CN110006884A - A kind of urine sulfhydryl compound mercury-free test strip and preparation method - Google Patents
A kind of urine sulfhydryl compound mercury-free test strip and preparation method Download PDFInfo
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- CN110006884A CN110006884A CN201910277662.6A CN201910277662A CN110006884A CN 110006884 A CN110006884 A CN 110006884A CN 201910277662 A CN201910277662 A CN 201910277662A CN 110006884 A CN110006884 A CN 110006884A
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- 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
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Abstract
The invention belongs to in-vitro diagnosis field technical fields, and in particular to urine sulfhydryl compound mercury-free test strip and preparation method.The test strips include the DTNB and EDTA on filter paper and the load filter paper.The preparation method includes the following steps: to configure buffer, the component for needing to be supported on filter paper is dissolved in the buffer, the mixed solution is applied on the filter paper by the method impregnated, is cut into small pieces after its drying, and is pasted on plastic strip stand-by.Technical solution provided by the invention uses DTNB as sulfydryl color developing agent and is successfully improved to the urine detection reagent of test strips formula, wherein no longer containing heavy metal ion such as mercury, also without containing corrosive substances such as trichloroacetic acids, improve safety in utilization, in addition detection rates are greatly speeded up, it was foreshortened within 5 minutes from least 15 minutes, to solve the problems, such as test strips detection to developing time requirement.
Description
Technical field
The invention belongs to in-vitro diagnosis field technical fields, and in particular to a kind of urine sulfhydryl compound is without mercury investigating test paper
Item and preparation method.
Background technique
In the malignant tumour that the whole world endangers women's health, cervix cancer is listed in third position, according to statistics, 2008 years
It is new to send out ratio shared by cervix cancer as 9% (529800) in global all new cancers of women, it is only second to breast cancer and knot
The intestines carcinoma of the rectum.And in the lethal patient of female cancer, it is 8% (275100) that female cancer patients, which die of cervical carcinoma, position
Occupy the 4th.And the patient for dying of cervix cancer has about 230,000 people to belong to developing country in the middle, accounts about total number of persons
83%.It is annual according to statistics newly to send out Patients with Cervical Cancer about example 130000 in China.
Thiol molecule is irreversible during the entire process of precancerous lesions of uterine cervix and cervical cancer pathogenesis falls off and body is discharged through urine
Outside.Therefore the sulfhydryl compound in detection urine can be used for the screening and auxiliary diagnosis of cervical carcinoma.
Detection, such as Chinese patent are mainly realized using phosphotungstic acid method in sulfydryl detection technique in existing urine:
Detection reagent of sulfhydryl compound and preparation method thereof and purposes, CN105115966A urine in a kind of urine of CN107576652A
Middle thiol compound detection kit and preparation method and application and a kind of cervical carcinoma urine detection reagent of CN106468715A
And its preparation method and application, this method has following defect: 1) all containing mercury ion, this manual operation sample-adding in detection method
Product for, the presence of mercury has great toxicity, in addition, the heavy metal ion such as mercury, nickel also can be to environment in detection waste liquid
It impacts, needs special processing；2) there are the corrosive goods confrontation operators such as trichloroacetic acid also to have to a certain degree in reagent
Influence, 3) mercury ion is easy to be influenced by the high concentration uric acid in urine in reagent, generate and greatly interfere, cause false negative knot
Fruit, 4) at present detection kit be using ampoule bottle flexible package punch, then with urine hybrid detection, this packaging volume is big,
Transport is inconvenient, waste is more.
Summary of the invention
The present invention provides a kind of urine sulfhydryl compound mercury-free test strip and preparation methods, urinate at present to solve
A series of problems existing for liquid sulfhydryl compound detection reagent.
In order to solve the above-mentioned technical problem, the technical scheme is that urine sulfhydryl compound mercury-free detection examination
Paper slip comprising filter paper and DTNB (5,5'- bis- thiobis (2- nitrobenzoic acid)) and EDTA (second two on the load filter paper
Amine tetraacethyl disodium), -2,4 pentanediol of 2- methyl, silico-tungstic acid and surfactant.
Colourless bis- thiobis of 5,5'- (2- nitrobenzoic acid) (5,5'-Dithiobis- (2-nitrobenzoic
Acid, DTNB) -2 nitrobenzoic acid of 5- sulfydryl for being transformed into yellow is reacted with sulfhydryl compound, can according to the yellow depth come into
The judgement of row sxemiquantitative.
Can sometimes there are the reducing substances such as albumen, vitamin C, uric acid in urine, the presence of these substances will affect
Detection, causes the result of false positive.2- methyl -2,4 pentanediol can effectively be dropped as protein precipitant, the oxidisability of silico-tungstic acid
Low these substance brings adverse effect, and in order to cooperate test strips to produce and use, select it is safer, without special odor
With stable 2- methyl -2,4 pentanediol, in addition the acidic environment of silico-tungstic acid can also assist DTNB more stable.
