A sample processing liquid for centrifugal microfluidic disc biochemical analysis chip

Abstract

The present invention relates to the field of biochemical analysis technology, specifically a sample processing solution for a centrifugal microfluidic disc biochemical analysis chip. The sample processing solution includes the following concentration components: buffer: 50-100 mmol / L, monovalent salt: 300-600 mmol / L, composite solubilizer: 2-20 g / L, EMULGEN B-66: 10-50 g / L, 3-[N,N-dimethyl (3-palmitamidopropyl) ammonium]-propanesulfonic acid inner salt: 1-15 g / L, flufenamic acid butyl ester: 0.1-2 mmol / L, N,N-diethylformamide: 0.5-6 mmol / L, ascorbic acid oxidase: 1-2 KU / L, alcohol: 5-10 mL / L, defoamer 204: 1-10 g / L. The sample processing solution prepared by the present invention has the effect of resisting the interference of fat turbidity, bilirubin, heparin and vitamin C, does not interfere with the content of the items that need to be detected by the sample, and is more suitable for clinical application.

Description

Technical Field

[0001] The invention relates to the technical field of biochemical analysis, in particular to a sample processing liquid used for a centrifugal microfluidic disc biochemical analysis chip. Background Art

[0002] Microfluidic technology can be divided into disc, card, sheet and droplet types according to the carrier form. Compared with the other three platforms, disc microfluidic technology can easily realize whole blood separation and multi-index detection. The disc chip design can be operated in parallel, that is, microfluidic control and detection of multiple detection units can be realized at the same time. Therefore, it is fast, high-throughput and easy to automate, suitable for multi-index determination. At present, the centrifugal microfluidic disc biochemical analysis chip can integrate up to a dozen items on a disc, greatly improving the detection efficiency of samples. However, the biochemical analysis method commonly used in clinical practice is to use a fully automatic biochemical analyzer for double-reagent determination. One of the reasons for using the double-reagent form is that in the process of clinical biochemical determination, in addition to the substance to be tested, the blood sample also contains various enzymes, organic matter, inorganic salts and other substances. These substances will interfere with or participate in the test reaction and cause non-specific reaction interference. Currently, the most common interfering factors in clinical practice include lipid turbidity, vitamin C, bilirubin, heparin, etc. One of the main purposes of the design of the dual-reagent test kit is to overcome these interfering reactions. During the measurement process, reagent I is first allowed to react with the interfering substances in the specimen. After a period of reaction, reagent II is used to start the actual test reaction, thereby ensuring the accuracy of the measurement results.

[0003] Since the centrifugal microfluidic disc biochemical analysis chip integrates many test items on a disc, and the reagents of these test items are pre-installed in the test holes in the form of freeze-dried beads, distilled water or physiological saline is generally used as the sample treatment liquid during the test to ensure that the sample treatment liquid will not interfere with the project detection. However, distilled water or physiological saline can only play the role of sample dilution during the test process, and cannot achieve the anti-interference function of reagent I in the dual-reagent kit design. Therefore, the centrifugal microfluidic disc biochemical analysis chip has the defect of weak anti-interference ability, which seriously affects its clinical application. Summary of the invention

[0004] In view of the shortcomings of the prior art, the present invention provides a sample processing liquid that can be used for centrifugal microfluidic disc biochemical analysis chip detection and has anti-lipid turbidity, bilirubin, heparin and vitamin C interference effects and does not interfere with many biochemical detection items, so as 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 sample processing liquid for a centrifugal microfluidic disc biochemical analysis chip comprises the following concentration components: buffer solution: 50-100 mmol / L, pH=7.0-7.5, monovalent salt: 300-600 mmol / L, composite solubilizer: 2-20 g / L, EMULGEN B-66: 10-50 g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 1-15 g / L, flufenamic acid butyl ester: 0.1-2 mmol / L, N,N-diethylformamide: 0.5-6 mmol / L, ascorbic acid oxidase: 1-2 KU / L, alcohol: 5-10 mL / L, and defoamer 204: 1-10 g / L.

[0007] More optimally, the sample processing liquid for a centrifugal microfluidic disc biochemical analysis chip comprises the following components, and the content of each component is: buffer: 100mmol / L, pH=7.5, monovalent salt: 500mmol / L, composite solubilizer 20g / L, EMULGEN B-66: 50g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 15g / L, flufenamic acid butyl ester: 2mmol / L, N,N-diethylformamide: 6mmol / L, ascorbic acid oxidase: 2KU / L, alcohol: 10mL / L, defoamer 204: 10g / L.

