Sampling device and sampling method thereof

Abstract

The invention provides a sampling device and a sampling method thereof, and relates to the technical field of fermentation broth sampling, a main pipeline is connected with a plurality of sampling assemblies, the plurality of sampling assemblies are independently opened and closed to ensure the sterility of each time of sampling, when sampling is needed, one sampling assembly is opened, and the other sampling assemblies are closed, so that the sampling efficiency is improved. A sample in the fermentation tank enters the sample receiving component through the sampling assembly in the open state, when sampling needs to be carried out again, the residual sample in the sampling assembly opened in the last time of sampling flows into the waste liquid receiving bottle, the waste liquid of the last time of sampling is effectively prevented from influencing the sampling result, then another sampling assembly is opened for sampling operation, and the sampling efficiency is improved. Therefore, multiple independent sampling is achieved, fermentation dynamic changes are monitored in real time, and the technical problems that in the prior art, a traditional fermentation tank sampling device is difficult to ensure sterile operation, and multiple sampling cannot be achieved are solved.

Description

Technical Field

[0001] The present invention relates to the technical field of fermentation broth sampling, and particularly to a sampling device and a sampling method thereof. Background Art

[0002] With the continuous development of biotechnological fermentation, real-time monitoring and control during the fermentation process have become key factors in improving product quality and efficiency. Fermentation is an important process step in microbial pharmaceutics. By monitoring various parameters during the fermentation process, the dynamic changes of fermentation can be better understood, thereby optimizing the fermentation conditions. In this context, sampling and analysis of fermentation broth have become a key link in monitoring the fermentation process and quality. The devices and methods for real-time sampling largely determine the quality of samples, which is of great significance for omics analysis, fermentation result prediction, and precise regulation of key metabolic nodes and pathways during the fermentation process.

[0003] In the prior art, sampling and analysis during the reaction process and storage process are important means for monitoring reaction substrates, reaction products, reaction processes, and stored substances. For the biological fermentation process, implementing online sampling can not only monitor fermentation processes such as whether there is contamination by miscellaneous bacteria, the state of bacteria, the material environment, substrate consumption, and product generation, but also provide a basis for omics analysis, fermentation result prediction, and real-time online dynamic control during the fermentation process. Therefore, the quality of real-time online sampling directly affects the effects of the aforementioned monitoring and control.

[0004] However, in practical applications, there are some deficiencies in the current sampling devices for fermentation tanks. On the one hand, traditional sampling methods are difficult to ensure aseptic operation, easily leading to contamination of the fermentation broth in the fermentation tank by miscellaneous bacteria, thereby affecting the accuracy of fermentation results. On the other hand, most of the existing sampling devices can only perform single sampling and cannot meet the requirement of multiple samplings during the fermentation process to monitor the dynamic changes of fermentation in real time. Especially in a Class D clean area or a general area, how to achieve aseptic sampling has become an important challenge because the inability to perform aseptic sampling on the fermentation tank will affect the judgment of the fermentation culture state. Summary of the Invention

[0005] The purpose of the present invention is to provide a sampling device and a sampling method thereof to alleviate the technical problems in the prior art that the traditional sampling device for a fermentation tank is difficult to ensure aseptic operation and cannot perform multiple samplings.

[0006] In a first aspect, the sampling device provided by the present invention for a fermentation tank includes: a main pipeline, a sampling assembly, a waste liquid receiving bottle, and a sample receiving member;

[0007] One end of the main pipeline is communicated with the fermentation tank, and the other end of the main pipeline is communicated with the waste liquid receiving bottle;

[0008] A plurality of the sampling components are provided, one end of each sampling component is communicated with the main pipeline, and the other end of each sampling component is connected with a sample receiving member;

[0009] Each of the sampling components has an open state and a closed state;

[0010] When sampling for the first time, one of the sampling components is in the open state, and the remaining sampling components are in the closed state, so that the sample in the fermentation tank enters the sample receiving member through the sampling component in the open state;

[0011] When sampling subsequently, the sample remaining in the sampling component that was opened during the previous sampling flows into the waste liquid receiving bottle, and then another sampling component is opened for sampling operation.

[0012] In an alternative embodiment,

[0013] The sampling component includes a sampling pipeline;

[0014] One end of the sampling pipeline is communicated with the main pipeline, and the other end of the sampling pipeline is communicated with the sample receiving member. The sample in the fermentation tank can enter the sample receiving member along the main pipeline and the sampling pipeline.

