CN111198173A - Preparation method of fluorescent test strip for environmental monitoring - Google Patents
Preparation method of fluorescent test strip for environmental monitoring Download PDFInfo
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- CN111198173A CN111198173A CN202010039143.9A CN202010039143A CN111198173A CN 111198173 A CN111198173 A CN 111198173A CN 202010039143 A CN202010039143 A CN 202010039143A CN 111198173 A CN111198173 A CN 111198173A
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Images
Classifications
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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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
- G01N21/274—Calibration, base line adjustment, drift correction
Abstract
The invention provides a preparation method of a fluorescent test strip for environmental monitoring, and relates to the technical field of environmental detection. A preparation method of a fluorescent test strip for environmental monitoring comprises the steps of firstly selecting a bottom plate, then preparing a sample pad, a nano material pad, a reaction membrane and a water absorption pad, and finally assembling the sample pad, the nano material pad, the reaction membrane and the water absorption pad on one side of the bottom plate in sequence. The invention can realize the determination of arsenic element in soil.
Description
Technical Field
The invention relates to the technical field of environmental detection, in particular to a preparation method of a fluorescent test strip for environmental monitoring.
Background
With legislation on soil control, supervision on soil pollution has become a new focus. When project environmental assessment and site investigation are carried out, soil investigation and monitoring are required.
Soil is an environment on which organisms live, and in recent years, a lot of environmental pollution poses an unprecedented challenge on the safety of soil. In order to enhance the stability of soil and prevent soil pollution, the ministry of ecological environment announces two new soil environment quality standards in 2018, 6 and 22 months. In the same year, 8 and 31 days, the state came out of the soil pollution prevention and treatment law of the people's republic of China. With legislation on soil control, supervision on soil pollution has become a new focus.
According to the relevant national regulations, when project environment influence evaluation and site environment investigation are carried out, soil investigation and monitoring are required. In the process, monitoring of various factors can be involved, and first-hand data of the soil environment of the project site can be mastered through monitoring. And mercury and arsenic monitoring is also an important content of soil environment monitoring. In 2008, the ministry of environmental protection originally issued atomic fluorescence method for measuring total mercury, total arsenic and total lead of soil quality, and specified the measuring requirements of mercury, arsenic and other elements in soil in detail.
The fluorescence analysis method is concerned with due to high analysis sensitivity, strong selectivity and simple and convenient use, the fluorescence test strip is an important monitoring tool in the fluorescence analysis method, and the fluorescence test strip as a simple and quick analysis tool is widely used in clinical diagnosis, food safety and environmental monitoring. There are no fluorescent test strips on the market for monitoring arsenic in soil.
Disclosure of Invention
The invention aims to provide a preparation method of a fluorescent test strip for environmental monitoring, which can realize the determination of arsenic element in soil.
The embodiment of the invention is realized by the following steps:
a preparation method of a fluorescent test strip for environmental monitoring comprises the following steps,
selecting a substrate, namely preparing a substrate for preparing the fluorescent test strip;
preparing a sample pad: one side of the bottom plate is provided with a sample pad, and the preparation steps of the sample pad are as follows: placing the sample pad in arsenic standard treatment liquid, soaking for about 40-100 minutes, taking out, and placing in a 36 ℃ forced air drying oven to dry for at least more than 12 hours;
preparing a nano-material pad, wherein one end of the sample pad is butted with the nano-material pad, the nano-material pad is connected with the bottom plate, and the preparation method of the nano-material pad comprises the following steps: spraying the graphene oxide suspension solution on a glass cellulose membrane, drying for 6-10 h at 100-140 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 15-25 min, and drying at 30-35 ℃;
preparing a reaction film: the end of the nano material pad far away from the sample pad is butted with a reaction membrane, the reaction membrane is connected with the bottom plate, and the preparation method of the reaction membrane comprises the following steps: a detection line is arranged at one end, close to the nano material pad, of the reaction membrane, a calibration line is arranged at one end, far away from the nano material, of the reaction membrane, the potassium borohydride solution and the avidin are incubated for 25-35 min and sprayed to the detection line, and an anti-fluorescein antibody corresponding to free fluorescein is sprayed on the calibration line;
preparing a water absorption pad: one end of the reaction membrane, which is far away from the nano material, is butted with a water absorption pad, the water absorption pad is connected with the bottom plate, and the water absorption pad is made of water absorption material.
In some embodiments of the present invention, the bottom plate is a rectangular plate, and the bottom plate is a PVC plate.
In some embodiments of the invention, the floor is made of a water impermeable material.
In some embodiments of the present invention, the nanomaterial pad is a porous nanomaterial membrane or a cellulose membrane modified by a nanomaterial, and the nanomaterial is at least one of graphene oxide, reduced graphene, gold nanoparticles, molybdenum disulfide, and carbon nanotubes.
