Patent Application
20240230614
This application relates to detection of soil organic matter, and more particularly to a device and method for on-site detection of soil organic matter, and a microfluidic chip.
Scientific management of fertilization and improvement of food production are two major challenges in the modern agricultural production. Organic matter is an important component of soil, and is an important indicator of soil fertility, providing crops with essential nutrients. The soil fertility can be estimated by detecting the content of soil organic matter. The soil fertility can be improved by supplementing soil nutrients through precise fertilization, so as to improve the crop yield and quality.
The soil organic matter content is generally detected through traditional chemical methods, such as potassium dichromate volumetric method, dry combustion method, and loss-on-ignition method. These traditional methods require complex and cumbersome manual operations for soil samples, and thus are inefficient, costly, and time-consuming. Besides, the involved detection equipment is expensive and bulky, and requires regular maintenance. Therefore, such traditional methods are not suitable for rapid on-site detection and analysis. With the development of science and technology, various advanced techniques, such as near-infrared spectroscopy, remote sensing and hyperspectral technique, have been widely used in the rapid estimation of soil nutrients in recent years. These methods involve the construction of a model for estimating the soil organic matter content, and for various soil samples, multiple estimation models are required, failing to achieve the rapid and accurate model construction.
Microfluidics, as a new analytical platform, has the advantages of miniaturization, automation, integration, simplification, and rapid detection, and thus have been widely applied in the practical detection. However, the centrifugal microfluidic analysis technology, in which several samples in the form of micro fluid are simultaneously detected in a disc-shaped chip under the drive of centrifugal force, has still not be reported in the on-site rapid detection of soil organic matter.
Therefore, there is an urgent need for a low-cost, rapid, and reliable method for on-site detection of soil organic matter and related chips.
An object of the present disclosure is to provide a device and method for on-site detection of soil organic matter, and a microfluidic chip, so as to overcome the deficiencies in the prior art, and achieve the on-site detection of soil organic matter with less time consumption, higher detection efficiency, and simplified operation.
The technical solutions of the present disclosure are described below.
In a first aspect, this application provides device for on-site detection of soil organic matter, comprising:
In some embodiments, the device further comprises:
In some embodiments, the pre-processing module is configured to process a solvent solution to obtain the extraction solvent.
In some embodiments, the centrifugal system is a centrifugal detector.
In some embodiments, the microfluidic chip comprises a channel layer and a cover layer arranged above the channel layer;
In some embodiments, the cover layer is provided with a first mounting hole, a plurality of sample injection holes, a plurality of second solvent injection ports and a plurality of second air holes; a middle of the channel layer main body is provided with a second mounting hole corresponding to the first mounting hole; the plurality of sample injection holes, the plurality of second solvent injection ports, the plurality of second air holes and the plurality of channel branches are the same in number, and the plurality of channel branches are in one-to-one correspondence with the plurality of sample injection holes, the plurality of second solvent injection ports and the plurality of second air holes; the plurality of sample injection holes are in one-to-one correspondence with sample inlets of the plurality of channel branches; the plurality of second air holes are in one-to-one correspondence with first air holes of the plurality of channel branches; and the plurality of second solvent injection ports are in one-to-one correspondence with first solvent injection ports of the plurality of channel branches.
In some embodiments, the cover layer is provided with an observation window; and the observation window comprises a penetration hole arranged on the cover layer and an optically-permeable film mounted in the penetration hole.
In some embodiments, a base plate layer is provided below the channel layer; and a middle of the base plate layer is provided with a third mounting hole.
In some embodiments, the filter pad is configured as at least one layer; and the filter pad is a metal filter screen, a non-metallic filter cloth, a filter membrane, or a combination thereof.
In a second aspect, this application provides a detection method using the aforementioned on-site detection device, comprising:
In a third aspect, this application provides a centrifugal microfluidic chip, comprising:
In some embodiments, the extraction well is provided with a plurality of heating columns.