[0015] In an alternative embodiment,

[0016] A first sampling control member and a second sampling control member are sequentially arranged on the sampling pipeline along the liquid flow direction, and the first sampling control member and the second sampling control member are used to control the switching of the sampling pipeline between the open state and the closed state.

[0017] In an alternative embodiment,

[0018] An interface member is arranged at one end of the sampling pipeline far from the main pipeline, and the sampling pipeline is communicated with the sample receiving member through the interface member.

[0019] In an alternative embodiment,

[0020] An air filter is arranged on the sample receiving member, and the air filter is used to exclude the gas in the bag when the sample receiving member receives the sample.

[0021] In an alternative embodiment,

[0022] A first pipeline control member and a second pipeline control member are sequentially arranged on the main pipeline along the liquid flow direction, and the first pipeline control member and the second pipeline control member are used to control the switching of the main pipeline between the open state and the closed state.

[0023] In an alternative embodiment,

[0024] the sampling device further includes a first connection member and a second connection member;

[0025] The first connection member and the second connection member are spaced apart on the main pipeline, and both the first connection member and the second connection member are used to connect at least two of the sampling assemblies to the main pipeline.

[0026] In an alternative embodiment,

[0027] A third pipeline control member is provided between the first connection member and the second connection member.

[0028] Second aspect, a sampling method based on the sampling device, comprising the following steps:

[0029] Installation step: Connect the main pipeline to the fermentation tank, calculate the number of experimental sampling times, and connect a plurality of sampling assemblies to the main pipeline according to the number of experimental sampling times;

[0030] Cleaning step: Rinse the connection part between the main pipeline and the sampling assembly;

[0031] First sampling step: Open one of the sampling assemblies, and the sample in the fermentation tank enters the sample receiving member through the opened sampling assembly;

[0032] Re - sampling step: After flowing the sample in the previously sampled sampling assembly into the waste liquid receiving bottle, open another sampling assembly.

[0033] In an alternative embodiment,

[0034] The sampling assembly includes a sampling pipeline;

[0035] One end of the sampling pipeline is connected to the main pipeline, the other end of the sampling pipeline is connected to the sample receiving member, and a first sampling control member and a second sampling control member are sequentially arranged on the sampling pipeline along the liquid flow direction;

[0036] The first sampling step includes the following steps:

[0037] Open the first sampling control member and the second sampling control member, the sample in the fermentation tank passes through the main pipeline and the sampling pipeline and enters the waste liquid receiving bottle. After sampling is completed, close the first sampling control member and the second sampling control member, and cut the sampling pipeline between the first sampling control member and the second sampling control member.

[0038] The sampling device provided by the present invention is connected with multiple sampling components on the main pipeline. The multiple sampling components can be independently opened and closed to ensure the sterility of each sampling. When sampling is required, one of the sampling components is opened and the rest are closed. The sample in the fermenter enters the sample receiving member through the opened sampling component. When sampling is required again, the residual sample in the sampling component opened during the previous sampling flows into the waste liquid receiving bottle, effectively preventing the waste liquid from the previous sampling from affecting the sampling result. Subsequently, another sampling component is opened for sampling operation, thereby realizing multiple independent samplings to monitor the dynamic changes of fermentation in real time, and alleviating the technical problems in the prior art that the traditional sampling device of the fermenter is difficult to ensure aseptic operation and cannot perform multiple samplings. Description of the Drawings

[0039] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0040] Figure 1 It is a schematic diagram of the overall structure of the sampling device provided by the embodiment of the present invention.

[0041] Reference numerals: 10 - fermenter; 100 - main pipeline; 110 - first pipeline control member; 120 - second pipeline control member; 130 - third pipeline control member; 200 - sampling component; 210 - sampling pipeline; 220 - first sampling control member; 230 - second sampling control member; 240 - interface member; 300 - waste liquid receiving bottle; 400 - sample receiving member; 500 - air filter; 600 - first connection member; 700 - second connection member. Specific Embodiments

[0042] The technical solutions of the present invention will be clearly and completely described below with reference to the drawings. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0043] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be construed as a limitation to the present invention. In addition, the terms "first", "second", "third" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance.

[0044] In the description of the present invention, it should be noted that unless otherwise clearly specified and limited, the terms "mounted", "connected", "coupled" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the communication inside two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0045] The following will describe in detail the specific embodiments of the present invention with reference to the drawings. It should be understood that the specific embodiments described herein are only for the purpose of illustrating and explaining the present invention, and are not used to limit the present invention.

