CN112945948A - Application of nitrogen-oxygen free radical compound as stabilizer in preparation of urine occult blood test paper - Google Patents
Application of nitrogen-oxygen free radical compound as stabilizer in preparation of urine occult blood test paper Download PDFInfo
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- 210000002700 urine Anatomy 0.000 title claims abstract description 37
- 239000003381 stabilizer Substances 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 238000009534 blood test Methods 0.000 title claims abstract description 14
- -1 nitrogen-oxygen free radical compound Chemical class 0.000 title claims abstract description 9
- 238000012360 testing method Methods 0.000 claims description 44
- 239000000758 substrate Substances 0.000 claims description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 28
- 210000004369 blood Anatomy 0.000 claims description 22
- 239000008280 blood Substances 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 21
- 238000007654 immersion Methods 0.000 claims description 20
- 239000004094 surface-active agent Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 14
- 238000002791 soaking Methods 0.000 claims description 14
- STZCRXQWRGQSJD-GEEYTBSJSA-M methyl orange Chemical compound [Na+].C1=CC(N(C)C)=CC=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 STZCRXQWRGQSJD-GEEYTBSJSA-M 0.000 claims description 9
- 229940012189 methyl orange Drugs 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- FRIBMENBGGCKPD-UHFFFAOYSA-N 3-(2,3-dimethoxyphenyl)prop-2-enal Chemical compound COC1=CC=CC(C=CC=O)=C1OC FRIBMENBGGCKPD-UHFFFAOYSA-N 0.000 claims description 8
- YRNWIFYIFSBPAU-UHFFFAOYSA-N 4-[4-(dimethylamino)phenyl]-n,n-dimethylaniline Chemical group C1=CC(N(C)C)=CC=C1C1=CC=C(N(C)C)C=C1 YRNWIFYIFSBPAU-UHFFFAOYSA-N 0.000 claims description 8
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- HFDLDPJYCIEXJP-UHFFFAOYSA-N 6-methoxyquinoline Chemical compound N1=CC=CC2=CC(OC)=CC=C21 HFDLDPJYCIEXJP-UHFFFAOYSA-N 0.000 claims description 7
- 239000007853 buffer solution Substances 0.000 claims description 7
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 6
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims description 6
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- ZLKGGEBOALGXJZ-UHFFFAOYSA-N 8-methoxyquinoline Chemical compound C1=CN=C2C(OC)=CC=CC2=C1 ZLKGGEBOALGXJZ-UHFFFAOYSA-N 0.000 claims description 5
- MPVDXIMFBOLMNW-ISLYRVAYSA-N 7-hydroxy-8-[(E)-phenyldiazenyl]naphthalene-1,3-disulfonic acid Chemical compound OC1=CC=C2C=C(S(O)(=O)=O)C=C(S(O)(=O)=O)C2=C1\N=N\C1=CC=CC=C1 MPVDXIMFBOLMNW-ISLYRVAYSA-N 0.000 claims description 4
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 4
- 238000005520 cutting process Methods 0.000 claims description 4
- GVGUFUZHNYFZLC-UHFFFAOYSA-N dodecyl benzenesulfonate;sodium Chemical compound [Na].CCCCCCCCCCCCOS(=O)(=O)C1=CC=CC=C1 GVGUFUZHNYFZLC-UHFFFAOYSA-N 0.000 claims description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 4
- 238000005096 rolling process Methods 0.000 claims description 4
- 238000010008 shearing Methods 0.000 claims description 4
- 229940080264 sodium dodecylbenzenesulfonate Drugs 0.000 claims description 4
- 239000000872 buffer Substances 0.000 claims description 3
- SPTHWAJJMLCAQF-UHFFFAOYSA-M ctk4f8481 Chemical compound [O-]O.CC(C)C1=CC=CC=C1C(C)C SPTHWAJJMLCAQF-UHFFFAOYSA-M 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 2
- 239000007979 citrate buffer Substances 0.000 claims description 2
- 239000000975 dye Substances 0.000 claims 3
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical class ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 abstract description 12
- 150000001875 compounds Chemical class 0.000 abstract description 12
- 150000003254 radicals Chemical class 0.000 abstract description 12
- 238000001514 detection method Methods 0.000 abstract description 11
- 239000000126 substance Substances 0.000 abstract description 6
- 238000005353 urine analysis Methods 0.000 abstract description 4
- 230000002255 enzymatic effect Effects 0.000 abstract description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 15
- 239000000463 material Substances 0.000 description 10
- 229920006316 polyvinylpyrrolidine Polymers 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 208000006750 hematuria Diseases 0.000 description 7
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 7
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 5
- 239000008213 purified water Substances 0.000 description 5
- 239000012086 standard solution Substances 0.000 description 5
- 229960005070 ascorbic acid Drugs 0.000 description 4
- 239000011668 ascorbic acid Substances 0.000 description 3
- 235000010323 ascorbic acid Nutrition 0.000 description 3