In some embodiments, the filtration well is provided with a microarray and a plurality of microbeads; the plurality of microbeads are located above the microarray; and a filter pad is provided at the outlet of the filtration well.
In some embodiments, the cover layer is provided with a first mounting hole, a plurality of sample injection holes, a plurality of first solvent injection ports, and a plurality of first air holes.
In some embodiments, each of the plurality of channel branches further comprises a second air hole connected to the waste collection well and a second solvent injection port connected to the inlet of the extraction well; a middle of the channel layer main body is provided with a second mounting hole corresponding to the first mounting hole; the plurality of sample injection holes, the plurality of first solvent injection ports, the plurality of first air holes, and the plurality of channel branches are the same in number, and the plurality of channel branches are in one-to-one correspondence with the plurality of sample injection holes, the plurality of first solvent injection ports and the plurality of first air holes; the plurality of sample injection holes are in one-to-one correspondence with sample inlets of the plurality of channel branches; the plurality of first air holes are in one-to-one correspondence with second air holes of the plurality of channel branches; and the plurality of first solvent injection ports are in one-to-one correspondence with second solvent injection ports of the plurality of channel branches.
In some embodiments, the cover layer is provided with a observation window; and the observation window comprises a penetration hole arranged on the cover layer and an optically-permeable film mounted in the penetration hole.
In some embodiments, a middle of the base plate layer is provided with a third mounting hole.
In some embodiments, the channel layer comprises a first channel layer and a second channel layer arranged in sequence; an upper portion of each of the plurality of channel branches is located in the first channel layer, and a lower portion of each of the plurality of channel branches is located in the second channel layer; and the upper portion is through in the first channel layer and is communicated with the lower portion in the second channel layer.
In some embodiments, the filter pad is configured as at least one layer; and the filter pad is a metal filter screen, a non-metal filter cloth, a filter membrane, or a combination thereof.
In a fourth aspect, this application provides a method for preparing the centrifugal microfluidic chip, comprising:
Compared to the prior art, this application has the following beneficial effects.
In the drawings, 1, microprocessor module; 2, centrifugal system; 3, pre-processing module; 4, photoelectric detection module; 5, temperature control module; 6, display; 7, power supply module; 8, communication module; 9, microfluidic chip; 10, heating plate; 11, drive module; 901, cover layer; 902, channel layer; 903, channel branch; 904; first air hole; 905, sample injection hole; 906, first solvent injection port; 907, sample inlet; 908, second solvent injection port; 909, heating column; 910, extraction well; 911, microchannel; 912, microarray; 913, microbead; 914, filter pad; 915, filtration well; 916, detection well; 917, waste collection well; and 918, second air hole.
The present disclosure will be further described below with reference to the embodiments and accompanying drawings.
As shown in
The pre-processing module 3 is configured to process a soil sample into a soil sample solution and process an extraction solvent into an extraction solvent solution. The pre-processing module 3 includes a through-valve, a quantification ring, a soil sample processing device, a solvent processing device, a scale, and a waste liquid processing device. The scale is configured for weighing. The through-valve is configured for feeding a sample. The quantification ring is configured for quantifying. The soil sample processing device is configured for converting a soil sample into a soil sample solution. The solvent processing device is configured for preparing a solvent solution. The waste liquid processing device is configured for treating the waste liquid.
The centrifugal system 2 is configured to generate a centrifugal force. The centrifugal system is a centrifugal detector. The drive module is configured to drive the centrifugal system to work. The centrifugal system and drive module are configured to achieve precise manipulation and transfer of the liquid on the microfluidic chip by using a rotating tray and a centrifugal microfluidic solvent tray, driven by centrifugal force.
The microfluidic chip 9 is configured for the flowing and mixing of the soil sample solution and the extraction solvent within the microfluidic chip for extraction under the action of the centrifugal force generated by the centrifugal system.
