CN104359904A - 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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- CN104359904A CN104359904A CN201410671941.8A CN201410671941A CN104359904A CN 104359904 A CN104359904 A CN 104359904A CN 201410671941 A CN201410671941 A CN 201410671941A CN 104359904 A CN104359904 A CN 104359904A
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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 being detected liquid adds with in polycarbonate (PC) and dimethyl silicone polymer (PDMS) microchannel that is material, again this microchannel is put in the detection zone of 39 yards "+", barcode scanning software on smart mobile phone is utilized to scan this " bio-barcode ", and screenshotss obtain the bar code number picture of perform region, the Y value of ultimate analysis digital picture detection zone realizes the quantitative test of residues of organophosphate pesticides detection.
Background technology
Along with the powerful gradually of smart mobile phone function and extensively popularize, the medical care diagnostic technology based on smart mobile phone causes to be paid close attention to and research widely.The people such as Ozcan increase microscopic system and modification on mobile phone are become simple microscope, fluorescent microscope on mobile phone, achieve 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) people such as Ericson utilizes chip and the software of particular design; the camera function of combined with intelligent mobile phone; achieve detection (the Oncescu V based on pH value in the saliva of colourimetry and sweat; O'Dell D, Erickson D. Lab Chip, 2013; 12:3232-3238.) and the detection of cholesterol (Oncescu V, Mancuso M, Erickson D. Lab Chip, 2014; 14:759-763. the people such as Shen (Li Shen.; Joshua A. Hagen.; Ian Papautsky. LabChip. 2012,12,4240 – 4243) utilize the camera function of smart mobile phone and achieve pH value in conjunction with particular color analysis software and detect; 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.) use the camera function of smart mobile phone, carrying out imaging to reacting complete testing sample in microchannel, utilizing computer software to carry out to obtained digital picture the detection that quantitative analysis achieves glucose in urine.The people such as Wang (Wang, S.; Zhao, X.; Khimiji, I.; Akbas, R.; Qiu, W.; Edwards, D.; Cramer, D.W.; Ye, B.; Demirci, U. Lab Chip. 2011,11,3411.), use IgG Antibody with Enzyme Linked Immunosorbent to detect the content of oophoroma biomarker HE4 in urine, by smart mobile phone imaging, utilize the software that mobile phone is installed to carry out corresponding analysis to the digital picture obtained and finally obtain testing result.
Above achievement in research, people are allowed to see smart mobile phone potentiality huge in following medical diagnosis, but owing to adding the Signal reception analytic system needing precision machined optical accessory and particular design, this smart mobile phone that utilizes in fact lost towards domestic consumer carries out real meaning and the value of instant detection.
One-dimensional bar code (barcode) refers to one group of regularly arranged bar and sky, and bar refers to that the part that reflectivity is little, sky refer to the part that reflectivity is large.Bar and empty different arrangement modes represent different meanings, the bar of the bar code of traditional sense and empty secret note and the informal voucher being respectively printing, the biological detection bar code that the application's patent proposes utilizes the colour developing result in biology, chemical detection as the bar of bar code with empty, and what is more important utilizes the free bar code scanning APP of common smart mobile phone just can realize the decoding of bar code.
It is to be noted Mirkin(Nam, J.-M. in 2003; Thaxton, C.S.; Mirkin, C.A. Science. 2003,301,1884-1886) etc. people invention bio-barcode detection technique completely different from the biological detection bar code that the application's patent proposes.What the application's patent proposed is the Structure of Bar-code of real meaning, utilize existing barcode encoding decoding principle just can realize quick diagnosis and the detection of biomolecule, and the bio-barcode detection technique that the people such as Mirkin propose is in conjunction with a series of DNA chain on nanogold particle probe, just make use of the figure notation concept of bar code, associate without any with the barcode design in practical application and reading.
