kinase 100~600U / L.

[0010] A method for preparing a whole blood composite quality control product for a microfluidic biochemical analysis chip comprises the following steps: S1: collecting lithium heparin anticoagulated pig whole blood, and separating plasma from blood cells by centrifugation;

[0011] S2: Add an equal volume of Aldrich solution to the separated blood cells, mix evenly by inversion to obtain blood cell preservation solution, and store at 2-8°C;

[0012] S3: The collected plasma is divided into two equal parts, and a buffer and an antioxidant are added to one of the plasma parts in sequence, and the mixture is stirred until completely dissolved. After mixing evenly, sodium cholesterol sulfate is added, and the mixture is heated to 55-70°C. After the mixture is completely dissolved, the mixture is cooled to room temperature, and potassium dihydrogen phosphate is added, and the mixture is stirred until completely dissolved and mixed evenly.

[0013] S4: add ammonium ferric sulfate dodecahydrate, calcium chloride, magnesium sulfate heptahydrate, urea, sodium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, glycocholic acid, glucose, creatinine, uric acid, triolein, bovine serum albumin, conjugated bilirubin, lactic acid and β-hydroxybutyric acid to another portion of plasma in sequence, add the next substance after the previous substance is completely dissolved and mixed evenly, stir until completely dissolved and mixed evenly;

[0014] S5: Evenly mix the solutions obtained in steps S3 and S4, add biocytin hydrochloride, levodopa, and xanthan gum in sequence, stir until completely dissolved, mix evenly, adjust the pH of the solution to 7.0-7.2, add alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipase, lactate dehydrogenase, γ-glutamyl transferase, amylase, adenosine deaminase, α-fucosidase, and creatine kinase in sequence, stir until completely dissolved, and mix evenly; add dabigatran, stir until completely dissolved, slowly add 4 times the volume of blood cell preservation solution, stir at a low speed, mix well, add freeze-dried additives, stir until completely dissolved, divide and freeze-dry to obtain freeze-dried powder of whole blood composite quality control product.

[0015] Preferably, the buffer is one of piperazine-1,4-diethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, and N,N-bis(hydroxyethyl)glycine.

[0016] Preferably, the antioxidant is one of tris(2-carboxyethyl)phosphine hydrochloride and sodium dithionate.

[0017] Preferably, the freeze-drying additive is one or more combinations of trehalose, glucose, sucrose and mannitol.

[0018] Preferably, the concentration of the buffer is 50-200 mmol / L, the concentration of dabigatran is 1-10 g / L, the concentration of the antioxidant is 10-200 μmol / L, and the concentration of the lyophilized additive is 20-100 g / L.

[0019] Compared with the prior art, the beneficial effects achieved by the present invention are: (1) providing a whole blood composite quality control product for a microfluidic biochemical analysis chip and a preparation method thereof, using pig whole blood as a matrix, which is closest to the matrix of the sample to be tested compared with the prior art, and can better and more comprehensively perform quality control on the centrifugal microfluidic disc biochemical analysis chip;

[0020] (2) Biocytin hydrochloride, levodopa and xanthan gum, which are relatively mild to red blood cells, are selected as stabilizers and protective agents for the whole blood composite quality control product, which can play a good stabilizing and protective role on the quality control product, and the performance of the quality control product of the present invention can meet the clinical use requirements; wherein, biocytin hydrochloride and levodopa are mild in nature, and both have acid and alkaline properties, and can effectively inhibit the pH change of the composite quality control product solution, thereby achieving the purpose of protecting the quality control product, and the mixed use of the two has a higher efficacy than when used alone; xanthan gum is stable to heat, acid and alkali, and salt, and has extremely strong antioxidant and anti-enzymatic capabilities. In the whole blood composite quality control product, it can reduce the content of free water to achieve the purpose of increasing the viscosity of the solution, thereby stabilizing and protecting the quality control product from being destroyed and degraded. DETAILED DESCRIPTION

[0021] The following is a preferred implementation of the embodiment of the present invention. Obviously, the described embodiment is only a part of the embodiment of the present invention, not all of the embodiments. For ordinary technicians in this technical field, all other embodiments obtained by ordinary technicians in this field without creative work without departing from the principle of the embodiment of the present invention are within the scope of protection of the present invention.

