Aseptic microsampling system

Abstract

The utility model relates to aseptic sampling technical field, concretely relates to a kind of aseptic trace sampling system, including aseptic sampling probe, aseptic sampling probe intercommunication fermentation tank, another end of aseptic sampling probe connects sampling pipeline, another end of sampling pipeline connects liquid-collecting container;Sampling pipeline includes first sampling pipeline and second sampling pipeline;First sampling pipeline and second sampling pipeline are communicated by valve group between;First valve is set on second sampling pipeline near the position of liquid-collecting container, second sampling pipeline is connected backflow branch, and the communication point of backflow branch and second sampling pipeline is located the front end of first valve, backflow branch intercommunication sterilization air source, liquid-collecting container is made of transparent material, and the diameter of lower half of liquid-collecting container is less than the diameter of upper half, and the outer surface of lower half has scale;The upper end of upper half is closed and is provided with breathing filter.The utility model can avoid the waste of sample, greatly reduce sampling cost.

Description

Technical Field

[0001] This utility model relates to the field of aseptic sampling technology, specifically to an aseptic micro-sampling system. Background Technology

[0002] Bio-fermentation is a bioengineering technology that utilizes microorganisms to obtain industrial products. The fermentation process using microorganisms is time-varying, nonlinear, and uncertain, involving the growth, reproduction, and metabolism of living organisms, making its mechanisms highly complex. Therefore, real-time monitoring of the bioreaction process within the fermenter is necessary to understand the metabolic state of the organisms, providing a reference for optimizing the bioreaction process, and simultaneously avoiding the adverse effects of sudden changes in biochemical indicators on the fermentation process.

[0003] Currently, there are two main methods for monitoring biological reaction processes: one is to use a bioreactor. A bioreactor can provide controllable environmental conditions for biological reactions in a biological system. Bioreactors can achieve online detection of conditions such as temperature, pH, and dissolved oxygen. However, its monitoring of biological reaction conditions is usually used when online detection of biochemical parameters cannot be obtained in real time.

[0004] Another method is detection and control through sampling. Currently, domestic and international sampling devices fall into two main categories: one is primarily pipe and valve devices, which sample the original fermentation broth containing raw materials and microbial cells; the other is membrane filtration sampling. However, with this method, the membrane pore size prevents the passage of microorganisms, leading to easy clogging of the filter. Furthermore, this method is unsuitable for situations requiring analysis of microorganisms within the original fermentation broth sample.

[0005] The first type of sampling method involves connecting the sampling valve and the collection container through a valve assembly. However, traditional sampling methods result in the waste of residual fermentation broth in the sampling pipeline at the rear end of the valve assembly, and the pipeline disassembly process is prone to leakage, causing environmental pollution. Furthermore, the front end of the valve assembly cannot effectively transfer mass and heat with the fermentation broth inside the tank, thus affecting the fermentation quality.

[0006] Chinese patent CN105890923A discloses a sterile micro-volume online sampling device and method. The sampling device consists of a sterile micro-volume sampling probe, an adjustable-speed sampling pump system, a pipeline cleaning and sterilization system, a quantitative sampling loop, and a sterile micro-volume sampling probe anti-clogging backflush system. Following the sampling procedure, it enables micro-volume online sampling in fermenters. The device features sterile sampling, reduced dead volume, extremely small sample volume, minimal impact on the fermentation system, continuous sampling thanks to the probe anti-clogging backflush system, and prevention of contamination and liquid residue by the pipeline cleaning and sterilization system. It is suitable for micro-volume online sampling in bioreactor online monitoring systems for microbial fermentation, cell culture, and other bioreactors.

[0007] However, the aseptic sampling operation of this patent is cumbersome, relies on a complex main control system, and is costly. At the same time, after sampling, the existing technology leaves some sample liquid remaining in the aseptic micro-sampling probe and sampling tubing, resulting in a large dead volume and sample waste; moreover, the sampling tubing is directly connected to the analytical instrument, and the tubing connected to the analytical instrument cannot be sterilized, thus posing a risk of sample contamination.

