CN115372605A - Urine sulfhydryl compound detection reagent, detection test paper and preparation method of detection kit - Google Patents
Urine sulfhydryl compound detection reagent, detection test paper and preparation method of detection kit Download PDFInfo
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- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 109
- 238000001514 detection method Methods 0.000 title claims abstract description 62
- 210000002700 urine Anatomy 0.000 title claims abstract description 36
- -1 sulfhydryl compound Chemical class 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000000243 solution Substances 0.000 claims abstract description 33
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 32
- 229920002261 Corn starch Polymers 0.000 claims abstract description 25
- 239000008120 corn starch Substances 0.000 claims abstract description 25
- 239000008363 phosphate buffer Substances 0.000 claims abstract description 22
- 238000005303 weighing Methods 0.000 claims abstract description 22
- DHRLEVQXOMLTIM-UHFFFAOYSA-N phosphoric acid;trioxomolybdenum Chemical compound O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.OP(O)(O)=O DHRLEVQXOMLTIM-UHFFFAOYSA-N 0.000 claims abstract description 19
- JLKDVMWYMMLWTI-UHFFFAOYSA-M potassium iodate Chemical compound [K+].[O-]I(=O)=O JLKDVMWYMMLWTI-UHFFFAOYSA-M 0.000 claims abstract description 19
- 229940093930 potassium iodate Drugs 0.000 claims abstract description 19
- 235000006666 potassium iodate Nutrition 0.000 claims abstract description 19
- 239000001230 potassium iodate Substances 0.000 claims abstract description 19
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 18
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 18
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 17
- 239000012498 ultrapure water Substances 0.000 claims abstract description 17
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 16
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 14
- PGLTVOMIXTUURA-UHFFFAOYSA-N iodoacetamide Chemical compound NC(=O)CI PGLTVOMIXTUURA-UHFFFAOYSA-N 0.000 claims abstract description 12
- 238000009835 boiling Methods 0.000 claims abstract description 7
- 239000012916 chromogenic reagent Substances 0.000 claims abstract description 7
- 239000007864 aqueous solution Substances 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 10
- 238000002791 soaking Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 238000005520 cutting process Methods 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 5
- 239000008055 phosphate buffer solution Substances 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 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
- 241000701806 Human papillomavirus Species 0.000 description 8
- 125000003396 thiol group Chemical group [H]S* 0.000 description 8
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 4
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 4
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 4
- 229930003268 Vitamin C Natural products 0.000 description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- 229940116269 uric acid Drugs 0.000 description 4
- 235000019154 vitamin C Nutrition 0.000 description 4
- 239000011718 vitamin C Substances 0.000 description 4
- 208000009608 Papillomavirus Infections Diseases 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000700605 Viruses Species 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 208000003154 papilloma Diseases 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 210000004392 genitalia Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 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
- 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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- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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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
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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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
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Abstract
The invention discloses a urine sulfhydryl compound detection reagent, which consists of three reagents, namely a phosphate buffer reagent, a chromogenic reagent and a contrast reagent; the preparation method of the phosphate buffer reagent comprises the following steps: respectively preparing 0.1 +/-0.05 mol/L phosphoric acid solution and potassium dihydrogen phosphate solution, and adjusting the pH value of the potassium dihydrogen phosphate solution to 1.9-2.5 by using the phosphoric acid solution to obtain the phosphate buffer reagent, wherein the preparation method of the chromogenic reagent comprises the following steps: weighing 1-10g of corn starch, adding the corn starch into about 800mL of ultrapure water, boiling until the corn starch is completely dissolved, after the temperature is reduced to room temperature, weighing 1-20g of phosphomolybdic acid and 10-20g of potassium iodate, adding the phosphomolybdic acid and the potassium iodate to be completely dissolved, and metering the volume to 1000mL to obtain a color reagent, wherein the color reagent is an aqueous solution of the phosphomolybdic acid, the potassium iodate and the corn starch, and the contrast reagent contains an aqueous solution of iodoacetamide.
