CN109507176A - Dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide - Google Patents
Dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide Download PDFInfo
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- CN109507176A CN109507176A CN201811338531.6A CN201811338531A CN109507176A CN 109507176 A CN109507176 A CN 109507176A CN 201811338531 A CN201811338531 A CN 201811338531A CN 109507176 A CN109507176 A CN 109507176A
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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
- 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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Abstract
A kind of dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide, belong to biochemistry detection technical field, it is characterized in that: being coated with secondary heme hexapeptide mixture reagent layer on the mat polyester film of detection card, titanium dioxide diffusion layer is coated on reagent layer, reagent layer mixture is polymer microsphere, secondary heme hexapeptide, 4-AAP- chlorophenol, phosphate buffer, surfactant and water-soluble binder mixture.Beneficial effect is: having the principle of peroxidase function using DhHP-6, prepares a kind of dry chemical H2O2Detection method calculates H in liquid sample by the variation to diffuse after detection reaction2O2Amount.Have the characteristics that the speed of response is fast, high sensitivity, the range of linearity is wide, system bulk is small, is easy to carry about with one, does not need outside agent.The departments such as medical and health and the food enterprises, the market of farm produce, quality surveillance and environmental monitoring be can be widely applied to H in fluid sample2O2Measure the field being monitored.
Description
Technical field
The invention belongs to biochemistry detection technical fields.
Background technique
Hydrogen peroxide (H2O2) it is a kind of common oxidant, in food production, biological medicine, pharmacy, industry and environment
Field plays an important role.But certain density H2O2After contacting or eat by mistake with human skin, uncomfortable or danger can be caused
Evil health, it is serious to induce a variety of diseases.State food safety management department and environmental protection department are to H2O2It is eating
The content in residual quantity and rivers and lakes in product and the residual volume after Industrial Waste Water Treatments suffer from strict requirements, but
It is or has some illegal enterprises and the personal laws and regulations abuse H for ignoring country2O2, so in order to more effectively monitor
H2O2Use, establish measurement H quickly, accurate, economic2O2The method of concentration has great importance.
H2O2The detection method of concentration includes titration, spectrometry, chemistry hair measuring method, fluorimetry, liquid phase
Chromatography and electrochemical process etc., these detection methods are all based on the catalysis to peroxidase (Peroxidase, POD)
Realize, although POD has a specificity and sensitivity of height, the application of free native enzyme by its stability it is poor and at
This high limitation.And above method is required to relatively large instrument and measures in laboratory, is not suitable for carrying out
Live real-time test.
In recent years, the application of analogue enztme has attracted extensive attention, and compared with native enzyme, analogue enztme has low cost,
The advantages of easily stored high stability, in chemical detection, immunoassays exploitation, cancer diagnosis and treatment and environmental protection
Aspect has great development space.Currently, having had been built up a large amount of small molecule compound and nano particle (NP) to simulate day
Right enzyme, including with materials such as the active secondary heme hexapeptide of POD and POD sample, the iron-based NPs of oxidation, vanadic anhydrides.
Secondary heme hexapeptide (Deuterohemin-Ala His Thr Val Glu Lys, DhHP-6) is with ferroheme
For the analogies with peroxidase activity of prothetic group, molecular weight 1228.48, chemical property is stablized, and molecular weight is small, dissolution
Property it is good, storage, transport, using etc. links no property change.
Traditional chemical reaction be usually liquid reagent and sample in the reaction vessel with certain volume into
Capable, this is also referred to as wet-chemical.Dry chemical (Dry Chemistry) is its also known as dry examination for wet-chemical
Agent chemistry or solid state chemistry, refer to and pre-fix part or all of reagent needed for measurement in the carrier of certain structure, examine
Fluid sample is applied directly to secure on the carrier of reagent when survey, the moisture in sample as solvent, cause determinand with
Reagent in carrier occurs to chemically react and generate reaction signal, finally by visual qualitative or sxemiquantitative, or passes through certain instrument
Device detection is shown in reaction number variation on carrier, to measure the concentration of determinand in sample.The carrier for securing reagent is known as
Dry plate or dry plate carrier, dry reagent etc., dry plate carrier have collectively constituted desiccation credit side analysis system with corresponding detecting instrument.
