Reagent adding and gas collecting device for soil anaerobic methane oxidation culture
By using a combination of a wide-mouth bottle made of PP material, a three-way valve, and a syringe, the problems of poor anaerobic environment and gas leakage in traditional devices were solved, achieving high efficiency, accuracy, and resource conservation in anaerobic methane oxidation experiments.
Patent Information
- Application Number
- CN202511312541.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-15
- Publication Date
- 2025-11-07
AI Technical Summary
Traditional soil culture devices suffer from gas leakage and air ingress when creating an anaerobic environment and adding 13C-labeled methane gas, which affects the accuracy of the experiment. Furthermore, gas collection is inconvenient, leading to resource waste and inaccurate results.
Using a wide-mouth bottle made of PP material combined with a three-way valve, a perforated rubber stopper, and a syringe, the inert gas replacement and gas collection are achieved through the switching of the three-way valve and the cooperation of the syringe, ensuring the tightness of the anaerobic environment and the stability of the gas, and preventing gas leakage.
This method enables the precise creation of anaerobic conditions and the stable preservation of gases, improving the accuracy and efficiency of experiments, simplifying the gas collection process, and ensuring the reliability of experimental results and the conservation of resources.
Smart Images
Figure CN120908415A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of soil environmental science and technology, specifically to a reagent addition and gas collection device for anaerobic methane oxidation culture in soil. Background Technology
[0002] Methane plays a vital role in Earth's biological evolution and human life and production. It is an important fossil fuel, a significant greenhouse gas, an abundant hydrocarbon in nature, and also serves as a substrate for microbial growth. Methane's contribution to the greenhouse effect is second only to carbon dioxide, reaching 20%. Changes in the metabolic pathways of microorganisms under anaerobic conditions in ecosystems lead to the production of different types and quantities of greenhouse gases, such as carbon dioxide, methane, and nitrous oxide. Methane's global warming potential is 25 times that of carbon dioxide, and its changes are particularly pronounced in flooded paddy fields, wetlands, and landfill soils. Anaerobic methane oxidation can be divided into three categories, one of which involves SO42-... 2－ As the final electron acceptor, it is called sulfate-reduced anaerobic methane oxidation; one type uses NO2 – NO3 – As the final electron acceptor, it is called denitrifying anaerobic methane oxidation; the third type is Fe 3+ Mn 4+ Cr 5+ MnO4 - Anaerobic methane oxidation uses metal ions as electron acceptors. Therefore, a deeper understanding and study of anaerobic methane oxidation processes in different environments is of great significance for further understanding the global carbon cycle and controlling methane emissions.
[0003] However, traditional soil culture devices have certain limitations for anaerobic methane oxidation experiments. Firstly, anaerobic methane oxidation experiments require the elimination of oxygen interference and the creation of a strictly anaerobic environment to study the influence of other electron acceptors on methane oxidation. Traditional soil culture devices are costly to create such an anaerobic environment and cannot guarantee against gas leakage during the closed culture process. Secondly, studying the anaerobic methane oxidation process requires the addition of... 13 Using C-labeled methane gas as a substrate, leakage can occur during the addition of methane to the bottle, affecting the experimental results and accuracy. Finally, gas samples need to be collected for analysis after the incubation period. CO2, a product of methane oxidation, is also present in the environment; therefore, it is crucial to prevent air from being collected during gas sample collection. However, traditional wide-mouth glass bottles cannot prevent air ingress, leading to significant resource consumption and reduced accuracy for experiments with large sample volumes. Summary of the Invention
[0004] The present application aims to provide a soil anaerobic methane oxidation culture reagent adding and gas collecting device to solve the problems of soil environmental science and technology in the background art.
[0005] To achieve the above object, the present application provides the following technical solution: a soil anaerobic methane oxidation culture reagent adding and gas collecting device, comprising a PP material wide-mouth bottle, a punch rubber plug movably connected at the bottle opening of the PP material wide-mouth bottle, a tee valve movably connected at the top of the punch rubber plug, a syringe one movably connected at one end of the tee valve, and a syringe two movably connected at the other end of the tee valve, a soil sample placed in the PP material wide-mouth bottle, and a screw cap provided at one end of the tee valve.
[0006] Preferably, the tee valve is fixedly connected with pipes at the upper end, lower end and left end, and the tee valve is provided with three holes, and the three holes are in communication with the three pipes.
[0007] Preferably, the injection end of the syringe one is tightly connected with the upper pipe of the tee valve, and the injection end of the syringe two is tightly connected with the left pipe of the tee valve.
[0008] Preferably, the punch rubber plug is provided with a through hole at the center, and the punch rubber plug is conical, and the side surface of the punch rubber plug is tightly attached to the inner wall of the bottle opening of the PP material wide-mouth bottle, and the bottom end of the lower pipe of the tee valve extends to the inside of the PP material wide-mouth bottle through the through hole of the punch rubber plug, and the outer surface of the lower pipe of the tee valve is tightly attached to the inner wall of the through hole of the punch rubber plug.