In some embodiments of the present invention, the absorbent pad is absorbent paper.
In some embodiments of the present invention, the reaction membrane is a nitrocellulose membrane.
In some embodiments of the present invention, the sample pad is made of a cellulose membrane.
In some embodiments of the present invention, the sides of the sample pad, the nanomaterial pad, the reaction membrane, and the water absorption pad facing the bottom plate are all convex, the bottom plate is provided with notches correspondingly matching with the convex portion of the sample pad, the convex portion of the nanomaterial pad, the convex portion of the reaction membrane, and the convex portion of the water absorption pad, and the sample pad, the nanomaterial pad, the reaction membrane, and the water absorption pad are connected with the bottom plate through glue.
The embodiment of the invention at least has the following advantages or beneficial effects:
the invention provides a preparation method of a fluorescent test strip for environmental monitoring, which comprises the following steps,
selecting a substrate, namely preparing a substrate for preparing the fluorescent test strip;
preparing a sample pad: one side of the bottom plate is provided with a sample pad, and the preparation steps of the sample pad are as follows: placing the sample pad in arsenic standard treatment liquid, soaking for about 40-100 minutes, taking out, and placing in a 36 ℃ forced air drying oven to dry for at least more than 12 hours;
preparing a nano-material pad, wherein one end of the sample pad is butted with the nano-material pad, the nano-material pad is connected with the bottom plate, and the preparation method of the nano-material pad comprises the following steps: spraying the graphene oxide suspension solution on a glass cellulose membrane, drying for 6-10 h at 100-140 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 15-25 min, and drying at 30-35 ℃;
preparing a reaction film: the end of the nano material pad far away from the sample pad is butted with a reaction membrane, the reaction membrane is connected with the bottom plate, and the preparation method of the reaction membrane comprises the following steps: a detection line is arranged at one end, close to the nano material pad, of the reaction membrane, a calibration line is arranged at one end, far away from the nano material, of the reaction membrane, the potassium borohydride solution and the avidin are incubated for 25-35 min and sprayed to the detection line, and an anti-fluorescein antibody corresponding to free fluorescein is sprayed on the calibration line;
preparing a water absorption pad: one end of the reaction membrane, which is far away from the nano material, is butted with a water absorption pad, the water absorption pad is connected with the bottom plate, and the water absorption pad is made of water absorption material.
When the content of arsenic in soil is detected, a soil aqueous solution of arsenic is dripped onto a sample pad, the sample can dialyze under the action of capillary action and moves towards the nano material pad, when the sample enters the nano material pad, the arsenic in the sample adsorbs thiourea in the nano material pad and drives the thiourea to move towards a detection line, a mixed solution of the arsenic and the thiourea enters the detection line of the nano material pad and reacts with potassium borohydride at the detection line to reduce the arsenic in the sample into arsine, and the detection line shows fluorescence when the device is placed under the irradiation of a special arsenic hollow cathode lamp. And simultaneously, the sample drives the free fluorescein in the nano material pad to move towards the calibration line, so that the free fluorescein is combined with the anti-fluorescein antibody, the fluorescence is shown at the calibration line, and the content of the arsenic element is determined by measuring the ratio of the fluorescence intensity at the detection line to the fluorescence intensity at the calibration line (the larger the ratio, the more the arsenic content is, and the smaller the arsenic content is, the less arsenic content is detected by a specific fluorescence intensity detection device, which is the existing device, and the detailed description is not given here). The sample pad is placed in the arsenic standard treatment solution to be soaked for about 40-100 minutes, and is taken out and then placed in a 36-DEG C blast drying oven to be dried for at least more than 12 hours, so that the immunity of the fluorescent test strip to other elements can be improved, and the precision of the fluorescent test strip is improved. If the sample does not contain the arsenic element to be detected, no fluorescence exists at the detection line, and no matter whether the sample is detected or not, the free fluorescein on the nano material pad is combined by the anti-fluorescein antibody at the calibration line when the free fluorescein goes up to the calibration line, so that the fluorescence appears on the calibration line. If no fluorescence appears at the calibration line, the test strip is invalid, and the test strip is replaced for retesting. The calibration line can timely know whether the test strip is invalid or not, and the detection accuracy is improved.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a schematic structural diagram of a fluorescent test strip according to an embodiment of the present invention;
icon: 100-bottom plate, 101-sample pad, 102-nano material pad, 103-reaction membrane, 104-water absorption pad, 1001-detection line, 1002-calibration line.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to specific examples.