[0022] Example 1: S1: Take lithium heparin anticoagulated pig whole blood, and separate plasma from blood cells by centrifugation;

[0023] S2: Add an equal volume of Aldrich solution to the separated blood cells, invert and mix evenly to obtain blood cell preservation solution;

[0024] S3: The collected plasma was divided into two equal parts, and 500mmol / L piperazine-1,4-diethanesulfonic acid and 500μmol / L tri(2-carboxyethyl)phosphine hydrochloride were added to one of the plasma parts in sequence, and stirred until completely dissolved. After mixing evenly, 20mmol / L cholesterol sulfate sodium salt was added, and the mixture was heated to 60°C. After completely dissolved, the mixture was cooled to room temperature, and then 10mmol / L potassium dihydrogen phosphate was added to the obtained solution, and the mixture was stirred until completely dissolved and mixed evenly.

[0025] S4: add 100 μmol / L ammonium ferric sulfate dodecahydrate, 20 mmol / L calcium chloride, 9 mmol / L magnesium sulfate heptahydrate, 60 mmol / L urea, 800 mmol / L sodium chloride, 200 μmol / L zinc sulfate heptahydrate, 150 μmol / L copper sulfate pentahydrate, 250 μmol / L glycocholic acid, 50 mmol / L glucose, 800 μmol / L creatinine, 2500 μmol / L uric acid, 10 mmol / L triolein, 400 g / L bovine serum albumin, 150 μmol / L conjugated bilirubin, 15 mmol / L lactic acid, and 2 mmol / L β-hydroxybutyric acid to another portion of plasma in sequence, add the next substance after the previous substance is completely dissolved and mixed evenly, stir until completely dissolved, and mix evenly;

[0026] S5: The solutions obtained in S3 and S4 were mixed evenly, 5 g / L biocytin hydrochloride, 100 mmol / L levodopa and 25 g / L xanthan gum were added in sequence, stirred until completely dissolved, mixed evenly, the pH of the solution was adjusted to 7.0, 200 U / L alanine aminotransferase, 200 U / L aspartate aminotransferase, 750 U / L alkaline phosphatase, 100 U / L lipase, 750 U / L lactate dehydrogenase, 200 U / L γ-glutamyl transferase, 250 U / L amylase, 100 U / L adenosine deaminase, 100 U / L α-fucosidase and 500 U / L creatine kinase were added in sequence, stirred until completely dissolved, mixed evenly, 5 g / L dabigatran was added, stirred After it is completely dissolved and mixed evenly, slowly add 4 times the volume of blood cell preservation solution, stir at low speed, mix well, add 20g / L sucrose and 50g / L mannitol, stir until it is completely dissolved and mixed evenly; package into 1ml / bottle, pre-freeze at -55℃ for 4 hours, slowly warm up to -40℃ and stabilize for 2 hours, then cool to -55℃ and stabilize for 3 hours to make it completely frozen, then gradually heat up at a heating rate of 2℃, stabilize for 2 hours after reaching equilibrium, and then gradually heat up to -35℃ and continue for 24 hours. The heating rate of the analytical drying stage is 3℃ / hour, and the highest temperature is heated to 15℃ and stabilized for 12 hours to prepare the freeze-dried powder of the whole blood composite quality control product. When used, the freeze-dried powder can be reconstituted.

[0027] The final concentrations of the raw materials in Example 1 are 50 mmol / L of piperazine-1,4-diethanesulfonic acid, 50 μmol / L of tris(2-carboxyethyl)phosphine hydrochloride, 2 mmol / L of cholesterol sulfate sodium salt, 1 mmol / L of potassium dihydrogen phosphate, 10 μmol / L of ammonium ferric sulfate dodecahydrate, 2 mmol / L of calcium chloride, 0.9 mmol / L of magnesium sulfate heptahydrate, 6 mmol / L of urea, 80 mmol / L of sodium chloride, 20 μmol / L of zinc sulfate heptahydrate, 15 μmol / L of copper sulfate pentahydrate, 25 μmol / L of glycocholic acid, 5 mmol / L of glucose, 80 μmol / L of creatinine, 250 μmol / L of uric acid, and 1 mmol / L of triolein. L, bovine serum albumin 40g / L, conjugated bilirubin 15μmol / L, lactic acid 1.5mmol / L, β-hydroxybutyric acid 0.2mmol / L, biocytin hydrochloride 1g / L, levodopa 20mmol / L, xanthan gum 5g / L, alanine aminotransferase 40U / L, aspartate aminotransferase 40U / L, alkaline phosphatase 150U / L, lipase 20U / L, lactate dehydrogenase 150U / L, γ-glutamyl transferase 40U / L, amylase 50U / L, adenosine deaminase 20U / L, α-fucosidase 20U / L, creatine kinase 100U / L, dabigatran 1g / L, sucrose 20g / L, mannitol 50g / L.

