CN114755195A - Preparation method of reagent for quickly detecting urine protein - Google Patents
Preparation method of reagent for quickly detecting urine protein Download PDFInfo
- Publication number
- CN114755195A CN114755195A CN202210663996.9A CN202210663996A CN114755195A CN 114755195 A CN114755195 A CN 114755195A CN 202210663996 A CN202210663996 A CN 202210663996A CN 114755195 A CN114755195 A CN 114755195A
- Authority
- CN
- China
- Prior art keywords
- reagent
- protein
- urine
- urine protein
- detection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000002700 urine Anatomy 0.000 title claims abstract description 136
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 122
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 122
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 98
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 238000001514 detection method Methods 0.000 claims abstract description 73
- 238000000034 method Methods 0.000 claims abstract description 60
- OHJMTUPIZMNBFR-UHFFFAOYSA-N biuret Chemical compound NC(=O)NC(N)=O OHJMTUPIZMNBFR-UHFFFAOYSA-N 0.000 claims description 30
- UXHUJQMSTNOTRT-ZIAGYGMSSA-N dicyclohexyl (2r,3r)-2,3-dihydroxybutanedioate Chemical compound O=C([C@H](O)[C@@H](O)C(=O)OC1CCCCC1)OC1CCCCC1 UXHUJQMSTNOTRT-ZIAGYGMSSA-N 0.000 claims description 29
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims description 29
- YZYASTRURKBPPS-UHFFFAOYSA-N C(CCC(=O)OCCCCCC(C)C)(=O)OCCCCCC(C)C.[Na] Chemical compound C(CCC(=O)OCCCCCC(C)C)(=O)OCCCCCC(C)C.[Na] YZYASTRURKBPPS-UHFFFAOYSA-N 0.000 claims description 27
- 238000003756 stirring Methods 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 238000002156 mixing Methods 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 15
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 12
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 11
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 10
- WUGCLPOLOCIDHW-UHFFFAOYSA-N 2-aminoethanol;benzoic acid Chemical compound [NH3+]CCO.[O-]C(=O)C1=CC=CC=C1 WUGCLPOLOCIDHW-UHFFFAOYSA-N 0.000 claims description 8
- NOOLISFMXDJSKH-UTLUCORTSA-N (+)-Neomenthol Chemical compound CC(C)[C@@H]1CC[C@@H](C)C[C@@H]1O NOOLISFMXDJSKH-UTLUCORTSA-N 0.000 claims description 4
- NOOLISFMXDJSKH-UHFFFAOYSA-N DL-menthol Natural products CC(C)C1CCC(C)CC1O NOOLISFMXDJSKH-UHFFFAOYSA-N 0.000 claims description 4
- 239000008367 deionised water Substances 0.000 claims description 4
- 229910021641 deionized water Inorganic materials 0.000 claims description 4
- 229940041616 menthol Drugs 0.000 claims description 4
- 239000004094 surface-active agent Substances 0.000 claims description 4
- 230000035945 sensitivity Effects 0.000 abstract description 13
- 230000004044 response Effects 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 47
- ONCZQWJXONKSMM-UHFFFAOYSA-N dialuminum;disodium;oxygen(2-);silicon(4+);hydrate Chemical group O.[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[Na+].[Na+].[Al+3].[Al+3].[Si+4].[Si+4].[Si+4].[Si+4] ONCZQWJXONKSMM-UHFFFAOYSA-N 0.000 description 18
- 229940080314 sodium bentonite Drugs 0.000 description 17
- 229910000280 sodium bentonite Inorganic materials 0.000 description 17
- 238000012360 testing method Methods 0.000 description 16
- 238000002835 absorbance Methods 0.000 description 15
- 238000005259 measurement Methods 0.000 description 15
- 230000008569 process Effects 0.000 description 13
- 230000008859 change Effects 0.000 description 12
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 11
- 239000004202 carbamide Substances 0.000 description 11
- 239000007864 aqueous solution Substances 0.000 description 10
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- 239000012496 blank sample Substances 0.000 description 9
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 8
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 8
- 239000011574 phosphorus Substances 0.000 description 8
- 229910052698 phosphorus Inorganic materials 0.000 description 8
- 239000011593 sulfur Substances 0.000 description 8
- 229910052717 sulfur Inorganic materials 0.000 description 8
- 238000001914 filtration Methods 0.000 description 7
- 238000002331 protein detection Methods 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 108010046334 Urease Proteins 0.000 description 6
- OSZCTNHVFQDBBY-UHFFFAOYSA-N 1-ethoxy-2-(isocyanatomethyl)benzene Chemical compound CCOC1=CC=CC=C1CN=C=O OSZCTNHVFQDBBY-UHFFFAOYSA-N 0.000 description 5
- 238000007696 Kjeldahl method Methods 0.000 description 5
- 238000000862 absorption spectrum Methods 0.000 description 4
- 230000002485 urinary effect Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 230000003252 repetitive effect Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 230000002195 synergetic effect Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 239000003463 adsorbent Substances 0.000 description 2
- 229940092782 bentonite Drugs 0.000 description 2
- 229910000278 bentonite Inorganic materials 0.000 description 2
- 239000000440 bentonite Substances 0.000 description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 2
- MQKXWEJVDDRQKK-UHFFFAOYSA-N bis(6-methylheptyl) butanedioate Chemical compound CC(C)CCCCCOC(=O)CCC(=O)OCCCCCC(C)C MQKXWEJVDDRQKK-UHFFFAOYSA-N 0.000 description 2
- 238000003370 dye binding method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 201000001474 proteinuria Diseases 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 229940048106 sodium lauroyl isethionate Drugs 0.000 description 2