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 2
- 238000007705 chemical test Methods 0.000 description 2
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- 230000002485 urinary effect Effects 0.000 description 2
- OBHRVMZSZIDDEK-UHFFFAOYSA-N urobilinogen Chemical compound CCC1=C(C)C(=O)NC1CC1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(CC3C(=C(CC)C(=O)N3)C)N2)CCC(O)=O)N1 OBHRVMZSZIDDEK-UHFFFAOYSA-N 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 239000002211 L-ascorbic acid Substances 0.000 description 1
- 235000000069 L-ascorbic acid Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 208000000733 Paroxysmal Hemoglobinuria Diseases 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 208000005374 Poisoning Diseases 0.000 description 1
- 206010048960 Streptococcal sepsis Diseases 0.000 description 1
- 208000003441 Transfusion reaction Diseases 0.000 description 1
- 206010064921 Urinary tract inflammation Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003167 anti-vitamin Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000010866 blackwater Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229940109239 creatinine Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 201000001505 hemoglobinuria Diseases 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000010339 medical test Methods 0.000 description 1
- 201000008383 nephritis Diseases 0.000 description 1
- 201000003045 paroxysmal nocturnal hemoglobinuria Diseases 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 206010038534 renal tuberculosis Diseases 0.000 description 1
- 238000009666 routine test Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 208000000143 urethritis Diseases 0.000 description 1
- 238000002562 urinalysis Methods 0.000 description 1
- 208000014001 urinary system disease Diseases 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Classifications
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- 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
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- 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
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- General Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Plasma & Fusion (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides an application of a nitrogen-oxygen free radical containing compound as a stabilizer in preparation of a urine occult blood test paper, belonging to the field of urine analysis and detection. The structural formula of the compound containing the nitroxide free radicals is shown as a formula 1, the compound containing the nitroxide free radicals is a non-enzymatic substance and has nitroxide free radicals, and the compound is used as a stabilizer in preparation of the urine occult blood test paper.
Description
Technical Field
The invention belongs to the field of urine analysis and detection, and particularly relates to an application of a nitrogen-oxygen free radical compound as a stabilizer in preparation of a urine occult blood test paper.
Background
Urinalysis is one of routine tests in medical laboratories, plays an important role in medical tests, and has important guidance for diagnosis and treatment of related diseases. The urine analysis can perform qualitative and quantitative analysis on various components in the urine, including a plurality of items such as urobilinogen, bilirubin, ketone bodies, blood, protein, nitrite, leucocyte, glucose, creatinine, calcium, specific gravity, pH value, ascorbic acid and the like, wherein the detection of the occult blood in the urine is one of the main detection items of the urine analysis. Urine hemoglobin (red blood cell) detection is one of the important test indicators for diagnosing urinary system diseases, particularly kidney diseases. The appearance of red blood cells in urine is an early signal reflecting pathological changes of the urinary system, which means the formation of hematuria, and clinically, the red blood cells are divided into macroscopic hematuria and microscopic hematuria according to the number of the red blood cells. The bleeding position can be classified into renal hematuria and non-renal hematuria. When more than 0.1% of blood is mixed in urine, the urine is characterized by visible hematuria, and the amount of blood below the amount can be proved only by occult blood reaction or urinary sediment microscopic examination. Hematuria is commonly seen in urinary tract inflammation (acute nephritis, renal tuberculosis, urethritis, etc.), tuberculosis, tumor, etc. Hemoglobinuria is manifested by paroxysmal hemoglobinuria, and also by various cases of poisoning, infection, streptococcal septicemia, malaria (black water heat), burn, hemolytic blood transfusion reaction, etc.