[0028] Example 2: S1: Take lithium heparin anticoagulated pig whole blood, and separate plasma from blood cells by centrifugation;

[0029] S2: Add an equal volume of Aldrich solution to the separated blood cells, invert and mix evenly to obtain blood cell preservation solution;

[0030] S3: The collected plasma was divided into two equal parts, and 1000mmol / L N,N-bis(hydroxyethyl)glycine and 1000μmol / L sodium dithionite were added to one of the plasma parts in sequence, and stirred until completely dissolved. After mixing evenly, 70mmol / L cholesterol sulfate sodium salt was added, and the mixture was heated to 65°C. After completely dissolved, the mixture was cooled to room temperature, and then 30mmol / L potassium dihydrogen phosphate was added to the obtained solution, and stirred until completely dissolved and mixed evenly.

[0031] S4: add 400 μmol / L ammonium ferric sulfate dodecahydrate, 35 mmol / L calcium chloride, 17 mmol / L magnesium sulfate heptahydrate, 220 mmol / L urea, 1200 mmol / L sodium chloride, 400 μmol / L zinc sulfate heptahydrate, 300 μmol / L copper sulfate pentahydrate, 500 μmol / L glycocholic acid, 160 mmol / L glucose, 4000 μmol / L creatinine, 7000 μmol / L uric acid, 40 mmol / L triolein, 800 g / L bovine serum albumin, 900 μmol / L conjugated bilirubin, 60 mmol / L lactic acid and 20 mmol / L β-hydroxybutyric acid to another portion of plasma in sequence, add the next substance after the previous substance is completely dissolved and mixed evenly, stir until completely dissolved and mixed evenly;

[0032] S5: The solutions obtained in step 3 and step 4 are mixed evenly, 50 g / L biocytin hydrochloride, 250 mmol / L levodopa and 100 g / L xanthan gum are added in sequence, stirred until completely dissolved, mixed evenly, the pH of the solution is adjusted to 7.2, 650 U / L alanine aminotransferase, 750 U / L aspartate aminotransferase, 2000 U / L alkaline phosphatase, 450 U / L lipase, 1500 U / L lactate dehydrogenase, 1000 U / L γ-glutamyl transferase, 1500 U / L amylase, 300 U / L adenosine deaminase, 500 U / L α-fucosidase and 3000 U / L creatine kinase are added in sequence, stirred until completely dissolved, mixed evenly, 50 g / L L dabigatran, stir until completely dissolved, mix evenly, slowly add 4 times the volume of blood cell preservation solution, stir at low speed, mix well, add 100g / L trehalose, stir until completely dissolved, mix evenly, and package into 1ml / bottle, pre-freeze at -55℃ for 4 hours, slowly warm up to -40℃ and stabilize for 2 hours, then cool to -55℃ and stabilize for 3 hours to make it completely frozen, then gradually heat up at a heating rate of 2℃, stabilize for 2 hours after reaching equilibrium, and then gradually heat up to -35℃ and continue for 24 hours. The heating rate in the analytical drying stage is 3℃ / hour, and the highest temperature is heated to 15℃ and stabilized for 12 hours to prepare the freeze-dried powder of the whole blood composite quality control product. When used, the freeze-dried powder can be reconstituted.

[0033] The final concentrations of the raw materials in Example 2 are 100 mmol / L of N,N-di(hydroxyethyl)glycine, 100 μmol / L of sodium dithionite, 7 mmol / L of cholesterol sulfate sodium salt, 3 mmol / L of potassium dihydrogen phosphate, 40 μmol / L of ammonium ferric sulfate dodecahydrate, 3.5 mmol / L of calcium chloride, 1.7 mmol / L of magnesium sulfate heptahydrate, 22 mmol / L of urea, 120 mmol / L of sodium chloride, 40 μmol / L of zinc sulfate heptahydrate, 30 μmol / L of copper sulfate pentahydrate, 50 μmol / L of glycocholic acid, 16 mmol / L of glucose, 400 μmol / L of creatinine, 700 μmol / L of uric acid, and 4 mm triolein. ol / L, bovine serum albumin 80g / L, conjugated bilirubin 90μmol / L, lactic acid 6mmol / L, β-hydroxybutyric acid 2mmol / L, biocytin hydrochloride 10g / L, levodopa 50mmol / L, xanthan gum 20g / L, alanine aminotransferase 130U / L, aspartate aminotransferase 150U / L, alkaline phosphatase 400U / L, lipase 90U / L, lactate dehydrogenase 300U / L, γ-glutamyl transferase 200U / L, amylase 300U / L, adenosine deaminase 60U / L, α-fucosidase 100U / L, creatine kinase 600U / L, dabigatran 10g / L, trehalose 100g / L.