[0008] Based on this, the present invention provides a sterile micro-sampling system. Utility Model Content

[0009] The purpose of this invention is to provide a sterile micro-sampling system to solve the technical problems in the prior art, such as the waste of residual fermentation liquid in the sampling pipeline of the sampling device, the environmental pollution caused by leakage during pipeline disassembly, and the complexity of operation / equipment.

[0010] A sterile micro-sampling system is provided, wherein a valve group is connected to one end of a sterile sampling probe, and a collection container is connected to the other end of the valve group. A first valve is installed on the pipeline near the collection container. A reflux branch is connected in parallel to the front end of the first valve. The reflux branch is connected to a sterile air source. After sampling, sterile air is used to push the sample in the pipeline back into the fermenter.

[0011] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0012] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of this utility model in any way. In this utility model, the singular is used in conjunction with the plural unless otherwise specified. It should also be noted that, unless otherwise specified, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0013] Front end, back end: Based on the sample flow path during sampling, the valve inlet is the front end, and the valve outlet is the back end.

[0014] To achieve the above objectives, the technical solution of this utility model is as follows: A sterile micro-sampling system includes a sterile sampling probe connected to a fermenter, the other end of the sterile sampling probe connected to a sampling pipeline, and the other end of the sampling pipeline connected to a collection container; the sampling pipeline includes a first sampling pipeline and a second sampling pipeline, the first sampling pipeline and the second sampling pipeline are connected in series, and the other end of the sterile sampling probe is connected to the first sampling pipeline. The first sampling pipeline and the second sampling pipeline are connected by a valve assembly; A first valve is installed on the second sampling pipeline near the collection container. The second sampling pipeline is connected to a reflux branch in parallel, and the connection point between the reflux branch and the second sampling pipeline is located at the front end of the first valve. The reflux branch is connected to a sterilization air source. The liquid collection container is made of transparent material and includes a lower part and an upper part. The diameter of the lower part is smaller than that of the upper part, and the outer surface of the lower part has graduations. The upper part is closed and equipped with a breathing filter.

[0015] Furthermore, a second valve is installed on the return branch.

[0016] Furthermore, the second sampling pipeline and the return branch are connected by a tee connector.

[0017] Furthermore, the breathing filter is also connected to a vacuum pump.

[0018] Furthermore, the smallest scale unit is 0.1ml, 0.2ml, 0.5ml, or 1ml.

[0019] Furthermore, the valve group includes a third valve, a fourth valve, a fifth valve, and a sixth valve. One end of the third valve is connected to the first sampling pipeline, and the other end of the third valve is connected in parallel to the first bypass and the connecting pipeline. The other end of the connecting pipeline is connected to the fifth valve, and the other end of the fifth valve is connected in parallel to the second bypass and the second sampling pipeline. The first bypass is connected to a steam source, and a fourth valve is installed on the first bypass; The second bypass is connected to the sewage pipe, and a sixth valve is installed on the second bypass.

[0020] Furthermore, the third, fourth, fifth, and sixth valves are all diaphragm valves.

[0021] Furthermore, the connecting pipe and the fifth valve are connected by clamp connectors, and the second bypass and the sewage pipe are connected by clamp connectors.

[0022] Furthermore, both the first and second sampling lines are made of silicone tubing or stainless steel tubing.

[0023] Furthermore, the first valve is a needle valve.

[0024] Compared with the prior art, the present invention has the following beneficial effects: The sterile micro-sampling system provided by this utility model has a valve installed near the liquid inlet of the collection container in the sampling pipeline, and a sterile air source is connected to the front end of the valve. After sampling, the sterile air source is used to push the sample in the pipeline back into the fermenter, thereby avoiding sample waste and reducing sampling costs. Moreover, the purpose of micro-sampling can be achieved by using a valve assembly and pipeline of conventional size combined with a collection container of a specific shape. At the same time, since the liquid in the pipeline is pushed back into the fermenter, environmental pollution caused by liquid residue is avoided during pipeline disassembly.

[0025] In addition, this utility model is based on the clamp connector to connect various pipelines and valves. The clamp connector can be used to remove the liquid collection container and return branch at the rear end for disinfection. The disinfected liquid collection container and return branch can then be connected to the connecting pipeline and sewage pipe using the clamp connector. After disinfection of the pipeline at the connection point, repeated sampling of the fermenter can be achieved under aseptic conditions. Attached Figure Description

[0026] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0027] Figure 1 This is a connection diagram of the present invention.