A preparation method of a fluorescent test strip for environmental monitoring comprises the following steps,
selecting a substrate, namely preparing a substrate 100 for preparing the fluorescent test strip;
preparation of sample pad 101: a sample pad 101 is disposed on one side of the base plate 100, and the preparation steps of the sample pad 101 are as follows: placing the sample pad 101 in arsenic standard treatment liquid to be soaked for about 40-100 minutes, taking out the sample pad, and placing the sample pad in a 36-DEG C forced air drying oven to be dried for at least more than 12 hours;
preparing the nano-material pad 102, wherein the nano-material pad 102 is arranged at one end of the sample pad 101 in a butt joint mode, the nano-material pad 102 is connected with the bottom plate 100, and the preparation method of the nano-material pad 102 comprises the following steps: spraying the graphene oxide suspension solution on a glass cellulose membrane, drying for 6-10 h at 100-140 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 15-25 min, and drying at 30-35 ℃;
preparation of the reaction film 103: one end of the nano-material pad 102 far away from the sample pad 101 is butted with a reaction membrane 103, the reaction membrane 103 is connected with the bottom plate 100, and the preparation method of the reaction membrane 103 comprises the following steps: a detection line 1001 is arranged at one end, close to the nano material pad 102, of the reaction membrane 103, a calibration line 1002 is arranged at one end, far away from the nano material, of the reaction membrane 103, the potassium borohydride solution and the avidin are incubated for 25-35 min and sprayed to the detection line 1001, and an anti-fluorescein antibody corresponding to free fluorescein is sprayed on the calibration line 1002;
preparation of absorbent pad 104: the end of the reaction membrane 103 far away from the nano material is butt-jointed with a water absorption pad 104, the water absorption pad 104 is connected with the bottom plate 100, and the water absorption pad 104 is made of water absorption material.
When the content of arsenic in soil is detected, a soil aqueous solution of arsenic is dripped onto a sample pad 101, the sample can dialyze under the action of capillary and moves towards a nano material pad 102, when the sample enters the nano material pad 102, the arsenic in the sample adsorbs thiourea in the nano material pad 102 and drives the thiourea to move towards a detection line 1001, a mixed solution of the arsenic and the thiourea enters the detection line 1001 of the nano material pad 102 and reacts with potassium borohydride at the detection line 1001 to reduce the arsenic in the sample into arsine, and the detection line shows fluorescence when the device is placed under the irradiation of a special arsenic hollow cathode lamp. Meanwhile, the sample drives the free fluorescein in the nano material pad 102 to move towards the calibration line 1002, so that the free fluorescein is combined with the anti-fluorescein antibody, fluorescence appears at the calibration line 1002, and the content of arsenic element is determined by measuring the ratio of the fluorescence intensity at the detection line 1001 to the fluorescence intensity at the calibration line 1002 (the larger the ratio is, the more the arsenic content is, and conversely, the less the arsenic content is, the more the arsenic content is detected by a specific fluorescence intensity detection device, which is an existing device, and is not described in detail here). The sample pad 101 is placed in the arsenic standard treatment solution to be soaked for about 40-100 minutes, and is taken out and then placed in a 36-DEG C blast drying oven to be dried for at least more than 12 hours, so that the immunity of the fluorescent test strip to other elements can be improved, and the precision of the fluorescent test strip is improved. If the sample does not contain the arsenic element to be detected, no fluorescence is present at the detection line 1001, and no matter whether the sample is to be detected or not, the free fluorescein on the nano material pad 102 is bound by the anti-fluorescein antibody at the calibration line 1002 when the free fluorescein moves upwards to the calibration line 1002, so that the calibration line 1002 fluoresces. If no fluorescence appears at the position of the calibration line 1002, the test strip is invalid, and the test strip is replaced for retesting. The calibration line 1002 can timely know whether the test strip is invalid or not, and the detection accuracy is improved.
The base plate 100 is a rectangular plate, and the base plate 100 is a PVC plate. The PVC plate is a plate having a honeycomb-shaped mesh structure in cross section, which is made of PVC as a raw material, and has characteristics of corrosion resistance, moisture resistance, mold resistance, non-water absorption, drillability, sawing ability, planing ability, easy thermoforming, hot bending processing, and the like, so that the selected bottom plate 100 is durable.
The base plate 100 is made of a watertight material. When the arsenic solution is dripped on the sample pad 101, the arsenic solution is adsorbed on the bottom plate 100 when the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the water absorption pad 104 move, and the bottom plate 100 is made of a water-impermeable material, so that the bottom plate 100 is prevented from absorbing a part of the solution, and the measurement error of the arsenic solution is increased.
The nano material pad 102 is a porous nano material film or a cellulose film modified by nano material, and the nano material is at least one of graphene oxide, reduced graphene, nano gold, molybdenum disulfide and carbon nanotubes.