[0008] Preferably, the buffer includes but is not limited to any one of 3-(N-morpholine)-2-hydroxypropanesulfonic acid sodium salt, piperazine-1,4-diethanesulfonic acid and hydroxyethylpiperazineethanesulfonic acid.

[0009] Preferably, the monovalent salt includes but is not limited to any one of sodium chloride and sodium sulfate.

[0010] More optimally, the composite solubilizer is polyoxyethylene lauryl ether, including but not limited to a combination of two or three of the models EMULGEN 103, EMULGEN 107, EMULGEN 109, and EMULGEN 109P.

[0011] Preferably, the alcohol includes but is not limited to any one of n-butanol, isobutanol, ethanol and methanol.

[0012] More optimally, the preparation method of the sample processing solution for the centrifugal microfluidic disc biochemical analysis chip includes the following steps:

[0013] Step 1: Under light-shielded conditions, add flufenamic acid butyl ester and N,N-diethylformamide to alcohol in sequence, stir and mix evenly, and let stand for 4 to 6 hours to obtain solution A;

[0014] Step 2: Add buffer, monovalent salt, composite solubilizer, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 to distilled water in sequence, stir and mix evenly, adjust the pH value to 7.0-7.5, add ascorbic acid oxidase, stir and mix evenly, and obtain solution B;

[0015] Step 3: In a dark environment, mix solution A and solution B evenly to obtain a mixed solution; at the same time, continuously introduce nitrogen gas into the mixed solution for 30 to 40 minutes to obtain a sample treatment solution, which is stored for later use.

[0016] Preferably, the sample processing solution is stored in a temperature range of 2 to 8° C. away from light.

[0017] Compared with the prior art, the beneficial effects achieved by the present invention are:

[0018] (1) Centrifugal microfluidic disc biochemical analysis chips currently generally use distilled water or physiological saline as sample processing liquid, which can only play the role of sample dilution. The present invention prevents the aggregation of lipid particles in the sample through the synergistic effect of the composite solubilizer, co-emulsifier EMULGENB-66 and emulsion stabilizer 3-[N,N-dimethyl (3-palmitamidopropyl) ammonium]-propane sulfonic acid inner salt in the sample processing liquid, and redisperses the already aggregated lipid particles to eliminate the interference of fat turbidity on detection;

[0019] (2) By adding the stable oxidant N,N-diethylformamide and the oxidation promoter flufenamic acid butyl ester to the sample processing solution, the interference of bilirubin on the detection can be removed to a certain extent;

[0020] (3) Through the synergistic effect of appropriate buffer, high concentration of monovalent salt ions and composite solubilizer, the interference of heparin anticoagulant on detection is effectively avoided; at the same time, the sample processing solution of the present invention itself does not interfere with many biochemical detection items. DETAILED DESCRIPTION

[0021] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0022] In the present embodiment, it should be noted that the purchase manufacturers of all raw materials involved in the present invention are not subject to any special restrictions, and exemplary examples include: 3-(N-morpholine)-2-hydroxypropanesulfonic acid sodium salt buffer was purchased from Shanghai Yuanye Biotechnology Co., Ltd., piperazine-1,4-diethanesulfonic acid buffer was purchased from Shanghai Yuanye Biotechnology Co., Ltd., hydroxyethylpiperazine ethanesulfonic acid buffer was purchased from Shanghai Yuanye Biotechnology Co., Ltd., sodium chloride was purchased from Shanghai Yuanye Biotechnology Co., Ltd., sodium sulfate was purchased from Shanghai Yuanye Biotechnology Co., Ltd., EMULGEN 103 was purchased from Xingtianwai Chemical (Shanghai) Co., Ltd., EMULGEN 107 was purchased from Xingtianwai Chemical (Shanghai) Co., Ltd., EMULGEN 109 was purchased from Xingtianwai Chemical (Shanghai) Co., Ltd., EMULGEN109P was purchased from Xingtianwai Chemical (Shanghai) Co., Ltd., EMULGEN B-66 was purchased from Xingtianwai Chemical (Shanghai) Co., Ltd., 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt was purchased from Aibixin (Shanghai) Biotechnology Co., Ltd., flufenamic acid butyl ester was purchased from Shanghai Yuanye Biotechnology Co., Ltd., N,N-diethylformamide was purchased from Shanghai Yuanye Biotechnology Co., Ltd., ascorbic acid oxidase was purchased from Shanghai Yuanye Biotechnology Co., Ltd., isobutanol was purchased from Sinopharm Chemical Reagent Co., Ltd., ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd., n-butanol was purchased from Sinopharm Chemical Reagent Co., Ltd., and defoamer 204 was purchased from Jiangsu Meg Biotechnology Co., Ltd.