[0034] Comparative Example 1: Based on Example 2, biocytin hydrochloride, levodopa and xanthan gum were not added.

[0035] Comparative Example 2: Based on Comparative Example 1, 50 g / L biocytin hydrochloride was added.

[0036] Comparative Example 3: Based on Comparative Example 1, 250 mmol / L levodopa was added.

[0037] Comparative Example 4: Based on Comparative Example 1, 100 g / L xanthan gum was added.

[0038] Comparative Example 5: Based on Comparative Example 1, 50 g / L biocytin hydrochloride and 250 mmol / L levodopa were added.

[0039] In the above embodiments, the raw materials used are all derived from commercially available products, including lithium anticoagulated porcine whole blood, Aldrich solution, piperazine-1,4-diethanesulfonic acid, tri(2-carboxyethyl)phosphine hydrochloride, cholesterol sulfate sodium salt, potassium dihydrogen phosphate, ammonium ferric sulfate dodecahydrate, calcium chloride, magnesium sulfate heptahydrate, urea, sodium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, glycocholic acid, glucose, creatinine, uric acid, triolein, bovine serum albumin, conjugated bilirubin, lactic acid, β-hydroxybutyric acid, levodopa Xanthan gum, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipase, lactate dehydrogenase, γ-glutamyl transferase, amylase, adenosine deaminase, α-fucosidase, creatine kinase, dabigatran, sucrose, mannitol, N,N-bicine, and trehalose were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; biocytin hydrochloride was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; sodium dithionate was purchased from Shanghai Yien Chemical Technology Co., Ltd.

[0040] The performance of the whole blood composite quality control products obtained in Examples 1 and 2 of the present invention and Comparative Examples 1 to 5 is described below in conjunction with the table: (I) Uniformity of the whole blood composite quality control product: 10 bottles of the quality control products prepared in Examples 1 and 2 above were randomly selected, reconstituted, mixed by inversion 3 times, placed at 2 to 8°C for 30 minutes, and gently shaken before use. The corresponding project detection reagents of the fully automatic biochemical analyzer produced by our company were used, and each bottle was measured once on the Hitachi H7100 instrument, and the mean of the 10 data was calculated. Standard deviation S1 and coefficient of variation CV1, take one bottle, test 20 times, and calculate the mean of the first 10 data The standard deviation S2 is calculated according to the formula and Calculation, if the measured precision CV1 and CV 瓶间 If the difference between each bottle of the batch of quality control products is less than 10.0%, it is considered that the difference between each bottle of the batch of quality control products is acceptable; otherwise, the batch of quality control products is unusable;

[0041] Table 1 Homogeneity of whole blood composite quality control products

[0042]

[0043]

[0044] From the experimental results in Table 1, it can be seen that the uniformity of the whole blood composite quality control products of Example 1 and Example 2 meets the requirements, indicating that the difference between each bottle of this batch of quality control products is acceptable and can be further used for verification of other properties.

[0045] (II) Stability of whole blood composite quality control product: (1) Accelerated destruction of stability at 37°C: The whole blood composite quality control product prepared in Examples 1 and 2 and Comparative Examples 1, 2, 3, 4, and 5 was placed at 37°C, taken out on day 0 and day 7, re-dissolved, mixed by inversion 3 times, placed at 2-8°C for 30 min, gently shaken, and tested on a Hitachi H7100 instrument using the corresponding test reagents of the fully automatic biochemical analyzer produced by our company. The test was repeated 2 times, and the average value was taken. The test results of the quality control product on day 7 were compared with the results on day 0. If the deviation did not exceed 10%, it was considered stable.