[0009] Preferably, the screw cap is provided with two, and one end of one of the screw caps is movably connected to one end of the upper pipe of the tee valve, and one end of the other screw cap is movably connected to one end of the left pipe of the tee valve.
[0010] Compared with the prior art, the present application has the following advantages:
[0011] 1. The device can effectively replace the air in the wide-mouth bottle with inert gas through the tee valve and the syringe one and the syringe two, thereby creating a strict anaerobic environment. In use, the residual air in the bottle can be almost completely replaced through multiple gas extraction and injection operations of the syringe one and the syringe two in cooperation with the tee valve, thereby providing precise anaerobic conditions for soil anaerobic methane oxidation culture, greatly ensuring the accuracy and reliability of the experimental conditions, and making the experimental results more truly reflect the oxidation of methane by soil under anaerobic conditions.
[0012] 2、The device can avoid the outside air entering the wide-mouth bottle through the accurate control of the three-way valve, effectively guaranteeing that the anaerobic condition is not destroyed, at the same time, the parts are tightly connected, such as the punched rubber plug and the inner wall of the wide-mouth bottle mouth are tightly attached, the three-way valve pipeline and the syringe and the punched rubber plug are tightly connected, and the sealing film is wound and sealed, the tight connection mode ensures that the methane gas will not leak, thereby maintaining a stable gas environment during the methane adding and subsequent culture process, improving the accuracy of the experiment, and making the research on the anaerobic methane oxidation process of soil more accurate.
[0013] 3、Through the flexible switching of the three-way valve and the cooperation of the syringe, the gas collection work can be efficiently completed. In operation, the steps are clear and definite, and the experimental personnel can easily master. Moreover, the overall structure of the device is stable, the parts are reliably connected, and the device can be stably operated for a long time. During the culture process, the sampling time can be designed according to the test, the screw cap can be conveniently removed at any time, the gas can be extracted by the syringe for detection, which provides great convenience for subsequent analysis work, helps to obtain experimental data in time, and tracks the change of the gas in the anaerobic methane oxidation process of soil. BRIEF DESCRIPTION OF DRAWINGS
[0014] Fig. 1 It is a structural schematic diagram of the present application;
[0015] Fig. 2 It is a connection structure schematic diagram of the syringe, the three-way valve and the punched rubber plug of the present application;
[0016] Fig. 3 It is a schematic diagram during the culture of the present application.
[0017] In the figure: 1, PP material wide-mouth bottle; 2, punched rubber plug; 3, three-way valve; 4, syringe 1; 5, syringe 2; 6, soil sample; 7, screw cap. DETAILED DESCRIPTION
[0018] The technical solutions in the embodiments of the present application will be described clearly and completely in combination with the drawings in the embodiments of the present application.
[0019] Please refer to Figs. 1-3 The present application provides a reagent adding and gas collecting device for anaerobic methane oxidation culture of soil, which comprises a PP material wide-mouth bottle 1, a punched rubber plug 2 movably connected at the bottle mouth of the PP material wide-mouth bottle 1, a three-way valve 3 movably connected at the top of the punched rubber plug 2, a syringe 1 4 movably connected at one end of the three-way valve 3, and a syringe 2 5 movably connected at the other end of the three-way valve 3, a soil sample 6 placed in the PP material wide-mouth bottle 1, and a screw cap 7 provided at one end of the three-way valve 3.
[0020] Further, the upper, lower and left ends of the three-way valve 3 are fixedly connected with pipes, and the three-way valve 3 is internally provided with three holes, and the three holes are communicated together with the three pipes respectively.
[0021] Further, the injection end of the injector one 4 is tightly connected with the upper pipe of the three-way valve 3, and the injection end of the injector two 5 is tightly connected with the left pipe of the three-way valve 3.
[0022] Further, the center of the punching rubber plug 2 is provided with a through hole, and the punching rubber plug 2 is conical, and the side surface of the punching rubber plug 2 is tightly attached to the inner wall of the bottle opening of the PP material wide-mouth bottle 1, and the bottom end of the lower pipe of the three-way valve 3 extends to the inside of the PP material wide-mouth bottle 1 through the through hole of the punching rubber plug 2, and the outer surface of the lower pipe of the three-way valve 3 is tightly attached to the inner wall of the through hole of the punching rubber plug 2.
[0023] Further, two screw caps 7 are arranged, and one end of one of the screw caps 7 is movably connected to one end of the upper pipe of the three-way valve 3, and one end of the other screw cap 7 is movably connected to one end of the left pipe of the three-way valve 3.
