CN212504873U - Sterile inoculation device - Google Patents
Sterile inoculation device Download PDFInfo
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- CN212504873U CN212504873U CN202020045404.3U CN202020045404U CN212504873U CN 212504873 U CN212504873 U CN 212504873U CN 202020045404 U CN202020045404 U CN 202020045404U CN 212504873 U CN212504873 U CN 212504873U
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Abstract
The invention discloses an aseptic inoculation device, which comprises an inoculation bottle and a piston, wherein the piston is arranged in the inoculation bottle; the inoculation bottle is provided with three openings, the first opening is a liquid outlet, the second opening is a liquid injection opening, the third opening is an exhaust opening, the liquid injection opening is sealed by a screw cap, and an air filter is arranged on the exhaust opening; the piston comprises a piston cylinder, a limiting pin and a piston column, and the piston column is arranged in the piston cylinder; one end of the piston cylinder is an open end and is a piston column insertion end, and the other end of the piston cylinder is a first quick connector which is a flange or a chuck with an opening in the middle; the length direction of the piston cylinder is provided with a liquid injection port close to the first quick connector, and the liquid outlet of the inoculation bottle and the liquid injection port can be combined together through a hose or the piston cylinder and the inoculation bottle are integrally processed and formed. The sterile inoculation device has the advantages of simple design, easy carrying, convenient operation and the like.
Description
Technical Field
The invention relates to an aseptic inoculation device, in particular to a disposable inoculation device capable of sterilizing on line, and belongs to the technical field of biology.
Background
The inoculation of strains into a sterile culture medium is a necessary link in the biological process, and the process requires a strict sterile environment, and if the process is careless, environmental microorganisms can pollute the whole fermentation process, thereby causing huge economic loss. Currently, two types of common modes are used for inoculating fermentation tanks: one is flame inoculation, i.e. the seeds are poured into a reactor under the protection of flame, and this operation requires cooperation of many people and risks of scalding and catching fire. The other type is a differential pressure inoculation method, namely, high-temperature steam is used for simultaneously sterilizing an inoculation pipeline on a reactor and a liquid outflow port of an inoculation bottle, and then the pressure of the inoculation bottle is enabled to be larger than that of the reactor by controlling the opening and closing of a plurality of valves, so that seed liquid can be transferred to the reactor under the pressure effect. The method is safer than flame inoculation.
However, the widely used differential pressure inoculation device, such as the authorized CN201820776124.2 utility model, has to be connected with a three-way diaphragm valve with high temperature sterilization on the inoculation bottle, and the inoculation bottle and the three-way diaphragm valve need to be carefully cleaned after the inoculation is completed and sterilized in the high pressure sterilization device before use, so that the inoculation device has the technical problems of inconvenient carrying, long operation time, difficult cleaning, etc.
Disclosure of Invention
The device combines the inoculation bottle and the piston to manufacture disposable consumables, and the consumables are filled into aseptic bags after sterilization, or the consumables are filled into packaging bags before sterilization. Therefore, compared with the patent CN201820776124.2, the utility model not only solves the technical problems of inconvenient carrying, long operation time, difficult cleaning and the like.