[0023] Example 1: A sample processing solution for a centrifugal microfluidic disc biochemical analysis chip:

[0024] Step 1: under light-proof conditions, add flufenamic acid butyl ester and N,N-diethylformamide to isobutyl alcohol in sequence, stir and mix evenly, and let stand for 4 hours to obtain solution A;

[0025] Step 2: 3-(N-morpholine)-2-hydroxypropanesulfonic acid sodium salt buffer, sodium chloride, EMULGEN 103, EMULGEN 109P, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 are sequentially added into distilled water, stirred and mixed evenly, and the pH value thereof is adjusted to 7.0, and ascorbic acid oxidase is added, stirred and mixed evenly to obtain solution B;

[0026] Step 3: Under light-proof conditions, solution A and solution B are mixed evenly to obtain a mixed solution; nitrogen is continuously introduced into the mixed solution for 30 minutes to obtain a sample treatment solution, and the solution is stored in a light-proof temperature range of 2 to 8°C;

[0027] The sample treatment solution includes the following concentration components: 3-(N-morpholine)-2-hydroxypropanesulfonic acid sodium salt buffer (pH=7.0): 100mmol / L, sodium chloride: 300mmol / L, EMULGEN 103: 1g / L, EMULGEN 109P: 1g / L, EMULGEN B-66: 10g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 10g / L, flufenamic acid butyl ester: 1mmol / L, N,N-diethylformamide: 0.5mmol / L, ascorbic acid oxidase: 1KU / L, isobutanol: 5mL / L, defoaming agent 204: 1g / L.

[0028] Example 2: A sample processing solution for a centrifugal microfluidic disc biochemical analysis chip:

[0029] Step 1: Under light-proof conditions, add flufenamic acid butyl ester and N,N-diethylformamide to ethanol in sequence, stir and mix evenly, and let stand for 5 hours to obtain solution A;

[0030] Step 2: piperazine-1,4-diethanesulfonic acid buffer, sodium sulfate, EMULGEN 103, EMULGEN 107, EMULGEN 109, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 are sequentially added into distilled water, stirred and mixed evenly, and the pH value thereof is adjusted to 7.2, ascorbic acid oxidase is added, stirred and mixed evenly, and solution B is obtained;

[0031] Step 3: Under light-proof conditions, solution A and solution B are mixed evenly to obtain a mixed solution; nitrogen is continuously introduced into the mixed solution for 30 minutes to obtain a sample treatment solution, and the solution is stored in a light-proof temperature range of 2 to 8°C;

[0032] The sample treatment solution includes the following concentration components: piperazine-1,4-diethanesulfonic acid buffer (pH=7.2): 100mmol / L, sodium sulfate: 600mmol / L, EMULGEN 103: 5g / L, EMULGEN 107: 5g / L, EMULGEN 109: 5g / L, EMULGEN B-66: 30g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 15g / L, flufenamic acid butyl ester: 2mmol / L, N,N-diethylformamide: 6mmol / L, ascorbic acid oxidase: 2KU / L, ethanol: 7mL / L, defoaming agent 204: 5g / L.