- BRMSVEGRHOZCAM-UHFFFAOYSA-M sodium;2-dodecanoyloxyethanesulfonate Chemical group [Na+].CCCCCCCCCCCC(=O)OCCS([O-])(=O)=O BRMSVEGRHOZCAM-UHFFFAOYSA-M 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- MGSRCZKZVOBKFT-UHFFFAOYSA-N thymol Chemical compound CC(C)C1=CC=C(C)C=C1O MGSRCZKZVOBKFT-UHFFFAOYSA-N 0.000 description 2
- WXHLLJAMBQLULT-UHFFFAOYSA-N 2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-n-(2-methyl-6-sulfanylphenyl)-1,3-thiazole-5-carboxamide;hydrate Chemical compound O.C=1C(N2CCN(CCO)CC2)=NC(C)=NC=1NC(S1)=NC=C1C(=O)NC1=C(C)C=CC=C1S WXHLLJAMBQLULT-UHFFFAOYSA-N 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- 239000005844 Thymol Substances 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 238000007705 chemical test Methods 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 229920005552 sodium lignosulfonate Polymers 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229960000790 thymol Drugs 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Plasma & Fusion (AREA)
- Food Science & Technology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a preparation method of a reagent for quickly detecting urine protein, belonging to the technical field of medical detection, and the method comprises the following steps: the rapid detection reagent for the urine protein has the characteristics of high specificity, good sensitivity, high response rate and good stability, is wide in detection range, can rapidly detect trace urine protein, is simple and convenient to operate, and can realize quantitative detection of the urine protein.
Description
Technical Field
The invention belongs to the technical field of medical detection, and particularly relates to a preparation method of a rapid urine protein detection reagent.
Background
Urine is a non-invasive biological fluid, an ideal analyte for monitoring kidney damage, and is commonly used in medical diagnostics and clinical research. Clinical urine detection can help doctors to know the changes of physiological and pathological functions of the urinary system of patients and can also indirectly reflect the functional conditions of the whole body system.
Proteinuria refers to a condition in which urine contains abnormally high concentrations of proteins, and normally most of the proteins do not enter the urine, but in the case of impaired kidney function, some of the proteins enter the urine, resulting in proteinuria. The detection of urine protein in urine is the basis of kidney disease diagnosis, the detection of urine protein is divided into qualitative detection and quantitative detection, the widely applied qualitative detection methods at present comprise a sulfosalicylic acid method, a heating acetic acid method and a dry chemical test strip method, the methods can quickly detect the existence of urine protein in urine, but the tests usually lack accuracy and sensitivity, and the quantitative detection methods mainly comprise a dye binding method, chip electrophoresis, capillary electrophoresis, high performance liquid chromatography, enzymatic immunoassay and turbidimetric inhibition immunoassay, but most of the methods need long sample detection time. The biuret method in the dye binding method is characterized in that peptide bonds in protein react with a copper sulfate solution to generate a purple macromolecular complex, the color depth of the macromolecular complex is in direct proportion to the protein content in urine, and the protein content can be quantitatively detected according to the change of the absorbance of a product. The biuret method has the advantages of simple operation, good repeatability and the like, can quickly determine the protein content, but has certain defects, such as poor detection sensitivity, certain difficulty in measuring a sample with low protein content, and higher requirements on quantitative detection of urine protein along with improvement of medical conditions. Therefore, it is not always easy to improve the conventional biuret method and provide a rapid, simple, economical and practical method for detecting urine protein.
Disclosure of Invention
Aiming at the problems, the invention provides a preparation method of a urine protein rapid detection reagent, and the urine protein rapid detection reagent has the characteristics of high specificity, good sensitivity, high response rate and good stability, has a wide detection range, can rapidly detect trace urine protein, is simple and convenient to operate, and can realize quantitative detection of urine protein.
The technical solution to solve the above problems is as follows.
In a first aspect, the present invention provides a method for preparing a reagent for rapid detection of urine protein, the method comprising:
mixing a surfactant and menthol according to a mass ratio of 1: 0.1-0.3, adding deionized water to dissolve the mixture to obtain a mixed solution, mixing the mixed solution with a biuret reagent according to a mass ratio of 1: 0.2-0.4, and stirring the mixed solution at the temperature of 30-40 ℃ for 0.5-2 hours to obtain the rapid urine protein detection reagent.
As a preferred embodiment, the surfactant is sodium lauroyl isethionate or sodium lignosulfonate.
As a preferred embodiment, the biuret reagent is prepared by the following method:
1) dissolving potassium hydroxide in water to prepare a solution of 0.05-0.25 g/mL as a reagent A;
2) sequentially dissolving monoethanolamine benzoate, sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in water, stirring for 10-40 min at 30-40 ℃, adding hydrated copper sulfate, and continuously stirring for 0.5-2 h to obtain a reagent B;
3) And uniformly mixing the reagent A and the reagent B according to the volume ratio of 5-8: 1 to obtain the biuret reagent.