Technical scheme of the invention
An aseptic inoculation device comprises an inoculation bottle and a piston;
the inoculation bottle is provided with three openings, the first of the three openings is a liquid outlet, the second of the three openings is a liquid injection opening, the third of the three openings is an exhaust opening, the liquid injection opening is sealed by a screw cap, an air filter is arranged on the exhaust opening, and the liquid outlet extends to the bottom of the inoculation bottle at one end of the inner side of the inoculation bottle. The volume of the inoculation bottle is 5mL-5000 mL;
the piston comprises a piston cylinder, a limiting pin and a piston column, the piston column is arranged in the piston cylinder, two limiting holes are formed in the piston cylinder, and the position of the piston column in the piston cylinder is controlled by switching the limiting pin in the two limiting holes;
one end of the piston cylinder is an open end and is a piston column insertion end, the other end of the piston cylinder is a first quick connector, and the first quick connector is a flange or a chuck with an opening in the middle;
the piston column is provided with a handle close to the open end of the piston cylinder and used for pushing and pulling the piston column, a first plunger is fixed on the piston column close to the middle position, and a second plunger is fixed on the piston column close to one end of the first quick connector;
a liquid injection port is arranged at a position, close to the first quick connector, in the length direction of the piston cylinder, and the size of the liquid injection port is consistent with that of a liquid outlet of the inoculation bottle;
the piston cylinder and the inoculation bottle are combined in two modes, one mode is that the piston cylinder and the inoculation bottle are respectively processed and formed, then a liquid injection opening of the piston cylinder is connected with a liquid outlet of the inoculation bottle through a hose, the other mode is that the upper wall of the inoculation bottle is used as the lower wall of the piston cylinder, and the piston cylinder and the inoculation bottle are integrally processed and formed;
two limiting holes are arranged above the piston cylinder, a limiting pin is arranged in the second limiting hole before the inoculation bottle and the piston are combined and sterilized, and the liquid injection port of the piston cylinder is positioned between the two layers of plungers, so that the inoculation bottle is prevented from being polluted by the external environment after the combination is sterilized;
the sterile inoculation device is characterized by further comprising a connecting piece besides an inoculation bottle and a piston, wherein the connecting piece is a four-way joint, a first way of the four-way joint is connected with an inoculation port of a device to be inoculated, a second way of the four-way joint is used for connecting a steam inlet, a third way of the four-way joint is connected with a first quick-connection port of a piston cylinder, and a fourth way of the four-way joint is used for connecting a discharge valve;
the inoculation bottle and the piston are made of glass, silica gel, resin, rubber, a paperboard or plastic with a moisture-proof layer, and the connecting piece is made of metal, silica gel or plastic;
the sterile inoculation device, the piston and the inoculation bottle are combined for sterilization and then are filled into a sterile packaging bag, or the sterile packaging bag is filled into the sterile packaging bag for sterilization. When in use, the packaging bag is disassembled in the sterile room, then the seed liquid is transferred into the inoculation bottle through the liquid injection port, then the seed liquid is sealed by the screw cap and then is packaged in the packaging bag, the packaging bag is reopened beside the device to be inoculated, and after the piston is connected with the connecting piece connected to the inoculation port, the inoculation pipeline is sterilized by steam;
the sterile inoculation device is used for inoculation in the microbial fermentation process, inoculation in the cell culture process, liquid supplementation in a sterile container and the like.
The use method of the sterile inoculation device is specifically shown in fig. 5:
firstly, a first through in a connecting piece, namely a four-way pipe, is connected with an inoculation port of a device to be inoculated through a clamping sleeve, a clamping hoop, a flange or welding;
then the connecting piece, namely a second cross in the cross is connected with the steam pipeline in a clamping sleeve, a clamping hoop, a flange or welding mode and the like; then the connecting piece, namely the fourth of the four-way is connected with a discharge valve in the ways of clamping sleeve, clamping hoop, flange or welding;
then, the connecting piece, namely a third tee in the four-way joint, is connected with the first quick joint of the piston cylinder through a hoop or a flange, then the discharge valve 7 at the lower end of the connecting piece is opened, and then the steam valve 6 on the inoculation pipe of the device to be inoculated is opened. At the moment, steam enters the connecting piece from top to bottom along the pipeline and diffuses to the outer side of the first quick connector of the piston cylinder of the sterile inoculation device, and the steam sterilizes the outer side of the first quick connector at high temperature. After 20 minutes, the steam valve 6 is closed, the inoculation valve 5 of the device to be inoculated is opened, at this time, the gas of the device to be inoculated flows out of the discharge valve 7 along the pipeline, and the gas can convert the steam in the pipeline on one hand and cool the pipeline on the other hand. When the temperature of the pipeline is reduced to normal temperature, the discharge valve 7 is closed. And then the piston limiting pin is pulled outwards, the piston limiting pin is moved to the first limiting hole from the second limiting hole, and then the piston column is pulled outwards until the piston column is clamped by the limiting pin of the first limiting hole. The bottle is then connected to the device to be inoculated and the liquid in the bottle can then be transferred to the device to be inoculated by means of a peristaltic pump or by applying compressed air to an air filter. When the inoculation volume meets the requirement, the piston column is pushed to the initial position in the piston cylinder, and the piston limiting pin is inserted into the second limiting hole. At this point, the inoculation valve 5 of the device to be inoculated is closed again and the discharge valve 7 is opened. The steam valve 6 is opened again, then the residual seed liquid in the pipeline can be removed by the steam, and then the steam valve 6 is closed again. And finally, opening the piston cylinder and a clamp or a flange of the connecting piece, and taking down the sterile inoculation device.
