CN109254001A - A kind of preparation and application of starch quick measuring card - Google Patents
A kind of preparation and application of starch quick measuring card Download PDFInfo
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- CN109254001A CN109254001A CN201811265774.1A CN201811265774A CN109254001A CN 109254001 A CN109254001 A CN 109254001A CN 201811265774 A CN201811265774 A CN 201811265774A CN 109254001 A CN109254001 A CN 109254001A
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- 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
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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 present invention relates to nano material, catalysis and analytical chemistry fields, specifically include the preparation and application of a kind of starch quick measuring card.This method utilizes and obtains the quick measuring card quickly detected on cobaltosic oxide nano ball, carbohydrase, glucose oxidase and developer-modified filter paper with imitative peroxidase activity, realizes the detection of simple, quick starch.This novel colorimetric method for detecting starch is practical, and therefore, this method has broad application prospects in food inspection, analytical chemistry, environmental project and agriculture field.
Description
Technical field
The present invention relates to nano material, catalysis and analytical chemistry field, specifically include a kind of starch quick measuring card preparation and
Using.
Background technique
Starch (starch) is a kind of polysaccharide, and molecular formula is (C6H10O5) n, it is a kind of mode of plant storing energy.It forms sediment
Powder can be regarded as the high polymer of glucose.Starch is commercially used for paste producing essence, maltose, glucose, alcohol in addition to edible
Deng being also used for modulation printing paste, the starching of textile, the gluing of paper, compacting of medicinal tablet etc..A variety of methods have been reported
For detecting starch, for example, Chattopadhyay et al. is in " Co-immobilization of glucoamylase and
glucose oxidase for electrochemical sequential enzyme electrode for starch
Biosensor and biofuel cell " utilize method detection starch (the Biosensors and of electrochemistry
Bioelectronics 2014,51,158-163).Beckles et al. is in " Structural Investigations and
Morphology of Tomato Fruit Starch " utilize the starch (Journal in the method detection tomato of gel electrophoresis
Of Agricultural and Food Chemistry 2009,57,282-291).However, all there is operation in these methods
The disadvantages of process is complicated and higher cost, therefore, it is necessary to develop a kind of sensitive, simple, efficiently method for detecting starch.
Test strips have been widely used in life as a kind of portable rapid detection method, such as blood sugar test paper
Item, urinalysis test strips etc..Therefore, many documents have also done certain research to test strips.Yan et al. exists
By the colloid in traditional test strip in " Nanozyme-strip for rapid local diagnosis of Ebola " text
Gold replaces with magnetic Nano enzyme, detects Ebola virus by the method for colour developing, and the sensitivity of test strips is made to improve 100
Again (Biosensors and Bioelectronics 2015,74,134-141).Cai et al. is in " A simple and
rapid colloidal gold-based immunochromatogarpicstriptest for detection of
FMDV derotype A " immunoassay is combined with test strips for detecting foot and mouth disease virus (ACS Applied in a text
Materials & Interfaces 2014,6,21624-21631).Although test strips have a degree of application,
Existing method manufacturing process is more complicated and higher cost, and the effect machine of the above-mentioned test strips mechanism of action and starch detection
Reason has essential distinction.Therefore, it is necessary to develop the new method of one kind to realize that simple, quickly preparation and starch detect.
Summary of the invention
The object of the present invention is to provide the preparations and application of a kind of starch quick measuring card.
A kind of preparation of starch quick measuring card, it is characterised in that: the starch quick measuring card by substrate, compound urge material and colour developing
Agent is constituted, and composite catalyzing material is the cobaltosic oxide nano ball (Co for being modified with carbohydrase and glucose oxidase (GOx)3O4
NPs), composite catalyzing material and color developing agent are fixed in substrate.