The urine erythrocyte detection method mainly adopts the methods of microscopic examination, dry chemical urine analyzer, flow cytometry urine analyzer, microscope type urine analyzer and the like for detection. The dry chemical detection is to perform qualitative and semi-quantitative determination on the sample through the color change, and compared with other methods, the dry chemical detection has the advantages of simple operation, low cost and high speed. The main principle of dry chemical detection is to use a peroxidase method, only the substrate peroxide and the chromogen are different, but the dry chemical test paper method has the defects of low stability, poor VC interference resistance and the like.
Disclosure of Invention
The invention aims to solve the problems of interference of a large amount of ascorbic acid and poor stability of test paper in the conventional dry chemical test paper method for detecting the occult blood of urine, and provides an application of a nitrogen-oxygen free radical compound as a stabilizer in preparation of the occult blood test paper.
In order to achieve the above object, the present invention provides the following technical solutions.
The invention provides an application of a compound containing nitroxide free radicals as a stabilizer in preparation of a urine occult blood test paper, wherein the structural formula of the compound containing nitroxide free radicals is shown as a formula 1:
in the formula 1, R1 is-H, -OH, -NH2or-COOH.
Preferably, the application comprises: a method for preparing a test paper for detecting occult blood in urine, which comprises a substrate and a filter paper fixed on the substrate, wherein the filter paper is obtained by immersing the substrate in a soaking solution, and comprises the following steps:
the method comprises the following steps: completely soaking filter paper in the soaking solution A, and drying at 75-85 deg.C for 20-25 min;
the immersion liquid A comprises a buffer solution, a first surfactant and a stabilizer;
step two: completely soaking the filter paper dried in the step one into the immersion liquid B, and drying at 75-85 ℃ for 10-15 minutes;
the immersion liquid B comprises a solvent, a second surfactant, a first substrate, a second substrate, a sensitizer and a dye;
step three: and D, shearing the dried filter paper in the step two, combining and fixing the cut filter paper with a substrate, and rolling and cutting the cut filter paper to prepare the test paper for detecting the occult blood in the urine.
Preferably, the buffer preferably comprises a citrate buffer or a phosphate buffer.
Preferably, the concentration of the stabilizer is 0.1% to 1%.
Preferably, the solvent comprises ethanol or N-N-dimethylformamide.
Preferably, the first surfactant and the second surfactant respectively comprise one or a mixture of more of polyvinylpyrrolidone, sodium dodecyl sulfate, sodium dodecyl benzene sulfonate or tween 20, and the concentration of the first surfactant is 2% -8% and the concentration of the second surfactant is 1% -4%.
Preferably, the first substrate comprises cumene hydroperoxide or diisopropylbenzene hydroperoxide, and the concentration of the first substrate is 0.5-1.5%;
preferably, the second substrate is tetramethylbenzidine and the concentration of the second substrate is 0.5 to 1.5%.
Preferably, the sensitizer comprises 6-methoxyquinoline or 8-methoxyquinoline, and the concentration of the sensitizer is 0.5-1.5%.
Preferably, the dye includes orange G and methyl orange, and the concentration of the dye is 0.005% -0.04%.
The invention has the advantages of
The invention provides an application of a compound containing nitroxide free radicals as a stabilizer in preparation of a urine occult blood test paper, wherein the structural formula of the compound containing nitroxide free radicals is shown as a formula 1. The compound containing nitroxide free radicals is a non-enzymatic substance and has nitroxide free radicals, and is applied to preparation of the urine occult blood test paper as a stabilizer, so that on one hand, interference of VC on test paper detection is eliminated through the action of the nitroxide free radicals and VC, the sensitivity and accuracy of the test paper are improved, and on the other hand, the high-temperature stability and the normal-temperature storage stability of the test paper are improved through the improvement of the sensitivity of the test paper.
Detailed Description
The invention provides an application of a compound containing nitroxide free radicals as a stabilizer in preparation of a urine occult blood test paper, wherein the structural formula of the compound containing nitroxide free radicals is shown as a formula 1:
in the formula 1, R1 is-H, -OH, -NH2or-COOH, preferably-OH.
