KR101821093B1 - Paper Sensor for Diagnostic of Environmental disorders - Google Patents

Abstract

The present invention relates to a paper sensor for diagnosing environmental disorder, enabling diagnosis on environmental disorder by detecting proline via color development reactions of proline reacting with ninhydrin. More specifically, the present invention relates to a paper sensor for diagnosing environmental disorder. By sealing microfluidic channels where proline and ninhydrin in a measurement sample flow, and then evenly coating ninhydrin, it is possible to precisely and accurately detect proline.

Description

[0001] The present invention relates to a paper sensor for diagnosing an environmental disorder,

The present invention relates to a paper sensor for diagnosing an environmental disorder, and more particularly, to a paper sensor for diagnosing an environmental disorder, in which a microfluidic channel in which a proline and a ninhydrin flow in a measurement sample is closed and a ninhydrin is uniformly coated, To a paper sensor for environmental fault diagnosis.

Generally, plants and crops (hereinafter referred to as "crops") are not able to move, so they are affected by various environmental factors throughout their growth.

In particular, among environmental factors, environmental problems such as drought, high salinity, heavy metals, cold weather, high temperature and ozone, that is, crops are faced with environmental stress, and such environmental disorder is a limiting factor of crop growth and development .

It is known that when crops are affected by environmental disturbances, proline is accumulated in the cytoplasm of crop cells, which is an amino acid. Such proline is very important for the growth of crops against environmental disorder, Is known to be a very important signal molecule for photosynthetic efficiency, flowering time, and endosperm development.

Therefore, we could know the effect of environmental disturbance on the growth of crops by using the concentration change of proline in the cytoplasm according to environmental disorder.

However, these measurements can only be made if the harvested crops are taken to the laboratories. It takes a long time to analyze and not only slows the growth and analysis of the crops, but also monitors the growth status of the crops There was no disadvantage.

A description of the effect of the environmental disorder on the growth of crops as described above is described in detail in Patent Document 1, so that a detailed description thereof will be omitted.

On the other hand, the inventor of the present invention has developed a technology capable of directly analyzing the plant, which has been collected after a lot of investment and research in order to solve the above problems, without moving the plant to the laboratory, Patent application, and it is registered as patent registration No. 10-1579045 (hereinafter referred to as "Prior Art 1").

Prior art 1 is a diagnostic kit for real-time measurement of the growth status of crops due to environmental disturbances. The diagnostic kit includes a paper sensor for absorbing a sample of crops collected in the field; A main body having paper sensors inserted therein; A heater provided in the body for heating the paper sensor; And a smart phone having a camera capable of photographing a color reaction of a paper sensor provided in the main body and being heated.

Accordingly, in the prior art 1, the crop can be measured and monitored in real time on site without any additional analysis, so that the growth status of crops due to environmental disturbances can be monitored quickly in the field, It is possible to minimize the cost of managing the crops and improve the production amount.

In addition, the present inventor has found that, in the prior art document 1, when the paper sensor is inserted into the main body, the paper sensor is not wrinkled or fixed, In order to solve the problem that the thermal efficiency is low and the diagnosis is not smoothly proceeded because it is not transmitted, in order to quickly measure the growth and growth condition of crops due to environmental problems such as drought and low temperature or moisture stress phenomenon Developed a diagnostic kit for preventing environmental disorder with a detachable sensor module and applied for a patent. This patent was registered as a patent registration No. 10-1699667 (hereinafter referred to as "Prior Art 2").

Prior art 2 is a diagnostic kit for real-time measurement of the growth status of crops due to environmental disturbances. The diagnostic kit includes a paper sensor for absorbing a sample of crops collected at the site; A sensor module assembly including a sensor module integrated with a heater and a temperature sensor, the sensor module assembly restricting the paper sensor; And a smartphone having a camera which is provided in the main body and is capable of photographing a coloring reaction of a paper sensor to be heated, wherein the paper sensor comprises a sensor module assembly, And is configured to cause a color reaction when energized.

Accordingly, in the prior art document 2, there is no need to form a separate stand in the main body, and it is possible to simplify the structure and to easily attach and detach the main body while maintaining the compatibility effect regardless of the type and size of the smartphone, There is no derailment or wrinkle in the position when the mobile phone is installed inside the main body, so that it can be easily measured through the smartphone.