[0033] Example 3: A sample processing solution for a centrifugal microfluidic disc biochemical analysis chip:

[0034] Step 1: under light-proof conditions, add flufenamic acid butyl ester and N,N-diethylformamide to n-butanol in sequence, stir and mix evenly, and let stand for 6 hours to obtain solution A;

[0035] Step 2: Add hydroxyethylpiperazine ethanesulfonic acid buffer, sodium chloride, EMULGEN 107, EMULGEN 109P, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 to distilled water in sequence, stir and mix evenly, adjust the pH value to 7.5, add ascorbic acid oxidase, stir and mix evenly, and obtain solution B;

[0036] Step 3: Under light-proof conditions, solution A and solution B are mixed evenly to obtain a mixed solution; nitrogen is continuously introduced into the mixed solution for 40 minutes to obtain a sample treatment solution, and the solution is stored in a light-proof temperature range of 2 to 8°C;

[0037] The sample treatment solution includes the following concentration components: hydroxyethylpiperazine ethanesulfonic acid buffer (pH=7.5): 100mmol / L, sodium chloride: 500mmol / L, EMULGEN 107: 10g / L, EMULGEN 109P: 10g / L, EMULGEN B-66: 50g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 15g / L, flufenamic acid butyl ester: 2mmol / L, N,N-diethylformamide: 6mmol / L, ascorbic acid oxidase: 2KU / L, n-butanol: 10mL / L, defoamer 204: 10g / L.

[0038] The effects of the sample treatment solutions and distilled water prepared in Examples 1 to 3 on the detection items of the centrifugal microfluidic disc biochemical analysis chip were as follows:

[0039] Detection method: (1) Take high and low concentration serum samples without hemolysis, bilirubin, lipid turbidity, vitamin C, and heparin, add the samples to the sample tank through the sample addition hole, add the sample treatment solution to the sample treatment solution tank through the sample treatment solution hole, start the measurement and then perform the detection; (2) Each sample is measured twice using a centrifugal microfluidic disc biochemical analysis chip, and the mean is calculated; the measurement results of Examples 1 to 3 are compared with the measurement results of distilled water, and the deviation does not exceed ±10%, which means that there is no effect on the detection results; the specific data are shown in Table 1:

[0040] Table 1

[0041]

[0042]

[0043] It can be seen from Table 1 above that the sample processing solution for centrifugal microfluidic disc-type biochemical analysis chip detection of the present invention does not interfere with biochemical detection items such as TP, ALB, and ALT.

[0044] Based on the results obtained from the data in Table 1 above, with Example 3 as a control, comparative examples 1 to 3 are set, as follows:

[0045] Comparative Example 1: Taking Example 3 as a control, the composite solubilizer EMULGEN 107, EMULGEN 109P, the co-emulsifier EMULGEN B-66, and the emulsion stabilizer 3-[N,N-dimethyl (3-palmitamidopropyl) ammonium]-propane sulfonic acid inner salt are removed from Comparative Example 1, and the other processes remain unchanged; specifically:

[0046] Step 1: under light-proof conditions, add flufenamic acid butyl ester and N,N-diethylformamide to n-butanol in sequence, stir and mix evenly, and let stand for 6 hours to obtain solution A;

[0047] Step 2: Add hydroxyethylpiperazine ethanesulfonic acid buffer, sodium chloride, and defoamer 204 to distilled water in sequence, stir and mix evenly, adjust the pH value to 7.5, add ascorbic acid oxidase, stir and mix evenly, and obtain solution B;

[0048] Step 3: Under light-proof conditions, solution A and solution B are mixed evenly to obtain a mixed solution; nitrogen is continuously introduced into the mixed solution for 40 minutes to obtain a sample treatment solution, and the solution is stored in a light-proof temperature range of 2 to 8°C;

[0049] The sample treatment solution includes the following concentration components: hydroxyethylpiperazine ethanesulfonic acid buffer (pH=7.5): 100mmol / L, sodium chloride: 500mmol / L, flufenamic acid butyl ester: 2mmol / L, N,N-diethylformamide: 6mmol / L, ascorbic acid oxidase: 2KU / L, n-butanol: 10mL / L, defoamer 204: 10g / L.

