CN102384971A - Spittle alcoholicity test strip and production method thereof - Google Patents
Spittle alcoholicity test strip and production method thereof Download PDFInfo
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- CN102384971A CN102384971A CN2011103148261A CN201110314826A CN102384971A CN 102384971 A CN102384971 A CN 102384971A CN 2011103148261 A CN2011103148261 A CN 2011103148261A CN 201110314826 A CN201110314826 A CN 201110314826A CN 102384971 A CN102384971 A CN 102384971A
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Abstract
The invention provides a spittle alcoholicity test strip and a production method thereof. The test strip comprises a substrate and a reaction test strip which is adhered to the surface at one side surface of the substrate; and the reaction test strip is dry filter paper which is soaked in a color-developing agent solution and a bi-enzymatic reaction liquid in sequence. The production method comprises the following steps of: soaking the filter paper in the color-developing agent solution and then drying the filter paper by means of being protected from light at a room temperature to obtain developing filter paper; and soaking the developing filter paper into the bi-enzymatic reaction liquid and then drying the developing filter paper by means of being protected from the light at the room temperature to obtain the reaction filter paper. The bi-enzymatic reaction liquid contains alcohol oxidase with the activity unit of 80 U/ml, peroxidase with the activity unit of 80 U/ml, bovine serum albumin which accounts for 1.6% of the total weight of the bi-enzymatic reaction liquid, trehalose which accounts for 1.5% of the total weight of the bi-enzymatic reaction liquid, and the balance of 0.3 M phosphate buffer with the pH value of 7.5. The test strip has the advantages of fastness and accuracy in detection, high sensitivity and good stability. Furthermore, the production method is simple, practical and very reliable.
Description
Technical field
The present invention relates to a kind of saliva alcohol content test-strips, specifically, related to a kind of saliva alcohol content test-strips and preparation method thereof, belong to the biosensor technology field.
Background technology
In recent years, along with developing rapidly of transportation, motor vehicle quantity increases with the personnel amount that has a driving license fast, and meanwhile, traffic hazard quantity is also increasing, wherein, and because of human pilot is drunk and the drunk traffic hazard that causes just accounts for 20%; Therefore, accurate detection driver's human body alcohol concentration receives the many concerns of People more and more.
Alcohol detection has important effect in medical jurisprudence and clinical examination, and detects blood alcohol concentration and can be used for diagnosing acute alcoholism and some syndromes relevant with alcohol abuse; At present, the alcohol content measuring technology mainly contains type of respiration, AAS and vapor-phase chromatography, and wherein, the type of respiration equipment price is expensive; Poor specificity exists interference many simultaneously, and it is poor to measure fiduciary level; Highly sensitive, good reproducibility, the range of linearity of AAS are wide, but sample need dilute, and pre-service is loaded down with trivial details; The gas chromatography determination time is long, and testing cost is high, is fit to batch detection.
Can know that according to relevant report alcohol concentration in the saliva and blood alcohol concentration have extremely similarity, the ratio of general blood and saliva is 1:1～1.04; So, it is worthy of note that alcohol concentration not only is sampling easily in the saliva to measuring, the alcohol concentration that also is to detect in the saliva can directly reflect the alcohol concentration in the blood.
Existing alcohol screen test bar all is subject to the influence of temperature and humidity; And cause test-strips preservation condition and holding time limited; Temperature and humidity is one of key factor that influences enzyme reaction efficient and storage, and this is to cause the reason that enzymatic reaction reagent can not long preservation; The enzyme activation composition that existing alcohol screen test bar is adopted is toxic, harmful, and environment is had pollution, and can't the active balance enzyme active and stable; In addition, there is part background color abnormality in the test-strips that prior art is made, is prone to cause test, contrast to occur than mistake.
Summary of the invention
The objective of the invention is deficiency to prior art; Thereby provide a kind of simple in structure, detection is easy, quick, accurate, efficient, use cost is low, highly sensitive, good stability and the high saliva alcohol content test-strips of reliability, and a kind of saliva alcohol content test-strips and preparation method thereof also is provided.
To achieve these goals; The technical scheme that the present invention adopted is: a kind of saliva alcohol content test-strips; It comprises substrate and sticks on the reaction strip of said substrate one side surface that said reaction strip is the dry filter paper that soaked chromogenic reagent solution and two enzyme reaction solutions successively.
Based on above-mentioned, said substrate is any in PVC, polycarbonate, the polystyrene.
