CN104359904B - Organophosphorus pesticide residue detection method based on readable bar code type pattern of smart phone - Google Patents
Organophosphorus pesticide residue detection method based on readable bar code type pattern of smart phone Download PDFInfo
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Abstract
The invention relates to an organophosphorus pesticide residue detection method based on a readable bar code type pattern of a smart phone. The method comprises the following steps: preparing an organophosphorus pesticide residue channel; printing and manufacturing a 39 code+code; adding a pesticide residue quick detection reagent into the organophosphorus pesticide residue channel; scanning a bar code to obtain a scanning result by using the smart phone; scanning the bar code to screen shot to obtain number picture of the scanning region for colorimetric analysis. Based on the readable bar code type pattern of the smart phone, the smart phone and a one-dimensional bar code are applied to quantitative and qualitative detection of biological and chemical molecules. As the widely applied smart phone is used for detecting and phone bar code scanning software and mobile phone image analyzing software can be downloaded freely from the Internet, the method has the remarkable advantages of quick detection speed, low cost and easiness in popularization.
Description
Technical field
The present invention relates to a kind of residues of organophosphate pesticides method for quick, specifically residues of organophosphate pesticides detection liquid is added
With makrolon（PC）And dimethyl silicone polymer（PDMS）In for the microchannel of material, then this microchannel is put into 39 yards of "+"
Detection zone in, this " bio-barcode " is scanned using barcode scanning software on smart mobile phone, and screenshotss obtain work
Make the bar code number picture in region, the Y value of ultimate analysis digital picture detection zone realizes quantifying for residues of organophosphate pesticides detection
Analysis.
Background technology
With gradually powerful and widely available, the medical care diagnostic technology based on smart mobile phone of smart mobile phone function
Cause extensive concern and research.Ozcan et al. increase on mobile phone microscopic system by modification on mobile phone turn into simple microscope,
Fluorescence microscope, realizes haemocyte technology（Isa Navruz.; Ahmet F. Coskun.; Justin Wong.;
Saqib Mohammad.; Derek Tseng.; Richie Nagi.; Stephen Phillipsand Aydogan
Ozcan. Lab Chip. 2013, 13, 4015-4023）And detection (the Ahmet F. Coskun. in food hypersenstivity source;
Justin Wong.; Delaram Khodadadi.; Richie Nagi.; Andrew Tey.; and Aydogan
Ozcan. Lab Chip. 2013,13,636-640) Ericson et al. using particular design chip and software, with reference to
The camera function of smart mobile phone, realizes detection (the Oncescu V, O'Dell of pH value in saliva and sweat based on colorimetric method
D, Erickson D. Lab Chip, 2013; 12:3232-3238.) and cholesterol detection（Oncescu V,
Mancuso M, Erickson D. Lab Chip, 2014; 14:759-763. Shen et al.（Li Shen.; Joshua
A. Hagen.;Ian Papautsky. LabChip. 2012, 12, 4240–4243）Using the camera function of smart mobile phone
And realize pH value detection with reference to particular color analysis software；G.M.Whitesides（Martinez, A.W.;
Phillips, S.T.; Carrilho, E.M.; Thomas, S.W.; Sindi, H.; Whitesides, G.M.
Anal. Chem. 2008, 80, 3699-3707.）Using the camera function of smart mobile phone, finished to being reacted in microchannel
Testing sample be imaged, quantitative analysis is carried out to acquired digital picture using computer software realizes grape in urine
The detection of sugar.Wang et al.（Wang, S.; Zhao, X.; Khimiji, I.; Akbas, R.; Qiu, W.;
Edwards, D.; Cramer, D.W.; Ye, B.; Demirci, U. Lab Chip. 2011, 11, 3411. ）, make
The content of oophoroma biomarker HE4 in urine is detected with IgG Antibody with Enzyme Linked Immunosorbent, is imaged by smart mobile phone, using on mobile phone
The software of installation carries out the last acquisition testing result of corresponding analysis to the digital picture for obtaining.
Above achievement in research, allows it is seen that smart mobile phone huge potentiality in following medical diagnosis, but due to
Increased needs the signal of precision machined optical accessory and particular design to receive analysis system, and this actually has been out
Towards the real meaning and value that instant detection is carried out using smart mobile phone of domestic consumer.
One-dimensional bar code（barcode）Refer to one group of regularly arranged bar and sky, bar refers to the small part of reflectivity, and sky is
The big part of digital reflex rate.The different arrangement modes of bar and sky represent different meanings, the bar and sky of the bar code of traditional sense
The secret note and informal voucher for respectively printing, the biological detection bar code that the application patent is proposed is using in biological, chemical detection
Colour developing result as bar code bar with it is empty, what is more important using common smart mobile phone free bar code scanning APP just
The decoding of bar code can be realized.