In addition, a paper sensor is provided on the upper part of the sensor module, and only when the sensor module assembly is inserted into the body, electric energy is prevented from being wasted by the heater.

In the above-mentioned prior art documents 1 and 2, a paper sensor for detecting proline by using a ninhydrin reaction is disclosed. The paper sensor detects a proline by a sample through the open channel and by a colorimetric method.

Herein, the ninhydrin reaction indicates that the amino acid of the protein reacts with ninhydrin to develop a bluish purple to red purple color, but in the case of proline contained in the crop, it reacts with ninhydrin to develop color. Therefore, it is possible to quantitatively detect the amount of proline by comparing the colors shown in the sensor unit.

However, in the conventional paper sensor, since the sample flows through the open channel, contaminants are mixed from the external environment during the sample analysis, and the ninhydrin solution and the reagent sulfosalicylic acid solution And the measurement sample is evaporated during the flow of the sample through the open channel, thereby making it difficult to perform accurate measurement.

When the ninhydrin is previously loaded on the sample rod and coated to reduce the number of reagents, the ninhydrin flows through the microfluidic channel and flows naturally along the sample rod and the channel. Resulting in a difference in concentration. Accordingly, when a proline is detected by loading a measurement sample, there is a problem that it is difficult to quantitatively detect proline because a heterogeneous reaction occurs in which the color of the sensor portion is not uniform and the color appears in a band shape as shown in Fig. have.

KR 10-1361149 B1 KR 10-1579045 B1 KR 10-1699667 B1

SUMMARY OF THE INVENTION Accordingly, the present invention has been made to solve the above-mentioned problems occurring in the prior art, and it is an object of the present invention to provide a paper sensor, which is used to measure a growing condition of a crop in real- The object of the present invention is to provide a paper sensor for environmental defect diagnosis and a method of manufacturing the paper sensor, which can accurately detect proline by preventing contamination of contamination and evaporation of a measurement sample.

In addition, the ninhydrin ninhydrin which is constituted independently is uniformly coated and loaded so that the on-site detection is facilitated, the sensor sensitivity is formed, and the predetermined color is detected according to the concentration of proline, and a method of manufacturing the paper sensor The purpose is to provide.

To achieve the above object, according to the present invention, there is provided a paper sensor for environmental disorder diagnosis, which detects proline through a ninhydrin reaction on a sample to be loaded in a sample loading part, A sample loading unit 11 formed on one side of the main body 10 formed of paper and coated with ninhydrin; A ninhydrin 12 coated with ninhydrin, which is located at a lower part of the sample loading part 11 in a different layer; A sensor unit 13 formed to a predetermined distance from the sample loading unit 11 to a ninhydrin sub-layer so as to detect proline of a sample to be measured using the color development reaction of proline by the ninhydrin reaction; The sensor unit 13 and the ninhydrin unit 12 are connected to each other so that the proline of the measurement sample and the ninhydrin of the ninhydrin unit are mixed with the sensor unit 13 while mixing the sample loading unit 11 and the ninhydrin unit, The sample loading part 11 and the sensor part 13 are exposed to the outside and the rest are placed inside the main body 10 to be shielded from the outside .

Further, the microfluidic channel 15 is characterized by forming an S-shaped channel so that proline and ninhydrin of the measurement sample can be sufficiently mixed.

A waterproof layer 14 is provided at a lower portion of the main body 10 to prevent proline and ninhydrin from flowing below the microfluidic channels 15. [