More preferably, in the reagent B, the mass fraction of the sodium diisooctyl succinate sulfonate is 0.2-0.45%, the mass fraction of the dicyclohexyl tartrate is 0.03-0.05%, the mass fraction of the monoethanolamine benzoate is 0.08-0.14%, and the mass fraction of the hydrated copper sulfate is 0.1-0.3%.
More preferably, in the reagent B, the mass ratio of the diisooctyl succinate sodium sulfonate to the dicyclohexyl tartrate is 1: 0.1-0.2.
The urine protein rapid detection reagent based on the biuret reagent is light blue, after the reagent is mixed with a urine sample to be detected, peptide bonds in protein and copper ions are combined to form a purple macromolecular complex, the aim of quantitative detection of urine protein can be realized through a standard curve, the detection limit reaches 15mg/L, and the reagent has the advantages of high sensitivity, simplicity in operation and good repeatability, and can be used for rapidly determining the content of protein; compared with the traditional biuret reagent, the sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in a certain mass ratio are added into the reagent provided by the invention, and have a certain synergistic effect, so that the dissolution of insoluble protein in a sample to be detected can be increased to a certain extent, and the measurement error caused by the insoluble protein is reduced, thereby improving the detection sensitivity and accuracy.
In a second aspect, the reagent for rapid detection of urine protein according to the first aspect is used for detection of urine protein.
In a third aspect, a method for detecting urine protein, the method comprising:
step one, taking water as a blank sample, dissolving bovine serum albumin in the water to prepare a 1g/mL aqueous solution, and then diluting to obtain a series of solutions with the protein concentration within the range of 15-6000 mg/L, wherein the solutions are used as standard protein samples;
step two, respectively mixing the urine protein rapid detection reagent with a blank sample and a standard protein sample, adding the mixture into a biochemical automatic instrument, stirring the mixture for 0.5 to 2 hours at the temperature of 30 to 40 ℃, taking the blank sample as a reference, measuring the absorbance of the standard protein sample at the position of 540nm of the dominant wavelength, and fitting a standard curve according to the concentration and the absorbance of the standard protein sample;
and step three, after pretreating the urine sample to be detected, adding a urine protein rapid detection reagent into the sample, stirring for 0.5-2 h at the temperature of 30-40 ℃, measuring absorbance, and substituting the absorbance into a standard curve to calculate the protein content.
In the second and third steps, the volume ratio of the sample to the urine protein rapid detection reagent is 1: 1-20.
As a preferred embodiment, the urine sample to be tested in the third step is fresh urine, urine stored at 2-8 ℃ for 24h or urine stored at room temperature for 24 h.
Preferably, a certain amount of thymol is added into the urine stored for 24 hours at room temperature as a preservative, so that bacterial proliferation in the urine standing process at room temperature can be inhibited, the influence of protein decomposition on a measurement result is reduced, and the measurement accuracy is improved.
In a preferred embodiment, in the third step, the pretreatment step of the urine sample to be tested comprises:
adding a urease aqueous solution into the urine sample to be detected, standing and incubating, and removing urea in the urine sample to be detected; and then adding an adsorbent, oscillating for 20-50 min, and filtering to remove sulfur and phosphorus in the urine.
Preferably, in the pretreatment process, the incubation temperature is 30-40 ℃ and the time is 10-30 min.
Preferably, the adsorbent is activated sodium bentonite during the pretreatment.
As a further preference, the active sodium bentonite is prepared by the following method:
the method comprises the steps of crushing sodium bentonite to 200-300 meshes, cleaning with 10-20% by mass of sodium chloride solution, dispersing the bentonite in dichloromethane, adding 2-ethoxy benzyl isocyanate, ultrasonically stirring at 75-90 ℃ for 20-50 min, filtering, washing with water, and drying. According to the invention, by utilizing the steps, firstly, the urea is decomposed by treating the urine sample to be detected by using urease, then the phosphorus and the sulfur are removed by using active sodium bentonite, and after the sodium bentonite is treated by 2-ethoxy benzyl isocyanate, the adsorbability of the bentonite can be improved, the removal efficiency of the urea and the phosphorus and the sulfur is improved, so that the problems of poor pH stability of the sample and the like caused by the existence of the urea and the like are reduced, and the detection accuracy is further improved.
The invention provides a method for detecting urine protein by using a urine protein rapid detection reagent, which comprises the steps of firstly pretreating a urine sample to be detected to remove urea, phosphorus, sulfur and the like, reducing the influence on the determination of the urine protein content and improving the accuracy, and then quantitatively detecting the urine sample to be detected by using the urine protein rapid detection reagent provided by the invention, and the method is simple and has high accuracy.
In a fourth aspect, the invention also provides application of sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in improving sensitivity and stability of a urine protein rapid detection reagent.