[0046] Table 2 Accelerated destruction stability at 37℃

[0047]

[0048]

[0049] From the experimental results in Table 2, it can be seen that after the whole blood composite quality control products of Comparative Examples 1, 2, 3, 4, and 5 were stored at 37°C for accelerated destruction for 7 days, the deviations of 10 items, 7 items, 7 items, 6 items, and 5 items exceeded 10%, respectively, indicating that biocytin hydrochloride, levodopa, and xanthan gum used alone have a certain promoting effect on the stability of the whole blood composite quality control products, and the mixed use of biocytin hydrochloride and levodopa is better than using them alone; after the quality control products of Example 1 and Example 2 were stored at 37°C for accelerated destruction for 7 days, the deviations of all items did not exceed 10%, and the stability met the requirements, indicating that the addition of biocytin hydrochloride, levodopa, and xanthan gum can play a good stabilizing and protective role on the quality control products, and the 37°C accelerated destruction stability of the quality control products can meet the requirements;

[0050] (2) Long-term storage stability at 2-8°C: The quality control products prepared in the above Examples 1 and 2 were stored at 4°C, taken out on day 0, 15 days, 1 month, 3 months, 6 months, 9 months and 12 months, redissolved, mixed by inversion 3 times, placed at 2-8°C for 30 min, gently shaken, and tested on a Hitachi H7100 instrument using the corresponding test reagents of the fully automatic biochemical analyzer produced by our company. The test was repeated 2 times and the average value was taken. The test results of the quality control products stored for 15 days, 1 month, 3 months, 6 months, 9 months and 12 months were compared with the results on day 0. If the deviation did not exceed 10%, it was considered stable.

[0051] Table 3 Example 1 Whole blood composite quality control product 2~8℃ long-term storage stability

[0052]

[0053]

[0054] Table 4 Example 2 Whole blood composite quality control product 2~8℃ long-term storage stability

[0055]

[0056]

[0057] It can be seen from the experimental results in Table 3 and Table 4 that the long-term storage stability of the whole blood quality control products of Example 1 and Example 2 at 2-8°C meets the requirements.

[0058] (III) Assignment of the value of the whole blood composite quality control product: randomly select one bottle of the whole blood composite quality control product prepared in the above-mentioned Example 1 and Example 2, reconstitute, mix by inversion three times, place at 2-8°C for 30 minutes, shake gently, and then test on the Hitachi H7100 instrument using the corresponding project detection reagent of the fully automatic biochemical analyzer produced by our company, test 20 times continuously, calculate the average value, and take the average value as the measured value, and take the (110%) average value as the acceptable range;

[0059] Table 5 Whole blood composite quality control values ​​of Example 1 and Example 2

[0060]

[0061]

[0062] (IV) Detection of centrifugal microfluidic disc biochemical analysis chip: Randomly select one bottle of the whole blood composite quality control product prepared in Example 1 and Example 2, reconstitute, mix by inversion three times, place at 2-8°C for 30 minutes, shake gently, and then measure with a centrifugal microfluidic disc biochemical analysis chip, repeat twice, calculate the average value, and the result should be within the acceptable range;

[0063] Table 6 Whole blood composite quality control product centrifugal microfluidic disc biochemical analysis chip measurement values

[0064]

[0065]

[0066] The results in Table 6 show that the measured values ​​of the whole blood composite quality control products of Example 1 and Example 2 detected by the centrifugal microfluidic disc biochemical analysis chip are all within the acceptable range, indicating that the whole blood composite quality control products prepared in Example 1 and Example 2 can be applied to the quality control of the centrifugal microfluidic disc biochemical analysis chip.

[0067] Finally, it should be noted that the above is only a preferred embodiment of the present invention, but the protection scope of this application is not limited thereto. Any changes or substitutions that can be easily thought of by any technician familiar with the technical field within the spirit and principle of the present invention and within the technical scope disclosed in this application should be included in the protection scope of this application; in the absence of conflict, the embodiments of this application and the features in the embodiments can be combined with each other. Therefore, the protection scope of this application shall be based on the protection scope of the claims.

Claims

1. A whole blood composite quality control product for microfluidic biochemical analysis chip, characterized in that: The raw materials of the whole blood composite quality control product include porcine whole blood, quality control item additives, Aldrich solution, buffer, antioxidant, biocytin hydrochloride, levodopa, xanthan gum, dabigatran and freeze-dried additives; the concentration of the biocytin hydrochloride is 1-10 g / L, the concentration of levodopa is 20-50 mmol / L, the concentration of xanthan gum is 5-20 g / L; the concentration of the buffer is 50-200 mmol / L, the concentration of dabigatran is 1-10 g / L, the concentration of the antioxidant is 10-200 μmol / L, the concentration of the freeze-dried additive is 20-100 g / L, and the rest is porcine whole blood and Aldrich solution; The added substances for the quality control items include cholesterol sulfate sodium salt, potassium dihydrogen phosphate, ammonium ferric sulfate dodecahydrate, calcium chloride, magnesium sulfate heptahydrate, urea, sodium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, glycocholic acid, glucose, creatinine, uric acid, triolein, bovine serum albumin, conjugated bilirubin, lactic acid, β-hydroxybutyric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipase, lactate dehydrogenase, γ-glutamyl transferase, amylase, adenosine deaminase, α-fucosidase and creatine kinase.