[0028] Explanation of reference numerals in the attached figures: 1. Fermenter; 2. Aseptic sampling probe; 3. Collection container; 4. First sampling line; 5. Second sampling line; 6. First valve; 7. Second valve; 8. Third valve; 9. Fourth valve; 10. Fifth valve; 11. Sixth valve; 12. Return branch; 13. First bypass; 14. Second bypass; 15. Breathing filter; 16. Clamp connector; 17. Connecting line; 18. Vacuum pump. Detailed Implementation

[0029] The technical solution of this utility model will be clearly described below with reference to the accompanying drawings. Obviously, the described embodiments are not all embodiments of this utility model. All other embodiments obtained by those skilled in the art without creative effort are within the protection scope of this utility model.

[0030] It should be noted that, unless otherwise specifically stated, the relative arrangement and numerical expressions of the components and steps described in these embodiments should not be construed as limiting the scope of this utility model.

[0031] The following description of exemplary embodiments is merely illustrative and is not intended to limit the present invention or its application or use in any way. Techniques, methods, and apparatus known to those skilled in the art may not be discussed in detail herein, but where applicable, such techniques, methods, and apparatus should be considered part of this specification.

[0032] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to the present invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0033] For ease of description, the words "up," "down," "left," and "right" appearing in this utility model only indicate that they are consistent with the up, down, left, and right directions of the accompanying drawings. They do not limit the structure and are merely for the purpose of facilitating the description of this utility model and simplifying the description. They do not indicate or imply that the device or component referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0034] Terminology Explanation: The terms "installation," "connection," "linking," and "fixing" in this utility model should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal connection of two components or the interaction relationship between two components. For those skilled in the art, the specific meaning of the above terms in this utility model can be understood according to the specific circumstances.

[0035] This embodiment provides a sterile micro-sampling system, such as Figure 1 As shown, it includes a sterile sampling probe 2, which is connected to the fermenter 1. The other end of the sterile sampling probe 2 is connected to a sampling pipeline, and the other end of the sampling pipeline is connected to a collection container 3. The sampling pipeline includes a first sampling pipeline 4 and a second sampling pipeline 5, which are connected in series. The other end of the sterile sampling probe is connected to the first sampling pipeline 4.

[0036] The first sampling pipeline 4 and the second sampling pipeline 5 are connected by a valve group; a first valve 6 is provided on the second sampling pipeline 5 near the liquid collection container 3; the second sampling pipeline 5 is connected to a reflux branch 12 in parallel, and the connection point between the reflux branch 12 and the second sampling pipeline 5 is located at the front end of the first valve 6; the reflux branch 12 is connected to a sterilization air source.

[0037] The collection container 3 is made of transparent material and is cylindrical in shape, comprising a lower half and an upper half. The diameter of the lower half is smaller than that of the upper half. Specifically, the lower half is a slender cylindrical structure with a volume ranging from 2ml to 5ml, thus enabling the collection of trace samples. The outer surface of the lower half has graduations for visual observation of the sample volume; the smallest graduation unit is 0.1ml, 0.2ml, 0.5ml, or 1ml. The upper end of the upper half is sealed and equipped with a breather filter 15.

[0038] The fermenter 1 is equipped with a sampling valve, and the inside of the fermenter 1 is under high pressure. The sterile sampling probe 2 is connected to the fermenter 1 through the sampling valve. When the sampling valve is opened, the high pressure inside the fermenter 1 is used to sample the liquid through the sterile sampling probe 2.

[0039] Both the first sampling line 4 and the second sampling line 5 are made of silicone tubing or stainless steel tubing. The valve group installed between the first sampling line 4 and the second sampling line 5 includes a third valve 8, a fourth valve 9, a fifth valve 10, and a sixth valve 11. One end of the third valve 8 is connected to the first sampling line 4, and the other end of the third valve 8 is connected in parallel to a first bypass 13 and a connecting line 17. The other end of the connecting line 17 is connected to the fifth valve 10, and the other end of the fifth valve 10 is connected in parallel to a second bypass 14 and a second sampling line 5. The first bypass 13 is connected to a steam source, which can be a steam generator, and a fourth valve 9 is installed on the first bypass 13; the second bypass 14 is connected to a sewage pipe, and a sixth valve 11 is installed on the second bypass 14.