Invention Ren is the people such as Li Xiaochun, publication number is the microchannel that 103674856A patent of invention " microchannel based on scanner uni colorimetric analysis is used for residues of organophosphate pesticides fast determining method technical field " proposes a kind of applicable scanning, and utilizing quick, the quantitative detecting method analyzing chromatic value and then realize residues of organophosphate pesticides, the scanner that this patent utilization is not easy to be subject to external light source interference obtains digital picture and carries out quantitative test to it.The weak point scanner volume of this technology is more very much not convenient for carrying and the digital picture obtained needs the professional color analysis softwares of Photoshop of importing computer to analyze, and is therefore unfavorable for extensive popularization and application.
Application people is the Zhang Yi of State Nanometer Science Center, and the publication number of Zhang Xing, Jiang Xingyu is that CN103272656A patent of invention " Bar code micro fluidic chip and uses thereof " has the difference of essence with the application's patent.Barcode encoding rule is dissolved in the design of micro-fluidic chip by the singularity of ": " and " 1 " of this patent utilization Codabar bar code, " 0 " and " 3 " structure of EAN code, each district of bar code is set to passage area by the method, each passage is reversibly fitted by substrate and the part with depression and formed microchannel, microchannel utilizes and passes into pipeline and discharge line to realize passing into and discharging of sample or reagent, produces bar pattern after directly distinguishing biochemistry detection reaction finally by bar-code reading device.Preparation technology's more complicated of the sunk part required for weak point passage of this technology and more thus to need to prepare number of channels more as the bar district quantity of passage area in force, need the bar code reading device of specialty simultaneously, thus be unfavorable for extensive popularization and application.
" bio-barcode " that relate in the application's patent be using the bar of 39 yards as detection zone, utilize the APP software scans bar code on smart mobile phone, the difference of detection zone testing concentration and obtain different results.The one-dimensional bar code used in this experiment is 39 yards, 39 yards be a kind of can the bar code of the information such as representative digit, letter, be mainly used in the automatic management of industry, books and ticket.Each character of 39 yards by 5 bars and 4 skies totally 9 unit form, these 9 unit only have two kinds of width, and wherein 3 wide unit, 9 narrow unit, are therefore called 39 yards.39 yards of standard are made up of starting character, coding symbol, full stop, and the starting character of 39 yards and full stop are identical by " * " representation.39 yards selected is out applied in this experiment is because its coding rule is simple and do not have check code.This experiment utilizes "+" code and "-" code of 39 representation, starting character, the full stop of these two bar codes are identical, and its code area only has rear four unit differences, utilize the characteristic of "+" code and "-" code structure, at the dotted line position of "+" code as detection zone (Fig. 1), after completion of the reaction, the barcode scanning software installed with smart mobile phone is to this bar code scan in detection zone, if scanning result is "-", represent result aobvious negative; Scanning result is "+", represents result aobvious positive.
Summary of the invention
The concrete technical problems that the present invention will solve is that the residual quick detection colourimetry of existing agriculture can only carry out half-quantitative detection, there is more false positive results, and Quantitative detection rule needs the spectrometer of specialty, there is testing cost high, is unfavorable for the shortcoming promoted.The invention provides a kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style, its objective is and testing sample is made into special " bio-barcode ", utilize bar code scanning function and the image analysis function of smart mobile phone, realize carrying out Quantitative detection to residues of organophosphate pesticides.
A kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style that the present invention is above-mentioned provided, method described in it follows these steps to carry out:
For the preparation of residues of organophosphate pesticides microchannel;
For adding the residual fast check reagent of agriculture in described residues of organophosphate pesticides microchannel;
For the printing and making of 39 yards of "+" codes;
Scanning result is obtained for smart mobile phone scanning bar code;
Obtain scanning area digital picture for smart mobile phone scanning bar code and carry out colorimetric analysis.
Based on technique scheme, the further technical scheme of the present invention is as follows.
The described preparation for residues of organophosphate pesticides microchannel is combined by physisorption with transparent covering layer by transparent base layer, transparent channel layer and is formed.Wherein, transparent base layer is the one in transparent plastic film, thin transparent silicon chip and thin transparent piezoid; Transparent channel layer is the thickness be made up of PDMS polymkeric substance is the thin slice of 1mm, and the residual fast inspection microchannel pore of the agriculture offering at least one vertical passage on thin slice, the composition of PDMS polymkeric substance and content thereof are that poly dimethyl methyl vinyl silicone mixes with poly dimethyl methyl hydrogen siloxane=10:1 in mass ratio, then are placed in the drying box baking 50 ~ 100min of 60 ~ 80 DEG C; Transparent covering layer is PC plate and the length being shorter in length than two longitudinal passage holes embedded on PDMS of PC plate, makes two longitudinal passage holes two ends all form a rectangular opening for application of sample and ventilation.