To achieve the above object, the technical solution adopted by the present invention are as follows:
A kind of preparation of starch quick measuring card, it is characterised in that the preparation process of the starch quick measuring card is as follows:
(1) bovine serum albumin (BSA) solution that mass concentration is 1% is dripped in substrate, stands 1 hour and is dried at room temperature for；
(2) the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N- hydroxyl amber for being 2% by mass concentration
Amber acid imide (NHS) drip respectively it is described in step (1) be modified in the substrate of BSA, stand 5 hours, wash and at room temperature
It is dry；
(3) it by substrate obtained in the uniform drop coating of the solution of composite catalyzing material to step (2), at 4 DEG C overnight, washes and does
It is dry；
(4) it will be modified in the substrate of composite catalyzing material described in organic chromogenic agent solution drop coating to step (3), done at 4 DEG C
It is dry, obtain quick measuring card.
The preparation process of composite catalyzing material, it is characterised in that this method comprises the following steps:
(1) it is added a certain amount of sodium borohydride into the mixed solution containing BSA and divalent cobalt ion, it is reaction, centrifugation, dry
To Co3O4NPs；
(2) by a certain amount of cobaltosic oxide nano ball (Co3O4NPs) it is added to the t-butyl carbamate (BOC) of 0.2 mM
In solution, washing, drying after standing 2 hours；
(3) by step (2) resulting Co3O4NPs is added in EDC the and NHS mixed solution that mass concentration is 2%, and it is small to stand 5
When, washing, drying；
(4) by step (3) resulting Co3O4NPs is added to carbohydrase (Glucoamylase) and glucose oxidase (GOx)
Mixed liquor in, reaction overnight at 4 DEG C is washing, dry, and composite catalyzing material is made.
A kind of preparation and application of starch quick measuring card, the composite catalyzing material can be used for the detection of starch.
Preferably, the substrate can be nitrocellulose filter, cellulose acetate membrane, polyester fiber, glass fibre, nonwoven
Cloth etc.；The organicvisualization reagent be 2,2 '-connection nitrogen-bis- (3- ethyl benzo thiazole phenanthroline -6- sulfonic acid) diamine salts, 3,3 ', 5,5 '-four
Methyl biphenyl amine, dopamine or phenol.
It is more excellent to be, the detection application of a kind of starch quick measuring card, it is characterised in that application process is as follows:
(1) two quick measuring cards are taken, one adds a certain amount of starch solution on quick measuring card, starch is not added on another quick measuring card,
It places 5 minutes, reacts starch sufficiently with composite catalyzing material；
(2) color change of quick measuring card is observed to realize the qualitative detection to starch, is added if comparing with the quick measuring card that starch is not added
The quick measuring card of starch changes colour, then explanation can realize the detection to starch with quick measuring card；
(3) taking photograph of intelligent mobile phone is used, can be carried out when taking pictures under LED light irradiation to prevent blurring of photos.By using hand
The color identification software of machine reads the specific rgb value of quick measuring card in photo, by BNB is calculated in=B/ (R+G+B)N, substitute into BN-
Concentration working curve equation realizes the quantitative detection to starch to calculate the concentration of starch.
Effect of the invention is:
1. the present invention utilizes the cobaltosic oxide nano ball and carbohydrase and glucose oxidase with imitative peroxidase activity
Tandem reaction, construct quickly detection starch sensor.
2. composite catalyzing material and prepared test strips that the present invention synthesizes realize the detection of one kettle way starch, avoid point
The progress of step reaction.
3. the present invention have certain practical value in actual life, and extend to food fermentation, biological medicine,
Chemical industry, environment, the quantitative analysis of biotechnology field and catalyst etc..
Detailed description of the invention
Fig. 1 is the preparation flow figure of quick measuring card provided in an embodiment of the present invention.
Fig. 2 is the detection principle diagram of quick measuring card provided in an embodiment of the present invention.
Fig. 3 is the feasibility of quick measuring card provided in an embodiment of the present invention detection.
Fig. 4 is that the one kettle way pH of composite catalyzing material provided in an embodiment of the present invention optimizes.
Fig. 5 is the qualitative detection photo of quick measuring card provided in an embodiment of the present invention.
Fig. 6 is the standard working curve of starch provided in an embodiment of the present invention quantitative determination.