[0040] Moreover, preferably, the third valve 8, the fourth valve 9, the fifth valve 10, and the sixth valve 11 are all diaphragm valves. The first valve 6 is a needle valve, which is a needle-shaped valve with a fine-tipped stem, capable of precisely controlling small flow rates, and thus enabling precise sampling of trace samples.

[0041] In addition, the connecting pipe 17 and the fifth valve 10 are connected by a clamp connector 16, and the second bypass 14 and the sewage pipe are connected by a clamp connector 16.

[0042] The second sampling line 5 and the return branch line 12 are connected by a tee connector. One port of the tee connector is connected to the second sampling line 5, the second port is connected to the return branch line 12, and the third port is connected to the first valve 6. A second valve 7 is provided on the return branch line 12 to ensure the flow of sterilization air.

[0043] In addition, a breathing filter 15 is provided at the upper end of the collection container 3, which allows the sample from the fermenter 1 to flow into the collection container 3 without the need for a pump. To directly determine the volume of the collected sample, graduations are provided on the outer wall of the collection container 3.

[0044] In a preferred embodiment, the breathing filter 15 is also connected to a vacuum pump 18, specifically as follows: Figure 1 As shown, the upper end of the liquid collection container 3 is connected to the breathing filter 15, and the breathing filter 15 is connected to the vacuum pump 18. The vacuum pump can be used to extract the air in the pipeline during sampling, thereby providing power for the flow of liquid and avoiding sampling difficulties caused by insufficient power. In addition, the vacuum pump can also be used to adjust the speed of liquid flow.

[0045] The operation method and working principle of this utility model are as follows: S1. Before sampling, ensure that the valve group is correctly installed and connected to the sterile sampling probe 2 and the fermenter 1, and that all valves are closed. Sterilize the fermenter and the sterile sampling probe 2, the first sampling pipeline 4, the third valve 8, the fourth valve 9, the first bypass 13 and the connecting pipeline 17 connected to the fermenter. S2. Connect the liquid collection container 3 with the breathing filter 15 and the reflux branch 12 (with the second valve 7 and in the closed state) to the fifth valve 10. At this time, the fifth valve 10 is in the closed state. Then, remove the fifth valve 10, the second bypass 14 (with the sixth valve 11), the reflux branch 12 and the liquid collection container 3, and put them into the sterilizer for sterilization. S3. Connect the sterilized components in S2. Specifically, connect the fifth valve 10 to the connecting pipe 17 through the clamp connector 16, and connect the sixth valve 11 to the sewage pipe through the clamp connector. S4. Open the fourth valve 9 and the sixth valve 11. High-temperature steam is introduced through the fourth valve 9 to sterilize the first bypass 13, the connecting pipe 17 and the second bypass 14. The sterilized steam and sewage are discharged through the sewage pipe. In this step, the third valve 8 can also be opened to sterilize the inside of the first sampling pipeline 4; S5. After sterilization, close the fourth valve 9 and the sixth valve 11, and open the third valve 8, the fifth valve 10, and the first valve 6. S6. Open the sampling valve on the fermenter 1. Under the high pressure drive in the fermenter and the action of the breathing filter 15, the sample flows directly into the collection container 3. As a preferred embodiment, if you want to speed up the sampling speed, you can start the vacuum pump 18 connected to the breathing filter 15 and use the vacuum pump 18 to provide the power for the sample flow.

[0046] S7. When the sample volume in the collection container 3 reaches the required volume, close the first valve 6. S8. Open the second valve 7 and introduce sterile air into the return branch 12. Use the sterile air to push the residual sample in the pipeline back to the fermenter 1. The time for introducing sterile air can be set. After the set time is reached, close the second valve 7, the third valve 8, the fifth valve 10 and the sampling valve to complete the sampling. The sterile air source has a certain high pressure, such as 3MPa, which is sufficient to push the residual sample back to the fermentation tank. S9. After sampling is completed, the fifth valve 10, the second sampling pipeline 5 and the downstream components can be removed by using the clamp connector 16 on the connecting pipeline 17 and the clamp connector between the sixth valve 11 and the fifth valve 10, and then cleaned and disinfected for future use.

