CN116550402B - 3D paper-based microfluidic device and method for rapidly detecting malathion - Google Patents
3D paper-based microfluidic device and method for rapidly detecting malathion Download PDFInfo
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- CN116550402B CN116550402B CN202310802135.9A CN202310802135A CN116550402B CN 116550402 B CN116550402 B CN 116550402B CN 202310802135 A CN202310802135 A CN 202310802135A CN 116550402 B CN116550402 B CN 116550402B
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- 239000005949 Malathion Substances 0.000 title claims abstract description 54
- JXSJBGJIGXNWCI-UHFFFAOYSA-N diethyl 2-[(dimethoxyphosphorothioyl)thio]succinate Chemical compound CCOC(=O)CC(SP(=S)(OC)OC)C(=O)OCC JXSJBGJIGXNWCI-UHFFFAOYSA-N 0.000 title claims abstract description 54
- 229960000453 malathion Drugs 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 23
- 238000001514 detection method Methods 0.000 claims abstract description 52
- 102000004190 Enzymes Human genes 0.000 claims abstract description 20
- 108090000790 Enzymes Proteins 0.000 claims abstract description 20
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 6
- YRWWOAFMPXPHEJ-OFBPEYICSA-K sodium L-ascorbic acid 2-phosphate Chemical compound [Na+].[Na+].[Na+].OC[C@H](O)[C@H]1OC(=O)C(OP([O-])([O-])=O)=C1[O-] YRWWOAFMPXPHEJ-OFBPEYICSA-K 0.000 claims abstract description 6
- 230000007062 hydrolysis Effects 0.000 claims abstract description 5
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 20
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 claims description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 8
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 5
- 239000007853 buffer solution Substances 0.000 claims description 5
- 238000011084 recovery Methods 0.000 claims description 4
- 229920000557 Nafion® Polymers 0.000 claims description 3
- 102000003992 Peroxidases Human genes 0.000 claims description 3
- 238000006555 catalytic reaction Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 claims description 3
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 3
- 239000002105 nanoparticle Substances 0.000 claims description 3
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 3
- 238000009210 therapy by ultrasound Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 description 20
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000000575 pesticide Substances 0.000 description 4
- 239000000447 pesticide residue Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
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- 238000010146 3D printing Methods 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 229940123973 Oxygen scavenger Drugs 0.000 description 1
- 101150110933 PHY gene Proteins 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000000895 acaricidal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
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- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
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- 230000001788 irregular Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
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- 230000001404 mediated effect Effects 0.000 description 1
- 238000004452 microanalysis Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000003987 organophosphate pesticide Substances 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
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- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N21/76—Chemiluminescence; Bioluminescence
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0874—Three dimensional network
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
- B01L2300/126—Paper
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The application provides a 3D paper-based microfluidic device for detecting malathion, which comprises a paper layer i (a), a paper layer ii (b) and a paper layer iii (C), wherein the paper layer ii (b) is positioned between the paper layer i (a) and the paper layer iii (C) and can be folded to fix the paper layer i (a) and the paper layer iii (C), a hydrophobic channel is drawn on the paper layer i (a) and the paper layer iii (C) by an oily mark pen to form a hydrophilic area, and the hydrophilic area is reserved on the paper layer ii (b), wherein the paper layer i (a) is a sample adding layer, the hydrophilic area is preloaded with ALP, the paper layer ii (b) is an enzyme hydrolysis layer, the hydrophilic area is fixed with AA2P, the paper layer iii (C) is a chemiluminescent layer, and the hydrophilic area is fixed with Pt-Co-N-C nano enzyme. The application also provides a method for detecting malathion by using the device. The application does not need any instrument, and has simple operation, light weight and portability; has higher sensitivity compared with other rapid detection methods.
Description
Technical Field
The application belongs to the technical field of rapid food detection, and particularly relates to a 3D paper-based microfluidic device and method for rapidly detecting malathion.