The photoelectric detection module 4 is configured to detect the extract to determine the organic matter content in the soil sample solution. The photoelectric detection module 4 includes a light source and a photoelectric sensor. By measuring the absorbance of the detection zone (color development zone) on the microfluidic chip 9, the photoelectric detection module 4 inputs the detected signals into the microprocessor module 1 for data processing to obtain test results. The soil organic matter content is determined according to the test results, and the precise test results are displayed on the display 6, or stored and printed through the communication module 8.
The heating plate 10 is arranged below the microfluidic chip 9 and is configured to heat the microfluidic chip 9. The temperature control module 5 is configured to control the heating temperature of the heating plate 10. The temperature control module 5 is configured to control the overall temperature of the microfluidic chip 9 by arranging the heating plate 10 and the temperature sensor at the bottom of the microfluidic chip 9.
The power supply module 7 is configured for supplying power to the microprocessor module 1.
The communication module 8 is configured for realizing communication of the microprocessor module 1 with other modules.
The display 6 is configured for displaying detection results of the photoelectric detection module.
The microprocessor module 1 is configured to receive and analyze the detection results of the photoelectric detection module 4 and display analysis results on the display 6, and control the drive module 11 and the temperature control module 5. The microprocessor module 1, the communication module 8 and the display 6 together achieve the control on the whole device, data acquisition, detection analysis, result display and printing, and transmission processing.
Through microfluidics technology, basic operation units, such as sample preparation, reaction, separation, and detection, can be made into micron and nanometer-scale components, and these components can be integrated into a tiny chip, thereby realizing the whole process of analysis and testing. As the microfluidic chip requires a small amount of sample and solvent and has high detection efficiency, it has been widely used in testing-related fields as a new analytical platform with the advantages of miniaturization, automation, integration, convenience, and rapidity. In this application, the microfluidic chip technology is applied to soil organic matter detection to develop a fully automated, fast, and simple detection platform and analysis method, which significantly saves time for sample processing and minimizes the cost of solvents and instruments. Moreover, it can also achieve intelligent rapid detection in the form of “sample in, result out” while ensuring accurate detection for the whole process. Therefore, it is significant to perform rapid and accurate analysis and determination on the soil organic matter and solve the problem of on-site rapid measurement of soil organic matter.
The centrifugal microfluidic chip includes a channel layer, a cover layer arranged above the channel layer, and a bottom layer arranged below the channel layer.
The channel layer includes a channel layer main body and multiple channel branches arranged on the channel layer main body. Each channel branch includes a sample inlet, an extraction well, a microchannel, a filtration well, a detection well, and a waste collection well. An inlet of the extraction well is connected to the sample inlet, and an outlet of the extraction well is connected to an inlet of the microchannel. An outlet of the microchannel is connected to an inlet of the filtration well. An outlet of the filtration well is connected to an inlet of the detection well, and an outlet of the detection well is connected to an inlet of the waste collection well.
In an embodiment, a microfluidic chip 9 as shown in
In another embodiment, a microfluidic chip 9 as shown in
As shown in
As shown in
The channel branch 903 has an upper portion located in channel layer I 9021 and a lower portion located in channel layer II 9022. The upper portion of the channel branch 903 is through in the channel layer I, and is connected to the channel branch in the channel layer II. The channel branch 903 includes a second solvent injection port 908, a sample inlet 907, an extraction well 910, a microchannel 911, a filtration well 915, a detection well 916, a waste collection well 917, and a second air hole 918. An inlet of the extraction well 910 is connected to the second solvent injection port 908 and the sample inlet 907, respectively. The outlet of extraction well 910 is connected to the inlet of microchannel 911. The outlet of the microchannel 911 is connected to the inlet of the filtration well 915. The outlet of the filtration well 915 is connected to an inlet of the detection well 916. The outlet of the detection well 916 is connected to the inlet of the waste collection well 917, and the waste collection well 917 is connected to the second air hole 918. Under the control of centrifugal force, the soil sample solution and the extraction solvent can be precisely manipulated and transferred inside the microfluidic chip to achieve mixing, reaction, separation and color development of the soil sample solution and the extraction solvent.