It is to be noted that Mirkin in 2003（Nam, J.-M.; Thaxton, C.S.; Mirkin, C.A.
Science. 2003, 301, 1884-1886）Et al. bio-barcode detection technique and the application patent of invention propose
Biological detection bar code is entirely different.What the application patent was proposed is the Structure of Bar-code of real meaning, using existing bar shaped
Code coding and decoding principle can just realize the quick diagnosis of biomolecule and detect, and the bio-barcode that Mirkin et al. is proposed
Detection technique is that a series of DNAs are combined on nanogold particle probe, just with the numeral mark concept of bar code, and
Barcode design and reading in practical application is without any association.
Inventor Li Xiaochun et al., Publication No. 103674856A patent of invention are " micro- based on scanner uni colorimetric analysis
Passage is used for residues of organophosphate pesticides fast determining method technical field " a kind of microchannel of suitable scanning is proposed, and colourity is analyzed in utilization
It is worth and then realizes quick, the quantitative detecting method to residues of organophosphate pesticides, the patent utilization is not readily susceptible to external light source interference
Scanner obtains digital picture and carries out quantitative analysis to it.The weak point scanner volume of the technology is larger to be inconvenient to carry
And the digital picture for obtaining needs importing computer specialty color analysis softwares of Photoshop to be analyzed, therefore is unfavorable for
Widely available application.
Apply for the Zhang Yi of artificial State Nanometer Science Center, the Publication No. CN103272656A inventions of Zhang Xing, Jiang Xingyu
Patent " Bar code micro fluidic chip and application thereof " has the difference of essence with the application patent.Patent utilization Codabar bar codes
"：" barcode encoding rule is dissolved into the design of micro-fluidic chip with " 1 ", the particularity of EAN yards " 0 " and " 3 " structure,
Each area of bar code is set to passage area by the method, and each passage is by substrate and the part with depression is reversible
Ground laminating and form microchannel, microchannel realized using pipeline and discharge line is passed through sample or reagent be passed through with
Discharge, directly distinguishes after biochemistry detection is reacted finally by bar code reading device and produces bar pattern.The weak point of the technology
The preparation technology of the sunk part required for passage it is more complicated and in force as passage area bar area quantity it is more because
And need to prepare that number of channels is more, while the bar code reading device of specialty is needed, so as to be unfavorable for widely available application.
" bio-barcode " being related in the application patent is using 39 yards of bar as detection zone, using on smart mobile phone
APP software scans bar codes, detection zone testing concentration difference and obtain different results.That is used in this experiment is one-dimensional
Bar code is 39 yards, and 39 yards is a kind of bar code for representing the information such as numeral, letter, is mainly used in industry, books and ticket
Automatic management.By 5 bars and 4 skies, totally 9 units are constituted 39 yards each character, and this 9 units only have two kinds of width,
9 narrow units of wherein 3 units wide, therefore referred to as 39 yards.39 yards of standard are made up of starting character, coding symbol, full stop, 39
The starting character and full stop of code are identicals by " * " representation.39 yards be selected out be applied in this experiment because it
Coding rule is simple and without check code.This experiment utilizes the "+" code and "-" code of 39 representations, the starting of the two bar codes
Symbol, full stop it is identical, and its code area only after four unit differences, using "+" code and "-" code structure spy
Property, at the dotted line position of "+" code as detection zone（Fig. 1）, detection zone after completion of the reaction, sweep by the bar code installed with smart mobile phone
Software is retouched to this bar code scan, if scanning result is "-", the aobvious feminine gender of result is represented；Scanning result is "+", represents result and shows
It is positive.
The content of the invention
The invention solves the problems that concrete technical problems be that the residual quick detection colorimetric method of existing agriculture can only carry out half-quantitative detection,
There are more false positive results, and Quantitative detection rule needs the spectrometer of specialty, it is high to there is testing cost, is unfavorable for
The shortcoming of popularization.The present invention provides a kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style, its mesh
Be that testing sample is fabricated to special " bio-barcode ", bar code scanning function and graphical analysis using smart mobile phone
Function, realization carries out Quantitative detection to residues of organophosphate pesticides.
A kind of above-mentioned the provided residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style of the present invention,
Its methods described is to follow these steps to carry out：
For the preparation of residues of organophosphate pesticides microchannel；
For adding the residual fast check reagent of agriculture in the residues of organophosphate pesticides microchannel；
For 39 yards of printing and makings of "+" code；
Scanning result is obtained for smart mobile phone scanning bar code；
Obtaining scanning area digital picture for smart mobile phone scanning bar code carries out colorimetric analysis.