Background
The micro laboratory on paper is a micro analysis system established by taking paper as a substrate, and utilizes a recognition element fixed on the surface of the paper to react with target molecules in a recognition way, so that the concentration of the analyte is converted into optical, electrochemical or other signals for qualitative, semi-quantitative or quantitative detection. In recent years, a micro laboratory on paper has become a research hot spot in the technical field of rapid food detection because of the advantages of rapidness, portability, sensitivity, economy and the like.
The pesticide is an important means for preventing and controlling plant diseases and insect pests and regulating plant growth in agricultural production, and plays a vital role in protecting the growth of crops and improving the quality of agricultural products. However, irregular use of pesticides causes serious food and environmental pollution, and damages human health through accumulation of food chains. Malathion is an organophosphorus pesticide widely used in the world, has low toxicity, good insecticidal and acaricidal effects and wide control range. However, malathion has toxicity of inhibiting neurotransmitters, and can cause toxicity in human body by skin or mucous membrane contact, inhalation or misfeeding. At present, the existing detection method for pesticide residues has complicated sample pretreatment, needs to rely on a large-scale precise analysis instrument, has higher detection cost, needs professional technicians to operate, and cannot meet the field rapid detection requirement of a large number of samples. Therefore, in order to overcome the existing difficulty, the application designs a 3D paper-based microfluidic device for rapidly detecting malathion, which is applied to actual detection.
Disclosure of Invention
The application aims to design a 3D paper-based microfluidic device and method for rapidly detecting malathion.
In order to meet the requirements of accurate and portable on-site rapid detection of food containing malathion, the application controls the reaction step through the position of the sample area, thereby realizing the portable detection of the food containing malathion.
The application provides a 3D paper-based microfluidic device for detecting malathion, which comprises a paper layer i a, a paper layer ii b and a paper layer iii C, wherein the paper layer ii b is positioned between the paper layer i a and the paper layer iii C, and can be folded to fix the paper layer i a and the paper layer iii C, an oily Mark pen is used for drawing a hydrophobic channel on the paper layer i a and the paper layer iii C to form a hydrophilic area, and the hydrophilic area is reserved on the paper layer ii b, wherein the paper layer i a is a sample adding layer, the hydrophilic area is preloaded with ALP, the paper layer ii b is an enzyme hydrolysis layer, the hydrophilic area is fixed with AA2P, the paper layer iii C is a chemiluminescent layer, and the hydrophilic area is fixed with Pt-Co-N-C nano enzyme.
Further, the paper-based carrier is Woltmann 1 paper, and the Woltmann 1 paper is obtained by dripping Pt-Co-N-C nano enzyme solution with simulated peroxidase activity into the Woltmann 1 paper and drying the Pt-Co-N-C nano enzyme solution at room temperature.
Further, the Pt-Co-N-C nano enzyme is obtained by dissolving Pt-Co-N-C nano particles in a system of 99% ethanol and 1% 5% Nafion solution, wherein the concentration is 0.05 mu M, and performing ultrasonic treatment.
Further, the paper layer ii b extends outwards to form a paper layer iv d and a paper layer v e, the paper layer iv d and the paper layer v d are folded to form a cover and a bottom plate of the device respectively, and the paper layer iv d and the paper layer v e are provided with positioning points.
Further, the 3D paper-based microfluidic device was folded using whatman 1 paper.
The application also provides a method for detecting malathion by using the device, which comprises the following steps:
dripping malathion extract sample into the sample adding area of paper layer ia, and incubating at 25deg.C for 10 min; and/or the number of the groups of groups,
adding 35 mu L of Tris-HCl buffer solution with pH of 9.0 into the hydrophilic area of the paper layer i a, and reacting for a certain time; and/or the number of the groups of groups,
40. Mu.L of NaOH buffer with pH of 12.0 is added in the hydrophilic region of the paper layer i a; and/or the number of the groups of groups,
adding a luminol reagent A solution and a luminol reagent B solution into the detection area of the paper layer iii c; and/or the number of the groups of groups,
and (3) placing the paper layer iii c of the 3D paper-based microfluidic device upwards in a camera bellows, and measuring the B value of each detection area under the HSB color mode.