The beneficial effects of the invention are as follows:
the invention provides a preparation method of a rapid detection reagent for urine protein, and the prepared rapid detection reagent for urine protein has the advantages of high sensitivity, simple operation and good repeatability, can rapidly determine the content of protein, and has a wide application prospect in clinical urine detection; the sodium diisooctyl succinate sulfonate and the dicyclohexyl tartrate in a certain mass ratio are added into the urine protein rapid detection reagent, and have a certain synergistic effect, so that the solubility of insoluble protein in a sample to be detected can be increased, the measurement error caused by the insoluble protein is reduced, the detection sensitivity and accuracy are improved, and the stability of the urine protein rapid detection reagent is improved; before the urine sample to be detected is detected, urease is used for treating and decomposing urea, and active sodium bentonite is used for removing phosphorus and sulfur.
Drawings
FIG. 1 is a standard curve of a standard protein sample;
FIG. 2 is an absorption spectrum of the reagent for rapid detection of urine protein of example 1 together with a conventional biuret reagent; in the figure, A represents a urine protein rapid detection reagent, and B represents a traditional biuret reagent;
FIG. 3 is a graph showing the results of a stability test of a reagent for rapid detection of urine protein; in the figure, B represents the change rate of urine protein measured after 5 days of storage, C represents the change rate of urine protein measured after 10 days of storage, and D represents the change rate of urine protein measured after 20 days of storage.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
According to the invention, firstly, a preparation method of a reagent for quickly detecting urine protein is provided, which comprises the following steps:
dissolving potassium hydroxide in water to prepare a solution of 0.05-0.25 g/mL as a reagent A; dissolving monoethanolamine benzoate, sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in water in sequence, stirring for 10-40 min at 30-40 ℃, adding hydrated copper sulfate, and continuously stirring for 0.5-2 h to obtain a reagent B, wherein in the reagent B, the mass fraction of the sodium diisooctyl succinate sulfonate is 0.2-0.45%, the mass fraction of the dicyclohexyl tartrate is 0.03-0.05%, the mass fraction of the monoethanolamine benzoate is 0.08-0.14%, and the mass fraction of the hydrated copper sulfate is 0.1-0.3%; uniformly mixing the reagent A and the reagent B according to the volume ratio of 5-8: 1 to obtain a biuret reagent; wherein the mass ratio of the diisooctyl succinate sodium sulfonate to the dicyclohexyl tartrate is 1: 0.1-0.2; mixing a surfactant and menthol according to a mass ratio of 1: 0.1-0.3, adding deionized water to dissolve the mixture to obtain a mixed solution, mixing the mixed solution with a biuret reagent according to a mass ratio of 1: 0.2-0.4, and stirring the mixed solution at the temperature of 30-40 ℃ for 0.5-2 hours to obtain the rapid urine protein detection reagent.
The urine protein rapid detection reagent based on the biuret reagent has the detection range of 15-6000 mg/L, has the advantages of high sensitivity, simplicity in operation and good repeatability, and can be used for rapidly determining the protein content; compared with the traditional biuret reagent, the sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in a certain mass ratio are added into the reagent provided by the invention, and have a certain synergistic effect, so that the solubility of insoluble protein in a sample to be detected can be increased, the measurement error caused by the insoluble protein is reduced, the detection sensitivity and accuracy are improved, in addition, the effect of improving the stability of the reagent for rapidly detecting the urine protein is also realized, the higher accuracy is still realized after the reagent is placed for a long time, and the quality guarantee period is long.
According to the present invention, there is also provided a method for detecting urine protein, the method comprising:
step one, taking water as a blank sample, dissolving bovine serum albumin in the water to prepare a 1g/mL aqueous solution, and then diluting to obtain a series of solutions with the protein concentration within the range of 15-6000 mg/L, wherein the solutions are used as standard protein samples;
step two, mixing the urine protein rapid detection reagent with a blank sample and a standard protein sample according to a volume ratio of 5-20: 1, adding the mixture into a biochemical automatic instrument, stirring the mixture for 0.5-2 hours at 30-40 ℃, taking the blank sample as a reference, determining the absorbance of the standard protein sample at a dominant wavelength of 540nm, and fitting a standard curve according to the concentration and the absorbance of the standard protein sample;
Step three, pretreating the urine sample to be detected, and specifically comprising the following steps: adding 100u/mg of urease aqueous solution into a urine sample to be detected, standing, incubating for 10-30 min at 30-40 ℃, and removing urea in the urine sample; crushing sodium bentonite to 200-300 meshes, cleaning with a sodium chloride solution with the mass fraction of 10-20%, dispersing the sodium bentonite in dichloromethane, adding 2-ethoxybenzyl isocyanate with the mass of 0.2-0.35 times that of the sodium bentonite, ultrasonically stirring at 75-90 ℃ for 20-50 min, filtering, washing with water, and drying to obtain active sodium bentonite; adding active sodium bentonite into a urine sample to be tested, oscillating for 20-50 min, and filtering to remove sulfur and phosphorus in the urine;
and step four, adding a urine protein rapid detection reagent into the pretreated urine sample to be detected according to the volume ratio of 1: 1-20, stirring for 0.5-2 h at 30-40 ℃, measuring absorbance, and substituting into a standard curve to calculate the protein content.