A method of manufacturing a paper sensor for environmental defect diagnosis according to the present invention includes a sample loading unit 11 which is disposed on a piece of paper at a predetermined interval and coated with ninhydrin, a ninhydrin coated ninhydrin unit 12 An S-shaped microfluidic channel 15 connecting the sensor section 13 and the ninhydrin section 12 to the sensor section 13 and a pattern designing step S10 of designing a paper folding line to divide the paper into four sections, Wow; After the designed sensor pattern is printed on a paper with a wax printer, the wax is penetrated into the paper by using a hot plate, so that the sample loading part 11, the sensor part 13 and the microfluidic channel 15 are sealed with the wax Printing step S20; A ninhydrin coating step (S30) in which a ninhydrin solution of a predetermined concentration is loaded on the sample loading part (11) and a ninhydrin part (12) below the sample loading part (11) and dried so that ninhydrin is coated at a constant concentration; The sample loading unit 11 and the sensor unit 13 are formed by folding the wax-impregnated paper along the paper folding line and the ninhydrin unit 12 and the sensor unit 13, which are located below the sample loading unit 11, (S40) for allowing the lower part of the microfluidic channel (15) to be connected by the microfluidic channel (15); (S50) in which only the surface layer of the sample loading part (11) and the sensor part (13) is exposed to the outside and the remainder is positioned inside the body (10) ); ≪ / RTI >

The sensor pattern designed in the pattern designing step S10 includes a first layer pattern 10a designed such that the sample loading unit 11 and the sensor unit 13 are spaced apart from each other by a predetermined distance, A second layer pattern 10b designed such that the sensor unit 13 and the sensor unit 13 are positioned opposite to the first layer pattern 10a and the second layer pattern 10b designed to place the sample loading unit 11 and the sensor unit 13 on the microfluidic channel 15 A third layer pattern 10c designed to be connected to the first layer pattern 10a and a fourth layer pattern 10d having no pattern and the first layer pattern 10a and the second layer pattern 10b, 10c and the fourth layer pattern 10d are sequentially arranged with reference to the paper fold line.

The wax-printing step (S20) is characterized in that the wax-printed paper is placed on a hot plate and heated at a temperature of 120 ° C for 2 to 3 minutes so that the wax is completely penetrated into the paper.

In the paper sensor and the method of manufacturing the same according to the present invention, the surface layer of the sample loading part and the sensor part is exposed, and the lower part of the sample loading part and the sensor part and the microfluidic channel connecting the sample loading part and the sensor part are sealed Therefore, it is possible to prevent contamination of foreign substances from the external environment and evaporation of the measurement sample, thereby improving the reliability of the measurement result.

Further, as the microfluidic channel is formed in an S-shape and the length becomes longer, proline and ninhydrin of the measurement sample are sufficiently mixed to provide a uniform coloring reaction.

In addition, a waterproof layer is formed under the microfluidic channel, so that proline or ninhydrin flowing along the microfluidic channel is prevented from leaking downward, and the reliability of the detection result is improved.

In addition, since ninhydrin is previously coated on the sample loading part and the ninhydrin part below the sample loading part, only the sulfosylic acid as a reagent can be prepared without preparing a separate ninhydrin solution, so that proline can be detected in the field have.

In addition, ninhydrin can be independently and uniformly loaded in the ninhydrin portion under the sample loading portion, so that the detection sensitivity in the sensor portion is improved and the color according to the color reaction of proline is constant.

1 is a perspective view and a cross-sectional view showing a paper sensor for environmental fault diagnosis of the present invention
2 is a flowchart showing a method of manufacturing a paper sensor for environmental fault diagnosis according to the present invention
3 is a plan view showing a pattern design according to the present invention.
4 is a process chart showing a wax printing step according to the present invention
5 is a view showing a state in which wax is infiltrated into paper,
6 is a graph showing changes in green color intensity according to the concentration of proline
7 is a reference diagram showing the color change of the sensor part according to the concentration of proline in the measurement sample
FIG. 8 is a perspective view showing an origin stage according to the present invention; FIG.
9 is a view showing a coloring reaction of a sensor portion using a conventional open type microfluidic channel

Although the term used in the present invention selects the general term that is widely used at present, there is also a term selected by the applicant in a specific case. In this case, the term used in the present invention is not a name of a simple term, It is necessary to understand the meaning.

Hereinafter, the technical structure of the present invention will be described in detail with reference to preferred embodiments shown in the accompanying drawings.