Further, the method comprises a step of fitting a standard curve, the step comprising:
adding a malathion solution with standard concentration into a detection device, photographing to obtain a B value, and fitting the concentration of malathion and the obtained B value into a standard curve.
Further, before dropping the extract sample, the method further comprises the steps of: the hydrophobic channels were drawn with an oily mark pen on the paper layers i a and iii c, leaving hydrophilic areas of the same size on the paper layer ii b.
Further, the B value is obtained through camera shooting of the smart phone, shooting parameters are that shutter time is set to be 8 s, ISO is set to be 500, exposure compensation is set to be 0 in a professional mode, chemiluminescence is measured when the reaction is carried out for 10 min through shooting, and the obtained photo is used for measuring the B value of each detection area in an HSB color mode through an application program.
Further, after the Tris-HCl buffer solution is dripped into the hydrophilic region of the paper layer i a, the method further comprises the steps of: the paper layer i a is pushed and pulled so that the rightmost side of the paper layer i a is aligned with the locating point on the paper layer ii b.
Further, the method comprises the step of verifying: the detection amount, recovery rate and RSD of the two methods were compared with the gold standard GC-MS as a reference.
By means of the technical scheme, the portable malathion-containing food rapid detection device provided by the application has at least one of the following advantages and beneficial effects:
(1) The 3D paper-based microfluidic detection device has the advantages of small volume, light weight and low cost;
(2) No instrument is needed, and the operation is simple, light and portable;
(3) Based on the device, a perfect on-site accurate, efficient and portable food detection method containing malathion can be constructed.
Drawings
FIG. 1 is a schematic diagram of a portable malathion-containing food rapid detection apparatus;
fig. 2 is a schematic diagram of the operation of the portable malathion-containing food rapid detection apparatus.
Fig. 3 shows the sensitivity of malathion detection based on the portable malathion-containing food rapid detection device.
Detailed Description
The following examples are illustrative of the application and are not intended to limit the scope of the application. Unless otherwise indicated, the technical means used in the examples are conventional means well known to those skilled in the art, and all raw materials used are commercially available.
Malathion is supplied by Sigma-Aldrich; the luminol reagent A solution and the luminol reagent B solution are provided by Phygene; sodium hydroxide, tris (hydroxymethyl) aminomethane, is supplied by microphone; hydrochloric acid is supplied by national pharmaceutical group chemical company, inc.
The application provides a 3D paper-based microfluidic device for detecting malathion, which mainly comprises three pieces of filter paper and a piece of plastic film, and can be split into two parts through bonding and folding. The device comprises a paper layer i a, a paper layer ii b and a paper layer iii c, the paper layer ii b being located between the paper layer i a and the paper layer iii c. The paper layer ii b extends to two sides to form a paper layer iv d and a paper layer v d, namely, the paper layer ii b, the paper layer iv d and the paper layer v e belong to the same piece of filter paper and are formed by folding, and the paper layer iv d and the paper layer v d respectively form a cover and a bottom plate of the device and are used for fixing the paper layer i a and the paper layer iii c. Paper layers i a and iii c are drawn with black circular hydrophobic channels by an oily mark pen to form hydrophilic areas, and paper layer ii b leaves circular hydrophilic areas with the same size. Two positioning points are arranged on the paper layer iv d and the paper layer v e, so that hydrophilic areas are aligned with each other when the paper layer is pushed and pulled, and dislocation is prevented. Paper layer i a is a sample addition layer, and the hydrophilic area is preloaded with ALP; paper layer ii b is an enzyme hydrolysis layer, and the hydrophilic area is fixed with AA2P; paper layer iii C is a chemiluminescent layer, and Pt-Co-N-C nano enzyme is immobilized on the hydrophilic region.