[0050] Comparative Example 2: Using Example 3 as a control, Comparative Example 2 removes the oxidant N,N-diethylformamide and the oxidation promoter flufenamic acid butyl ester, and the other processes remain unchanged; specifically:

[0051] Step 1: Add hydroxyethylpiperazine ethanesulfonic acid buffer, sodium chloride, EMULGEN 107, EMULGEN 109P, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 to distilled water in sequence, stir and mix evenly, adjust the pH value to 7.5, add ascorbic acid oxidase, stir and mix evenly, and obtain solution B;

[0052] Step 2: In a light-proof condition, n-butanol and solution B are mixed evenly to obtain a mixed solution; nitrogen is continuously introduced into the mixed solution for 40 minutes to obtain a sample treatment solution, and the solution is stored in a light-proof condition at a temperature range of 2 to 8°C;

[0053] The sample treatment solution includes the following concentration components: hydroxyethylpiperazine ethanesulfonic acid buffer (pH=7.5): 100mmol / L, sodium chloride: 500mmol / L, EMULGEN 107: 10g / L, EMULGEN 109P: 10g / L, EMULGEN B-66: 50g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 15g / L, ascorbic acid oxidase: 2KU / L, n-butanol: 10mL / L, defoaming agent 204: 10g / L.

[0054] Comparative Example 3: Taking Example 3 as a control, the sodium chloride and the composite solubilizers EMULGEN 107 and EMULGEN 109P were removed from Comparative Example 3, and the other processes remained unchanged; specifically:

[0055] Step 1: under light-proof conditions, add flufenamic acid butyl ester and N,N-diethylformamide to n-butanol in sequence, stir and mix evenly, and let stand for 6 hours to obtain solution A;

[0056] Step 2: Add hydroxyethylpiperazine ethanesulfonic acid buffer, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 to distilled water in sequence, stir and mix evenly, adjust the pH value to 7.5, add ascorbic acid oxidase, stir and mix evenly, and obtain solution B;

[0057] Step 3: Under light-proof conditions, solution A and solution B are mixed evenly to obtain a mixed solution; nitrogen is continuously introduced into the mixed solution for 40 minutes to obtain a sample treatment solution, and the solution is stored in a light-proof temperature range of 2 to 8°C;

[0058] The sample treatment solution includes the following concentration components: hydroxyethylpiperazine ethanesulfonic acid buffer (pH=7.5): 100mmol / L, EMULGEN B-66: 50g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 15g / L, flufenamic acid butyl ester: 2mmol / L, N,N-diethylformamide: 6mmol / L, ascorbic acid oxidase: 2KU / L, n-butanol: 10mL / L, defoamer 204: 10g / L.

[0059] Detection experiment: the sample treatment solution prepared in Example 3 and Comparative Example 1 was subjected to an anti-lipid interference test; the sample treatment solution prepared in Example 3 and Comparative Example 2 was subjected to an anti-bilirubin interference test; the sample treatment solution prepared in Example 3 and Comparative Example 3 was subjected to an anti-heparin interference test; the sample treatment solution prepared in Example 3 was subjected to an anti-vitamin C interference test; and the samples were compared with distilled water as the sample treatment solution. The specific tests are as follows:

[0060] 1. Anti-fat turbidity interference test:

[0061] (1) First, the lipid interfering substance (14500 FTU, defined as 100%) was prepared into a series of concentration solutions with distilled water, namely 100%, 75%, 50%, 25% and 0%; then, the solutions were mixed with the serum samples at a dilution ratio of 10 times to obtain a series of lipid interfering samples, corresponding to lipid concentrations of 1450, 1087.5, 725, 362.5 and 0 FTU, respectively;

[0062] (2) adding the lipid interference sample into the sample tank through the sample adding hole, and adding distilled water, the sample processing solution of Example 3 and the sample processing solution of Comparative Example 1 into the sample processing solution tank through the sample processing solution hole, respectively, and starting the determination for detection;

[0063] (3) Each sample was measured three times using a centrifugal microfluidic disc total protein biochemical analysis chip, and the mean was calculated; the measurement results of the 1450, 1087.5, 725, and 362.5 FTU lipid mixed specimens were compared with the measurement results of 0 FTU. If the deviation did not exceed ±10%, it was considered that the interfering substance had no interfering effect on the measurement results. The specific data are shown in Table 2:

[0064] Table 2

[0065]

[0066]

[0067] It can be seen from Table 2 above that when distilled water and comparative example 1 are used as sample processing liquids, when the fat turbidity concentration is higher than 725 FTU, there is interference with the determination result of total protein; when Example 3 of the present invention is used as the sample processing liquid, when the fat turbidity concentration is as high as 1450 FTU, there is still no interference with the determination result of total protein, indicating that by adding composite solubilizers EMULGEN 107 and EMULGEN 109P, co-emulsifier EMULGEN B-66 and emulsion stabilizer 3-[N,N-dimethyl (3-palmitamidopropyl) ammonium]-propane sulfonic acid inner salt to the sample processing liquid, the interference of fat turbidity on reagent detection can be eliminated to a certain extent.