Optionally, the surfactant is selected from Triton X-100 (Triton X-100) and/or poly- second two
Alcohol.DTNB as color developing agent there is also a storage stabilities the problem of, by adding suitable Triton X- in buffer
100 can solve the problems, such as its commercialization with/polyethylene glycol, the test strips of configuration with long-term preservation.
Optionally, brilliant green dyestuff is also loaded on the filter paper.Brilliant green addition so that the color change of the test strips more
Obviously, it is more advantageous to naked eyes interpretation.
The present invention also provides the preparation methods of urine sulfhydryl compound mercury-free test strip comprising following steps:
Buffer is configured, the component for needing to be supported on filter paper is dissolved in the buffer, by the mixed solution by impregnating
Method is applied on the filter paper, is cut into small pieces after its drying, and is pasted on plastic strip stand-by.
Optionally, the concentration of DTNB is 5-8g/L in the mixed solution.
Optionally, the concentration of EDTA is 200mg-1000mg/L in the mixed solution.
Optionally, the concentration of 2- methyl -2,4 pentanediol is 10-20g/L in the mixed solution.
Optionally, the concentration of silico-tungstic acid is 5-10g/L in the mixed solution.
Optionally, the buffer is phosphate aqueous solution, pH value 7.0-10.0.
Optionally, the phosphate is sodium dihydrogen phosphate and disodium hydrogen phosphate dodecahydrate.
The present invention also provides a kind of kits of urine sulfhydryl compound mercury-free detection comprising above-mentioned urine sulfhydrylation
Close object mercury-free test strip and standard color comparison card.
The determination of standard color comparison card is predefined using normal concentration test sample.
Technical solution provided by the invention is qualitative to sulfydryl progress in urine as sulfydryl color developing agent using DTNB, and successfully
It is improved to the urine detection reagent of test strips formula, detection reagent component is environmental-friendly, wherein no longer containing heavy metals such as mercury
Ion, the waste after use not will cause environmental pollution, also no longer containing corrosive substances such as trichloroacetic acids, improve using peace
Quan Xing, in addition detection rates are greatly speeded up, and are foreshortened within 5 minutes from least 15 minutes, to solve test strips detection pair
The problem of developing time requires；In conclusion the test strip provided has strong operability, the spy easy to spread used
Point, therefore wide market.
Specific embodiment
In order to make it easy to understand, illustrating the urine sulfhydryl compound mercury-free test strip and its system below with reference to embodiment
Preparation Method, it should be appreciated that these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention.
All raw material is the general chemistry preparation for analyzing pure rank, and preparation process also all carries out under room temperature, normal pressure.
Embodiment 1
1) it configures buffer: weighing 1.5g sodium dihydrogen phosphate and 70.22g disodium hydrogen phosphate dodecahydrate, water is added to be settled to
1L, the pH value of the buffer are 7.8.
2) bis- thiobis of 5,5'- (2- nitrobenzoic acid) (DNTB) (6.14g/L) of 6.14g, the ethylenediamine of 0.32g are weighed
Tetraacethyl disodium (EDTA) (320mg/L), -2,4 pentanediol of 2- methyl (15g/L) of 10g, 10g silico-tungstic acid (10g/L), 14g
Polyethylene glycol and 1.7g Triton X-100, will said components be added configuration buffer in；
3) component to configure in step 2) is completely dissolved, and filter paper is impregnated in the solution, is cut into small pieces after taking out drying,
And with sticking double faced adhesive tape on plastic strip.
Embodiment 2
1) it configures buffer: weighing 1.5g sodium dihydrogen phosphate and 70.22g disodium hydrogen phosphate dodecahydrate, water is added to be settled to
1L, the pH value of the buffer are 7.8.
2) bis- thiobis of 5,5'- (2- nitrobenzoic acid) (DNTB) (6.14g/L) of 6.14g, the ethylenediamine of 0.32g are weighed
Tetraacethyl disodium (EDTA) (320mg/L), -2,4 pentanediol of 2- methyl (15g/L) of 15g, 6g silico-tungstic acid (10g/L), 14g
Polyethylene glycol, 1.7g Triton X-100,2mL brilliant green (0.1%) coloring agent, by said components be added configuration buffer
In；
3) component to configure in step 2) is completely dissolved, and filter paper is impregnated in the solution, is cut into small pieces after taking out drying,
And with sticking double faced adhesive tape on plastic strip.