Based on above-mentioned technical proposal, the further technical scheme of the present invention is as follows.
The preparation for residues of organophosphate pesticides microchannel is led to transparent covering layer by transparent base layer, transparent channel layer
Physical absorption is crossed to combine and constitute.Wherein, during transparent base layer is transparent plastic film, transparent thin silicon wafer and thin transparent quartz plate
One kind；Transparent channel layer is that the thickness being made up of PDMS polymer is the thin slice of 1mm, and at least one is offered on thin slice
The residual fast inspection microchannel pore of agriculture of vertical passage, the composition and its content of PDMS polymer are poly dimethyl ethylene methacrylic in mass ratio
Radical siloxane and poly dimethyl methyl hydrogen siloxane=10：1 mixing, then it is placed in 50 ~ 100min of baking in 60 ~ 80 DEG C of drying box；
Transparent covering layer is that PC plate and PC plate are shorter in length than two length of longitudinal passage holes embedded on PDMS, leads to two longitudinal directions
Road hole two ends are respectively formed a rectangular opening for being loaded and ventilating.
The composition and its content of the residual fast check reagent of agriculture are by volume iodo thioacetyl Dan Jian ﹕ dithiobis
The Ben Jia Suan ﹕ Yi phatidylcholine Zhi Mei ﹕ fruit and vegetable residual pesticides ﹕ 20 of 1 ﹕ of liquid=1 ﹕ 1 to be checked, carry out mixing acquisition.Wherein, fruit and vegetable residual pesticide is to be checked
Liquid is a kind of residual extract solution of agriculture in apple fruits and vegetables or celery.
It is described to be used for 39 yards of printing and makings of "+" code, it is to print on A4 paper 39 yards of "+" code, in the detection of "+" code
Shear two rectangle hollow out shapes of 20mm × 3mm and hough transform area corresponding with detection zone relevant position in area.
Described is that bar code is scanned with smart mobile phone for smart mobile phone scanning bar code acquisition scanning result,
It is the positive that "+" represents result, and it is negative that "-" represents result.
The acquisition of the residues of organophosphate pesticides detection microchannel digital picture is that screenshotss obtain smart mobile phone scanning bar code work
Make the picture in region, and chromatic value Y is obtained using handset image analysis application program, so as to obtain organophosphor agriculture in measuring samples
Residual concentration.
A kind of above-mentioned the provided residues of organophosphate pesticides detection based on smart mobile phone bar code readable style of the present invention is provided
Method, compared with the residual detection technique of existing agriculture, has the following advantages that：（1）Self-control agriculture residual detection microchannel have many flux detections,
Reuse, it is with low cost the advantages of；（2）Mobile phone screenshotss obtain picture under bar code is in scan pattern；（3）Scanning bar code
When open mobile phone flashlight cause so as to avoid detection zone from shade occur mess code occur simultaneously ensure every time scanning bar shaped
The brightness value of code is consistent；（6）The colouring information of detection picture is directly read using handset image analysis software, it is convenient and swift；（7）
Smart mobile phone has been spread in the middle of each family, barcode scanning software and handset image analysis software can it is free from network under
Carry and install.
Brief description of the drawings
Fig. 1 is the passage design principle figure that the present invention is based on 39 yards of "+" and "-" structural particularity.
Fig. 2 is residues of organophosphate pesticides detection microchannel design of the present invention and residues of organophosphate pesticides smart mobile phone scan stripes of the present invention
Shape code procedure chart.
Fig. 3 is " bio-barcode " design principle figure of the invention.
Fig. 4 is that bar code scan working region bar code screenshot picture is surveyed in residues of organophosphate pesticides examination criteria product examine of the present invention.
Fig. 5 is residues of organophosphate pesticides standard curve of the present invention.
Specific embodiment
Implement a kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style, its methods described be with
PC is substrate, and the PDMS microchannels film and PC surface coatings designed according to certain size, shape are covered each by the above
The microchannel that solution adds hole and passage is left into transparent, many flux, each passage that thickness is several millimeters thicks.Will be organic
The residual detection liquid of phosphorus agriculture is added in microchannel, then microchannel is put into 39 yards of detection zones of "+" code, is swept with smart mobile phone bar code
Retouch software to be scanned this " bio-barcode ", writing scan result.While scanning, mobile phone screenshotss obtain working region
Bar code picture, finally using the chromatic value Y of handset image analysis software detection zone, the variable quantity quantitative determination of Y value
The residual concentration of agriculture.