The paper sensor for environmental fault diagnosis according to the present invention detects proline through a ninhydrin reaction on a measurement sample loaded in the sample loading section 11. As shown in Fig. 1, a measurement sample obtained from a crop is loaded A sample loading unit 11 formed on one side of the main body 10 formed of paper and coated with ninhydrin; A ninhydrin 12 coated with ninhydrin, which is located at a lower part of the sample loading part 11 in a different layer; A sensor unit 13 formed to a predetermined distance from the sample loading unit 11 to a ninhydrin sub-layer so as to detect proline of a sample to be measured using the color development reaction of proline by the ninhydrin reaction; The sensor unit 13 and the ninhydrin unit 12 are connected to each other so that the proline of the measurement sample and the ninhydrin of the ninhydrin unit are mixed with the sensor unit 13 while mixing the sample loading unit 11 and the ninhydrin unit, And a microfluidic channel (15) positioned differently from the microfluidic channel (15).

Only the surface layer of the sample loading unit 11 and the sensor unit 13 is exposed to the outside of the main body 10 and the remaining part of the sample loading unit 11 and the sensor unit 13 and the ninhydrin unit 12, And the microfluidic channel (15) are located inside the main body (10) and are sealed to be blocked from the outside.

Accordingly, foreign substances are prevented from being mixed with the measurement sample loaded on the sample loading unit 11 or the measurement sample flowing along the microfluidic channel 15 due to the external environment, the measurement sample is not evaporated, The reliability of the measurement result is improved.

In addition, it is preferable that the microfluidic channel 15 form an S-shaped channel so that proline and ninhydrin of the measurement sample can be sufficiently mixed. The proline of the measurement sample and the ninhydrin of the ninhydrin moiety are mixed while flowing along the microfluidic channel 15 in the closed state and moved to the sensor unit 13 so that the microfluidic channel 15 can be sufficiently mixed They should be as long as possible and free from angled parts so that they flow smoothly. In consideration of this point, the microfluidic channel 15 has an S-shaped channel.

In addition, it is preferable that a waterproof layer 14 is provided at a lower portion of the body 10 so that proline and ninhydrin do not flow under the microfluidic channel 15. [

When the waterproof layer is provided on the lower portion of the microfluidic channel 15, proline and ninhydrin are prevented from leaking to the lower portion of the body 10, thereby improving the measurement sensitivity of the sensor portion 13, The reliability is improved.

As shown in FIGS. 2 to 9, the method for manufacturing the paper sensor for diagnosing environmental disorder according to the present invention comprises a sample loading unit 11 which is disposed on a piece of paper at a predetermined interval and coated with ninhydrin, A microfluidic channel 15 for connecting the coated ninhydrin part 12, the sensor part 13, the ninhydrin part 12 and the sensor part 13 with each other, and three paper folding lines Designing a pattern designing step S10; After the designed sensor pattern is printed on a paper with a wax printer, the wax is infiltrated into the paper by using a hot plate, and the wax for sealing the ninth droplet portion 12, the sensor portion 13 and the microfluidic channel 15 is sealed by the wax Printing step S20; A ninhydrin coating step (S30) in which ninhydrin is coated at a predetermined concentration on the sample loading part (11) and the ninhydrin part (12) below the sample loading part (11); The sample loading unit 11 and the sensor unit 13 are formed by folding the wax-impregnated paper along the paper folding line and the ninhydrin unit 12 and the sensor unit 13, which are located below the sample loading unit 11, (S40) for allowing the lower part of the microfluidic channel (15) to be connected by the microfluidic channel (15); And a bonding step (S50) of bonding the layers folded by paper folding with an adhesive to form the main body (10).

3, the sensor pattern designed in the pattern designing step S10 includes a first layer pattern 10a designed such that the sample loading part 11 and the sensor part 13 are arranged at a predetermined interval, The feeding section 12 and the sensor section 13 are designed to be positioned symmetrically with respect to the paper folding line 10-1 on the sample loading section 11 and the sensor section 13 of the first layer pattern 10a The second layer pattern 10b and the ninhydrin part 12 and the sensor part 13 are formed on the ninhydrin part 12 of the second layer pattern 10b and the paper folding line 10- 2 and the third layer pattern 10c designed to be connected to each other by the microfluidic channel 15 and the fourth layer pattern 10d having no pattern.

At this time, the first layer pattern 10a, the second layer pattern 10b, the third layer pattern 10c, and the fourth layer pattern 10d are sequentially arranged with reference to the paper fold line, Wax printing is completed by the process alone. Since there is no pattern on the fourth layer pattern 10d, the wax is printed on the entire surface of the paper in the fourth layer in the wax printing process S20 to form the waterproof layer 14.