Through the steps, the urea is decomposed by using urease treatment, the phosphorus and the sulfur are removed by using the active sodium bentonite, and then the urea protein is detected by using a urine protein rapid detection reagent.
The present invention is explained in detail below with reference to examples.
Example 1:
the present embodiment provides a method for detecting urine protein, comprising the following steps:
1) dissolving potassium hydroxide in water to prepare a solution of 0.1g/mL as a reagent A; sequentially dissolving monoethanolamine benzoate, sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in water, stirring for 25min at 37 ℃, adding hydrated copper sulfate, and continuing stirring for 1h to obtain a reagent B, wherein in the reagent B, the mass fraction of the sodium diisooctyl succinate sulfonate is 0.3%, the mass fraction of the dicyclohexyl tartrate is 0.045%, the mass fraction of the monoethanolamine benzoate is 0.1%, and the mass fraction of the hydrated copper sulfate is 0.2%; uniformly mixing the reagent A and the reagent B according to the volume ratio of 6:1 to obtain a biuret reagent; mixing sodium lauroyl isethionate and menthol according to the mass ratio of 1:0.2, adding 10 times of deionized water for dissolving to obtain a mixed solution, mixing with a biuret reagent according to the mass ratio of 1:0.3, and stirring at 35 ℃ for 1h to obtain a rapid urine protein detection reagent;
2) taking water as a blank sample, dissolving bovine serum albumin in the water to prepare a 1g/mL aqueous solution, and then diluting to obtain a series of solutions with the protein concentration within the range of 15-6000 mg/L, wherein the solutions are used as standard protein samples;
3) Respectively mixing the urine protein rapid detection reagent with a blank sample and a standard protein sample according to a volume ratio of 5:1, adding the mixture into a biochemical automatic analyzer, stirring the mixture for 1h at 37 ℃, taking the blank sample as a reference, measuring the absorbance of the standard protein sample at a main wavelength of 540nm, fitting a standard curve by using the concentration and the absorbance of the standard protein sample, and obtaining a result shown in figure 1, wherein the concentration and the absorbance have a good linear relation, and the linear correlation degree R is20.9996, and the curve equation is y =0.0078 x-0.1926;
4) adding urease aqueous solution with the volume 0.1 time and 100U/mg of the sample into the urine sample to be detected, standing, incubating for 20min at 37 ℃, and removing urea in the urine sample; crushing sodium bentonite to 250 meshes, and cleaning with 15% sodium chloride solution by mass; dispersing sodium bentonite in 10 weight times of dichloromethane, adding 2-ethoxy benzyl isocyanate with the mass of 0.3 time of that of the sodium bentonite, ultrasonically stirring at 80 ℃ for 35min, filtering, washing with water, and drying to obtain active sodium bentonite; adding sodium bentonite into a urine sample to be tested, oscillating for 40min, and filtering to remove sulfur and phosphorus in the urine;
5) Adding a urine protein rapid detection reagent into the pretreated urine sample to be detected according to the volume ratio of 1:5, stirring for 1h at 37 ℃, measuring absorbance, and substituting into a standard curve to calculate the protein content.
Example 2:
this example provides another method for detecting urine protein, which has the same steps as in example 1, except that in the process of preparing the reagent for rapidly detecting urine protein, the mass fraction of sodium diisooctyl succinate sulfonate in reagent B is 0.3%, and the mass fraction of dicyclohexyl tartrate in reagent B is 0.03%, that is, the mass ratio of sodium diisooctyl succinate sulfonate to dicyclohexyl tartrate is 1: 0.1.
Example 3:
this example provides another method for detecting urine protein, which has the same steps as in example 1, except that in the process of preparing the reagent for rapidly detecting urine protein, the mass fraction of sodium diisooctyl succinate sulfonate in reagent B is 0.25%, and the mass fraction of dicyclohexyl tartrate in reagent B is 0.05%, that is, the mass ratio of sodium diisooctyl succinate sulfonate to dicyclohexyl tartrate is 1: 0.2.
Example 4:
this example provides another method for detecting urine protein, which has substantially the same steps as example 1, except that in the process of preparing a reagent for rapid detection of urine protein, the mass fraction of sodium diisooctyl succinate sulfonate in reagent B is 0.3%, and the mass fraction of dicyclohexyl tartrate in reagent B is 0.015%, that is, the mass ratio of sodium diisooctyl succinate sulfonate to dicyclohexyl tartrate is 1: 0.05.
Example 5:
this example provides another method for detecting urine protein, which has substantially the same procedure as in example 1, except that in the process of preparing a reagent for rapid detection of urine protein, the mass fraction of sodium diisooctyl succinate sulfonate in reagent B is 0.3%, and the mass fraction of dicyclohexyl tartrate in reagent B is 0.09%, that is, the mass ratio of sodium diisooctyl succinate sulfonate to dicyclohexyl tartrate is 1: 0.3.
Example 6:
this example provides another method for detecting urine protein, which has substantially the same steps as example 1, except that dicyclohexyl tartrate is not added to the reagent B during the preparation of the reagent for rapid detection of urine protein, and the mass fraction of sodium diisooctyl succinate sulfonate added is 0.3%.