Below by taking the residual detection of parathion-methyl agriculture as an example, specific embodiment of the invention is made further instructions.
A kind of implementation for being used for residues of organophosphate pesticides fast determining method based on smart mobile phone scanning bar code,
（1）Bar code stencil design：Using 39 yards of "+" codes of Coreldraw Software for Design, the detection of this experiment parathion-methyl
Liquid is in finally yellow, therefore the bar of bar code designs Fill Color with CMYK passages, and C, M, K value are set to 0, and Y value is set to 100, obtains
To yellow "+" code, bar code is highly set to 20mm, and cell width wide is 6mm, and narrow cell width is 3mm；Use ink-jet printer
The bar code that will be designed is according to 1:1 ratio is printed on A4 paper；In the detection zone of "+" code（That is the 6th unit of code area
With the 9th latter half of unit）, and it is fabricated to two rectangle hollow out shapes of 20mm × 3mm.
（2）The preparation of organophosphor microchannel：It is thin to prepare PDMS from Dow Corning Corporation's product 184 silicon rubber of Sylgard
Film, comprises the concrete steps that：（1）From square culture dish as the carrier for displaying PDMS liquid mixtures.Preparation PDMS institutes are taken respectively
With matrix of materials and curing agent according to 10:1 ratio is well mixed in being put into above-mentioned culture dish.(2) mould as described in above-mentioned steps 1
The detection zone of plate, to the PDMS passages required for detection zone shear obtaining the transparent logical of two 22mm × 4mm of interval 3mm
Channel layer, it is the transparent PC of 50mm × 35mm to cut length, and transparent channel layer is adsorbed on PC by physisorption, is cut
Transparent PC layers of 28mm × 20mm is covered in transparent channel layer above by physisorption, and the two ends of coating PC are stayed respectively
There are well and passage, that is, complete the preparation of microchannel.
（3）The residual examination criteria curve plotting of agriculture：Using the parathion-methyl standard sample of pesticide in 100 μ g/mL acetone as mother
Liquid is dissolved in buffer solution, be configured to concentration be respectively 0,0.005,0.008,0.01,0.05,0.1,0.5,1,2,3,4,5,6, μ
The parathion-methyl agricultural chemicals of g/mL, the μ L of various concentrations parathion-methyl agricultural chemicals 200 are placed in centrifuge tube, then take respectively 10 μ L show
Color liquid and 10 μ L enzyme liquids are placed in each centrifuge tube, at 30 ~ 40 DEG C, incubate 20 ~ 30min.Take 10 μ L substrates and be separately added into each
In centrifuge tube, after 5 ~ 10min of reaction, the residual detection liquid of 50 ~ 100 μ L agricultures is taken from centrifuge tube and adds microchannel described in above-mentioned steps 2
It is interior.
（4）It is prepared by " bio-barcode "：By the micro- logical of the residues of organophosphate pesticides detection liquid for adding reaction to finish in above-mentioned steps 3
Under road is as rectangle hollow out detection zone " 1 " " 2 " in above-mentioned steps 1, bar code reverse side glues one layer tape（Scratch and remove detection zone glue
Band）, avoid scanning that mess code is obtained because there is gap shade occur microchannel and bar code bonding, so that it is also ensured that bar shaped
Code can be with Reusability, and " bio-barcode " is prepared and completed.
（5）Scanning obtains microchannel detection digital picture：By mobile phone level as on optical bracket, with what is installed on mobile phone
As a result barcode scanning software there is the bar code that preceding screenshotss obtain scanning area to the bar code scanning being placed on optical table
Picture.
（6）Colorimetric analysis prepares standard curve：Handset image analysis software opens the picture preserved in above-mentioned steps 5, point
Not Du Qu under various concentrations detection zone chromatic value Y.Result shows：The variable quantity of Y value detects liquid concentration with parathion-methyl
Increase and reduce.Canonical plotting is made with this, as shown in Figure 5.