Preferably, the paper-based carrier of the 3D paper-based microfluidic detection device provided by the application is Woltmann 1 paper, and the paper-based is obtained by dripping Pt-Co-N-C nano enzyme solution with simulated peroxidase activity into the Woltmann 1 paper and drying at room temperature. The Pt-Co-N-C nano enzyme is obtained by dissolving Pt-Co-N-C nano particles in a system of 99% ethanol and 1% 5% Nafion solution, wherein the concentration is 0.05 mu M, and carrying out ultrasonic treatment.
In one embodiment, the 3D paper-based microfluidic detection device provided by the application is made of a polyacetic acid material through an FDM 3D printer, and has the advantages of small volume, light weight and low cost. In one embodiment, the 3D printing device is 80 mm in size60 mm/>5 mm, the mass is only 14 g, and the carrying is convenient.
In other embodiments, the D-paper based microfluidic device is folded using whatman 1 paper.
The 3D paper-based microfluidic detection device provided by the application can be used for detecting malathion, and as shown in figure 2A, the detection principle comprises two parts of identifying malathion and nano-enzyme catalyzed luminol chemiluminescence amplifying signals by an enzyme inhibition method. As one of organophosphorus pesticides, malathion has obvious inhibition effect on ALP enzyme activity. Under alkaline conditions, the substrate AA2P can be hydrolyzed by ALP, yielding AA with strong reducibility. On the other hand, the oxidation reaction of luminol and H2O2 is accompanied by blue chemiluminescence, and the catalysis of the Pt-Co-N-C nano enzyme can amplify the chemiluminescence signal by 73 times. When both reactions occur in the same system, AA acts as an active oxygen scavenger, quenching the chemiluminescent light of luminol, reducing its signal. Thus, the higher the concentration of malathion, the weaker the enzymatic activity of ALP, the less AA is produced by hydrolysis, and the stronger the luminol chemiluminescent signal is detected, i.e., malathion is positively correlated with the chemiluminescent signal.
Further, the application also provides a method for detecting malathion by using the 3D paper-based microfluidic device, which comprises the steps of fitting a standard curve: adding a malathion solution with standard concentration into a detection device, photographing to obtain a B value, and fitting the concentration of malathion and the obtained B value into a standard curve. As shown in fig. 2, the specific operation steps are as follows:
opening a 3D paper-based microfluidic paper layer iv D, taking 40 mu L of malathion standard solution or pesticide residue extracting solution, and dripping the solution into a round sample adding area of the 3D paper-based microfluidic paper layer i a, and incubating for 10 min at 25 ℃. Next, the paper layers i a and iii c are pushed and pulled so that the rightmost side of the paper layer i a is aligned with the locating point on the paper layer ii b. 35 mu LpH 9.0.9.0 Tris-HCl buffer is added dropwise to the circular hydrophilic zone of paper layer i a, reacted for a certain period of time, preferably 30min at 37 ℃, and then 40 mu L of pH12.0 NaOH buffer is added. Then, the 3D paper-based microfluidic device was flipped back and forth, and 5 μl of commercial luminol reagent a solution and 5 μl of luminol reagent B solution were added to the detection zone of paper layer iii c. Next, the device paper layer iii c was placed up in a camera box, the camera of the smartphone was turned on, the shutter time was set to 8 s, iso was 500, the exposure compensation was 0, and chemiluminescence was measured in a photographing for 10 min. Finally, the obtained photographs were imported into a Color Grab application program, and the B value in the HSB Color mode of each detection area was measured. The concentration of malathion and the obtained B value are input into the mobile phone Data Analysis Lite application program to fit a standard curve.