A kind of method for making of saliva alcohol content test-strips, this method for making may further comprise the steps:
Step 1, filter paper is dipped in the chromogenic reagent solution, takes out, under 25 ℃～35 ℃, lucifuge, vacuum drying 15～25 minutes, obtain the filter paper that develops the color;
Wherein, Said chromogenic reagent solution is the potpourri of developer and developer solvent; The volume ratio of the weight of said developer and said developer solvent is 6～10 milligrams: 5 milliliters; Said developer is a tetramethyl benzidine, and said developer solvent comprises water and acetone, and the volume ratio of water and acetone is 1:1～1.5;
Step 2, said colour developing filter paper is dipped in two enzyme reaction solutions, takes out, under 25 ℃～35 ℃, lucifuge, vacuum drying 15～25 minutes, obtain reacting strip;
The said pair of enzyme reaction solution comprise active unit be 60～90U/ml alcohol oxidase, peroxidase that active unit is 70～100U/ml, account for said pair of enzyme liquid general assembly (TW) 1.2～2.0% bovine serum albumin(BSA), account for said pair of enzyme liquid general assembly (TW) 1.0～1.5% stabilizing agent, all the other are phosphate buffer; Wherein, The concentration of said phosphate buffer is that 0.1～0.6M, pH value are 7～8, and said stabilizing agent is one or several the potpourri in trehalose, shitosan, sodium alginate, sodium citrate, the gelatin.
Based on above-mentioned, in step 2, said reaction strip is sticked on substrate one side surface; Obtain the strip plate; Again the strip plate is cut into the wide strip of 0.5cm with automatic control cutting cutter, then, the monolithic strip packed into to be included in the aluminium foil bag of sack drying agent; Vacuum Package promptly gets saliva alcohol content test-strips finished product.
Based on above-mentioned, in step 1, filter paper is dipped in the chromogenic reagent solution fully, kept 5～10 seconds.
Based on above-mentioned, in step 2, said colour developing filter paper is dipped in two enzyme liquid fully, kept 5～10 seconds.
Based on above-mentioned, said peroxidase is a horseradish peroxidase.
The relative prior art of the present invention has outstanding substantive distinguishing features and marked improvement, and specifically, the present invention has following outstanding advantage:
1, adopt trehalose etc. to make stabilizing agent; Can only keep the active of enzyme with other stabilizing agent and slow down enzyme and compare because of the situation of long-term storage inactivation; Trehaloses etc. are cooked stabilizing agent and can be made heat sensitive enzyme is at high temperature kept stability and plays the active effect of kinase, and it can suppress the inactivation of enzyme under the high temperature; In the dry run, then can replace water or, thereby can reduce the influence of temperature and humidity, make that the test-strips of making is stable, reliable, the resting period is long the test-strips quality as the hyaloid stabilizing agent;
2, this method for making is beneficial to and prepares and use chromogenic reagent solution more easily, simultaneously, has reduced the oxidized rate of developer, can better keep enzyme activity;
3, adopt this saliva alcohol content test-strips to need not saliva is carried out pre-service; Can directly contact human saliva; Reaction, variable color is rapid; Can make things convenient for, judge intuitively test result with colour code contrast, it has advantage simple in structure, that detection is easy, quick, accurate, efficient, use cost is low, highly sensitive, good stability and reliability are high and has highly application value;
4, the technology of this method for making is simple, practical, and processing and encapsulation are convenient, and processing cost is low, helps large-scale production, and adopts this method for making, and quality index such as the repeatability of test-strips, precision, difference between batch are significantly increased.
Description of drawings
Fig. 1 is the structural representation of said saliva alcohol content test-strips.
Embodiment
Through embodiment, technical scheme of the present invention is done further detailed description below.
As shown in Figure 1, a kind of saliva alcohol content test-strips, it comprises substrate 2 and the reaction strip 1 that sticks on said substrate 2 one side surfaces, said reaction strip 1 is the dry filter paper that soaked chromogenic reagent solution and two enzyme reaction solutions successively; Said substrate 2 is any in PVC, polycarbonate, the polystyrene.