[0046] As Figure 1 shown, the sampling device provided in this embodiment is used for the fermenter 10 and includes: a main pipeline 100, a sampling assembly 200, a waste liquid receiving bottle 300, and a sample receiving member 400; one end of the main pipeline 100 is connected to the fermenter 10, and the other end of the main pipeline 100 is connected to the waste liquid receiving bottle 300. When the main pipeline 100 is connected to the fermenter 10, a sealed connection needs to be ensured.

[0047] A plurality of sampling assemblies 200 are provided. The specific number of the sampling assemblies 200 is determined according to the calculated number of experimental sampling times. For example, if the number of experimental sampling times is N, the sampling assemblies 200 are set to N + 2, and the sampling assemblies 200 with the number of N + 2 are connected to the main pipeline 100.

[0048] One end of each sampling assembly 200 is connected to the main pipeline 100, and the other end of each sampling assembly 200 is connected to a sample receiving member 400.

[0049] Each sampling component 200 has an open state and a closed state; during the first sampling, one of the sampling components 200 is in the open state, and the remaining sampling components 200 are in the closed state, so that the sample in the fermenter 10 enters the sample receiving member 400 through the sampling component 200 in the open state. Subsequently, the sampling component 200 is cut off, transferred to a laminar flow hood, and in a sterile environment, the sample is poured into a sterile EP tube for subsequent experimental operations.

[0050] During subsequent sampling, the sample remaining in the sampling component 200 that was open during the previous sampling flows into the waste liquid receiving bottle 300. Subsequently, another sampling component 200 is opened for sampling operation to achieve multiple sampling operations.

[0051] The sampling device provided in this embodiment connects multiple sampling components 200 to the main pipeline 100. The multiple sampling components 200 are independently opened and closed to ensure the sterility of each sampling. When sampling is required, one of the sampling components 200 is opened and the remaining sampling components 200 are closed. The sample in the fermenter 10 enters the sample receiving member 400 through the sampling component 200 in the open state. When sampling is required again, the sample remaining in the sampling component 200 that was open during the previous sampling flows into the waste liquid receiving bottle 300, effectively preventing the waste liquid from the previous sampling from affecting the sampling result. Subsequently, another sampling component 200 is opened for sampling operation, thereby achieving multiple independent samplings to monitor the dynamic changes of fermentation in real time, alleviating the technical problems in the prior art that the traditional sampling device for the fermenter 10 is difficult to ensure sterile operation and cannot perform multiple samplings.

[0052] Regarding the structure and shape of the sampling component 200, specifically:

[0053] The sampling component 200 includes a sampling pipeline 210; one end of the sampling pipeline 210 is communicated with the main pipeline 100, and the other end of the sampling pipeline 210 is communicated with the sample receiving member 400. The sample in the fermenter 10 can enter the sample receiving member 400 along the main pipeline 100 and the sampling pipeline 210, and the sample receiving member 400 is used to collect the sample.

[0054] The sampling pipeline 210 is sequentially provided with a first sampling control member 220 and a second sampling control member 230 along the liquid flow direction. Both the first sampling control member 220 and the second sampling control member 230 are set as pipeline clamps, and the first sampling control member 220 and the second sampling control member 230 are used to control the switching of the sampling pipeline 210 between the open state and the closed state.

[0055] In addition, it should be noted that there is a certain distance between the first sampling control member 220 and the second sampling control member 230, and this distance is the cut-off point of the sampling pipeline 210.

[0056] In an alternative embodiment, an interface member 240 is provided at one end of the sampling pipeline 210 remote from the main pipeline 100. The sampling pipeline 210 communicates with the sample receiving member 400 through the interface member 240. Specifically, the interface member 240 is provided as a rotary interface, and the sample receiving member 400 is specifically provided as a rotary sample receiving bag. The rotary interface is rotatably connected to the rotary sample receiving bag. After sampling, in a sterile environment, the rotary interface and the rotary sample receiving bag are rotated and twisted open, and then the sample is poured into a sterile EP tube for subsequent experimental operations.

[0057] In an alternative embodiment, the sample receiving member 400 is provided with an air filter 500, and the air filter 500 is used to exhaust the gas in the bag when the sample receiving member 400 receives the sample, so that the liquid can smoothly enter the bag.

[0058] In an alternative embodiment, a first pipeline control member 110 and a second pipeline control member 120 are sequentially arranged on the main pipeline 100 along the liquid flow direction. Both the first pipeline control member 110 and the second pipeline control member 120 are provided as pipeline clamps, and the first pipeline control member 110 and the second pipeline control member 120 are used to control the switching of the main pipeline 100 between the open state and the closed state.