Description
Technical Field
The invention relates to the technical field of rapid in-vitro diagnosis, in particular to a urine sulfhydryl compound detection reagent, a detection reagent, detection test paper and a preparation method of a detection kit.
Background
Human Papilloma Virus (HPV) belongs to the genus of Papilloma vacuolatum Virus A of the family of papovaviridae, and is a small-molecular, non-enveloped, circular double-stranded DNA Virus. HPV infects humans by direct or indirect contact with contaminated articles or sexually transmitted. HPV viruses can infect human skin and mucosal epithelial cells, causing various papillomas or warts of human skin and proliferative lesions of the genital epithelium.
The HPV virus is continuously infected and can cause cervical cancer, and in 2017, partial subtypes of the human papilloma virus are listed in a category 3 carcinogen list published by the international cancer research institution of the world health organization.
The existing HPV infection detection method usually needs to collect the living tissue of a detected person, needs to be carried out in a formal hospital, is detected by professionals, and is not suitable for routine examination at home. The procedure of admission examination brings certain psychological pressure to the testee, so that many people do not want to examine in advance, and the treatment time can be delayed. Therefore, a rapid, convenient, efficient and safe detection means is imperative.
Researches prove that urine of HPV infected patients contains a large amount of sulfhydryl metabolites and urine of normal people contains few sulfhydryl metabolites, so that the content of the sulfhydryl metabolites in the urine can be used as an important index for whether an organism has HPV infection and cervical lesion, and the detection rate and treatment efficiency of HPV infection can be effectively improved by detecting the content of the sulfhydryl metabolites in the urine.
Disclosure of Invention
The invention aims to provide a urine sulfhydryl compound detection reagent, a detection reagent, detection test paper and a preparation method of a detection kit, and solves the problems.
In order to achieve the purpose, the invention is realized by the following technical scheme:
the invention provides a urine sulfhydryl compound detection reagent, which consists of three reagents, namely a phosphate buffer reagent, a chromogenic reagent and a contrast reagent.
Preferably, the preparation method of the phosphate buffer reagent comprises the following steps: respectively preparing 0.1 +/-0.05 mol/L phosphoric acid solution and potassium dihydrogen phosphate solution, and adjusting the pH value of the potassium dihydrogen phosphate solution to 1.9-2.5 by using the phosphoric acid solution to obtain a phosphate buffer reagent, wherein the concentration of phosphate ions in the phosphate buffer reagent is 0.05-0.15mol/L, the pH range is 1.9-2.5, and the color developing reagent has good color developing reaction in the region.
Preferably, the color reagent is aqueous solution of phosphomolybdic acid, potassium iodate and corn starch.
Preferably, the preparation method of the color reagent comprises the following steps: weighing 1-10g of corn starch, adding the corn starch into about 800mL of ultrapure water, boiling until the corn starch is completely dissolved, after the temperature is reduced to room temperature, weighing 1-20g of phosphomolybdic acid and 10-20g of potassium iodate, adding and completely dissolving, and metering to 1000mL to obtain a color reagent, wherein the concentration of phosphomolybdic acid in the color reagent is 0.1-2%, the concentration of potassium iodate is 1-2%, and the concentration of corn starch is 0.1-1%.
Preferably, the control reagent comprises an aqueous solution of iodoacetamide.
Preferably, the preparation method of the control reagent comprises the following steps: weighing 1-2g of iodoacetamide, dissolving in 800mL of ultrapure water, and diluting to 1000mL to obtain a contrast reagent, wherein the concentration of the iodoacetamide in the contrast reagent is 0.1-0.2%.