The absorbent pad 104 is absorbent paper. The main component of the water-absorbing paper is fiber which is a natural organic high molecular compound, the fiber in the paper is interlaced to form a net shape, a plurality of gaps are arranged among the fiber, and water can generate capillary effect through the gaps, so that the water-absorbing paper can greatly retain water and has the advantages of good water-absorbing performance and low manufacturing cost.
The above reaction membrane 103 employs a nitrocellulose membrane as the reaction membrane 103. The nitrocellulose membrane is also called NC membrane, is used as a carrier of a C/T line in the colloidal gold test paper, and is also a place for immunoreaction. Therefore, the NC film becomes the most important consumable in the present invention. Of course, the reaction membrane 103 is not limited to a nitrocellulose membrane, and other membranes that perform equivalent functions may be used herein.
The sample pad 101 is made of a cellulose film. Cellulose filter membranes are porous organic membranes. Generally, the chromatographic column is made of cellulose nitrate or cellulose acetate, and has the advantage of remarkable chromatographic effect.
The sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are convex on one side facing the bottom plate 100, the bottom plate 100 is provided with notches correspondingly matched with the convex part of the sample pad 101, the convex part of the nano-material pad 102, the convex part of the reaction membrane 103 and the convex part of the absorbent pad 104, and the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are connected with the bottom plate 100 through glue. The convex parts of the sample pad 101, the nano material pad 102, the reaction membrane 103 and the water absorption pad 104 are butted with the notch on the bottom plate 100, so that the connection stability of the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 is improved, and the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 are connected through glue, so that the advantages of quick connection and small connection gap are achieved.
Example 1
A preparation method of a fluorescent test strip for environmental monitoring comprises the following steps,
selecting a substrate, namely preparing a substrate 100 for preparing the fluorescent test strip;
preparation of sample pad 101: a sample pad 101 is disposed on one side of the base plate 100, and the preparation steps of the sample pad 101 are as follows: placing the sample pad 101 in arsenic standard treatment liquid to be soaked for about 40 minutes, taking out the sample pad, and placing the sample pad in a 36 ℃ forced air drying oven to be dried for at least more than 12 hours;
preparing the nano-material pad 102, wherein the nano-material pad 102 is arranged at one end of the sample pad 101 in a butt joint mode, the nano-material pad 102 is connected with the bottom plate 100, and the preparation method of the nano-material pad 102 comprises the following steps: spraying the graphene oxide suspension solution to a glass cellulose membrane, drying for 6 hours at 100 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 15min, and drying at 30 ℃;
preparation of the reaction film 103: one end of the nano-material pad 102 far away from the sample pad 101 is butted with a reaction membrane 103, the reaction membrane 103 is connected with the bottom plate 100, and the preparation method of the reaction membrane 103 comprises the following steps: a detection line 1001 is arranged at one end, close to the nano material pad 102, of the reaction membrane 103, a calibration line 1002 is arranged at one end, far away from the nano material, of the reaction membrane 103, the potassium borohydride solution and the avidin are incubated for 25min and sprayed to the detection line 1001, and an anti-fluorescein antibody corresponding to free fluorescein is sprayed on the calibration line 1002;
preparation of absorbent pad 104: the end of the reaction membrane 103 far away from the nano material is butt-jointed with a water absorption pad 104, the water absorption pad 104 is connected with the bottom plate 100, and the water absorption pad 104 is made of water absorption material.
The base plate 100 is a rectangular plate, and the base plate 100 is a PVC plate. The PVC plate is a plate having a honeycomb-shaped mesh structure in cross section, which is made of PVC as a raw material, and has characteristics of corrosion resistance, moisture resistance, mold resistance, non-water absorption, drillability, sawing ability, planing ability, easy thermoforming, hot bending processing, and the like, so that the selected bottom plate 100 is durable.
The base plate 100 is made of a watertight material. When the arsenic solution is dripped on the sample pad 101, the arsenic solution is adsorbed on the bottom plate 100 when the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the water absorption pad 104 move, and the bottom plate 100 is made of a water-impermeable material, so that the bottom plate 100 is prevented from absorbing a part of the solution, and the measurement error of the arsenic solution is increased.
The nano material pad 102 is a porous nano material film, and the nano material is graphene oxide or reduced graphene.
The absorbent pad 104 is absorbent paper. The main component of the water-absorbing paper is fiber which is a natural organic high molecular compound, the fiber in the paper is interlaced to form a net shape, a plurality of gaps are arranged among the fiber, and water can generate capillary effect through the gaps, so that the water-absorbing paper can greatly retain water and has the advantages of good water-absorbing performance and low manufacturing cost.