[0059] The first pipeline control member 110 and the second pipeline control member 120 cooperate with each other to form a strict sterile control system. Before sampling, only when all the pipeline clamps to be opened are in the correct positions can the fermentation broth smoothly flow into the sampling container. After sampling, by closing the corresponding pipeline clamps, it is possible to effectively prevent foreign bacteria from entering the sampling pipeline and the fermentation tank 10.

[0060] In an alternative embodiment, the sampling device further includes a first connecting member 600 and a second connecting member 700; the first connecting member 600 and the second connecting member 700 are spaced apart on the main pipeline 100, and both the first connecting member 600 and the second connecting member 700 are used to communicate at least two sampling assemblies 200 with the main pipeline 100. Both the first connecting member 600 and the second connecting member 700 are provided as cross four-way air pipe joints, and a plurality of sampling assemblies 200 are connected to the main pipeline 100 by using the first connecting member 600 and the second connecting member 700.

[0061] In an alternative embodiment, a third pipeline control member 130 is provided between the first connection member 600 and the second connection member 700. The third pipeline control member 130 is provided as a pipeline clamp. The on-off between the first connection member 600 and the second connection member 700 is controlled by the third pipeline control member 130. When it is necessary for the sample to enter the second connection member 700, the third pipeline control member 130 is opened. When the sample only needs to enter the first connection member 600, the third pipeline control member 130 is closed.

[0062] It should be noted that both the sampling pipe and the main pipeline 100 are made of materials resistant to high temperature and corrosion.

[0063] Based on the above sampling device, the sampling method of the above sampling device includes the following steps: Installation step: Connect the main pipeline 100 to the fermentation tank 10, calculate the number of experimental sampling times, and connect multiple sampling components 200 to the main pipeline 100 according to the number of experimental sampling times; Cleaning step: Rinse the connection part between the main pipeline 100 and the sampling component 200; First sampling step: Open one of the sampling components 200, and the sample in the fermentation tank 10 enters the sample receiving member 400 through the opened sampling component 200; Re-sampling step: After the sample in the sampling component 200 sampled last time flows into the waste liquid receiving bottle 300, open another sampling component 200.

[0064] Before sampling operations are carried out, all pipeline clamps must be in a closed state.

[0065] When sampling is required, the first pipeline control member 110, the first sampling control member 220, and the second sampling control member 230 are opened in sequence. At this time, the bacterial liquid sample will flow along the pipeline into the waste liquid receiving bottle 300 under the action of the pressure difference. After sampling is completed, promptly close the first pipeline control member 110, the first sampling control member 220, and the second sampling control member 230, and cut off the sampling pipeline 210. The sampling pipeline 210 taken off at this time is the sample obtained aseptically. After transferring it to the ultra-clean workbench, in a sterile environment, twist open the knob-type interface and the knob-type sample receiving bag, and then pour the sample into a sterile EP tube for subsequent experimental operations.

[0066] When re-sampling is required, first open the first pipeline control member 110, the second pipeline control member 120, and the third pipeline control member 130, so that the liquid remaining in the pipeline during the previous sampling flows into the waste liquid receiving bottle 300 under the action of gravity. Then close the first pipeline control member 110, the second pipeline control member 120, and the third pipeline control member 130. Next, open the pipeline clamp of the first pipeline control member 110 and another sampling pipeline 210, and the bacterial liquid sample will flow into the knob-type sample receiving bag along the pipeline again. The subsequent experimental operations are the same as the steps after the first sampling.

[0067] Before each sampling, it is necessary to open the first pipeline control member 110, the second pipeline control member 120, and the third pipeline control member 130 first, so that the liquid remaining in the pipeline during the previous sampling flows into the waste liquid receiving bottle 300. Subsequently, the first pipeline control member 110, the second pipeline control member 120, and the third pipeline control member 130 are closed, and then the subsequent sampling steps are carried out.

[0068] The sampling device provided by the present invention meets the requirement of monitoring the fermentation broth multiple times during the fermentation process. It can perform multiple aseptic sampling operations at any time according to needs. The device is simple to operate and reusable. It can not only perform multiple aseptic samplings, but also perform non-aseptic samplings; it provides more accurate data support for optimizing the fermentation process. The structure is simple, the operation is convenient, and it is easy to install and maintain; the sampling device occupies less space, and there is no risk of breakage caused by easy collision due to too many bottles, and it is convenient for high pressure.