3 standard color comparison card of embodiment makes and uses
1) 1mmol/L, 0.5mmol/L, 0.1mmol/L, cysteine (Cys) mark of 0.04mmol/L, 0mmol/L are configured
Quasi- solution；
2) 5 are immersed in the cysteine standard solution of each concentration of step 1) configuration and prepare stand-by test strips, then
It takes out；
3) colorimetric card is prepared according to the colour developing situation after 2 minutes；
4) it in use, test strips immersion sample to be tested is then taken out, will be added in reagent bottle according to sample to be tested, 5 points
Clock discoloration stablize after compared with standard color comparison card, with concentration corresponding to the immediate standard color comparison card of test specimens color be to
The concentration for surveying liquid, if estimating reading thiol concentration between two color ranges.
4 compliance test result test of embodiment
The aqueous cystein solution for configuring four various concentrations, is followed successively by 0mmol/L, 0.04mmol/L from low to high,
Test strips prepared by Examples 1 and 2 are successively immersed half Guang ammonia of various concentration by 0.1mmol/L, 0.5mmol/L and 1mmol/L
In aqueous acid, discoloration is observed after five minutes, and the results are shown in Table 1.
Table 1
By table 1 it can be found that occurring after test sample of the test strips provided by the invention immersion containing cysteine
Apparent color change, content is higher, and discoloration is higher.
4.2 stability experiment
The reagent of Examples 1 and 2 is saved one week in 50 DEG C of baking ovens, is then tested respectively, Examples 1 and 2 are equal
It can normally develop the color.
4.3 interference--free experiments
Configure various concentration cysteine solution (0mmol/L, 0.04mmol/L, 0.1mmol/L, 0.5mmol/L and
1mmol/L), it is separately added into sodium chloride (NaCl) 1%, uric acid (UA) 2mol/L, vitamin C (Vc) 0.5mg/ml, specially using this
Embodiment 2 can normally develop the color in benefit, and test result is as shown in table 2.
Table 2
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations.Although
Present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: it still may be used
To modify to technical solution documented by previous embodiment, or some or all of the technical features are equal
Replacement, and these modifications or substitutions, the model for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution
It encloses.
Claims (9)
1. a kind of urine sulfhydryl compound mercury-free test strip, which is characterized in that including filter paper and load on the filter paper
- 2,4 pentanediol of DTNB, EDTA, 2- methyl, silico-tungstic acid and surfactant.
2. urine sulfhydryl compound mercury-free test strip according to claim 1, which is characterized in that the surfactant
For Triton X-100 and/or polyethylene glycol.
3. urine sulfhydryl compound mercury-free test strip according to claim 1, which is characterized in that also negative on the filter paper
Brilliant green dyestuff is carried.
4. the preparation method of any urine sulfhydryl compound mercury-free test strip of claim 1-3, which is characterized in that packet
Include following steps: the component for needing to be supported on filter paper is dissolved in the buffer, by the mixed solution by configuration buffer
It is applied on the filter paper by the method for immersion, is cut into small pieces after its drying, and be pasted on plastic strip stand-by.
5. preparation method according to claim 4, which is characterized in that the concentration of DTNB is 5-8g/L in the mixed solution.
6. preparation method according to claim 4, which is characterized in that the concentration of EDTA is 200mg- in the mixed solution
1000mg/L, preferably 300-500mg/L.
7. preparation method according to claim 4, which is characterized in that the concentration of silico-tungstic acid is 5-10g/ in the mixed solution
L。
8. preparation method according to claim 4, which is characterized in that 2- methyl -2,4 pentanediol is dense in the mixed solution
Degree is 10-20g/L.
9. a kind of kit of urine sulfhydryl compound mercury-free detection, which is characterized in that including any urine of claim 1-3
Liquid sulfhydryl compound mercury-free test strip and standard color comparison card.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910277662.6A CN110006884A (en) | 2019-04-08 | 2019-04-08 | A kind of urine sulfhydryl compound mercury-free test strip and preparation method |
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| CN201910277662.6A CN110006884A (en) | 2019-04-08 | 2019-04-08 | A kind of urine sulfhydryl compound mercury-free test strip and preparation method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2021032103A1 (en) * | 2019-08-19 | 2021-02-25 | 杭州爱光医疗器械有限公司 | Sulfydryl compound detection reagent, test paper, reagent kit, test paper box and preparation thereof |
| CN115290904A (en) * | 2022-10-09 | 2022-11-04 | 山东三医生物技术有限公司 | In-situ hybridization HPV detection test strip, kit and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021032103A1 (en) * | 2019-08-19 | 2021-02-25 | 杭州爱光医疗器械有限公司 | Sulfydryl compound detection reagent, test paper, reagent kit, test paper box and preparation thereof |
| CN115290904A (en) * | 2022-10-09 | 2022-11-04 | 山东三医生物技术有限公司 | In-situ hybridization HPV detection test strip, kit and preparation method thereof |
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Application publication date: 20190712 |