The composition of the residual fast check reagent of described agriculture and content thereof are iodo thioacetyl Dan Jian ﹕ bis-sulfo-Er Xiao Ji Ben Jia Suan ﹕ Yi phatidylcholine Zhi Mei ﹕ fruit and vegetable residual pesticide liquid=1 ﹕ 1 to be checked ﹕ 1 ﹕ 20 by volume, carry out mixing and obtain.Wherein, fruit and vegetable residual pesticide liquid to be checked is the residual extract of a kind of agriculture in apple, vegetables or celery.
The described printing and making for 39 yards of "+" codes, by 39 yards of "+" code printings on A4 paper, shears the rectangle hollow out shape of two 20mm × 3mm and the hough transform district corresponding with relevant position, detection zone in the detection zone of "+" code.
Described is scan bar code with smart mobile phone for smart mobile phone scanning bar code acquisition scanning result, and "+" represents result for positive, and "-" represents result for negative.
The acquisition that described residues of organophosphate pesticides detects microchannel digital picture is the picture that screenshotss obtain smart mobile phone scanning bar code perform region, and utilizes handset image analysis application to obtain chromatic value Y, thus obtains the concentration of residues of organophosphate pesticides in measuring samples.
Implement a kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style that the present invention is above-mentioned provided, compared with the residual detection technique of existing agriculture, tool has the following advantages: (1) self-control agriculture residual detection microchannel has that many flux detect, reuse, the advantage such as with low cost; (2) under bar code is in scan pattern, mobile phone screenshotss obtain picture; (3) open mobile phone flashlight during scanning bar code thus avoid detection zone to occur shade and cause the appearance of mess code to ensure simultaneously the brightness value of the bar code of each scanning is consistent; (6) handset image analysis software is adopted directly to read the colouring information detecting picture, convenient and swift; (7) smart mobile phone has spread in the middle of each family, and barcode scanning software and handset image analysis software all can be installed from network in free download.
Accompanying drawing explanation
Fig. 1 be the present invention based on the channels designs schematic diagram of 39 yards "+" and "-" structural particularity.
Fig. 2 is that residues of organophosphate pesticides of the present invention detects microchannel design and residues of organophosphate pesticides smart mobile phone scanning bar code procedure chart of the present invention.
Fig. 3 is the present invention's " bio-barcode " design concept figure.
Fig. 4 is that residues of organophosphate pesticides examination criteria product of the present invention detect bar code scan perform region bar code screenshot picture.
Fig. 5 is residues of organophosphate pesticides typical curve of the present invention.
Embodiment
Implement a kind of residues of organophosphate pesticides detection method based on smart mobile phone bar code readable style, method described in it take PC as substrate, cover the PDMS microchannel film and PC surface coating that design according to certain size in the above respectively, form that thickness is transparent, many flux of a few millimeters thick, each passage all leaves the microchannel that solution adds hand-hole and air hole.Residues of organophosphate pesticides is detected liquid add in microchannel, then microchannel is put into the detection zone of 39 yards of "+" codes, with smart mobile phone barcode scanning software, this " bio-barcode " is scanned, writing scan result.While scanning, mobile phone screenshotss obtain the bar code picture of perform region, and finally utilize the chromatic value Y of handset image analysis software detection zone, the variable quantity of Y value quantitatively detects the residual concentration of agriculture.
Be detected as example so that parathion-methyl agriculture is residual below, the specific embodiment of the present invention is made further instructions.