The extraction well 910 is provided with a plurality of heating columns 909. The heating columns 909 are configured to heat the mixture of sample and extraction solvent in the extraction well 910 such that the sample and solvent can react more fully in the extraction well under heated conditions. The heating plate is configured to heat the entire microfluidic chip and the liquid therein to ensure that the mixture can react fully throughout the process.
The filtration well 915 is provided with a microarray 912 and a plurality of microbeads 913 varied in sizes. The microbeads 913 are located above the microarray 912, and the microarray 912 can be directly processed into a square microcolumn array in the filtration well 915. The filtration well 915 can be provided with the microarray, the plurality of microbeads, or a combination thereof. In addition, a filter pad 914 is provided at the outlet of the filtration well 915, and the filter pad 914 is a metal filter screen, a non-metal filter cloth, a filter membrane, or a combination thereof, which can be configured as a layer or multiple layers, depending on the situation. The number of the microsphere 913 is more than ore, and the plurality of microbeads 913 are randomly arranged in the filtration well 915 and are varied in size. The microbeads 913 are organic polymers, polystyrene (PS) microbeads, or silica microbeads synthesized in situ. The microarray 912, the microbeads 913 and the filter pad 914 are collaborative to effectively filter out fine particles from the liquid to be measured. The filtration well 915 can effectively remove particles with different sizes from the extract through the filtration structures of the microarray 912, microbeads 913 and the filter pad 914 for secondary filtration of the extract to be tested, thus improving the accuracy of the test results.
When being tested, the sample solution is introduced through the sample injection hole 905, and the extraction solvent is introduced through the first solvent injection port 906 and the second solvent injection port 908. Then the sample solution and the extraction solvent flow through the extraction well 910, the filtration well 915 and the detection well 916 along the microchannel 911 to complete the color development reaction of the solution. The solvent can be liquid or solid. If the solvent is liquid, it can be introduced under pressure in the pre-processing module 3 or encapsulated inside the microfluidic chip 9 in the form of a liquid capsule. If the solvent is solid, it can be powder or a block solvent sealed inside the microfluidic chip 9.
As shown in
The sample injection hole 905, the first solvent injection port 906, the first air hole 904, and the channel branch 903 are provided in equal numbers and in correspondence. The sample injection hole 905 and the sample inlet 907 are arranged in correspondence and are communicated with each other. The soil sample solution is added from the sample injection hole 905, and then flows into the sample inlet 907 along the sample injection hole 905. The first air hole 904 is provided in correspondence with the second air hole 918 to allow the interior of channel branch 903 to communicate with the outside atmosphere to maintain the pressure balance. The first solvent injection port 906 and the second solvent injection port 908 are corresponding and communicated with each other. The solvent is added from the first solvent injection port 906, and flows to the second solvent injection port 908 along the first solvent injection port 906.
Furthermore, the cover layer 901 is provided with an observation window. The observation window includes a penetration hole arranged on the cover layer and an optically-permeable film mounted in the penetration hole. The photoelectric detection module 4 emits detection light through the observation window to optically detect the reaction results in the detection well.
The following is a method for preparing and using the aforementioned centrifugal microfluidic chip, which includes the following steps.
The working process of the microfluidic chip provided in the present disclosure is described as follows.
When the microfluidic chip 9 is used for soil organic matter detection, the soil sample solution is injected into the microfluidic chip 9 through the sample injection hole 905, and then flows to the sample inlet 907 from the sample injection hole 905. The solvent is injected from the first solvent injection port 906 or is pre-built in the first solvent injection port 906 and the second solvent injection port 908.