A kind of method for making of saliva alcohol content test-strips, this method for making may further comprise the steps:
Step 1, the thick type filter paper of elder generation cut into the wide filter paper of 0.4～0.6cm, filter paper are dipped in the chromogenic reagent solution fully again, keep 5～10 seconds; Take out; Under 25 ℃～35 ℃, lucifuge, vacuum drying 15～25 minutes, obtain the filter paper that develops the color, and be saved in the exsiccator for use;
Wherein, Said chromogenic reagent solution is the potpourri of developer and developer solvent; The volume ratio of the weight of said developer and said developer solvent is 6～10 milligrams: 5 milliliters; Said developer is a tetramethyl benzidine, and said developer solvent comprises water and acetone, and the volume ratio of water and acetone is 1:1～1.5;
Step 2, said colour developing filter paper is dipped in two enzyme liquid fully, kept 5～10 seconds, taking-up under 25 ℃～35 ℃, lucifuge, vacuum drying 15～25 minutes, obtains reacting strip, and is saved in the exsiccator for use;
The said pair of enzyme reaction solution comprise active unit be 60～90U/ml alcohol oxidase, peroxidase that active unit is 70～100U/ml, account for said pair of enzyme liquid general assembly (TW) 1.2～2.0% bovine serum albumin(BSA), account for said pair of enzyme liquid general assembly (TW) 1.0～1.5% stabilizing agent, all the other are phosphate buffer; Wherein, Said peroxidase is a horseradish peroxidase; The concentration of said phosphate buffer is that 0.1～0.6M, pH value are 7～8, and said stabilizing agent is one or several the potpourri in trehalose, shitosan, sodium alginate, sodium citrate, the gelatin;
Step 3, said reaction strip is sticked on substrate one side surface, obtain the strip plate;
Step 4, the strip plate is cut into the wide strip of 0.5cm with automatic control cutting cutter, then, the monolithic strip packed into to be included in the aluminium foil bag of sack drying agent, and Vacuum Package promptly gets saliva alcohol content test-strips finished product.
The biology enzyme that catalysis saliva ethanol synthesis is arranged on the reaction strip; Like alcohol oxidase and peroxidase; After contact contains the saliva of ethanol, can react and produce the color of different depth, then; With saliva alcohol content test-strips after the variable color and colour code contrast, can judge the content of alcohol in the saliva again.
This test-strips is used for the fast detecting alcohol concentration of human saliva.
Should be noted that at last: above embodiment is only in order to technical scheme of the present invention to be described but not to its restriction; Although with reference to preferred embodiment the present invention has been carried out detailed explanation, the those of ordinary skill in affiliated field is to be understood that: still can specific embodiments of the invention make amendment or the part technical characterictic is equal to replacement; And not breaking away from the spirit of technical scheme of the present invention, it all should be encompassed in the middle of the technical scheme scope that the present invention asks for protection.
Claims (7)
1. saliva alcohol content test-strips is characterized in that: it comprises substrate and sticks on the reaction strip of said substrate one side surface that said reaction strip is the dry filter paper that soaked chromogenic reagent solution and two enzyme reaction solutions successively.
2. saliva alcohol content test-strips according to claim 1 is characterized in that: said substrate is any in PVC, polycarbonate, the polystyrene.
3. the method for making of the described saliva alcohol content of claim 1 test-strips, it is characterized in that: this method for making may further comprise the steps:
Step 1, filter paper is dipped in the chromogenic reagent solution, takes out, under 25 ℃～35 ℃, lucifuge, vacuum drying 15～25 minutes, obtain the filter paper that develops the color;
Wherein, Said chromogenic reagent solution is the potpourri of developer and developer solvent; The volume ratio of the weight of said developer and said developer solvent is 6～10 milligrams: 5 milliliters; Said developer is a tetramethyl benzidine, and said developer solvent comprises water and acetone, and the volume ratio of water and acetone is 1:1～1.5;
Step 2, said colour developing filter paper is dipped in two enzyme reaction solutions, takes out, under 25 ℃～35 ℃, lucifuge, vacuum drying 15～25 minutes, obtain reacting strip;
The said pair of enzyme reaction solution comprise active unit be 60～90U/ml alcohol oxidase, peroxidase that active unit is 70～100U/ml, account for said pair of enzyme liquid general assembly (TW) 1.2～2.0% bovine serum albumin(BSA), account for said pair of enzyme liquid general assembly (TW) 1.0～1.5% stabilizing agent, all the other are phosphate buffer; Wherein, The concentration of said phosphate buffer is that 0.1～0.6M, pH value are 7～8, and said stabilizing agent is one or several the potpourri in trehalose, shitosan, sodium alginate, sodium citrate, the gelatin.