Example 7:
this example provides another method for detecting urine protein, which has substantially the same steps as example 1, except that sodium diisooctyl succinate sulfonate is not added to the reagent B during the preparation of the reagent for rapid detection of urine protein, and the added mass fraction of dicyclohexyl tartrate is 0.045%.
Example 8:
this example provides another method for detecting urine protein, which comprises the same steps as example 1, except that dicyclohexyl tartrate and sodium diisooctyl succinate sulfonate are not added to reagent B during the preparation of the reagent for rapid detection of urine protein.
Example 9:
this example provides another method for detecting urine protein, which has the same steps as example 1, except that sodium bentonite is not treated with 2-ethoxybenzyl isocyanate in the pretreatment process of the urine sample to be detected.
Test example 1:
and (3) testing the accuracy:
the same urine sample to be tested is tested by a Kjeldahl method, a traditional double-biuret method and the methods of the embodiments 1, 4 to 9, each group is tested for 5 times, the average value is taken as the protein content, and the relative error is calculated by the following formula: relative error = (C)0-C)/C0X 100%, wherein, the C0C represents the measurement result of the traditional biuret method or the measurement results of the methods of the embodiments 1, 4 to 9, wherein the traditional biuret method is as follows: adding 3mL of 0.1g/L sodium hydroxide aqueous solution into 3mL of urine sample to be tested, after uniform oscillation, adding 2 drops of 0.01g/mL copper sulfate aqueous solution, and after uniform oscillation, measuring the absorbance.The results are shown in Table 1.
TABLE 1 test results
Table 1 shows the measurement result of the urine sample to be measured, and as shown in table 1, the relative error of the measurement result of the urine protein in example 1 is 0.17 compared with the measurement result of the kjeldahl method, and the relative error of the protein content measured by the conventional biuret method is 1.18% compared with the measurement result of the kjeldahl method, and the relative error of the detection method provided by the present invention is smaller, so that compared with the conventional biuret method, the protein content measured by the rapid urine protein detection reagent provided by the present invention is closer to the true content of the protein, and has higher accuracy; it can also be seen that, compared with the results of kjeldahl method, the results of examples 4 to 8, which have a relative error of 0.74 to 1.34% and are significantly higher than those of example 1, show that the addition of sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in a certain mass ratio to the reagent for rapid detection of urine protein contributes to the improvement of the detection accuracy of the reagent; it can be seen that the relative error of the measurement result of urine protein in example 9 is 0.94% compared with the protein measured by kjeldahl method, and thus it can be seen that the pretreatment of the urine sample to be tested can reduce the influence of urea and the like on the detection of urine protein, and improve the accuracy of the detection result.
Test example 2:
and (3) testing the sensitivity:
adding the traditional biuret reagent and the urine protein rapid detection reagent provided by the embodiment 1 of the invention into the same urine sample to be detected, uniformly oscillating, and testing an absorption spectrum; the traditional biuret reagent comprises: 0.1g/L of sodium hydroxide aqueous solution and 0.01g/mL of copper sulfate aqueous solution; the measured absorption spectrum is shown in FIG. 2.
FIG. 2 is an absorption spectrum of a urine protein rapid detection reagent and a conventional biuret reagent, wherein A represents the urine protein rapid detection reagent, and B represents the conventional biuret reagent; as can be seen from the observation of FIG. 2, the maximum value of the absorbance of the protein measured by the reagent for rapidly detecting urine protein provided by the invention is 0.317, while the maximum value of the absorbance of the protein measured by the traditional biuret reagent is 0.139, therefore, the method of the invention has obvious sensitization effect, and the sensitivity is obviously improved compared with the traditional biuret reagent under the same condition.
Test example 3:
and (3) repeatability test:
the method provided by the embodiment 1, 4-8 is utilized to repeatedly test the same urine sample to be tested for 10 times, the standard deviation and the variation coefficient are calculated, the variation coefficient is utilized to evaluate the repeatability, and the measurement result is shown in the table 2.
TABLE 2 repeatability test results
Table 2 shows the result of the repetitive tests of the reagent for rapid detection of urinary protein, and it can be seen from the observation of table 2 that the method provided in example 1 performs repetitive tests 10 times on the urine sample to be detected, the standard deviation is 1.2mg/L, the coefficient of variation is 0.61%, which indicates that the error of the repetitive tests 20 times is small, and the reagent for rapid detection of urinary protein provided by the present invention has good reproducibility and high accuracy; comparing examples 1 and 4-8, it can be seen that the standard deviation and the coefficient of variation of examples 4-8 are significantly higher than those of example 1, which illustrates that adding sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in a certain mass ratio to a rapid urine protein detection reagent helps to improve the repeatability and accuracy of the reagent.
Test example 4:
and (3) testing the stability:
the reagent for rapid detection of urine protein in examples 1, 4-8 is stored at 37 ℃, and the same urine sample to be detected is detected according to the method provided in example 1 at 0d, 5d, 10d, and 20d, respectively, and the change rate is calculated by using the following formula: rate of change (%) = (C)t-C0)/C0×100%；CtMeasured content of urine protein after storage for various periods, C0The time is the initial measurement content of urine protein, and the rate of change is used to evaluate stability, and the measurement results are shown in fig. 3.