The above reaction membrane 103 employs a nitrocellulose membrane as the reaction membrane 103. The nitrocellulose membrane is also called NC membrane, is used as a carrier of a C/T line in the colloidal gold test paper, and is also a place for immunoreaction. Therefore, the NC film becomes the most important consumable in the present invention. Of course, the reaction membrane 103 is not limited to a nitrocellulose membrane, and other membranes that perform equivalent functions may be used herein.
The sample pad 101 is made of a cellulose film. Cellulose filter membranes are porous organic membranes. Generally, the chromatographic column is made of cellulose nitrate or cellulose acetate, and has the advantage of remarkable chromatographic effect.
The sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are convex on one side facing the bottom plate 100, the bottom plate 100 is provided with notches correspondingly matched with the convex part of the sample pad 101, the convex part of the nano-material pad 102, the convex part of the reaction membrane 103 and the convex part of the absorbent pad 104, and the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are connected with the bottom plate 100 through glue. The convex parts of the sample pad 101, the nano material pad 102, the reaction membrane 103 and the water absorption pad 104 are butted with the notch on the bottom plate 100, so that the connection stability of the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 is improved, and the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 are connected through glue, so that the advantages of quick connection and small connection gap are achieved.
Example 2
A preparation method of a fluorescent test strip for environmental monitoring comprises the following steps,
selecting a substrate, namely preparing a substrate 100 for preparing the fluorescent test strip;
preparation of sample pad 101: a sample pad 101 is disposed on one side of the base plate 100, and the preparation steps of the sample pad 101 are as follows: placing the sample pad 101 in arsenic standard treatment liquid to be soaked for about 70 minutes, taking out the sample pad, and placing the sample pad in a 36 ℃ forced air drying oven to be dried for at least more than 12 hours;
preparing the nano-material pad 102, wherein the nano-material pad 102 is arranged at one end of the sample pad 101 in a butt joint mode, the nano-material pad 102 is connected with the bottom plate 100, and the preparation method of the nano-material pad 102 comprises the following steps: spraying the graphene oxide suspension solution to a glass cellulose membrane, drying for 8 hours at 120 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 20min, and drying at 33 ℃;
preparation of the reaction film 103: one end of the nano-material pad 102 far away from the sample pad 101 is butted with a reaction membrane 103, the reaction membrane 103 is connected with the bottom plate 100, and the preparation method of the reaction membrane 103 comprises the following steps: a detection line 1001 is arranged at one end, close to the nano material pad 102, of the reaction membrane 103, a calibration line 1002 is arranged at one end, far away from the nano material, of the reaction membrane 103, the potassium borohydride solution and the avidin are incubated for 30min and sprayed to the detection line 1001, and an anti-fluorescein antibody corresponding to free fluorescein is sprayed on the calibration line 1002;
preparation of absorbent pad 104: the end of the reaction membrane 103 far away from the nano material is butt-jointed with a water absorption pad 104, the water absorption pad 104 is connected with the bottom plate 100, and the water absorption pad 104 is made of water absorption material.
The base plate 100 is a rectangular plate, and the base plate 100 is a PVC plate. The PVC plate is a plate having a honeycomb-shaped mesh structure in cross section, which is made of PVC as a raw material, and has characteristics of corrosion resistance, moisture resistance, mold resistance, non-water absorption, drillability, sawing ability, planing ability, easy thermoforming, hot bending processing, and the like, so that the selected bottom plate 100 is durable.
The base plate 100 is made of a watertight material. When the arsenic solution is dripped on the sample pad 101, the arsenic solution is adsorbed on the bottom plate 100 when the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the water absorption pad 104 move, and the bottom plate 100 is made of a water-impermeable material, so that the bottom plate 100 is prevented from absorbing a part of the solution, and the measurement error of the arsenic solution is increased.
The nano-material pad 102 is a porous nano-material film, and the nano-material is graphene oxide.
The absorbent pad 104 is absorbent paper. The main component of the water-absorbing paper is fiber which is a natural organic high molecular compound, the fiber in the paper is interlaced to form a net shape, a plurality of gaps are arranged among the fiber, and water can generate capillary effect through the gaps, so that the water-absorbing paper can greatly retain water and has the advantages of good water-absorbing performance and low manufacturing cost.
The above reaction membrane 103 employs a nitrocellulose membrane as the reaction membrane 103. The nitrocellulose membrane is also called NC membrane, is used as a carrier of a C/T line in the colloidal gold test paper, and is also a place for immunoreaction. Therefore, the NC film becomes the most important consumable in the present invention. Of course, the reaction membrane 103 is not limited to a nitrocellulose membrane, and other membranes that perform equivalent functions may be used herein.
The sample pad 101 is made of a cellulose film. Cellulose filter membranes are porous organic membranes. Generally, the chromatographic column is made of cellulose nitrate or cellulose acetate, and has the advantage of remarkable chromatographic effect.
The sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are convex on one side facing the bottom plate 100, the bottom plate 100 is provided with notches correspondingly matched with the convex part of the sample pad 101, the convex part of the nano-material pad 102, the convex part of the reaction membrane 103 and the convex part of the absorbent pad 104, and the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are connected with the bottom plate 100 through glue. The convex parts of the sample pad 101, the nano material pad 102, the reaction membrane 103 and the water absorption pad 104 are butted with the notch on the bottom plate 100, so that the connection stability of the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 is improved, and the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 are connected through glue, so that the advantages of quick connection and small connection gap are achieved.
Example 3
A preparation method of a fluorescent test strip for environmental monitoring comprises the following steps,
selecting a substrate, namely preparing a substrate 100 for preparing the fluorescent test strip;
preparation of sample pad 101: a sample pad 101 is disposed on one side of the base plate 100, and the preparation steps of the sample pad 101 are as follows: placing the sample pad 101 in arsenic standard treatment liquid to be soaked for about 100 minutes, taking out the sample pad, and placing the sample pad in a 36 ℃ forced air drying oven to be dried for at least more than 12 hours;
preparing the nano-material pad 102, wherein the nano-material pad 102 is arranged at one end of the sample pad 101 in a butt joint mode, the nano-material pad 102 is connected with the bottom plate 100, and the preparation method of the nano-material pad 102 comprises the following steps: spraying the graphene oxide suspension solution to a glass cellulose membrane, drying for 10 hours at 140 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 25min, and drying at 35 ℃;
preparation of the reaction film 103: one end of the nano-material pad 102 far away from the sample pad 101 is butted with a reaction membrane 103, the reaction membrane 103 is connected with the bottom plate 100, and the preparation method of the reaction membrane 103 comprises the following steps: a detection line 1001 is arranged at one end, close to the nano material pad 102, of the reaction membrane 103, a calibration line 1002 is arranged at one end, far away from the nano material, of the reaction membrane 103, the potassium borohydride solution and the avidin are incubated for 35min and sprayed to the detection line 1001, and an anti-fluorescein antibody corresponding to free fluorescein is sprayed on the calibration line 1002;
preparation of absorbent pad 104: the end of the reaction membrane 103 far away from the nano material is butt-jointed with a water absorption pad 104, the water absorption pad 104 is connected with the bottom plate 100, and the water absorption pad 104 is made of water absorption material.
The base plate 100 is a rectangular plate, and the base plate 100 is a PVC plate. The PVC plate is a plate having a honeycomb-shaped mesh structure in cross section, which is made of PVC as a raw material, and has characteristics of corrosion resistance, moisture resistance, mold resistance, non-water absorption, drillability, sawing ability, planing ability, easy thermoforming, hot bending processing, and the like, so that the selected bottom plate 100 is durable.
The base plate 100 is made of a watertight material. When the arsenic solution is dripped on the sample pad 101, the arsenic solution is adsorbed on the bottom plate 100 when the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the water absorption pad 104 move, and the bottom plate 100 is made of a water-impermeable material, so that the bottom plate 100 is prevented from absorbing a part of the solution, and the measurement error of the arsenic solution is increased.
The nano-material pad 102 is a porous nano-material film, and the nano-material is reduced graphene.
The absorbent pad 104 is absorbent paper. The main component of the water-absorbing paper is fiber which is a natural organic high molecular compound, the fiber in the paper is interlaced to form a net shape, a plurality of gaps are arranged among the fiber, and water can generate capillary effect through the gaps, so that the water-absorbing paper can greatly retain water and has the advantages of good water-absorbing performance and low manufacturing cost.
The above reaction membrane 103 employs a nitrocellulose membrane as the reaction membrane 103. The nitrocellulose membrane is also called NC membrane, is used as a carrier of a C/T line in the colloidal gold test paper, and is also a place for immunoreaction. Therefore, the NC film becomes the most important consumable in the present invention. Of course, the reaction membrane 103 is not limited to a nitrocellulose membrane, and other membranes that perform equivalent functions may be used herein.
The sample pad 101 is made of a cellulose film. Cellulose filter membranes are porous organic membranes. Generally, the chromatographic column is made of cellulose nitrate or cellulose acetate, and has the advantage of remarkable chromatographic effect.
The sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are convex on one side facing the bottom plate 100, the bottom plate 100 is provided with notches correspondingly matched with the convex part of the sample pad 101, the convex part of the nano-material pad 102, the convex part of the reaction membrane 103 and the convex part of the absorbent pad 104, and the sample pad 101, the nano-material pad 102, the reaction membrane 103 and the absorbent pad 104 are connected with the bottom plate 100 through glue. The convex parts of the sample pad 101, the nano material pad 102, the reaction membrane 103 and the water absorption pad 104 are butted with the notch on the bottom plate 100, so that the connection stability of the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 is improved, and the sample pad 101, the nano material pad 102, the reaction membrane 103, the water absorption pad 104 and the bottom plate 100 are connected through glue, so that the advantages of quick connection and small connection gap are achieved.