（7）Residues of organophosphate pesticides detection in fruits and vegetables：With the apple in the apple in fruits and vegetables, pears, grape, spinach, Lettuce
As a example by carry out the residual detection of parathion-methyl agriculture.It is dissolved in as mother liquor using the parathion-methyl standard sample of pesticide in 100 μ g/mL acetone
In buffer solution, the parathion-methyl solution that concentration is 100,60,40,20 μ g/mL is configured to.Ultra-pure water is cleaned 3 times, epidermis
After globule natural air drying, by the parathion-methyl pesticide spraying of concentrations above on apple surface, after 3 ~ 10 hours of wait,
The residual natural air drying of apple epidermis agriculture.Cut and take 5 ~ 10g apple epidermises and be dissolved in the buffer solution of 5 ~ 10mL, quickly shake up 1 ~ 5min, carry
Above-mentioned solution is taken, in 1 ~ 3min is centrifuged in centrifuge, 200 μ L supernatant liquors is taken and is added in centrifuge tube as the residual liquid to be checked of agriculture.Again
10 μ L nitrite ions are taken respectively and 10 μ L enzyme liquids are placed in centrifuge tube, at 30 ~ 40 DEG C, incubate 20 ~ 30min.Take 10 μ L substrates difference
Add in centrifuge tube, after 5 ~ 10min of reaction, the residual fast check reagent of 50 ~ 100 μ L agricultures is taken from centrifuge tube and is added described in above-mentioned steps 2
It is to be scanned in any microchannel in microchannel etc..
Claims (1)
1. a kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style, its methods described is by following step
Suddenly carry out：
For the preparation of residues of organophosphate pesticides microchannel；
For adding the residual fast check reagent of agriculture in the residues of organophosphate pesticides microchannel；
For 39 yards of printing and makings of "+" code；
Scanning result is obtained for smart mobile phone scanning bar code；
Obtaining scanning area digital picture for smart mobile phone scanning bar code carries out colorimetric analysis；
The preparation for residues of organophosphate pesticides microchannel is to pass through thing with transparent covering layer by transparent base layer, transparent channel layer
Reason absorption is combined and constituted；
The transparent base layer is the one kind in transparent plastic film, transparent thin silicon wafer and thin transparent quartz plate；
The transparent channel layer is that the thickness being made up of PDMS polymer is the thin slice of 1mm, and at least one is offered on thin slice
The residual fast inspection microchannel pore of agriculture of bar vertical passage；
Wherein, the composition and its content of the PDMS polymer are 10 in mass ratio：1 is mixed to get dimethyl silicone polymer mixing
Thing, then it is placed in 50 ~ 100min of baking in 60 ~ 80 DEG C of drying box；
The transparent covering layer is that PC plate and PC plate are shorter in length than two length of longitudinal passage holes embedded on PDMS, makes two
Individual longitudinal passage holes two ends are respectively formed a rectangular opening for being loaded and ventilating；
The composition and its content of the residual fast check reagent of agriculture are by volume iodo thioacetyl Dan Jian ﹕ dithiobis benzene first
The Suan ﹕ Yi phatidylcholine Zhi Mei ﹕ fruit and vegetable residual pesticides ﹕ 20 of 1 ﹕ of liquid=1 ﹕ 1 to be checked, carry out mixing acquisition；
The fruit and vegetable residual pesticide liquid to be checked is a kind of residual extract solution of agriculture in apple fruits and vegetables or celery；
It is described to be used for 39 yards of printing and makings of "+" code, it is to print on A4 paper 39 yards of "+" code, cut in the detection zone of "+" code
Cut two rectangle hollow out shapes of 20mm × 3mm and hough transform area corresponding with detection zone relevant position；
Described is that bar code is scanned with smart mobile phone for smart mobile phone scanning bar code acquisition scanning result, "+" generation
Table result is the positive, and it is feminine gender that "-" represents result；
The acquisition of the residues of organophosphate pesticides detection microchannel digital picture is that screenshotss obtain smart mobile phone scanning bar code workspace
The picture in domain, and chromatic value Y is obtained using handset image analysis application program, so as to obtain residues of organophosphate pesticides in measuring samples
Concentration.
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| CN105784706B (en) * | 2016-03-09 | 2018-08-24 | 太原理工大学 | A kind of aflatoxin B1 fast determining method based on smart mobile phone detecting system |
| CN106645116B (en) * | 2016-09-29 | 2019-06-07 | 太原理工大学 | Biological rapid detection method based on smart phone two-dimensional code style |
| CN109116029A (en) * | 2018-07-21 | 2019-01-01 | 福建医科大学 | A method of paper substrate micro-fluidic chip is made based on inkjet printing technology controllability and is detected for glucose quantitation |
| CN112730367B (en) * | 2020-12-30 | 2022-07-15 | 四川农业大学 | Method and device for determining alkaline phosphatase by multi-signal spectrum sensing platform based on portable intelligent terminal |
| CN113567658B (en) * | 2021-07-27 | 2023-07-21 | 中国农业科学院农业质量标准与检测技术研究所 | Organophosphorus pesticide multi-residue biological bar code immunodetection kit based on hairpin self-assembly and application thereof |
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