4. the method for making of saliva alcohol content test-strips according to claim 3 is characterized in that: in step 2, said reaction strip is sticked on substrate one side surface; Obtain the strip plate; Again the strip plate is cut into the wide strip of 0.5cm with automatic control cutting cutter, then, the monolithic strip packed into to be included in the aluminium foil bag of sack drying agent; Vacuum Package promptly gets saliva alcohol content test-strips finished product.
5. according to the method for making of claim 3 or 4 described saliva alcohol content test-strips, it is characterized in that: in step 1, filter paper is dipped in the chromogenic reagent solution fully, kept 5～10 seconds.
6. according to the method for making of claim 3 or 4 described saliva alcohol content test-strips, it is characterized in that: in step 2, said colour developing filter paper is dipped in two enzyme liquid fully, kept 5～10 seconds.
7. according to the method for making of claim 3 or 4 described saliva alcohol content test-strips, it is characterized in that: said peroxidase is a horseradish peroxidase.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011103148261A CN102384971A (en) | 2011-10-18 | 2011-10-18 | Spittle alcoholicity test strip and production method thereof |
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| CN2011103148261A CN102384971A (en) | 2011-10-18 | 2011-10-18 | Spittle alcoholicity test strip and production method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267758A (en) * | 2013-05-17 | 2013-08-28 | 浙江东方基因生物制品有限公司 | Dry chemical method rapid diagnostic reagent strip for testing content of alcohol in saliva and preparation method thereof |
| CN104198686A (en) * | 2014-08-14 | 2014-12-10 | 苏州康铭诚业医用科技有限公司 | Compound stabilizer for uric acid assay kit |
| CN106073829A (en) * | 2016-05-26 | 2016-11-09 | 李宝 | The oral cavity wiping bar of detection alcohol content |
| CN104198686B (en) * | 2014-08-14 | 2017-01-04 | 苏州康铭诚业医用科技有限公司 | A kind of testing uric acid test kit |
| CN107607730A (en) * | 2017-10-26 | 2018-01-19 | 南通伊仕生物技术股份有限公司 | For detecting the reagent strip of alcohol content, preparation method and kit in saliva |
| CN110006885A (en) * | 2019-04-18 | 2019-07-12 | 南京师范大学 | A method for quantitative analysis of alcohol based on dual-enzyme-inorganic nanoflower composites |
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| WO2009083964A2 (en) * | 2007-12-27 | 2009-07-09 | Alexander Schoenfeld | Sobriety interlock device |
| CN101566637A (en) * | 2009-05-21 | 2009-10-28 | 浙江大学 | Biosensor used for detecting alcohol concentration of human saliva |
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| CN1904619A (en) * | 2006-08-01 | 2007-01-31 | 上海唯卓生物科技有限公司 | Reagent stripe for detecting alcohol content in saliva and box kit |
| WO2009083964A2 (en) * | 2007-12-27 | 2009-07-09 | Alexander Schoenfeld | Sobriety interlock device |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267758A (en) * | 2013-05-17 | 2013-08-28 | 浙江东方基因生物制品有限公司 | Dry chemical method rapid diagnostic reagent strip for testing content of alcohol in saliva and preparation method thereof |
| CN103267758B (en) * | 2013-05-17 | 2015-05-13 | 浙江东方基因生物制品有限公司 | Dry chemical method rapid diagnostic reagent strip for testing content of alcohol in saliva and preparation method thereof |
| CN104198686A (en) * | 2014-08-14 | 2014-12-10 | 苏州康铭诚业医用科技有限公司 | Compound stabilizer for uric acid assay kit |
| CN104198686B (en) * | 2014-08-14 | 2017-01-04 | 苏州康铭诚业医用科技有限公司 | A kind of testing uric acid test kit |
| CN106073829A (en) * | 2016-05-26 | 2016-11-09 | 李宝 | The oral cavity wiping bar of detection alcohol content |
| CN107607730A (en) * | 2017-10-26 | 2018-01-19 | 南通伊仕生物技术股份有限公司 | For detecting the reagent strip of alcohol content, preparation method and kit in saliva |
| CN110006885A (en) * | 2019-04-18 | 2019-07-12 | 南京师范大学 | A method for quantitative analysis of alcohol based on dual-enzyme-inorganic nanoflower composites |
| CN110006885B (en) * | 2019-04-18 | 2021-11-02 | 南京师范大学 | A method for quantitative analysis of alcohol based on dual-enzyme-inorganic nanoflower composites |
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Application publication date: 20120321 |