Specific embodiment
In order in depth illustrate the contents of the present invention, some embodiments will be further enumerated below, but the present invention does not limit to
In cited embodiment.Specific experiment condition or method are such as not specified in the following example, press the normal condition of this field
Or method carries out.
Embodiment 1
The preparation process of composite catalyzing material, it is characterised in that this method comprises the following steps:
(1) a certain amount of sodium borohydride is added into the mixed solution containing BSA and divalent cobalt ion, passes through going back for sodium borohydride
Former and oxygen oxidation obtains Co3O4NPs is centrifuged, is dried to obtain solid；
(2) by a certain amount of Co3O4NPs is added to the BOC solution of 0.2 mM to protect Co3O4It is small to stand 2 for amino on NPs
When after washing, dry；
(3) by step (2) resulting Co3O4NPs is added in EDC the and NHS mixed solution that mass concentration is 2% with activation
Co3O4Carboxyl on NPs stands 5 hours, washing, drying；
(4) by the Co after activated carboxyl obtained by step (3)3O4NPs is added in the mixed liquor of carbohydrase and GOx, carbohydrase and
Amino meeting and Co on GOx3O4 Amide reaction, which occurs, for the carboxyl on NPs makes it be fixed to Co3O4It is anti-overnight at 4 DEG C on NPs
It answers, washes, dries, composite catalyzing material is made.
Embodiment 2
The preparation flow (Fig. 1) of quick measuring card:
(1) the BSA solution that mass concentration is 1% is dripped on cellulose membrane, stands 1 hour and is dried at room temperature for；
(2) EDC and NHS that mass concentration is 2% cellulose membrane for being modified with BSA in step (1) is dripped respectively (to select
The nitrocellulose filter of good water absorption) on the carboxyl of activated b SA, stand 5 hours, wash and be dried at room temperature for；
(3) by cellulose membrane obtained in the uniform drop coating of the solution of composite catalyzing material to step (2), on composite catalyzing material
Amino can with the carboxyl on BSA occur amide react make its be fixed to cellulose membrane on, at 4 DEG C overnight, wash simultaneously drying；
(4) it will be modified on the cellulose membrane of composite catalyzing material described in organic chromogenic agent solution drop coating to step (3), 4 DEG C
Lower drying, obtains quick measuring card.
The testing principle (Fig. 2) of quick measuring card:
Starch quick measuring card is made of substrate, enzyme and color developing agent, substrate use nitrocellulose filter paper, will be connected with carbohydrase and
The Co of GOx3O4NPs and color developing agent are fixed in substrate, the basic principle of detection are as follows: when starch is added, starch exists first
Glucose is generated under the catalytic action of carbohydrase, the glucose continuation of generation is reacted with GOx generates H2O2, there is imitative peroxide later
The Co of compound enzymatic activity3O4NPs can be catalyzed H2O2ABTS is set to change colour, the card that tests the speed becomes green from colourless；Starch quick measuring card is not added
Color change it is smaller or non-discolouring, can according to the discoloration of quick measuring card realize starch quantitative analysis.
Embodiment 3
The feasibility of quick measuring card detection:
Catalystic converter system (a), which shreds colorimetric card, to be put into starch (1 mmol/L) solution (pH=5.0), and reaction takes after five minutes
Reaction solution carries out full wavelength scanner.It separately does one group of control experiment (b) and shreds colorimetric card and be put into buffer solution (pH=5.0), instead
Middle carry out full wavelength scanner should be extracted reaction solution after five minutes.
As shown in figure 3, catalystic converter system (a) has apparent ultraviolet absorption peak, catalystic converter system (b) is without UV absorption
Peak, it was demonstrated that only catalysis reaction just can be carried out in the presence of starch.
Embodiment 4
The one kettle way pH of composite catalyzing material optimizes:
Catalystic converter system includes the buffer (pH of composite catalyzing material (30 μ g/mL), starch (1 mmol/L) and different pH
1.0 2.0, glycine-HCI buffer；PH 3.0 6.0, acetic acid-acetic acid acid sodium buffer；PH 7.0 8.0, phosphate are slow
Fliud flushing；PH 9.0, Tris- hydrochloride buffer).It is reacted under room temperature (25 °C) after five minutes, surveys reaction solution at 650 nm
Absorbance.As shown in Figure 4 (a), optimal pH is 4.0 or so.