In recent years, dry chemical and dry analysis method achieve great progress, have partially reached as quantitative analysis method
The level for having arrived conventional chemical is a kind of important method of analytical chemistry.
Summary of the invention
The object of the present invention is to provide a kind of dry chemical H based on secondary heme hexapeptide2O2Detection method, it utilizes secondary
Ferroheme hexapeptide (DhHP-6) has the principle of peroxidase function, realizes and realizes quick, letter with dry analysis method
Just, H is accurately detected2O2。
Technical scheme is as follows: the mixing of secondary heme hexapeptide is coated on the mat polyester film of detection card
Object reagent layer, is coated with titanium dioxide diffusion layer on reagent layer, and reagent layer mixture is polymer microsphere, secondary heme six
Peptide (DhHP-6), 4-AAP- chlorophenol, phosphate buffer, surfactant and water-soluble binder mixture.
Polylactic acid polymer microsphere, secondary heme hexapeptide (DhHP-6), 4-AAP- chlorobenzene in reagent layer mixture
Phenol, phosphate buffer, surfactant and water-soluble binder proportion are 14-17:0.012-0.016:0.080-0.090:
0.2-0.4:0.05:0.4-0.6, above-mentioned material are weight ratio.
The beneficial effects of the present invention are: having the principle of peroxidase function using DhHP-6, peroxide is substituted
Enzyme prepares a kind of dry chemical H2O2Detection method calculates liquid sample by the variation to diffuse after detection reaction
Middle H2O2Amount.The present invention is fast with the speed of response, high sensitivity, the range of linearity is wide, system bulk is small, is easy to carry about with one, is not required to
The characteristics of wanting outside agent.It can be widely applied to medical and health and the food enterprises, the market of farm produce, quality surveillance and environment
The departments such as monitoring are to H in fluid sample2O2Measure the field being monitored.
Detailed description of the invention
Fig. 1 is H2O2Dry chemical detection lug (4) structure chart；
Fig. 2 is H2O2Dry chemical detection card (5) structure chart；
Fig. 3 is H2O2Dry chemical detection card (5) schematic diagram；
Fig. 4 is structure of the invention schematic block diagram；
Fig. 5 is dry chemical detection card detection various concentration H of the present invention2O2Reflectivity it is linear；
Table 1 is H2O230 various concentration H of dry chemical detection card2O2Testing result.
Specific embodiment
Embodiment 1
The present invention is described further with reference to the accompanying drawing:
As shown in Figure 1, 2, 3, wherein 1 be mylar, 2 be reagent layer, 3 be diffusion layer, 4 be H2O2Dry chemical detection
Piece, 5 are H2O2Dry chemical detection card, 5-1 are that dry chemical detects card bottom card, 5-2 is blocked on dry chemical detection card.
1, DhHP-6-4-AAP- chlorophenol colour reagent measures anti-H2O2Answer principle
H2O2Oxygen radical (O is generated under the catalysis of DhHP-6-), O-It is aobvious by oxidation 4-AAP- chlorophenol system
Color.
Developing concentration and H after reaction2O2Content is directly proportional, can carry out quantitative analysis by diffusing reflection photoelectric colorimetry.
2、H2O2The production process of dry chemical detection card
2.1 material
Titanium dioxide (TiO2)；Polylactic acid polymer microsphere；DhHP-6；4-AAP- chlorophenol；Mylar；Phosphoric acid
Salt buffer；Polyvinyl alcohol water-soluble binder；Surfactant；High-pressure spray gun.
2.2 production process
(1), in proportion by polylactic acid polymer microsphere, DhHP-6,4-AAP- chlorophenol, phosphate buffer, surface
Activator and water-soluble binder are mixed by weight 16:0.015:0.085:0.3:0.05:0.5, use high-pressure spray gun
This mixture is uniformly sprayed on mylar (1), is allowed to be formed with certain thickness, certain volume liquid can be accommodated
Reagent layer (2).All chemical reactions and colour developing are completed herein.