A preparation method of a urine sulfhydryl compound detection reagent is prepared by adopting the urine sulfhydryl compound detection reagent, and specifically comprises the following steps:
s1: respectively preparing 0.1 +/-0.05 mol/L phosphoric acid solution and potassium dihydrogen phosphate solution, and adjusting the pH value of the potassium dihydrogen phosphate solution to 1.9-2.5 by using the phosphoric acid solution to obtain a phosphate buffer reagent;
s2: weighing 1-10g of corn starch, adding the corn starch into about 800mL of ultrapure water, boiling until the corn starch is completely dissolved, after the temperature is reduced to room temperature, weighing 1-20g of phosphomolybdic acid and 10-20g of potassium iodate, adding the phosphomolybdic acid and the potassium iodate, completely dissolving, and carrying out constant volume treatment to 1000mL to obtain a color reagent;
s3: weighing 1-2g of iodoacetamide, dissolving in 800mL of ultrapure water, and diluting to 1000mL to obtain a contrast reagent;
s4: and (3) mixing the phosphate buffer reagent, the chromogenic reagent and the control reagent according to the proportion of 4.
A preparation method of urine sulfhydryl compound detection test paper is prepared by adopting the detection reagent prepared by the preparation method of the urine sulfhydryl compound detection reagent, and is characterized in that: the method comprises the following steps:
a: applying a detection reagent on the test paper by a soaking method, wherein the ultrasonic wave is carried out for more than 10 seconds during soaking;
b: and taking out the test paper after soaking, vacuumizing to dry the test paper, cutting the test paper into small pieces, and adhering the test paper to a plastic strip to obtain the test paper.
9. A method for preparing a urine thiol compound detection kit, which is prepared by using the detection reagent prepared by the urine thiol compound detection reagent preparation method of claim 7, and is characterized in that: the kit comprises a detection box, a detection strip, a drying sheet and a colorimetric strip, wherein the preparation method of the detection strip comprises the following steps:
(1): preparing a phosphate buffer reagent, a color development reagent and a control reagent according to claim 7 respectively;
(2): mixing a phosphate buffer reagent and a contrast reagent according to the proportion of 9;
(3): mixing a phosphate buffer solution and a color reagent according to the proportion of 5, soaking the test paper, drying, and cutting the test paper to obtain a test paper 2;
(4): connecting the test paper 1 and the test paper 2 with each other and sticking the test paper on a plastic strip to obtain a detection strip;
(5): and assembling the detection box, the detection strip, the drying sheet, the colorimetric strip and the colorimetric card to obtain the urine sulfydryl detection kit.
The invention has the beneficial effects that: the color reagent in the detection reagent can oxidize a sulfhydryl compound to generate a blue substance, the urine of a normal person also contains reducing substances such as vitamin C, uric acid and the like, the common color reagent can quickly develop color and turn blue when reacting with the vitamin C or the uric acid to cause false positive, the vitamin C and the uric acid preferentially react with the contrast reagent after the contrast reagent is added to avoid reaction with the color reagent, the urine of a patient infected by HPV contains a large amount of free sulfhydryl compounds, and the contrast reagent simultaneously reacts with the reducing substances in the urine to contain the vitamin C, the uric acid and the sulfhydryl compounds, although the contrast reagent also consumes a small amount of the sulfhydryl compounds, the residual sulfhydryl compounds can still react with the color reagent to develop color and turn blue to obtain a positive result.
Detailed Description
The following examples are provided to further illustrate embodiments of the present invention. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.
Example 1
Preparation of urine sulfhydryl compound detection reagent
Preparing a phosphate buffer solution: measuring 438ml of ultrapure water, and adding 3mL of 85% concentrated phosphoric acid to obtain 0.1mol/L phosphoric acid solution; weighing 13.6g of monopotassium phosphate, dissolving the monopotassium phosphate in ultrapure water, and fixing the volume to 1000mL to obtain 0.1mol/L potassium dihydrogen phosphate solution; the pH of the obtained potassium dihydrogen phosphate solution is adjusted to 2.0 by using 0.1mol/L phosphoric acid solution, and the phosphate buffer reagent with the concentration of 0.1mol/L is obtained.