FIG. 3 is a graph showing the results of stability test of the reagent for rapid detection of urine protein; in the figure, B represents the change rate of the urine protein after 5 days of storage, C represents the change rate of the urine protein after 10 days of storage, and D represents the change rate of the urine protein after 20 days of storage; as can be seen from the observation of FIG. 3, the reagent for rapid detection of urinary protein in the preferred embodiment 1 has high stability, does not generate much turbidity after being stored at 37 ℃ for 20 days, omits a pretreatment step of filtering the reagent before use, does not change the performance, changes only 1.13% of the protein detection result, and still has high accuracy; the urine protein rapid detection reagents of examples 4 to 8 are different in that the biuret reagent is different in composition, and the change rates of examples 4 to 8 are significantly higher than those of example 1, so that it is known that the addition of sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate has a large influence on the stability of the urine protein rapid detection reagent.
The applicant states that the present invention is illustrated by the above examples to show the detailed process equipment and process flow of the present invention, but the present invention is not limited to the above detailed process equipment and process flow, i.e. it does not mean that the present invention must rely on the above detailed process equipment and process flow to be implemented. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (5)
1. A preparation method of a reagent for quickly detecting urine protein is characterized by comprising the following steps:
mixing a surfactant and menthol according to the mass ratio of 1: 0.1-0.3, adding deionized water to dissolve the mixture to obtain a mixed solution, mixing the mixed solution with a biuret reagent according to the mass ratio of 1: 0.2-0.4, and stirring the mixed solution at the temperature of 30-40 ℃ for 0.5-2 hours to obtain the rapid detection reagent for the urine protein.
2. The method of claim 1, wherein the biuret reagent is prepared by the following method:
1) dissolving potassium hydroxide in water to prepare a solution of 0.05-0.25 g/mL as a reagent A;
2) sequentially dissolving monoethanolamine benzoate, sodium diisooctyl succinate sulfonate and dicyclohexyl tartrate in water, stirring for 10-40 min at 30-40 ℃, adding hydrated copper sulfate, and continuously stirring for 0.5-2 h to obtain a reagent B;
3) and uniformly mixing the reagent A and the reagent B according to the volume ratio of 5-8: 1 to obtain the biuret reagent.
3. The method as claimed in claim 2, wherein in the reagent B, the mass fraction of sodium diisooctyl succinate sulfonate is 0.2-0.45%, the mass fraction of dicyclohexyl tartrate is 0.03-0.05%, the mass fraction of monoethanolamine benzoate is 0.08-0.14%, and the mass fraction of hydrated copper sulfate is 0.1-0.3%.
4. The method according to claim 3, wherein the mass ratio of sodium diisooctyl succinate sulfonate to dicyclohexyl tartrate is 1: 0.1-0.2.
5. The use of the reagent for rapid detection of urine protein prepared by the method of any one of claims 1 to 4 in the detection of urine protein.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210663996.9A CN114755195A (en) | 2022-06-14 | 2022-06-14 | Preparation method of reagent for quickly detecting urine protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210663996.9A CN114755195A (en) | 2022-06-14 | 2022-06-14 | Preparation method of reagent for quickly detecting urine protein |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114755195A true CN114755195A (en) | 2022-07-15 |
Family
ID=82336287
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210663996.9A Pending CN114755195A (en) | 2022-06-14 | 2022-06-14 | Preparation method of reagent for quickly detecting urine protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114755195A (en) |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1459705A (en) * | 1974-04-02 | 1976-12-31 | Commw Scient Ind Res Org | Automated biuret analysis |
| CN1057525A (en) * | 1991-02-05 | 1992-01-01 | 中国人民解放军302医院 | A kind ofly measure the solid reagent that total serum protein is used |
| WO1996033162A1 (en) * | 1995-04-21 | 1996-10-24 | Rhone-Poulenc Limited | Compounds containing a substantially planar, fused ring system with at least 4 aromatic rings and their use as a chiral stationary phase in enantiomeric separation |
| US5693516A (en) * | 1995-11-27 | 1997-12-02 | Novo Nordisk Biotech, Inc. | Method for solubilizing proteins in organic solvents |
| EP1170593A2 (en) * | 2000-06-19 | 2002-01-09 | Felix Levine | Reagent, kit and method for the determination of total proteins and globulines such as hemoglobin in biological fluids |
| CN105486682A (en) * | 2015-12-28 | 2016-04-13 | 东北农业大学 | Rapid protein detection kit as well as detection method and application using same |
| CN106442352A (en) * | 2016-09-24 | 2017-02-22 | 济南中安生物技术服务有限公司 | Total serum protein detection kit with strong anti-interference capability |