[0068] 2. Anti-bilirubin interference test:

[0069] (1) First, a 5.76 g / L conjugated bilirubin solution was prepared with distilled water as a storage solution, and then a series of concentration gradient conjugated bilirubin solutions of 5.76, 4.32, 2.88, 1.44 and 0 g / L were prepared based on the storage solution, and then mixed with the serum sample at a dilution ratio of 20 times to obtain a series of concentration gradient conjugated bilirubin interference samples, corresponding to the conjugated bilirubin concentrations of 0.288, 0.216, 0.144, 0.072 and 0 g / L, respectively;

[0070] (2) adding the bilirubin interference sample into the sample tank through the sample adding hole, and respectively adding distilled water, the sample processing solution of Example 3 and the sample processing solution of Comparative 2 into the sample processing solution tank through the sample processing solution hole, and starting the determination for detection;

[0071] (3) Each sample was measured three times using a centrifugal microfluidic disc-type total protein biochemical analysis chip, and the mean was calculated; the measurement results of the mixed sample of 0.288, 0.216, 0.144, and 0.072 g / L combined bilirubin were compared with the measurement result of 0 g / L, and the deviation did not exceed ±10%, which means that the interfering substance had no interfering effect on the measurement result; the specific data are shown in Table 3:

[0072] Table 3

[0073]

[0074] It can be seen from Table 3 above that when distilled water and Comparative Example 2 are used as sample processing liquids, when the combined bilirubin concentration is higher than 0.144 g / L, there is interference with the determination result of total protein; when Example 3 of the present invention is used as the sample processing liquid, when the combined bilirubin concentration is as high as 0.288 g / L, there is still no interference with the determination result of total protein, indicating that by adding the oxidant N, N-diethylformamide and the oxidation promoter flufenamic acid butyl to the sample processing liquid, the interference of bilirubin on the reagent detection can be removed to a certain extent.

[0075] 3. Anti-heparin interference test:

[0076] (1) First, a 1750 U / mL heparin solution was prepared with distilled water as a storage solution, and then a series of concentration gradient heparin solutions of 1750, 1312.5, 875, 437.5 and 0 U / mL were prepared based on the storage solution, and then mixed with the serum sample at a dilution ratio of 20 times to obtain a series of concentration gradient heparin interference samples, corresponding to heparin concentrations of 87.5, 65.6, 43.8, 21.9 and 0 U / mL, respectively;

[0077] (2) adding the heparin interference sample into the sample tank through the sample adding hole, and respectively adding distilled water, the sample treatment solution of Example 3 and Comparative Example 3 into the sample treatment solution tank through the sample treatment solution hole, and starting the determination for detection;

[0078] (3) Each sample was measured three times using a centrifugal microfluidic disk-type α-fucosidase biochemical analysis chip, and the mean was calculated. The measurement results of 87.5, 65.6, 43.8, and 21.9 U / mL heparin mixed specimens were compared with the measurement result of 0 U / mL. If the deviation did not exceed ±10%, it was considered that the interfering substance had no interfering effect on the measurement result. The specific data are shown in Table 4:

[0079] Table 4

[0080]

[0081] It can be seen from Table 4 above that when distilled water and comparative example 3 are used as sample processing liquids, when the heparin concentration is higher than 21.9 U / ml, there is interference with the determination result of α-fucosidase; while when Example 3 of the present invention is used as the sample processing liquid, when the heparin concentration is as high as 87.5 U / ml, there is still no interference with the determination result of α-fucosidase, indicating that by adding high concentration of sodium chloride and composite solubilizers EMULGEN 107 and EMULGEN 109P to the sample processing liquid, the interference of heparin anticoagulant on reagent detection can be effectively avoided.