One group of control experiment separately is done, catalystic converter system includes Co3O4NPs(30 μ g/mL), carbohydrase (0.3mg/mL),
GOx(0.1mg/mL), buffer (pH 1.0 2.0, the glycine-HCI buffer of starch (1 mmol/L) and different pH；pH
3.0 6.0, acetic acid-acetic acid acid sodium buffer；PH 7.0 8.0, phosphate buffer；PH 9.0 10.0, Tris- hydrochloride buffer
Liquid).It is reacted under room temperature (25 °C) after five minutes, surveys absorbance of the reaction solution at 650 nm.As shown in Figure 4 (b), it is in pH
3.0 4.5 or so have greater activity, but activity of its activity less than 4(a).
Embodiment 5
The qualitative detection of quick measuring card:
(1) three quick measuring cards are taken, starch, quick measuring card (b) plus 1mM starch is not added in quick measuring card (a), and quick measuring card (c) adds 100 mM's
Starch is placed 5 minutes, makes have starch sufficiently to act on composite catalyzing material；
(2) taking photograph of intelligent mobile phone is used, can be carried out when taking pictures under LED light irradiation to prevent blurring of photos.As shown in figure 5,
The quick measuring card (a) that starch is not added is non-discolouring, adds the quick measuring card (b) of smaller concentration starch to darken compared with quick measuring card (a), adds larger
Concentration starch quick measuring card (c) darkens compared with quick measuring card (b), illustrates the starch concentration being added dropwise on quick measuring card (c) maximum, quick measuring card
(b) taking second place, quick measuring card (a) does not add starch, and this again demonstrate the feasibilities of this starch quick measuring card detection starch, therefore, this shallow lake
The detection to starch may be implemented in powder quick measuring card.
Embodiment 6
The standard working curve of starch quantitative determination:
(1) 12 quick measuring cards are taken, the starch (0-10 mM) of various concentration is added dropwise on quick measuring card, places 5 minutes, makes starch and answers
Catalysis material is closed sufficiently to act on；
(2) taking photograph of intelligent mobile phone is used, can be carried out when taking pictures under LED light irradiation to prevent blurring of photos.By using hand
The color identification software of machine reads the specific rgb value of quick measuring card in photo, by BNB is calculated in=B/ (R+G+B)N, marked
Directrix curve, for specific works curve as shown in fig. 6, the range of linearity is 0-0.3 mM, equation is y=0.6062x+0.0998(R2=
0.9918).
Claims (5)
1. a kind of preparation of starch quick measuring card, it is characterised in that: the starch quick measuring card by substrate, compound urge material and color developing agent
It constitutes, composite catalyzing material is the cobaltosic oxide nano ball (Co for being modified with carbohydrase and glucose oxidase (GOx)3O4
NPs), composite catalyzing material and color developing agent are fixed in substrate.
2. a kind of preparation of starch quick measuring card according to claim 1, it is characterised in that the preparation of the starch quick measuring card
Journey is as follows:
(1) bovine serum albumin (BSA) solution that mass concentration is 1% is dripped in substrate, stands 1 hour and is dried at room temperature for；
(2) the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N- hydroxyl amber for being 2% by mass concentration
Amber acid imide (NHS) drip respectively it is described in step (1) be modified in the substrate of BSA, stand 5 hours, wash and at room temperature
It is dry；
(3) it by substrate obtained in the uniform drop coating of the solution of composite catalyzing material to step (2), is stayed overnight at 4 DEG C, dry and water
It washes；
(4) it will be modified in the substrate of composite catalyzing material described in organic chromogenic agent solution drop coating to step (3), done at 4 DEG C
It is dry, obtain quick measuring card.