In the wax printing step S20, the paper on which the wax is printed is placed on a hot plate and heated at a temperature of 120 ° C for 2 to 3 minutes to completely penetrate the wax. For example, if the paper on which the wax is printed is placed on a hot plate and heated at a temperature of 120 ° C for about 160 seconds, the printed wax melts and permeates into the side of the paper to completely penetrate to the bottom of the paper.

The wax printed on paper melts at a temperature of 120 ° C and impregnates into the paper, which is impregnated deeper over time. Specifically, in the case of printing the wax on a paper sheet having a thickness of 800 μm, as shown in FIGS. 5 and 6, even if heated for 40 seconds under a temperature condition of 120 ° C., the wax is not completely penetrated into the paper, When heated, it penetrates to the side of the paper and completely penetrates to the bottom of the paper.

In the wax-printing step (S20), only the paper portion where the wax is not printed remains, and these portions form the sample loading portion (11), the sensor portion (13) and the microfluidic channel (15). Since the periphery of the sample loading part 11, the sensor part 13 and the microfluidic channel 15 is made of paper impregnated with wax, in the main body 10 formed by the sticking step S50, Only the surface layer of the loading part 11 and the sensor part 13 is exposed to the outside and the remaining part of the sample loading part 11 and the sensor part 13 and the ninhydrin part 12 and the microfluidic channel 15, (10) and is shut off from the outside.

In the ninhydrin coating step (S30), the ninhydrin solution of a predetermined concentration is loaded on the sample loading part 11 and the ninhydrin part 12 below the sample loading part 11, followed by drying, thereby coating the ninhydrin with a constant concentration.

As described above, when the ninhydrin is coated on the ninhydrin 12 at a predetermined concentration, proline can be detected easily in the field even if it contains only the sulfosilicic acid solution as a reagent. Accordingly, it is possible to eliminate the inconvenience of having two sulfosilic acid solutions and two ninhydrin solutions, which are reagents for extracting proline to detect proline in the field.

As the ninhydrin is uniformly coated on the ninhydrin 12 at the lower part of the sample loading part 11 at a constant concentration, the sensitivity of the sensor part 13 is improved and a certain color is detected according to the concentration of proline And the reliability of the detection result is improved.

On the other hand, conventionally, a paper sensor composed of a sample loading part, a microfluidic channel, and a sensor part is formed in a single layer structure, so that there is a difference in ninhydrin concentration depending on the position, and thus it is difficult to quantitatively detect proline. That is, in the conventional single layer structure, the ninhydrin coated on the sample loading portion flows through the microfluidic channel, resulting in a difference in ninhydrin concentration depending on the position of the sample loading portion and the microfluidic channel, It was difficult to quantitatively detect proline because the color appearing in the sensor part was not uniform when the probe was detected and the color appeared in a band shape.

When the ninhydrin coating is completed through the above process, an Origami step for folding the wax-coated paper is performed. In the origami step S40, each layer constituting the main body 10 is simply folded on the basis of the origami line without special alignment. When the origami step S40 is completed, the layers are adhered with an adhesive to form an integral Thereby completing the main body 10. Origami means folding a piece of paper by folding a piece of paper to form various shapes. In the step S40, the paper coated with wax is folded to form the shape of the main body 10.

The process of detecting the proline of the measurement sample using the paper sensor for diagnosing environmental disorder according to the present invention will now be described.

In order to diagnose environmental disorder, a measurement sample is sampled from the crop to be sampled, and the measurement sample is loaded into the sample loading section 11. [ The proline of the loaded measurement sample reacts and binds to the ninhydrin coated on the ninhydrin moiety 12 under the sample loading section 11. The bound proline and ninhydrin form S-shaped microfluidic channels 15 ). The proline and ninhydrin of the measurement sample are perfectly mixed during the flow of the microfluidic channel 15.

Approximately two minutes after the measurement sample is loaded into the sample loading section 11, the proline which is bound to the ninhydrin reaches the sensor section 13. When the ninhydrin coupled proline arrives at the sensor unit 13, it is heated for about 3 minutes at a temperature of about 100 to 110 DEG C using a hot plate. As a result, color development is performed in the sensor unit 13 by the proline-ninhydrin reaction, and the color of the sensor unit 13 is changed.