[0047] If repeated sampling is required, two sets of fifth valve 10, second sampling pipeline 5, and rear-end components can be prepared. During the first sampling, one set of fifth valve 10, second sampling pipeline 5, and rear-end components is connected to connecting pipeline 17 and sixth valve 11 via clamp connector 16, and the sampling operation is performed according to the above steps. After the first sampling is completed, the fifth valve 10, second sampling pipeline 5, and rear-end components are removed. The other set of fifth valve 10, second sampling pipeline 5, and rear-end components are disinfected (or disinfected in advance) and then connected to connecting pipeline 17 and sixth valve 11. The second sampling is then completed in the order of steps S4 to S8. The two sets of components are used alternately to complete the repeated sampling operation.

[0048] Alternatively, prepare only one set of the fifth valve 10, the second sampling pipeline 5, and the downstream components. After step S9, if repeated sampling is required, reconnect the disinfected fifth valve 10, the second sampling pipeline 5, and the downstream components to the sampling system, and complete the repeated sampling in the order of steps S4 to S9.

[0049] The above specific embodiments are only used to illustrate the technical solution of this utility model and are not intended to limit it. Although this utility model has been described in detail with reference to examples, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solution of this utility model without departing from the scope of the technical solution of this utility model, and all such modifications or substitutions should be covered within the scope of the claims of this utility model.

Claims

1. A sterile micro-sampling system, comprising a sterile sampling probe, wherein the sterile sampling probe is connected to a fermenter, characterized in that, The other end of the sterile sampling probe is connected to a sampling tube, and the other end of the sampling tube is connected to a collection container; the sampling tube includes a first sampling tube and a second sampling tube, the first sampling tube and the second sampling tube are connected in series, and the other end of the sterile sampling probe is connected to the first sampling tube. The first sampling pipeline and the second sampling pipeline are connected by a valve assembly; A first valve is installed on the second sampling pipeline near the collection container. The second sampling pipeline is connected to a reflux branch in parallel, and the connection point between the reflux branch and the second sampling pipeline is located at the front end of the first valve. The reflux branch is connected to a sterilization air source. The liquid collection container is made of transparent material and includes a lower half and an upper half. The diameter of the lower half is smaller than that of the upper half, and the outer surface of the lower half has graduations. The upper part of the upper half is closed and equipped with a breath filter. The breathing filter is also connected to a vacuum pump, which is used to extract air from the collection container and the sampling pipeline connected to it during sampling.

2. The aseptic micro-sampling system according to claim 1, characterized in that, A second valve is installed on the return branch.

3. The aseptic micro-sampling system according to claim 1, characterized in that, The second sampling pipeline and the return branch are connected by a tee connector.

4. The aseptic micro-sampling system according to claim 1, characterized in that, The smallest unit of the scale is 0.1 ml, 0.2 ml, 0.5 ml, or 1 ml.

5. The aseptic micro-sampling system according to claim 1, characterized in that, The valve group includes a third valve, a fourth valve, a fifth valve, and a sixth valve. One end of the third valve is connected to the first sampling pipeline, and the other end of the third valve is connected in parallel to the first bypass and the connecting pipeline. The other end of the connecting pipeline is connected to the fifth valve, and the other end of the fifth valve is connected in parallel to the second bypass and the second sampling pipeline. The first bypass is connected to a steam source, and a fourth valve is installed on the first bypass; The second bypass is connected to the sewage pipe, and a sixth valve is installed on the second bypass.

6. The aseptic micro-sampling system according to claim 5, characterized in that, The third, fourth, fifth, and sixth valves are all diaphragm valves.

7. The aseptic micro-sampling system according to claim 5, characterized in that, The connecting pipe and the fifth valve are connected by clamp connectors, and the second bypass and the sewage pipe are connected by clamp connectors.

8. The aseptic micro-sampling system according to claim 1, characterized in that, Both the first and second sampling lines are made of silicone tubing or stainless steel tubing.

9. The aseptic micro-sampling system according to claim 1, characterized in that, The first valve is a needle valve.

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