Test example 1
Collecting a part of soil containing arsenic element, preparing a sample reagent A1, collecting a part of soil without arsenic pattern, preparing a sample reagent B1, making only one part of the two parts of the collected soil contain arsenic, one part does not contain arsenic, and the other properties are the same, preparing two fluorescent test strips prepared by the invention, wherein one fluorescent test strip is defined as C1, the other fluorescent test strip is defined as D1, dropwise adding the sample reagent A1 to the fluorescent test strip C1, dropwise adding the sample reagent B1 to the fluorescent test strip D1, and after a period of time, making the fluorescent test strips C1 and D1 to observe the luminescence of the fluorescent test strips C1 and D1 under the irradiation of a D1 arsenic hollow cathode lamp, wherein the fluorescent test strips C1 and D1 are both prepared according to example 1.
TABLE 1
| Fluorescent test paper strip | Sample reagent | Detection line | Calibration line |
| C1 | A1 | Weak luminous intensity | Weak luminous intensity |
| D2 | B1 | Does not emit light | Weak luminous intensity |
As can be seen from the above test example 1, the fluorescent test strip prepared according to the present invention can detect whether the sample solution contains arsenic element through the detection line, and can detect arsenic element in soil. However, the fluorescence test strip C1 has weak light and relatively poor detection effect.
Test example 2
Collecting a part of soil containing arsenic element, preparing a sample reagent A2, collecting a part of soil without arsenic pattern, preparing a sample reagent B2, making only one part of the two parts of the collected soil contain arsenic, one part does not contain arsenic, and the other properties are the same, preparing two fluorescent test strips prepared by the invention, wherein one fluorescent test strip is defined as C2, the other fluorescent test strip is defined as D2, dropwise adding the sample reagent A2 to the fluorescent test strip C2, dropwise adding the sample reagent B2 to the fluorescent test strip D2, and after a period of time, making the fluorescent test strips C2 and D2 to observe the luminescence of the fluorescent test strips C2 and D2 under the irradiation of a D2 arsenic hollow cathode lamp, wherein the fluorescent test strips C2 and D2 are both prepared according to example 2.
TABLE 2
| Fluorescent test paper strip | Sample reagent | Detection line | Calibration line |
| C2 | A2 | General luminous intensity | General luminous intensity |
| D2 | B2 | Does not emit light | General luminous intensity |
As can be seen from the above test examples, the fluorescent test strip prepared by the invention can detect whether the sample solution contains arsenic element through the detection line, and can detect the arsenic element in the soil. However, the fluorescence test strip C2 shows a relatively general light and has a relatively general detection effect.
Test example 3
Collecting a part of soil containing arsenic element, preparing a sample reagent A3, collecting a part of soil without arsenic pattern, preparing a sample reagent B3, making only one part of the two parts of the collected soil contain arsenic, one part does not contain arsenic, and the other properties are the same, preparing two fluorescent test strips prepared by the invention, wherein one fluorescent test strip is defined as C3, the other fluorescent test strip is defined as D3, dropwise adding the sample reagent A3 to the fluorescent test strip C3, dropwise adding the sample reagent B3 to the fluorescent test strip D3, and after a period of time, making the fluorescent test strips C3 and D3 observe the luminescence of the fluorescent test strips C3 and D3 under the irradiation of a D3 arsenic hollow cathode lamp, wherein the fluorescent test strips C3 and D3 are both prepared according to example 3.
TABLE 3
| Fluorescent test paper strip | Sample reagent | Detection line | Calibration line |
| C3 | A3 | Weak luminous intensity | Weak luminous intensity |
| D3 | B3 | Does not emit light | Weak luminous intensity |
As can be seen from the above test examples, the fluorescent test strip prepared by the invention can detect whether the sample solution contains arsenic element through the detection line, and can detect the arsenic element in the soil. The fluorescent test strip C3 has strong light-emitting property and good detection effect.