As shown in FIG. 6, as the concentration of proline is increased, the degree of color development of a red color in the color of the sensor unit 10 is increased as the concentration of proline of the sensor unit 10 increases It appears dark. FIG. 6 shows the color of the sensor unit 13 when the paper sensor is heated at 150 ° C. for 10 minutes and the proline concentration is 50, 100, 150, or 200 μmol, respectively. As shown in FIG. 7, when the concentration of proline in the measurement sample is increased, the intensity of green color is decreased as shown in FIG. 7, because the degree of color development of red color is increased as the concentration of proline is increased.

As described above, since the color of the sensor unit 13 is changed according to the concentration of proline in the measurement sample, by using the colorimetric method in which the degree of discoloration of the sensor unit 13 is measured and compared with the reference color, The concentration can be detected.

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, Various modifications and variations will be possible without departing from the spirit of the invention. Therefore, the scope of the present invention should be construed as being covered by the scope of the appended claims, and technical scope within the scope of equivalents thereof should be construed as being included in the scope of the present invention.

10 ... body
11 .... Sample Loading Unit
12 ... Ninhiddrain
13 ... sensor unit
14 ... waterproof layer
15 ... microfluidic channel

Claims (9)

delete delete delete delete A ninhydrin coated ninhydrin portion 12, a sensor portion 13, a ninhydrin portion 12, and a sensor 14. The ninhydrin-coated sample loading portion 11, the ninhydrin-coated ninhydrin portion 12, An S-shaped microfluidic channel 15 connecting the first and second portions 13 to each other, and a pattern designing step S10 designing a paper folding line to divide the paper into four sections;
After the designed sensor pattern is printed on a paper with a wax printer, the wax is infiltrated into the paper by using a hot plate, and the wax for sealing the ninth droplet portion 12, the sensor portion 13 and the microfluidic channel 15 is sealed by the wax Printing step S20;
A ninhydrin coating step (S30) in which ninhydrin is coated at a predetermined concentration on the sample loading part (11) and the ninhydrin part (12) below the sample loading part (11);
The sample loading unit 11 and the sensor unit 13 are formed by folding the wax-impregnated paper along the paper folding line and the ninhydrin unit 12 and the sensor unit 13, which are located below the sample loading unit 11, (S40) for connecting the lower part of the microfluidic channel (15) with the S-shaped microfluidic channel (15);
(S50) of adhering each layer folded by paper folding with an adhesive to form a main body (10)
The sensor pattern designed in the pattern designing step S10 includes a first layer pattern 10a designed such that the sample loading unit 11 and the sensor unit 13 are spaced apart from each other by a predetermined distance, And the sensor unit 13 are disposed on the sample loading unit 11 and the sensor unit 13 of the first layer pattern 10a in such a manner that they are symmetrically positioned with respect to the paper folding line 10-1 10b of the second layer pattern 10b and the ninhydrin part 12 and the sensor part 13 are arranged on the ninhydrin part 12 and the sensor part 13 of the second layer pattern 10b with respect to the origami line 10-2 A third layer pattern 10c designed to be symmetrically positioned and connected to each other by the microfluidic channel 15 and a fourth layer pattern 10d formed as a waterproof layer without any pattern,
The first layer pattern 10a, the second layer pattern 10b, the third layer pattern 10c, and the fourth layer pattern 10d are sequentially arranged with reference to a paper fold line,
In the ninhydrin coating step S30, ninhydrin solution of a predetermined concentration is loaded on the sample loading part 11 and the ninhydrin part 12 below the sample loading part 11,
The adhesion step S50 is performed such that only the surface layer of the sample loading part 11 and the sensor part 13 is exposed to the outside of the body 10 and the remaining part of the sample loading part 11 and the sensor part 13 The ninhydrin portion 12 and the microfluidic channel 15 are located inside the main body 10 and adhered to be intercepted from the outside,
Wherein the wax-printing step (S20) comprises placing a paper on which a wax is printed on a hot plate and heating the paper for 2 to 3 minutes under a temperature condition of 120 ° C to completely penetrate the wax to the lower part of the paper A method of manufacturing a sensor. delete delete delete delete

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