Drawings
FIG. 1, schematic diagram of the general structure of the sterile inoculating device of example 1;
FIG. 2, cross-sectional view of an inoculum bottle;
FIG. 3, a cross-sectional view of the piston;
FIG. 4, a cross-sectional view of the connector;
FIG. 5 is a schematic view showing the structure of the connection of the sterile inoculating device and the bioreactor;
FIG. 6 is a schematic view showing the overall structure of the sterile inoculating device of example 2
Detailed description of the preferred embodiments
The invention is further illustrated, but not limited, by the following specific examples in connection with the accompanying drawings.
Example 1
A sterile inoculating device, as shown in figure 1, figure 2, figure 3 and figure 4, comprising an inoculating bottle 1, a piston 2 and a connecting piece 3;
in FIG. 2, the said inoculation bottle 1 is provided with three openings, the first is a liquid outlet 111, the liquid outlet is in the shape of a conduit, one end of the inner side of the inoculation bottle extends to the bottom of the inoculation bottle, the other end is sleeved with a flexible pipe 12 and is connected with the liquid injection port of the piston cylinder through the flexible pipe, the second is a liquid injection port 112 which is sealed by a screw cap 13, the third opening is an exhaust port 113 on which an air filter 14 is arranged, and the volume of the inoculation bottle is 50 mL;
in fig. 3, the piston 2 includes a piston cylinder 21, a limit pin 22 and a piston column 23, the piston column 23 is installed in the piston cylinder 21, the limit pin 22 is installed on the piston cylinder 21, the position of the piston column 23 in the piston cylinder 21 is controlled by the limit pin 22, when in use, the limit pin 22 is pulled out from a second limit hole 213 and moved into a first limit hole 212, the piston column 23 is pulled until being clamped, the piston column 23 is provided with a handle 231 near an open end of the piston cylinder for pulling and pushing the piston column, a first plunger 232 is fixed on the piston column near a middle position, and a second plunger 233 is fixed on the piston column near a first quick-connection end;
one end 211 of the piston cylinder 21 is an open end and is a piston column insertion end, the other end 212 is a first quick connector 216, the first quick connector is a sanitary chuck with a middle opening, and the middle opening of the first quick connector is a piston outlet 214;
a liquid injection port 215 is arranged at the position, close to the sanitary chuck, of the piston cylinder 21 in the length direction, the opening of the liquid injection port is circular, the outer diameter of the liquid injection port is 8mm, the inner diameter of the liquid injection port is 4mm, and the opening 215 is the same as the opening of the liquid outlet 111 of the inoculation bottle 1; the connection is made by means of a hose 12.
The connecting piece 3 is a four-way joint, a first passage 31 in the four-way joint is connected with an inoculation port of a device to be inoculated, a second passage 32 in the four-way joint is connected with a steam pipeline, a third passage 33 in the four-way joint is a second quick connector, the second quick connector is connected with a first quick connector 216 of the piston cylinder 23 in a sealing mode through a hoop when the connecting piece is used, and a fourth passage 34 in the four-way joint is connected with a discharge valve 7;
the piston cylinder 21 of the piston and the inoculation bottle 1 are respectively processed and molded; the inoculation bottle 1 and the piston 2 are made of polypropylene.
The method for using the sterile inoculation device to perform inoculation in the microbial fermentation process (refer to fig. 5) specifically comprises the following steps:
firstly, taking out an inoculation bottle and a piston combination in the aseptic inoculation device in an aseptic chamber, opening a liquid injection port on the inoculation bottle, introducing seed liquid into the inoculation bottle through a suction pipe, and sealing the liquid injection port by using a screw cap;
then connecting a connecting piece in the sterile inoculation device, namely a first through in the four-way connector, with an inoculation port of a fermentation tank to be inoculated through a hoop; the second of the four-way is connected with a steam pipeline of the fermentation tank through a hoop, and the fourth of the four-way is connected with a discharge valve through the hoop;
then, the connecting piece, namely a third tee in the four-way joint, is connected with the first quick joint of the piston cylinder through a clamping hoop, and then the discharge valve 7 at the lower end of the connecting piece is opened first, and then the steam valve 6 is opened. At the moment, steam enters the connecting piece from top to bottom along the pipeline and diffuses to the outer side of the first quick connector of the piston cylinder of the sterile inoculation device, and the steam sterilizes the outer side of the first quick connector at high temperature. After 20 minutes, the steam valve 6 is closed and the inoculation valve 5 of the fermenter is opened, whereupon the gas in the fermenter which is being held under pressure flows out of the discharge valve 7 along the line. When the temperature of the pipeline is reduced to normal temperature, the discharge valve 7 is closed. And then the piston limiting pin is pulled outwards, the piston limiting pin is moved to the first limiting hole from the second limiting hole, and then the piston column is pulled outwards until the piston column is clamped by the limiting pin of the first limiting hole. At this time, the inoculation bottle is communicated with the device to be inoculated, and the liquid in the inoculation bottle is transferred into the fermentation tank through the peristaltic pump. When the inoculation volume meets the requirement, the piston column is pushed to the initial position in the piston cylinder, and the piston limiting pin is inserted into the second limiting hole. At this point, the inoculation valve 5 of the fermenter is closed again and the discharge valve 7 is opened. The steam valve 6 is opened again, at this time, the liquid in the pipeline can be removed by the steam, and then the steam valve 6 is closed. And finally, opening the piston cylinder and a clamp or a flange of the connecting piece, and taking down the sterile inoculation device to finish inoculation.