According to the invention, said application comprises: a method for preparing a test paper for detecting occult blood in urine, which comprises a substrate and a filter paper fixed on the substrate, wherein the filter paper is obtained by immersing the substrate in a soaking solution, and comprises the following steps:
the method comprises the following steps: completely soaking filter paper in the soaking solution A, and drying at 75-85 deg.C for 20-25 min;
the immersion liquid A comprises a buffer solution, a first surfactant and a stabilizer;
the buffer solution preferably comprises a citric acid buffer solution or a phosphate buffer solution, more preferably a citric acid buffer solution, and the citric acid buffer solution comprises citric acid and sodium citrate; the phosphate buffer preferably comprises potassium dihydrogen phosphate and disodium hydrogen phosphate; the concentration of the buffer is preferably 20% to 80%, more preferably 40% to 60%.
The first surfactant preferably comprises: one or more of polyvinylpyrrolidone, sodium dodecyl sulfate, sodium dodecyl benzene sulfonate and tween 20, more preferably polyvinylpyrrolidone, and the concentration of the first surfactant is preferably 2% -8%.
The concentration of the stabilizer is preferably 0.1% to 1%, more preferably 0.4% to 0.6%.
Step two: completely soaking the filter paper dried in the step one into the immersion liquid B, and drying at 75-85 ℃ for 10-15 minutes;
the immersion liquid B comprises a solvent, a second surfactant, a first substrate, a second substrate, a sensitizer and a dye;
the solvent preferably comprises ethanol or N-dimethylformamide, more preferably ethanol.
The second surfactant preferably comprises one or a mixture of more of polyvinylpyrrolidone, sodium dodecyl sulfate, sodium dodecyl benzene sulfonate or tween 20, more preferably polyvinylpyrrolidone, and the concentration of the second surfactant is preferably 1% -4%.
The first substrate preferably comprises cumene hydroperoxide or diisopropylbenzene hydroperoxide, and the concentration of the first substrate is preferably 0.5-1.5%.
The second substrate is tetramethyl benzidine, and the concentration of the second substrate is preferably 0.5-1.5%.
The sensitizer preferably comprises 6-methoxyquinoline or 8-methoxyquinoline, more preferably 6-methoxyquinoline, preferably in a concentration of 0.5-1.5%.
The dye preferably comprises orange G and methyl orange, and the concentration of the dye is preferably 0.005% -0.04%. Wherein, the concentration of orange G is preferably 0.01-0.04%, and the concentration of methyl orange is preferably 0.005-0.015%.
Step three: and D, shearing the dried filter paper in the step two, combining and fixing the cut filter paper with a substrate, and rolling and cutting the cut filter paper to prepare the test paper for detecting the occult blood in the urine.
According to the invention, in the preparation process of the test paper, the environmental temperature requirement is as follows: 18-28 ℃, preferably 23-25 ℃, ambient humidity requirement: 10-40%, preferably 20-30%.
The test paper anti-vitamin C interference performance verification method comprises the following steps: 10 cells/mu L of the standard liquid of the occult blood in urine is prepared, 5.6mmol/L of ascorbic acid is added, test strips are tested, and the test strips are read by an eye test method and a urine analyzer, and the result is not negative.
The test paper stability verification method comprises the following steps: the prepared test paper is respectively placed at room temperature for 36 months and 50 ℃ for 45 days, standard solutions with different concentration gradients are prepared for testing, and the test result accords with a measured value corresponding to the gradient of the standard solution.
The present invention is further illustrated by the following specific examples, which are provided for the purpose of illustration only and are not to be construed as limiting the general technical solutions of the present invention.
The stabilizer in the following examples is the nitroxide radical containing compound of formula 1 of the present invention, wherein the R1 group is-H and is named stabilizer 1, the R1 group is-OH and is named stabilizer 2, the R1 group is-NH 2 and is named stabilizer 3, and the R1 group is-COOH and is named stabilizer 4.
Example 1
(1) Preparing an immersion liquid A:
purifying water: 1000ml
Sodium citrate: 58.8g
Citric acid: 12.6g
Polyvinylpyrrolidone K30: 50g
Stabilizer 1: 1.5g
Sodium lauryl sulfate: 1.5g
Adding the above materials into purified water, and dissolving to obtain extract A.