This centrifugal microfluidic chip is configured for detecting soil organic matter, which is a disc-shaped chip consisting of multiple layers of chips. Driven by the centrifugal force generated by rotation, the mixing, reaction, separation and color development process of the to-be-tested sample and the reaction reagent are realized. Finally, the organic matter content in the soil sample is quantitatively detected by UV-visible spectrophotometer. This centrifugal microfluidic chip for soil organic matter detection requires a small amount of sample and reagents and can process and test multiple samples in parallel, which is fast and convenient.
Under the action of centrifugal force generated by the centrifugal system, the soil sample solution in the sample inlet 907 flows into the extraction well 910 to mix with the extraction solvent. The heating column 909 in the extraction well 910 heats the mixture of the soil sample solution and the extraction solvent in the extraction well 910 to accelerate the extracting rate. The soil sample solution and the extraction solvent undergo an initial extraction in the extraction well, and continue to move forward under the centrifugal force of the centrifugal system 2 to flow in the spiral or round-trip bent-shaped microchannels 911 for extraction. By designing the microchannels 911 in a spiral or round-trip bend shape, the soil sample solution can be fully extracted with the solvent.
With centrifugal force as the driving force, the microfluid can be precisely manipulated and transferred to achieve the mixing, reaction, separation and color development of the sample and the reaction reagent on the chip. The content of organic matter in the sample is qualitatively or quantitatively detected by the photoelectric detector, which can realize the on-site fast detection of soil organic matter with short detection cycle, high detection efficiency, simple and convenient operations.
When the mixture of soil sample solution and solvent flows to the filtration well, the mixture moves forward along the horizontal direction to filter the excess particulate matter through microbeads 913 and microarray 912 while moving from top to bottom, during which the mixture passes through the microbeads 913 to filter the excess particulate matter therein and then passes through the microarray 912 to filter the particulate matter. When the mixture flows to the filter pad 914 at the outlet of the filtration well 915, the filter pad 914 filters the mixture once more to filter out the excess particulate matter therein. Through the multiple filtrations of the microarray 912, the microbeads 913 and the filter pad 914, the excess particulate matter in the mixture can be filtered out as much as possible to ensure the accuracy of the test results.
After multiple filtration processes, the mixture flows into the detection well 916, and the extracted liquid is tested by the photoelectric detection module 4 to determine the organic matter content in the soil. After the test is completed, the liquid flows to the waste collection well. During the flowing of the mixture, the extraction keeps going between the soil sample solution and the solvent.
This application also provides a detection method by using the above-mentioned detection device, which includes the following steps.
The centrifugal system is started such that the to-be-tested sample solution and the extraction solvent are mixed, reacted and separated in the extraction well under the centrifugal force. The centrifugal speed of the centrifugal system is changed such that the extract is transferred from the extraction well to the filtration well through a microvalve for solid-liquid separation, where the microvalve is installed between the extraction well and the filtration well. The centrifugal speed of the centrifugal system is changed again such that the clarified liquid after filtration is transferred into the detection well through a microvalve, where the microvalve is installed between the filtration well and the detection well.
The beneficial effects of the present disclosure are described below.
Described above are merely preferred embodiments of the present disclosure, which are not intended to limit the scope of the present disclosure. Without departing from the spirit of the present disclosure, all variations and improvements made to the technical solutions of the present disclosure by one of ordinary skill in the art shall fall within the scope of the disclosure defined by the appended claims.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202210783531.7 | Jul 2022 | CN | national |
| 202210787357.3 | Jul 2022 | CN | national |
This application is a continuation of International Patent Application No. PCT/CN2022/128072, filed on Oct. 27, 2022, which claims the benefit of priority from Chinese Patent Application Nos. 202210787357.3 and 202210783531.7, both filed on Jul. 5, 2022. The content of the aforementioned application, including any intervening amendments thereto, is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 20240133857 A1 | Apr 2024 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/CN2022/128072 | Oct 2022 | WO |
| Child | 18396388 | US |