[0082] 4. Anti-vitamin C interference test

[0083] (1) First, a 600 mg / L vitamin C solution was prepared with deionized water as a storage solution, and then a series of concentration gradient solutions of 600, 450, 300, 150 and 0 mg / L were prepared based on the storage solution, and then mixed with the serum sample at a dilution ratio of 20 times to obtain a series of concentration vitamin C interference samples, corresponding to vitamin C concentrations of 30, 22.5, 15, 7.5 and 0 mg / L, respectively;

[0084] (2) Add the vitamin C interference sample into the sample tank through the sample adding hole, add the sample processing solution into the sample processing solution tank through the sample processing solution hole, and start the measurement to perform the detection;

[0085] (3) Each sample was measured three times using a centrifugal microfluidic disc-type glucose biochemical analysis chip, and the mean was calculated. The measurement results of the mixed samples of 30, 22.5, 15, and 7.5 mg / L vitamin C were compared with the measurement results of 0 mg / L. If the deviation did not exceed ±10%, it was considered that the interfering substance had no interference effect on the measurement results. The specific data are shown in Table 5:

[0086] Table 5

[0087]

[0088] It can be seen from Table 5 above that when the sample processing liquid is only distilled water, when the vitamin C concentration is higher than 15 mg / L, there is interference with the glucose measurement result; however, when Example 3 of the present invention is used as the sample processing liquid, when the vitamin C concentration is as high as 30 mg / L, there is still no interference with the glucose measurement result.

[0089] In summary, from the above-mentioned anti-interference test results of the sample processing fluid, it can be seen that the sample processing fluid for the centrifugal microfluidic disc biochemical analysis chip prepared by the present invention has the effect of resisting the interference of lipid turbidity, bilirubin, heparin and vitamin C, and does not interfere with the specific items that need to be detected in the sample, and is more suitable for clinical application.

[0090] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A sample processing solution for a centrifugal microfluidic disc biochemical analysis chip, characterized in that: The invention comprises the following concentration components: buffer solution: 50-100 mmol / L, pH=7.0-7.5, monovalent salt: 300-600 mmol / L, composite solubilizer: 2-20 g / L, EMULGEN B-66: 10-50 g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 1-15 g / L, flufenamic acid butyl ester: 0.1-2 mmol / L, N,N-diethylformamide: 0.5-6 mmol / L, ascorbic acid oxidase: 1-2 KU / L, alcohol: 5-10 mL / L, defoamer 204: 1-10 g / L; Wherein, the buffer comprises any one of 3-(N-morpholine)-2-hydroxypropanesulfonic acid sodium salt, piperazine-1,4-diethanesulfonic acid and hydroxyethylpiperazineethanesulfonic acid; The monovalent salt includes any one of sodium chloride and sodium sulfate; The composite solubilizer is polyoxyethylene lauryl ether, including a combination of two or three of EMULGEN 103, EMULGEN 107, EMULGEN 109, and EMULGEN 109P; The alcohol includes any one of n-butanol, isobutanol, ethanol and methanol.

2. The sample processing liquid for centrifugal microfluidic disc biochemical analysis chip according to claim 1, characterized in that: The invention comprises the following concentration components: buffer: 100 mmol / L, pH=7.5, monovalent salt: 500 mmol / L, composite solubilizer 20 g / L, EMULGEN B-66: 50 g / L, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt: 15 g / L, flufenamic acid butyl ester: 2 mmol / L, N,N-diethylformamide: 6 mmol / L, ascorbic acid oxidase: 2KU / L, alcohol: 10 mL / L, defoamer 204: 10 g / L.

3. The sample processing solution for a centrifugal microfluidic disc biochemical analysis chip according to claim 1, characterized in that: The preparation method comprises the following steps: Step 1: Under light-shielded conditions, add flufenamic acid butyl ester and N,N-diethylformamide to alcohol in sequence, mix well, and let stand for 4 to 6 hours to obtain solution A; Step 2: Add buffer, monovalent salt, composite solubilizer, EMULGEN B-66, 3-[N,N-dimethyl(3-palmitamidopropyl)ammonium]-propanesulfonic acid inner salt, and defoamer 204 to distilled water in sequence, mix well, adjust the pH value to 7.0-7.5, add ascorbic acid oxidase, mix well, and obtain solution B; Step 3: In a dark environment, mix solution A and solution B evenly to obtain a mixed solution; at the same time, continuously introduce nitrogen gas into the mixed solution for 30 to 40 minutes to obtain a sample treatment solution, which is stored for later use.

4. The sample processing liquid for centrifugal microfluidic disc biochemical analysis chip according to claim 3, characterized in that: The sample processing solution is stored in a temperature range of 2 to 8° C. away from light.

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