(2) Preparing an immersion liquid B:
ethanol: 1000ml
Orange G0.23G
Methyl orange: 0.07g
Tetramethyl benzidine: 5g
Polyvinylpyrrolidone K30: 20g of
Cumene hydroperoxide: 5ml of
5.8ml of 8-methoxyquinoline
Adding above materials into ethanol in sequence, and dissolving to obtain extract B.
(3) Preparing test paper:
the method comprises the following steps: completely soaking filter paper in the soaking solution A, and drying at 80 deg.C for 25 min;
step two: completely immersing the filter paper dried in the step one into the immersion liquid B, and drying at 75 ℃ for 15 minutes;
step three: and D, shearing the dried filter paper in the step two, combining and fixing the cut filter paper with a substrate, and rolling and cutting the cut filter paper to prepare the test paper 1 for detecting the occult blood in the urine.
Example 2
(1) Preparing an immersion liquid A:
purifying water: 1000ml
Potassium dihydrogen phosphate: 68g of
Disodium hydrogen phosphate: 21.3g
Polyvinylpyrrolidone K30: 50g
Stabilizer 2: 5g
Sodium lauryl sulfate: 1.5g
Adding the above materials into purified water, and dissolving to obtain extract A.
(2) Preparing an immersion liquid B:
ethanol: 1000ml
Orange G0.23G
Methyl orange: 0.07g
Tetramethyl benzidine: 5g
Polyvinylpyrrolidone K30: 20g of
Cumene hydroperoxide: 5ml of
5.8ml of 6-methoxyquinoline
Adding above materials into ethanol in sequence, and dissolving to obtain extract B.
(3) Preparing test paper: test paper for occult blood 2 was prepared according to the preparation method of example 1.
Example 3
(1) Preparing an immersion liquid A:
purifying water: 1000ml
Potassium dihydrogen phosphate: 68g of
Disodium hydrogen phosphate: 21.3g
Polyvinylpyrrolidone K30: 50g
Stabilizer 3: 3.5g
Adding the above materials into purified water, and dissolving to obtain extract A.
(2) Preparing an immersion liquid B:
ethanol: 1000ml
Orange G0.23G
Methyl orange: 0.07g
Tetramethyl benzidine: 5g
Polyvinylpyrrolidone K30: 20g of
Cumene hydroperoxide: 5ml of
5.8ml of 8-methoxyquinoline
Adding above materials into ethanol in sequence, and dissolving to obtain extract B.
(3) Preparing test paper: a test strip for detecting occult blood 3 was prepared by the preparation method of example 1.
Example 4
(1) Preparing an immersion liquid A:
purifying water: 1000ml
Potassium dihydrogen phosphate: 68g of
Disodium hydrogen phosphate: 21.3g
Polyvinylpyrrolidone K30: 50g
Stabilizer 4: 4.5g
Sodium lauryl sulfate: 1.5g
Adding the above materials into purified water, and dissolving to obtain extract A.
(2) Preparing an immersion liquid B:
ethanol: N-N-dimethylformamide
Orange G0.23G
Methyl orange: 0.07g
Tetramethyl benzidine: 5g
Polyvinylpyrrolidone K30: 20g of
Cumene hydroperoxide: 5ml 6-Methoxyquinoline 5.8ml
Adding above materials into ethanol in sequence, and dissolving to obtain extract B.
(3) Preparing test paper: a test strip 4 for detecting occult blood in urine was prepared by the preparation method of example 1.
Comparative example 1
(1) Preparing an immersion liquid A:
purifying water: 1000ml
Potassium dihydrogen phosphate: 68g of
Disodium hydrogen phosphate: 21.3g
Polyvinylpyrrolidone K30: 50g
Sodium lauryl sulfate: 1.5g
Adding the above materials into purified water, and dissolving to obtain extract A.
(2) Preparing an immersion liquid B:
ethanol: N-N-dimethylformamide
Orange G0.23G
Methyl orange: 0.07g
Tetramethyl benzidine: 5g
Polyvinylpyrrolidone K30: 20g of
Cumene hydroperoxide: 5ml of
5.8ml of 6-methoxyquinoline
Adding above materials into ethanol in sequence, and dissolving to obtain extract B.
(3) Preparing test paper: a test strip for detecting occult blood in urine 5 was prepared by the preparation method of example 1.