3. by the preparation process of composite catalyzing material described in claim 1,2, it is characterised in that:
(1) it is added a certain amount of sodium borohydride into the mixed solution containing BSA and divalent cobalt ion, it is reaction, centrifugation, dry
To Co3O4NPs；
(2) by a certain amount of Co3O4NPs is added in t-butyl carbamate (BOC) solution of 0.2mM, water after standing 2 hours
It washes, dry；
(3) by step (2) resulting Co3O4NPs is added in EDC the and NHS mixed solution that mass concentration is 2%, and it is small to stand 5
When, washing, drying；
(4) by step (3) resulting Co3O4NPs is added in the mixed liquor of carbohydrase and GOx, reaction overnight at 4 DEG C, washing,
It is dry, composite catalyzing material is made.
4. a kind of starch quick measuring card described in -3 according to claim 1, it is characterised in that:
The substrate can be nitrocellulose filter, cellulose acetate membrane, polyester fiber, glass fibre, non-woven fabrics etc.；It is described to have
Machine color developing agent be 2,2 '-connection nitrogen-bis- (3- ethyl benzo thiazole phenanthroline -6- sulfonic acid) diamine salts, 3,3 ', 5,5 '-tetramethyl benzidines,
Dopamine or phenol.
5. a kind of detection application of starch quick measuring card described in -4 according to claim 1, it is characterised in that application process is as follows:
(1) two quick measuring cards are taken, one adds a certain amount of starch solution on quick measuring card, starch is not added on another quick measuring card,
It places 5 minutes, reacts starch sufficiently with composite catalyzing material；
(2) color change of quick measuring card is observed to realize the qualitative detection to starch, is added if comparing with the quick measuring card that starch is not added
The quick measuring card of starch changes colour, then explanation can realize the detection to starch with quick measuring card；
(3) taking photograph of intelligent mobile phone is used, can be carried out under LED light irradiation to prevent blurring of photos, by using hand when taking pictures
The color identification software of machine reads the specific rgb value of quick measuring card in photo, by BNB is calculated in=B/ (R+G+B)N, substitute into BN-
Concentration working curve equation realizes the quantitative detection to starch to calculate the concentration of starch.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811265774.1A CN109254001B (en) | 2018-10-29 | 2018-10-29 | Preparation and application of starch rapid test card |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811265774.1A CN109254001B (en) | 2018-10-29 | 2018-10-29 | Preparation and application of starch rapid test card |
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| CN109254001B CN109254001B (en) | 2021-06-04 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130052740A1 (en) * | 2011-08-24 | 2013-02-28 | Multisorb Technologies, Inc. | Copper, starch and iodide moisture indicator |
| CN105388150A (en) * | 2015-10-29 | 2016-03-09 | 大连理工大学 | Oxytetracycline test paper based on chromatic aberration comparison, using method and making method |
| CN105606604A (en) * | 2016-01-08 | 2016-05-25 | 昆明泊银科技有限公司 | Rapid hydrogen peroxide test paper and preparation method thereof |
| CN107857307A (en) * | 2017-11-20 | 2018-03-30 | 青岛农业大学 | A kind of new strategy for realizing one kettle way grape grape color developing detection |
-
2018
- 2018-10-29 CN CN201811265774.1A patent/CN109254001B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130052740A1 (en) * | 2011-08-24 | 2013-02-28 | Multisorb Technologies, Inc. | Copper, starch and iodide moisture indicator |
| CN105388150A (en) * | 2015-10-29 | 2016-03-09 | 大连理工大学 | Oxytetracycline test paper based on chromatic aberration comparison, using method and making method |
| CN105606604A (en) * | 2016-01-08 | 2016-05-25 | 昆明泊银科技有限公司 | Rapid hydrogen peroxide test paper and preparation method thereof |
| CN107857307A (en) * | 2017-11-20 | 2018-03-30 | 青岛农业大学 | A kind of new strategy for realizing one kettle way grape grape color developing detection |
Non-Patent Citations (1)
| Title |
|---|
| 巩发永 等: "《马铃薯食品加工技术与质量控制》", 31 July 2018, 西南交通大学出版社 * |
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