2. The whole blood composite quality control product for microfluidic biochemical analysis chip according to claim 1, characterized in that: The concentrations of each substance are as follows: cholesterol sulfate sodium salt 2~7mmol / L, potassium dihydrogen phosphate 1~3mmol / L, ammonium ferric sulfate dodecahydrate 10~40μmol / L, calcium chloride 2~3.5mmol / L, magnesium sulfate heptahydrate 0.9~1.7mmol / L, urea 6~22mmol / L, sodium chloride 80~120mmol / L, zinc sulfate heptahydrate 20~40μmol / L, copper sulfate pentahydrate 15~30μmol / L, glycocholic acid 25~50μmol / L, glucose 5~16mmol / L, creatinine 80~400μmol / L, uric acid 250~700μmol / L, triolein 1~4 mmol / L, bovine serum albumin 40~80g / L, conjugated bilirubin 15~90μmol / L, lactic acid 1.5~6mmol / L, β-hydroxybutyric acid 0.2~2mmol / L, alanine aminotransferase 40~130U / L, aspartate aminotransferase 40~150U / L, alkaline phosphatase 150~400U / L, lipase 20~90U / L, lactate dehydrogenase 150~300U / L, γ-glutamyl transferase 40~200U / L, amylase 50~300U / L, adenosine deaminase 20~60U / L, α-fucosidase 20~100U / L, creatine kinase 100~600U / L.

3. The whole blood composite quality control product for microfluidic biochemical analysis chip according to claim 1, characterized in that: The pig whole blood includes plasma and blood cells, and the ratio of the added amount of Aldrich solution to the volume of blood cells is 1:

1.

4. A method for preparing a whole blood composite quality control product for a microfluidic biochemical analysis chip, characterized in that: The following steps are involved: S1: Collect lithium heparin anticoagulated pig whole blood and separate plasma from blood cells by centrifugation; S2: Add an equal volume of Aldrich solution to the separated blood cells, mix evenly by inversion to obtain blood cell preservation solution, and store at 2-8°C; S3: The collected plasma is divided into two equal parts, and a buffer and an antioxidant are added to one of the plasma parts in sequence, and the mixture is stirred until completely dissolved. After mixing evenly, sodium cholesterol sulfate is added, and the mixture is heated to 55-70°C. After the mixture is completely dissolved, the mixture is cooled to room temperature, and potassium dihydrogen phosphate is added, and the mixture is stirred until completely dissolved and mixed evenly. S4: add ammonium ferric sulfate dodecahydrate, calcium chloride, magnesium sulfate heptahydrate, urea, sodium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, glycocholic acid, glucose, creatinine, uric acid, triolein, bovine serum albumin, conjugated bilirubin, lactic acid and β-hydroxybutyric acid to another portion of plasma in sequence, add the next substance after the previous substance is completely dissolved and mixed evenly, stir until completely dissolved and mixed evenly; S5: Evenly mix the solutions obtained in steps S3 and S4, add biocytin hydrochloride, levodopa, and xanthan gum in sequence, stir until completely dissolved, mix evenly, adjust the pH of the solution to 7.0-7.2, add alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipase, lactate dehydrogenase, γ-glutamyl transferase, amylase, adenosine deaminase, α-fucosidase, and creatine kinase in sequence, stir until completely dissolved, and mix evenly; add dabigatran, stir until completely dissolved, slowly add 4 times the volume of blood cell preservation solution, stir at a low speed, mix well, add freeze-dried additives, stir until completely dissolved, divide and freeze-dry to obtain freeze-dried powder of whole blood composite quality control product.

5. The method for preparing a whole blood composite quality control product for a microfluidic biochemical analysis chip according to claim 4, characterized in that: The buffer is one of piperazine-1,4-diethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, and N,N-di(hydroxyethyl)glycine; the antioxidant is one of tris(2-carboxyethyl)phosphine hydrochloride and sodium dithionite; and the freeze-drying additive is one or more combinations of trehalose, glucose, sucrose, and mannitol.

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