Example 5
The results of measurement of the occult blood test strips obtained in examples 1 to 4 and comparative example 1 on 10cells/ul of an occult blood standard solution to which 5.6mmol/L ascorbic acid was added and interpretation by a urine analyzer are shown in Table 1.
TABLE 1
| Test paper number | Qualitative results | Semi-quantitative results |
| Test paper 1 | ± | 10cells/ul |
| Test paper 2 | ± | 10cells/ul |
| Test paper 3 | ± | 10cells/ul |
| Test paper 4 | ± | 10cells/ul |
| Test paper 5 | Neg | 0cells/ul |
Table 1 shows that the test strips obtained in examples 1 to 4 have a better VC interference resistance than the test strip obtained in comparative example 1.
Example 6
The urine occult blood test strips obtained in examples 1 to 4 and comparative example 1 were stored in a sealed and dark environment at 20 to 25 ℃ for 36 months, and then the test strips were used to measure occult blood standard solutions of 10cells/ul, 25cells/ul, 80cells/ul and 200cells/ul, and the results were interpreted by a urine analyzer and are shown in table 2.
TABLE 2
Table 2 shows that the test paper obtained in examples 1 to 4 is superior to the test paper obtained in comparative example 1 in long-term stability.
Example 7
The urine occult blood test strips obtained in examples 1 to 4 and comparative example 1 were stored in a sealed and dark environment at 50 ℃ for 45 days, and then the test strips were used to measure occult blood standard solutions of 10cells/ul, 25cells/ul, 80cells/ul and 200cells/ul, and the results were interpreted by a urine analyzer and are shown in table 3.
TABLE 3
Table 3 shows that the test paper obtained in examples 1 to 4 is superior in high temperature stability to the test paper obtained in comparative example 1.
Claims (10)
1. The application of the nitrogen-oxygen free radical compound as a stabilizer in the preparation of the urine occult blood test paper is characterized in that the structural formula of the nitrogen-oxygen free radical compound is shown as the formula 1:
in the formula 1, R1 is-H, -OH, -NH2or-COOH.
2. The application according to claim 1, comprising: a method for preparing a test paper for detecting occult blood in urine, which comprises a substrate and a filter paper fixed on the substrate, wherein the filter paper is obtained by immersing the substrate in a soaking solution, and comprises the following steps:
the method comprises the following steps: completely soaking filter paper in the soaking solution A, and drying at 75-85 deg.C for 20-25 min;
the immersion liquid A comprises a buffer solution, a first surfactant and a stabilizer;
step two: completely soaking the filter paper dried in the step one into the immersion liquid B, and drying at 75-85 ℃ for 10-15 minutes;
the immersion liquid B comprises a solvent, a second surfactant, a first substrate, a second substrate, a sensitizer and a dye;
step three: and D, shearing the dried filter paper in the step two, combining and fixing the cut filter paper with a substrate, and rolling and cutting the cut filter paper to prepare the test paper for detecting the occult blood in the urine.
3. Use according to claim 2, wherein the buffer preferably comprises a citrate buffer or a phosphate buffer.
4. Use according to claim 2, wherein the concentration of the stabilizer is between 0.1% and 1%.
5. The use according to claim 2, wherein the solvent comprises ethanol or N-dimethylformamide.
6. The application of claim 2, wherein the first surfactant and the second surfactant respectively comprise one or more of polyvinylpyrrolidone, sodium dodecyl sulfate, sodium dodecyl benzene sulfonate or tween 20, the concentration of the first surfactant is 2% -8%, and the concentration of the second surfactant is 1% -4%.
7. The use according to claim 2, wherein the first substrate comprises cumene hydroperoxide or diisopropylbenzene hydroperoxide, and the concentration of the first substrate is 0.5-1.5%.
8. The use according to claim 2, wherein said second substrate is tetramethylbenzidine and the concentration of the second substrate is 0.5-1.5%.
9. Use according to claim 2, wherein the sensitizer comprises 6-methoxyquinoline or 8-methoxyquinoline, and the concentration of the sensitizer is 0.5-1.5%.
10. Use according to claim 2, wherein the dyes comprise orange G and methyl orange, the concentration of the dye being between 0.005% and 0.04%.
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