Preparing a color developing reagent: weighing 5g of corn starch, adding the corn starch into about 800mL of ultrapure water, boiling until the corn starch is completely dissolved, after the temperature is reduced to room temperature, weighing 10g of phosphomolybdic acid and 10g of potassium iodate, adding the phosphomolybdic acid and the potassium iodate, completely dissolving, and carrying out constant volume treatment to 1000mL to obtain a color reagent;
preparing a contrast reagent; weighing 2g of iodoacetamide, dissolving the iodoacetamide in 800mL of ultrapure water, and diluting to 1000mL to obtain a contrast reagent;
the phosphate buffer reagent, the chromogenic reagent and the control reagent are mixed according to the proportion of 4.
Example 2
Preparation of urine sulfhydryl compound detection kit
Preparing a phosphate buffer solution: measuring 438ml of ultrapure water, and adding 3mL of 85% concentrated phosphoric acid to obtain 0.1mol/L phosphoric acid solution; weighing 13.6g of monopotassium phosphate, dissolving the monopotassium phosphate in ultrapure water, and fixing the volume to 1000mL to obtain 0.1mol/L potassium dihydrogen phosphate solution; the pH of the obtained potassium dihydrogen phosphate solution is adjusted to 2.2 by using 0.1mol/L phosphoric acid solution, and the phosphate buffer reagent with the concentration of 0.1mol/L is obtained.
Preparing a color developing reagent: weighing 5g of corn starch, adding the corn starch into about 800mL of ultrapure water, and boiling until the corn starch is completely dissolved; and after the temperature is reduced to room temperature, weighing 20g of phosphomolybdic acid and 10g of potassium iodate, adding the phosphomolybdic acid and the potassium iodate into the solution, completely dissolving the phosphomolybdic acid and the potassium iodate, and carrying out constant volume to 1000mL to obtain the color developing reagent.
Preparing a contrast reagent; weighing 2g of iodoacetamide, dissolving the iodoacetamide in 800mL of ultrapure water, and carrying out constant volume treatment to 1000mL to obtain the contrast reagent.
Mixing a phosphate buffer reagent and a contrast reagent according to the proportion of 9.
Mixing a phosphate buffer solution and a color reagent according to the proportion of 5, soaking the test paper, drying, and cutting the test paper to obtain a test paper 2;
connecting the test paper 1 and the test paper 2 with each other and sticking the test paper on a plastic strip for standby;
and assembling the detection box, the detection strip, the drying sheet, the colorimetric strip and the colorimetric card to obtain the urine sulfydryl detection kit.
Example 3
Preparation of Standard colorimetric cards
Preparing cysteine standard solution of 0mmol/L,0.04mmol/L,0.2mmol/L,1mmol/L,5mmol/L and 25 mmol/L;
respectively taking 100uL of the prepared cysteine standard solution, and respectively dripping the prepared cysteine standard solution into the prepared urine sulfhydryl compound detection kit;
after standing at room temperature for 10 minutes, a color comparison card was prepared according to the color development.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (9)
1. A urine sulfhydryl compound detection reagent, which is characterized in that: the detection reagent consists of three reagents, namely a phosphate buffer reagent, a chromogenic reagent and a contrast reagent.
2. The urine thiol compound detection reagent according to claim 1, characterized in that: the preparation method of the phosphate buffer reagent comprises the following steps: respectively preparing 0.1 +/-0.05 mol/L phosphoric acid solution and potassium dihydrogen phosphate solution, and adjusting the pH value of the potassium dihydrogen phosphate solution to 1.9-2.5 by using the phosphoric acid solution to obtain the phosphate buffer reagent.
3. The urine thiol compound detection reagent according to claim 1, characterized in that: the color reagent is water solution of phosphomolybdic acid, potassium iodate and corn starch.