[0024] The application embodiment is used as follows: firstly, the soil to be tested 6 is placed in the PP material wide-mouth bottle 1, and the required reagent or water for the experiment is added and mixed. The bottle is flushed with inert gas, and then the connected three-way valve 3 is connected with the PP material wide-mouth bottle 1 through the punched rubber plug 2, the connection is sealed with sealing film, and at the same time, the three-way valve 3 is connected with the syringe one 4 and the syringe two 5 through the upper and left pipelines, and the connection is also sealed with sealing film. Then, the syringe one 4 contains inert gas, and the syringe two 5 does not contain any gas. Subsequently, the three-way valve 3 is rotated to point to the left pipeline and the lower pipeline, and after the gas in the bottle is extracted by the syringe two 5, the three-way valve 3 is rotated to point to the upper pipeline and the lower pipeline, and the inert gas in the syringe one 4 is sucked into the bottle by using the gas pressure balance. Then, the gas in the syringe two 5 is emptied and inserted into the left pipeline. After the above steps are repeated four to five times, the residual air in the bottle is completely replaced by inert gas, and the bottle reaches an anaerobic environment. Subsequently, the three-way valve 3 is rotated to point to the upper pipeline and the left pipeline to avoid air entering when preparing the subsequent steps. In this experiment, the isotope-labeled methane gas is needed to be introduced. After the methane gas is extracted by the syringe one 4, the syringe two 5 is inserted into the left pipeline. Subsequently, the three-way valve 3 is rotated to point to the left pipeline and the lower pipeline, and according to the required amount of methane, the same volume of inert gas in the bottle is extracted by the syringe two 5. Then, the three-way valve 3 is rotated to point to the upper pipeline and the lower pipeline, and the methane gas in the syringe one 4 in the upper pipeline is sucked into the bottle by using the gas pressure balance, so as to complete the replacement of the gas in the bottle. Finally, the three-way valve 3 is rotated to point to the upper pipeline and the left pipeline, the syringe is removed, and the upper and left pipeline openings are sealed with the screw cap 7. Then, the soil sample 6 is cultured according to the required experimental conditions. The methane oxidation gas accumulates in the wide-mouth bottle 1. According to the sampling time of the test design, the screw cap 7 on the upper pipeline is unscrewed, the syringe one 4 for emptying the gas is inserted, the three-way valve 3 is rotated to point to the upper pipeline and the lower pipeline, and the gas in the bottle is mixed uniformly by slowly extracting the gas three times by the syringe one 4. Then, a certain amount of gas is extracted by the syringe one 4, and the isotope ratio mass spectrometer is used to measure the CO2 concentration. Finally, the rubber plug is opened, and the soil in the bottle is taken out for soil sample detection. The gas is taken every 7 days, and after 28 days of culture, the collected gas in the experiment can be detected by the machine, the data is good, and it is proved that the airtightness of the device is good. 13 CO2 concentration. Finally, the rubber plug is opened, and the soil in the bottle is taken out for soil sample detection. The gas is taken every 7 days, and after 28 days of culture, the collected gas in the experiment can be detected by the machine, the data is good, and it is proved that the airtightness of the device is good.
[0025] Although the present application is described in detail with reference to the foregoing embodiments, those skilled in the art can modify the technical solutions recorded in the foregoing embodiments or make equivalent replacements to part of the technical features, and any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.
Claims
1. A reagent adding and gas collecting device for soil anaerobic methane oxidation culture, comprising a PP material wide-mouth bottle (1), characterized in that: The PP material wide-mouth bottle (1) is movably connected with a punching rubber plug (2) at the bottle mouth, the top of the punching rubber plug (2) is movably connected with a three-way valve (3), one end of the three-way valve (3) is movably connected with a syringe (4), and the other end of the three-way valve (3) is movably connected with a syringe (5), the inside of the PP material wide-mouth bottle (1) is placed with a soil sample (6), and one end of the three-way valve (3) is provided with a screw cap (7).
2. The reagent addition and gas collection apparatus for soil anaerobic methanotrophic culture according to claim 1, characterized in that: The upper, lower and left ends of the three-way valve (3) are fixedly connected with pipelines, and the inside of the three-way valve (3) is provided with three holes, and the three holes are communicated together.
3. The reagent addition and gas collection apparatus for soil anaerobic methanotrophic culture according to claim 2, characterized in that: The injection end of the syringe (4) is tightly connected with the upper pipeline of the three-way valve (3), and the injection end of the syringe (5) is tightly connected with the left pipeline of the three-way valve (3).
4. The reagent addition and gas collection apparatus for soil anaerobic methanotrophic culture according to claim 2, characterized in that: The center of the punching rubber plug (2) is provided with a through hole, the punching rubber plug (2) is conical, the side surface of the punching rubber plug (2) is tightly attached to the inner wall of the bottle mouth of the PP material wide-mouth bottle (1), the bottom end of the lower pipeline of the three-way valve (3) extends to the inside of the PP material wide-mouth bottle (1) through the through hole of the punching rubber plug (2), and the outer surface of the lower pipeline of the three-way valve (3) is tightly attached to the inner wall of the through hole of the punching rubber plug (2).
5. The reagent addition and gas collection apparatus for soil anaerobic methanotrophic culture according to claim 2, characterized in that: The screw cap (7) is provided with two, one end of one of the screw caps (7) is movably connected with one end of the upper pipeline of the three-way valve (3), and one end of the other screw cap (7) is movably connected with one end of the left pipeline of the three-way valve (3).