An enforcement for residues of organophosphate pesticides fast determining method is used for based on smart mobile phone scanning bar code,
(1) bar code stencil design: utilize Coreldraw Software for Design 39 yards of "+" codes, this experiment parathion-methyl detects liquid finally in yellow, therefore the bar of bar code CMYK channels designs Fill Color, C, M, K value is all set to 0, Y value is set to 100, obtains yellow "+" code, and bar code height is set to 20mm, wide cell width is 6mm, and narrow cell width is 3mm; With ink-jet printer, the bar code designed is printed on A4 paper according to 1:1 ratio; In the detection zone (i.e. the 6th unit of code area and the latter half of the 9th unit) of "+" code, and be made into the rectangle hollow out shape of two 20mm × 3mm.
(2) preparation of organophosphorus microchannel: select Dow Corning Corporation product Sylgard 184 silicon rubber to prepare PDMS film, concrete steps are: (1) selects square double dish as the carrier displaying PDMS liquid mixture.Get preparation PDMS material therefor matrix respectively to put into above-mentioned double dish with hardening agent according to 10:1 ratio and mix.(2) by the detection zone of template described in above-mentioned steps 1, PDMS passage required for detection zone is sheared to the transparent channel layer of the 22mm × 4mm obtaining two interval 3mm, cutting length is the transparent PC of 50mm × 35mm, transparent channel layer is adsorbed on PC by physisorption, the transparent PC layer of cutting 28mm × 20mm covers above transparent channel layer by physisorption, the two ends of overlayer PC leave well and air hole respectively, namely complete the preparation of microchannel.
(3) the residual examination criteria curve plotting of agriculture: be dissolved in damping fluid as mother liquor using the parathion-methyl standard sample of pesticide in 100 μ g/mL acetone, be mixed with concentration be respectively 0,0.005,0.008,0.01,0.05,0.1,0.5,1,2,3,4,5,6, the parathion-methyl agricultural chemicals of μ g/mL, variable concentrations parathion-methyl agricultural chemicals 200 μ L is placed in centrifuge tube, get 10 μ L nitrite ions more respectively and 10 μ L enzyme liquid are placed in each centrifuge tube, at 30 ~ 40 DEG C, incubation 20 ~ 30min.Getting 10 μ L substrates adds in each centrifuge tube respectively, after reaction 5 ~ 10min, gets the residual detection liquid of 50 ~ 100 μ L agriculture and add in microchannel described in above-mentioned steps 2 in centrifuge tube.
(4) " bio-barcode " preparation: detect the microchannel of liquid as under rectangle hollow out detection zone " 1 " " 2 " in above-mentioned steps 1 by adding the complete residues of organophosphate pesticides of reaction in above-mentioned steps 3, bar code reverse side glues one layer tape (scratching except surveyed area adhesive tape), microchannel and bar code are bonded to avoid scanning and obtains mess code because there being gap to occur shade, thus also can ensure that bar code can Reusability, prepared by " bio-barcode ".
(5) scanning obtains microchannel and detects digital picture: by mobile phone level as on optical bracket, with the barcode scanning software that mobile phone is installed to the bar code scanning be placed on optical table, and before result occurs, screenshotss obtain the bar code picture of scanning area.
(6) colorimetric analysis preparation standard curve: handset image analysis software opens the picture preserved in above-mentioned steps 5, reads the chromatic value Y of surveyed area under variable concentrations respectively.Result shows: the variable quantity of Y value detects the increase of liquid concentration with parathion-methyl and reduces.With this production standard curve map, as shown in Figure 5.
(7) residues of organophosphate pesticides in fruits and vegetables detects: carry out the residual detection of parathion-methyl agriculture for the apple in the apple in fruits and vegetables, pears, grape, spinach, Lettuce.Be dissolved in damping fluid using the parathion-methyl standard sample of pesticide in 100 μ g/mL acetone as mother liquor, being mixed with concentration is 100,60, the parathion-methyl solution of 40,20 μ g/mL.Ultrapure water cleans 3 times, after epidermis globule natural air drying, by the parathion-methyl pesticide spraying of above concentration on apple surface, waits for after 3 ~ 10 hours, the residual natural air drying of apple epidermis agriculture.Cut and get 5 ~ 10g apple epidermis and be dissolved in the damping fluid of 5 ~ 10mL, shake up 1 ~ 5min fast, extract above-mentioned solution, centrifugal 1 ~ 3min in hydro-extractor, get 200 μ L supernatant liquors and add in centrifuge tube as the residual liquid to be checked of agriculture.Get 10 μ L nitrite ions more respectively and 10 μ L enzyme liquid are placed in centrifuge tube, at 30 ~ 40 DEG C, incubation 20 ~ 30min.Get 10 μ L substrates to add respectively in centrifuge tube, after reaction 5 ~ 10min, to get in arbitrary microchannel that the residual fast check reagent of 50 ~ 100 μ L agriculture adds microchannel described in above-mentioned steps 2 etc. to be scanned in centrifuge tube.