4. The reagent for detecting a thiol compound in urine according to claim 1, wherein: the preparation method of the color reagent comprises the following steps: weighing 1-10g of corn starch, adding the corn starch into about 800mL of ultrapure water, boiling until the corn starch is completely dissolved, after the temperature is reduced to room temperature, weighing 1-20g of phosphomolybdic acid and 10-20g of potassium iodate, adding the phosphomolybdic acid and the potassium iodate into the solution, completely dissolving the phosphomolybdic acid and the potassium iodate, and carrying out constant volume treatment to 1000mL to obtain the color reagent.
5. The urine thiol compound detection reagent according to claim 1, characterized in that: the control reagent comprises an aqueous solution of iodoacetamide.
6. The urine thiol compound detection reagent according to claim 1, characterized in that: the preparation method of the contrast reagent comprises the following steps: weighing 1-2g of iodoacetamide, dissolving in 800mL of ultrapure water, and diluting to 1000mL to obtain the contrast reagent.
7. A method for producing a urine thiol compound detection reagent according to any one of claims 1 to 6, comprising: the method specifically comprises the following steps:
s1: respectively preparing 0.1 +/-0.05 mol/L phosphoric acid solution and potassium dihydrogen phosphate solution, and adjusting the pH value of the potassium dihydrogen phosphate solution to 1.9-2.5 by using the phosphoric acid solution to obtain a phosphate buffer reagent;
s2: weighing 1-10g of corn starch, adding the corn starch into about 800mL of ultrapure water, boiling until the corn starch is completely dissolved, after the temperature is reduced to room temperature, weighing 1-20g of phosphomolybdic acid and 10-20g of potassium iodate, adding the phosphomolybdic acid and the potassium iodate, completely dissolving, and performing constant volume treatment to 1000mL to obtain a color developing reagent;
s3: weighing 1-2g of iodoacetamide, dissolving in 800mL of ultrapure water, and diluting to 1000mL to obtain a contrast reagent;
s4: and (3) mixing the phosphate buffer reagent, the chromogenic reagent and the control reagent according to the proportion of 4.
8. A method for preparing a test paper for detecting urine thiol compounds, which is prepared by using the detection reagent prepared by the method for preparing a detection reagent for urine thiol compounds according to claim 7, and is characterized in that: the method comprises the following steps:
a: applying a detection reagent on the test paper by a soaking method, wherein the ultrasonic wave is carried out for more than 10 seconds during soaking;
b: and taking out the test paper after soaking, vacuumizing to dry the test paper, cutting the test paper into small pieces, and adhering the test paper to a plastic strip to obtain the test paper.
9. A method for preparing a urine thiol compound detection kit, which is prepared by using the detection reagent prepared by the urine thiol compound detection reagent preparation method of claim 7, and is characterized in that: the kit comprises a detection box, a detection strip, a drying sheet and a colorimetric strip, wherein the preparation method of the detection strip comprises the following steps:
(1): preparing a phosphate buffer reagent, a color development reagent and a control reagent according to claim 7 respectively;
(2): mixing a phosphate buffer reagent and a contrast reagent according to the proportion of 9;
(3): mixing a phosphate buffer solution and a color reagent according to the proportion of 5, soaking the test paper, drying, and cutting the test paper to obtain a test paper 2;
(4): connecting the test paper 1 and the test paper 2 with each other and sticking the test paper on a plastic strip to obtain a detection strip;
(5): and assembling the detection kit, the detection strip, the drying sheet, the colorimetric strip and the colorimetric card to obtain the urine sulfydryl detection kit.
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| CN110672593A (en) * | 2019-08-19 | 2020-01-10 | 杭州爱光医疗器械有限公司 | Sulfhydryl compound detection reagent, preparation method and kit thereof |
| CN114441523A (en) * | 2022-02-10 | 2022-05-06 | 梅宗泉 | Preparation method of kit for detecting sulfhydryl metabolites in urine |
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