In summary, the invention provides a preparation method of a fluorescent test strip for environmental monitoring, which has at least the following beneficial effects:
when the content of arsenic in soil is detected, a soil aqueous solution of arsenic is dripped onto a sample pad 101, the sample can dialyze under the action of capillary and moves towards a nano material pad 102, when the sample enters the nano material pad 102, the arsenic in the sample adsorbs thiourea in the nano material pad 102 and drives the thiourea to move towards a detection line 1001, a mixed solution of the arsenic and the thiourea enters the detection line 1001 of the nano material pad 102 and reacts with potassium borohydride at the detection line 1001 to reduce the arsenic in the sample into arsine, and the detection line shows fluorescence when the device is placed under the irradiation of a special arsenic hollow cathode lamp. Meanwhile, the sample drives the free fluorescein in the nano material pad 102 to move towards the calibration line 1002, so that the free fluorescein is combined with the anti-fluorescein antibody, fluorescence appears at the calibration line 1002, and the content of arsenic element is determined by measuring the ratio of the fluorescence intensity at the detection line 1001 to the fluorescence intensity at the calibration line 1002 (the larger the ratio is, the more the arsenic content is, and conversely, the less the arsenic content is, the more the arsenic content is detected by a specific fluorescence intensity detection device, which is an existing device, and is not described in detail here). The sample pad 101 is placed in the arsenic standard treatment solution to be soaked for about 40-100 minutes, and is taken out and then placed in a 36-DEG C blast drying oven to be dried for at least more than 12 hours, so that the immunity of the fluorescent test strip to other elements can be improved, and the precision of the fluorescent test strip is improved. If the sample does not contain the arsenic element to be detected, no fluorescence is present at the detection line 1001, and no matter whether the sample is to be detected or not, the free fluorescein on the nano material pad 102 is bound by the anti-fluorescein antibody at the calibration line 1002 when the free fluorescein moves upwards to the calibration line 1002, so that the calibration line 1002 fluoresces. If no fluorescence appears at the position of the calibration line 1002, the test strip is invalid, and the test strip is replaced for retesting. The calibration line 1002 can timely know whether the test strip is invalid or not, and the detection accuracy is improved.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A preparation method of a fluorescent test strip for environmental monitoring is characterized by comprising the following steps,
selecting a substrate, namely preparing a substrate for preparing the fluorescent test strip;
preparing a sample pad: arranging a sample pad on one side of the bottom plate, wherein the preparation steps of the sample pad are as follows: placing the sample pad in arsenic standard treatment liquid, soaking for about 40-100 minutes, taking out, and placing in a 36 ℃ forced air drying oven to dry for at least more than 12 hours;
preparing a nano-material pad, wherein one end of the sample pad is butted with the nano-material pad, the nano-material pad is connected with the bottom plate, and the preparation method of the nano-material pad comprises the following steps: spraying the graphene oxide suspension solution on a glass cellulose membrane, drying for 6-10 h at 100-140 ℃ to obtain a nano material, dropwise adding a thiourea reagent and free fluorescein to the nano material, reacting for 15-25 min, and drying at 30-35 ℃;
preparing a reaction film: the end, far away from the sample pad, of the nano material pad is in butt joint with a reaction membrane, the reaction membrane is connected with the bottom plate, and the preparation method of the reaction membrane comprises the following steps: a detection line is arranged at one end, close to the nano material pad, of the reaction membrane, a calibration line is arranged at one end, far away from the nano material, of the reaction membrane, a potassium borohydride solution and avidin are incubated for 25-35 min and sprayed to the detection line, and an anti-fluorescein antibody corresponding to the free fluorescein is sprayed on the calibration line;
preparing a water absorption pad: one end of the reaction membrane, which is far away from the nano material, is butted with a water absorption pad, the water absorption pad is connected with the bottom plate, and the water absorption pad is made of a water absorption material.
2. The method for preparing a fluorescent test strip for environmental monitoring according to claim 1, wherein the base plate is a rectangular plate, and the base plate is a PVC plate.
3. The method for preparing a fluorescent test strip for environmental monitoring according to claim 1, wherein the base plate is made of a water-impermeable material.
4. The method for preparing a fluorescent test strip for environmental monitoring according to claim 1, wherein the nano-material pad is a porous nano-material film or a cellulose film modified by nano-material, and the nano-material is at least one of graphene oxide, reduced graphene, nanogold, molybdenum disulfide and carbon nanotubes.
5. The method for preparing a fluorescent test strip for environmental monitoring according to claim 1, wherein the absorbent pad is absorbent paper.
6. The method for preparing a fluorescent test strip for environmental monitoring according to claim 1, wherein the reaction membrane is a nitrocellulose membrane.
7. The method for preparing a fluorescent test strip for environmental monitoring according to claim 1, wherein the sample pad is made of a cellulose membrane.
8. The method for preparing a fluorescent test strip for environmental monitoring as claimed in claim 1, wherein the sample pad, the nano-material pad, the reaction membrane and the absorbent pad are convex on their sides facing the bottom plate, the bottom plate is provided with notches corresponding to the convex portions of the sample pad, the nano-material pad, the reaction membrane and the absorbent pad, and the sample pad, the nano-material pad, the reaction membrane and the absorbent pad are connected to the bottom plate by glue.
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