In one specific example of malathion-containing food product testing, as shown in fig. 2, residual malathion is first extracted from the surface of a food product sample. The cotton swab is firstly used for uniformly wiping the surface of the food to fully extract the residual pesticide on the food, so that the damage to the texture and the nutritional ingredients of the food and the influence on the physical and chemical properties of the food are avoided. The extracted sample is dissolved in Tris-HCl buffer solution, and is dripped into a hydrophilic sample adding area of the 3D paper-based microfluidic paper layer i a, and incubated with ALP for 10 min at 25 ℃, so that the enzyme activity of the ALP is inhibited. And then adding Tris-HCl buffer solution dropwise to bring ALP into an enzymolysis reaction zone of the paper layer ii b, and hydrolyzing the preloaded AA2P serving as a substrate of the ALP at 37 ℃ to generate AA. The addition of NaOH buffer, the strongly alkaline environment not only provides the best reaction conditions for luminol chemiluminescence, but also terminates the hydrolysis reaction of ALP, bringing the resulting AA into the detection zone of paper layer ii b. As shown in fig. 2, the device was turned over and the added luminol reagent was blue-colored under the catalysis of the pre-immobilized Pt-Co-N-C nanoenzyme while a portion was quenched by AA. The more pesticide residue, the less AA is produced, the brighter the chemiluminescence, and vice versa. Because chemiluminescent signals are difficult to capture by the naked eye in bright places, 3D paper-based microfluidics is placed in a dark field, and the chemiluminescent signals are converted into B values in HSB mode by utilizing the advantage of slow-gate photography technology that optical signals can be collected for a longer time.
As shown in FIG. 3, 0-160 μg/kg malathion was measured using 3D paper-based microfluidic, the luminescence signal was recorded with a mobile phone and Color analysis was performed using Color Grab application. As malathion concentration increases, the chemiluminescent signal captured by the cell phone gradually increases, and the higher the B value in HSB color mode, as shown in fig. 3A. Calculating inhibition ratio (inhibition ratio) = (B1-B0)/B0 by using a formula100, wherein B0 is the B value of the blank group and B1 is the B value of the test group. As shown in fig. 3B, whenWhen the concentration of malathion is in the range of 60-130 mug/kg, the concentration of malathion has a linear relation with the inhibition ratio: y=0.415 x-18.8 (r2=0.996). The detection limit was 10.7. Mu.g/kg calculated according to the formula 3.3 ɛ/S.
Table 1 shows the results of the practical detection of the portable malathion-containing food rapid detection device, and in order to verify the feasibility of the Pt-Co-N-C nano enzyme-mediated chemiluminescence 3D paper-based microfluidic in the practical sample detection, negative and positive standard adding samples are set as samples to be detected as shown in Table 1. Meanwhile, the detection amounts of the two detection methods, the recovery rate and the RSD thereof are compared by taking a gold standard GC-MS as a reference. Wherein, no negative sample was detected. The recovery rate is 89.8-99.2%, the RSD is 1.9-6.5%, and the on-site rapid detection requirement of malathion can be met.
TABLE 1
("-" means undetected)
Because the detection device provided by the application has two small blue triangles as positioning points, when three layers of paper layers i a, ii b and iii c are aligned, the two positioning points are on the midverticals of three circular sample areas, and then the sample adding step is completed; when the paper layer i a is pulled leftwards to the rightmost side to be aligned with the positioning point, the circular hydrophilic areas of the paper layer i a, the paper layer ii b and the paper layer iii c are aligned, and the two reactions are completed. Therefore, the detection device provided by the application can realize the accurate and portable on-site rapid detection requirement of the food containing malathion, and the portable detection of the food containing malathion is realized by controlling the reaction step through the position of the sample area.
The detection device and the detection method provided by the application do not need any independent instrument, and are simple to operate, light and portable; has higher sensitivity compared with other rapid detection methods. The method has great application prospect in the field detection of pesticide residues.
The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the application, and is not meant to limit the scope of the application, but to limit the application to the particular embodiments, and any modifications, equivalents, improvements, etc. that fall within the spirit and principles of the application are intended to be included within the scope of the application.