| CN107525774A (en) * | 2016-06-21 | 2017-12-29 | 山东博科生物产业有限公司 | Potassium hydroxide method total protein diagnostic test kits |
| CN108014753A (en) * | 2017-11-02 | 2018-05-11 | 铜陵博锋实业有限公司 | A kind of preparation method of porous modified alta-mud granule adsorbent |
| CN110632325A (en) * | 2019-09-27 | 2019-12-31 | 昆山迪安医学检验实验室有限公司 | Total protein detection reagent and preparation method thereof |
| CN111077148A (en) * | 2019-12-19 | 2020-04-28 | 苏州浚惠生物科技有限公司 | Method for capturing and detecting abnormal metabolic cells of urine |
-
2022
- 2022-06-14 CN CN202210663996.9A patent/CN114755195A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1459705A (en) * | 1974-04-02 | 1976-12-31 | Commw Scient Ind Res Org | Automated biuret analysis |
| CN1057525A (en) * | 1991-02-05 | 1992-01-01 | 中国人民解放军302医院 | A kind ofly measure the solid reagent that total serum protein is used |
| WO1996033162A1 (en) * | 1995-04-21 | 1996-10-24 | Rhone-Poulenc Limited | Compounds containing a substantially planar, fused ring system with at least 4 aromatic rings and their use as a chiral stationary phase in enantiomeric separation |
| US5693516A (en) * | 1995-11-27 | 1997-12-02 | Novo Nordisk Biotech, Inc. | Method for solubilizing proteins in organic solvents |
| EP1170593A2 (en) * | 2000-06-19 | 2002-01-09 | Felix Levine | Reagent, kit and method for the determination of total proteins and globulines such as hemoglobin in biological fluids |
| CN105486682A (en) * | 2015-12-28 | 2016-04-13 | 东北农业大学 | Rapid protein detection kit as well as detection method and application using same |
| CN107525774A (en) * | 2016-06-21 | 2017-12-29 | 山东博科生物产业有限公司 | Potassium hydroxide method total protein diagnostic test kits |
| CN106442352A (en) * | 2016-09-24 | 2017-02-22 | 济南中安生物技术服务有限公司 | Total serum protein detection kit with strong anti-interference capability |
| CN108014753A (en) * | 2017-11-02 | 2018-05-11 | 铜陵博锋实业有限公司 | A kind of preparation method of porous modified alta-mud granule adsorbent |
| CN110632325A (en) * | 2019-09-27 | 2019-12-31 | 昆山迪安医学检验实验室有限公司 | Total protein detection reagent and preparation method thereof |
| CN111077148A (en) * | 2019-12-19 | 2020-04-28 | 苏州浚惠生物科技有限公司 | Method for capturing and detecting abnormal metabolic cells of urine |
Non-Patent Citations (5)
| Title |
|---|
| 张玲等: "响应面法优化反胶束提取花生粕蛋白前萃工艺", 《粮食与油脂》 * |
| 张鹏等: "酒石酸衍生物作为手性选择剂在对映体的HPLC拆分中的应用", 《化学世界》 * |
| 蒋善庆等: "黄土颗粒吸附水解尿液中磷酸盐特性及机制", 《化工进展》 * |
| 袁水斌等: "稳定型总蛋白(TP)试剂的研制", 《江西医学检验》 * |
| 陆强等: "用反胶束萃取法提取与分离蛋白质和酶", 《生物工程进展》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Walters et al. | An ultramicromethod for the determination of conjugated and total bilirubin in serum or plasma | |
| Yatzidis | New method for direct determination of" true" creatinine | |
| RU2247393C2 (en) | Method and cell for carrying out analysis | |
| CN111638106B (en) | Urine dry chemical analysis quality control object | |
| NO300853B1 (en) | Assay methods, reagent composition and use thereof in glucose determination | |
| US4485176A (en) | Turbidimetric method for measuring protein in urine and cerebrospinal fluid | |
| Chromy et al. | Photometric determination of total protein in lipemic sera | |
| Datta et al. | Interference by IgG paraproteins in the Jaffe method for creatinine determination | |
| CN114755195A (en) | Preparation method of reagent for quickly detecting urine protein | |
| Hess | Polarographic Estimation of Chloroamphenicol (Chloromycetin) | |
| Barbour et al. | Studies on measurement of plasma magnesium: application of the Magon dye method to the" Monarch" centrifugal analyzer. | |
| CN113376150A (en) | Urine microalbumin dry chemical detection test paper and preparation method thereof | |
| CN104673878A (en) | Kit for measuring concentration ratio of glycated albumin and albumin by virtue of single system | |
| US5633169A (en) | Measurement of carbon dioxide in blood | |
| CN108007922B (en) | A kind of kit detecting glucose using luminol chemiluminescence analysis | |
| CN111455020A (en) | 1, 5-sorbitan detection kit and detection method | |
| US4278440A (en) | Reagent and method for direct determination of chloride in serum | |
| CN113655006B (en) | Urinary system knot Dan Chengdan risk factor detection and test system | |
| US5776780A (en) | Method for quantitatively measuring white blood cells esterase activity in urine | |
| CA1324066C (en) | Simultaneous assay for calcium and phosphorus | |
| Liedtke et al. | Automated calmagite compleximetric measurement of magnesium in serum, with sequential addition of EDTA to eliminate endogenous interference. | |
| JP4602595B2 (en) | Total protein quantification method and quantification reagent | |
| Li et al. | Fixed-time kinetic assay of plasma ammonia, with NADPH as cofactor, with a centrifugal analyzer. | |
| RU2593361C1 (en) | Spectrophotometric method of determination of protein in biological fluids | |
| CN115980331B (en) | Kit for detecting heparin binding protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220715 |
|
| RJ01 | Rejection of invention patent application after publication |