[0069] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A sampling device for a fermentation tank (10), characterized in that: include: A main pipeline (100), a sampling assembly (200), a waste liquid receiving bottle (300) and a sample receiving member (400); One end of the main pipeline (100) is in communication with the fermentation tank (10), and the other end of the main pipeline (100) is in communication with the waste liquid receiving bottle (300); A plurality of sampling assemblies (200) are provided, one end of each sampling assembly (200) is in communication with the main pipeline (100), and the other end of each sampling assembly (200) is connected to a sample receiving component (400); Each of the sampling components (200) has an open state and a closed state; During the first sampling, one of the sampling components (200) is in an open state, and the other sampling components (200) are in a closed state, so that the sample in the fermentation tank (10) enters the sample receiving component (400) through the sampling component (200) in the open state; During subsequent sampling, the sample remaining in the sampling assembly (200) opened during the last sampling flows into the waste liquid receiving bottle (300), and then another sampling assembly (200) is opened to perform the sampling operation.

2. The sampling device according to claim 1, characterized in that: The sampling assembly (200) comprises a sampling pipeline (210); One end of the sampling line (210) is connected to the main line (100), and the other end of the sampling line (210) is connected to the sample receiving component (400), so that the sample in the fermentation tank (10) can enter the sample receiving component (400) along the main line (100) and the sampling line (210).

3. The sampling device according to claim 2, characterized in that: The sampling pipeline (210) is provided with a first sampling control component (220) and a second sampling control component (230) in sequence along the liquid flow direction, and the first sampling control component (220) and the second sampling control component (230) are used to control the sampling pipeline (210) to switch between an open state and a closed state.

4. The sampling device according to claim 3, characterized in that: An interface component (240) is provided at one end of the sampling pipeline (210) away from the main pipeline (100), and the sampling pipeline (210) is connected to the sample receiving component (400) through the interface component (240).

5. The sampling device according to claim 4, characterized in that: The sample receiving component (400) is provided with an air filter (500), and the air filter (500) is used to remove the gas in the bag when the sample receiving component (400) receives the sample.

6. The sampling device according to claim 1, characterized in that: The main pipeline (100) is provided with a first pipeline control component (110) and a second pipeline control component (120) in sequence along the liquid flow direction, and the first pipeline control component (110) and the second pipeline control component (120) are used to control the main pipeline (100) to switch between an open state and a closed state.

7. The sampling device according to claim 6, characterized in that: The sampling device further comprises a first connecting member (600) and a second connecting member (700); The first connecting member (600) and the second connecting member (700) are arranged on the main pipeline (100) at intervals, and the first connecting member (600) and the second connecting member (700) are both used to connect at least two sampling components (200) with the main pipeline (100).

8. The sampling device according to claim 7, characterized in that: A third pipeline control component (130) is provided between the first connecting component (600) and the second connecting component (700).

9. A sampling method based on the sampling device according to any one of claims 1 to 8, characterized in that: The following steps are involved: Installation steps: connecting the main pipeline (100) to the fermentation tank (10), calculating the number of experimental samplings, and connecting a plurality of sampling components (200) to the main pipeline (100) according to the number of experimental samplings; Cleaning step: flushing the connection between the main pipeline (100) and the sampling assembly (200); A sampling step: opening one of the sampling components (200), and the sample in the fermentation tank (10) enters the sample receiving component (400) through the sampling component (200) in the opened state; Re-sampling step: After the sample in the sampling component (200) of the previous sampling flows into the waste liquid receiving bottle (300), another sampling component (200) is opened.

10. The sampling method according to claim 9, characterized in that: The sampling assembly (200) comprises a sampling pipeline (210); One end of the sampling pipeline (210) is in communication with the main pipeline (100), and the other end of the sampling pipeline (210) is in communication with the sample receiving component (400). The sampling pipeline (210) is provided with a first sampling control component (220) and a second sampling control component (230) in sequence along the liquid flow direction; The one-time sampling step comprises the following steps: The first sampling control component (220) and the second sampling control component (230) are opened, and the sample in the fermentation tank (10) passes through the main pipeline (100) and the sampling pipeline (210) and enters the waste liquid receiving bottle (300). After the sampling is completed, the first sampling control component (220) and the second sampling control component (230) are closed, and the sampling pipeline (210) between the first sampling control component (220) and the second sampling control component (230) is cut off.

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