Example 2
An aseptic inoculation device, the upper wall of an inoculation bottle is used as the lower wall of a piston cylinder, the piston cylinder and the inoculation bottle are integrally processed and molded, the rest is the same as the embodiment 1, and the specific structure is shown in figure 6.
In conclusion, compared with the existing device, the inoculation device capable of sterilizing online provided by the invention reduces 1 three-way diaphragm valve, has the advantages of convenience in carrying, simple structure, easiness in operation and the like, and is very suitable for online sterile inoculation operation in microbial fermentation and cell culture processes.
The foregoing is merely an example of the embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Claims (6)
1. An aseptic inoculation device comprises an inoculation bottle and is characterized by also comprising a piston;
the inoculation bottle is provided with three openings, wherein the first opening is a liquid outlet, the second opening is a liquid injection opening, the third opening is an exhaust opening, the liquid injection opening is sealed by a screw cap, and an air filter is arranged on the exhaust opening;
the piston comprises a piston cylinder, a limiting pin and a piston column, the piston column is arranged in the piston cylinder, two limiting holes are formed in the piston cylinder, and the position of the piston column in the piston cylinder is controlled by switching the limiting pin in the two limiting holes;
one end of the piston cylinder is an open end and is a piston column insertion end, and the other end of the piston cylinder is a first quick connector which is a flange or a chuck with an opening in the middle;
the piston column is provided with a handle close to the open end of the piston cylinder and used for pushing and pulling the piston column, a first plunger is fixed on the piston column close to the middle position, and a second plunger is fixed on one end of the piston column close to the first quick connector;
and a liquid injection opening is formed in the piston cylinder in the length direction and is close to the first quick connector, and the size of the liquid injection opening is consistent with that of a liquid outlet of the inoculation bottle.
2. An aseptic inoculating device as claimed in claim 1, wherein the inoculating bottle is provided with a liquid outlet which extends to the bottom of the bottle at one end of the inside of the bottle.
3. An aseptic inoculating device as claimed in claim 1, wherein the piston cylinder is formed separately from the inoculating bottle, and the liquid inlet of the piston cylinder is connected to the liquid outlet of the inoculating bottle by a flexible tube.
4. An aseptic inoculating device as claimed in claim 1, in which the upper wall of the inoculating bottle is provided as the lower wall of a piston cylinder, the piston cylinder being formed integrally with the inoculating bottle.
5. A sterile inoculating device as claimed in claim 1 wherein the volume of the inoculating bottle is 5-5000 mL.
6. An aseptic inoculating device as claimed in claim 1, wherein the inoculating bottle and piston are made of glass, silica gel, resin, rubber, cardboard or plastic with a moisture-proof layer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020045404.3U CN212504873U (en) | 2020-01-09 | 2020-01-09 | Sterile inoculation device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020045404.3U CN212504873U (en) | 2020-01-09 | 2020-01-09 | Sterile inoculation device |
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| Publication Number | Publication Date |
|---|---|
| CN212504873U true CN212504873U (en) | 2021-02-09 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202020045404.3U Active CN212504873U (en) | 2020-01-09 | 2020-01-09 | Sterile inoculation device |
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| Country | Link |
|---|---|
| CN (1) | CN212504873U (en) |
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2020
- 2020-01-09 CN CN202020045404.3U patent/CN212504873U/en active Active
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