(2), after candidate agent layer (2) is dry, in proportion by titanium dioxide (TiO2), surfactant and water soluble adhesive
Agent is mixed, this mixture is sprayed on reagent layer (2) using high-pressure spray gun, is allowed to be formed with certain thickness, be formed
Can rapidly diffused liquid diffusion layer (3).After diffusion layer (3) are dry, H is completed2O2The production of dry chemical detection lug (4).System
H after2O2Dry chemical detection lug (4) is a kind of card of white.
(3), by H2O2Dry chemical detection lug (4) is cut into the square of 7 × 7mm, and merging has 7 × 7mm square recessed
In sunken dry chemical detection card bottom card (5-1), then covers after card (5-2) is fixed on dry chemical detection card and complete H2O2Dry chemical
Detect the production (5) of card.
Measure H2O2Experimental procedure:
(1), DhHP-6 reagent has the function of peroxidase, can cause H2O2It decomposes and generates O-.Therefore difference can be used
The H of concentration2O2Carry out Germicidal efficacy.
(2), by 3.0%H2O2Be diluted to concentration be 100,50.0,25.0,12.5,6.25,3.13,1.56mmol/L it is standby
With.
(3), 15 μ L is taken to contain various concentration H respectively2O2Specimen liquid, it is uniformly added in H2O2Dry chemical detection
Block on the dry chemical detection lug (4) in (5), is placed on the diffusing reflection photoelectric color comparator with 37 DEG C of temperature incubation functions 5 minutes, contains
There is H2O2Liquid sample by diffusion layer (3) enter reagent layer (2), it is anti-with DhHP-6-4-AAP- chlorophenol colour reagent
It answers, generates color change (red), show H on diffusing reflection photoelectric color comparator2O2Concentration (mmol/L).Experimental result is such as
Shown in Fig. 4.H2O2The H of dry chemical detection card detection various concentration2O2Show preferable linear relationship.
(4), respectively in 30 H2O2The H of 50.0,12.5,6.25mmol/L are detected on dry chemical detection card2O2Sample,
Its average value (X) standard deviation (SD) and the coefficient of variation (CV%) are calculated, experimental result is as shown in table 1.
Table 1
H2O2The experimental result of measurement shows, H2O2The H of dry chemical detection card measurement various concentration2O2With preferable linear
And stability.
Claims (2)
1. a kind of dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide, it is characterized in that: the bottom plate in detection card is poly-
Ester gum on piece is coated with secondary heme hexapeptide mixture reagent layer, and titanium dioxide diffusion layer, reagent layer are coated on reagent layer
Mixture is polymer microsphere, secondary heme hexapeptide, 4-AAP- chlorophenol, phosphate buffer, surfactant and water-soluble
Property adhesive mixture.
2. a kind of dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide as described in claim 1, it is characterized in that:
Polylactic acid polymer microsphere, secondary heme hexapeptide, 4-AAP- chlorophenol, phosphate-buffered in the reagent layer mixture
Liquid, surfactant and water-soluble binder proportion are 14-17:0.012-0.016:0.080-0.090:0.2-0.4:0.05:
0.4-0.6, above-mentioned material are weight ratio.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811338531.6A CN109507176A (en) | 2018-11-12 | 2018-11-12 | Dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide |
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| CN201811338531.6A CN109507176A (en) | 2018-11-12 | 2018-11-12 | Dry chemical hydrogen peroxide detection method based on secondary heme hexapeptide |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110146499A (en) * | 2019-06-21 | 2019-08-20 | 长春汇力生物技术有限公司 | A kind of dry plate detection reagent card of quantitative detection concentration of hydrogen peroxide and preparation method thereof |
| CN110794147A (en) * | 2019-10-31 | 2020-02-14 | 无锡锦帛诚医疗器械科技有限公司 | Albumin quantitative determination dry tablet for in vitro diagnosis |
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| CN110146499A (en) * | 2019-06-21 | 2019-08-20 | 长春汇力生物技术有限公司 | A kind of dry plate detection reagent card of quantitative detection concentration of hydrogen peroxide and preparation method thereof |
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Application publication date: 20190322 |