Claims (10)
1., based on a residues of organophosphate pesticides detection method for smart mobile phone bar code readable style, method described in it follows these steps to carry out:
For the preparation of residues of organophosphate pesticides microchannel;
For adding the residual fast check reagent of agriculture in described residues of organophosphate pesticides microchannel;
For the printing and making of 39 yards of "+" codes;
Scanning result is obtained for smart mobile phone scanning bar code;
Obtain scanning area digital picture for smart mobile phone scanning bar code and carry out colorimetric analysis.
2. the method for claim 1 is be combined by physisorption with transparent covering layer by transparent base layer, transparent channel layer and formed for the preparation of residues of organophosphate pesticides microchannel described in it.
3. method as claimed in claim 2, transparent base layer described in it is the one in transparent plastic film, thin transparent silicon chip and thin transparent piezoid.
4. method as claimed in claim 2, transparent channel layer described in it is the thickness be made up of PDMS polymkeric substance is the thin slice of 1mm, and the residual fast inspection microchannel pore of the agriculture offering at least one vertical passage on thin slice;
Wherein, the composition of described PDMS polymkeric substance and content thereof in mass ratio for 10:1 is mixed to get polydimethylsiloxanemixture mixture, then are placed in the drying box baking 50 ~ 100min of 60 ~ 80 DEG C.
5. method as claimed in claim 2, transparent covering layer described in it is PC plate and the length being shorter in length than two longitudinal passage holes embedded on PDMS of PC plate, makes two longitudinal passage holes two ends all form a rectangular opening for application of sample and ventilation.
6. the method for claim 1, the composition of the residual fast check reagent of agriculture described in it and content thereof are iodo thioacetyl Dan Jian ﹕ bis-sulfo-Er Xiao Ji Ben Jia Suan ﹕ Yi phatidylcholine Zhi Mei ﹕ fruit and vegetable residual pesticide liquid=1 ﹕ 1 to be checked ﹕ 1 ﹕ 20 by volume, carry out mixing and obtain.
7. the method for claim 1, the liquid to be checked of fruit and vegetable residual pesticide described in it is the residual extract of a kind of agriculture in apple, vegetables or celery.
8. the method for claim 1, for the printing and making of 39 yards of "+" codes described in it, by 39 yards of "+" code printings on A4 paper, shear the rectangle hollow out shape of two 20mm × 3mm in the detection zone of "+" code and the hough transform district corresponding with relevant position, detection zone.
9. the method for claim 1, obtaining scanning result for smart mobile phone scanning bar code described in it is scan bar code with smart mobile phone, and "+" represents result for positive, and "-" represents result for negative.
10. the method for claim 1, the acquisition that residues of organophosphate pesticides described in it detects microchannel digital picture is the picture that screenshotss obtain smart mobile phone scanning bar code perform region, and utilize handset image analysis application to obtain chromatic value Y, thus obtain the concentration of residues of organophosphate pesticides in measuring samples.
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| CN112730367A (en) * | 2020-12-30 | 2021-04-30 | 四川农业大学 | Method and device for determining alkaline phosphatase by multi-signal spectrum sensing platform based on portable intelligent terminal |
| CN113567658A (en) * | 2021-07-27 | 2021-10-29 | 中国农业科学院农业质量标准与检测技术研究所 | Hairpin self-assembly-based organophosphorus pesticide multi-residue biological bar code immunoassay kit and application thereof |
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