Claims (8)
1. A method for rapid detection of malathion using a 3D paper-based microfluidic device, wherein the device comprises paper layers i (a), ii (b) and iii (c), with ii (b) being located between i (a) and iii (c) and being foldable to secure i (a) and iii (c) paper layers, ii (b) extending outwardly to form iv (D) and v (e) paper layers, iv (D) and v (e) paper layers being folded to form the lid and base plate of the device respectively, iv (D) and v (e) paper layers being provided with anchor points;
drawing a hydrophobic channel on the paper layer i (a) and the paper layer iii (C) by using an oily Mark pen to form a hydrophilic region, and reserving the hydrophilic region on the paper layer ii (b), wherein the paper layer i (a) is a sample adding layer, ALP is preloaded in the hydrophilic region, the paper layer ii (b) is an enzyme hydrolysis layer, AA2P is fixed on the hydrophilic region, the paper layer iii (C) is a chemiluminescent layer, pt-Co-N-C nano enzyme is fixed on the hydrophilic region, and the catalysis energy of the Pt-Co-N-C nano enzyme can amplify a chemiluminescent signal;
the method comprises the following steps: dripping a malathion extract sample into the sample adding area of the paper layer i (a), and incubating for 10 min at 25 ℃; and, adding 35 mu L of Tris-HCl buffer solution with pH of 9.0 into the hydrophilic area of the paper layer i (a), and reacting for 30min; and, adding 40. Mu.L of NaOH buffer with pH of 12.0 to the hydrophilic region of the paper layer i (a); and, adding a luminol reagent A solution and a luminol reagent B solution to the detection zone of paper layer iii (c); and, a step of, in the first embodiment,
and (3) placing the paper layer iii (c) of the 3D paper-based microfluidic device upwards in a camera bellows, and measuring the B value of each detection area under the HSB color mode.
2. The method for detecting malathion according to claim 1, comprising the step of fitting a standard curve, said step comprising: adding a malathion solution with standard concentration into a detection device, photographing to obtain a B value, and fitting the concentration of malathion and the obtained B value into a standard curve.
3. The method for detecting malathion according to claim 2, wherein the B value is obtained by camera shooting of a smart phone, shooting parameters are that shutter time is set to 8 s, iso is 500, exposure compensation is 0, chemiluminescence is measured when the reaction is performed for 10 min in a professional mode, and the obtained photo is used for measuring the B value in the HSB color mode of each detection area through an application program.
4. The method for detecting malathion according to claim 3, further comprising the step of, after the step of dropping the extraction liquid sample: pulling paper layers i (a) and iii (c) to align the rightmost sides of paper layers i (a) and iii (c) with the locating points on paper layer ii (b).
5. The method for detecting malathion of claim 4 further comprising the step of verifying: the detection amount, recovery rate and RSD of the two detection methods were compared with the gold standard GC-MS as a reference.
6. The method for detecting malathion according to claim 1, wherein the carrier of the 3D paper-based microfluidic device is whatman 1 paper, obtained by dropping Pt-Co-N-C nano enzyme solution having a simulated peroxidase activity on the whatman 1 paper and drying at room temperature.
7. The method for detecting malathion according to claim 1, wherein the Pt-Co-N-C nano-enzyme is obtained by dissolving Pt-Co-N-C nano-particles in a system of 99% ethanol and 1% 5% Nafion solution at a concentration of 0.05 μΜ and performing ultrasonic treatment.
8. The method of claim 1, wherein the 3D paper-based microfluidic device is folded using whatman 1 paper.
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Inventor after: Cheng Nan Inventor after: Luo Yunbo Inventor after: Chen Yang Inventor after: Zhang Qi Inventor after: Lu Mengyu Inventor after: Ke Ruiqing Inventor after: Ma Tianyi Inventor after: He Xiaoyun Inventor after: Xu Wentao Inventor after: Huang Kunlun Inventor before: Cheng Nan Inventor before: Zhang Junjie Inventor before: Xing Weiwei Inventor before: He Xiaoyun Inventor before: Xu Wentao Inventor before: Huang Kunlun Inventor before: Luo Yunbo |