JPH07151651A - Sampler and sampling method - Google Patents

Abstract

PURPOSE:To realize sampling in a closed system under perfectly sterilized state by simplifying replacement of sample tube and to automate sampling and replacement of sample tube without requiring any large scale facility such as a clean room. CONSTITUTION:The sampler comprises a drain pipe 4 having a port 48 for introducing a sterilizing material, a port 43 for draining unnecessary sterilizing material, and a port for introducing a sample, a sterilizing tube 8 having a port 7 for introducing the sterilizing material and one open end, an enclosed sampling pipe 10 previously subjected to vacuum sterilization and mounted detachably onto the sterilizing tube, and a hollow needle 5 previously mounted on the drain pipe in order to couple the drain pipe and the sterilizing pipe with the sampling pipe thus transferring the sample from the drain pipe into the sampling pipe.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sampling device and a sampling method, and in particular, the quality of products in which contamination by live bacteria (hereinafter simply referred to as bacteria) is a problem in the food industry, chemical industry, pharmaceutical industry, etc. BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a device and method for sampling in a sterile state which is useful for management.

[0002]

2. Description of the Related Art Products such as beer which undergo a fermentation process in the food industry, chemical industry, pharmaceutical industry, etc., are liable to be mixed with bacteria such as aerobic lactic acid bacteria in the fermentation tank or the pipes around the fermentation tank during fermentation. In some cases, these live germs grow in the fermenter or the pipes with time, and the quality of the product is deteriorated due to the germs and products produced by the germs, and the product cannot be finally shipped as a product.

Therefore, in order to control the quality of the product, it is necessary to periodically analyze the alcohol concentration and the amount of lactic acid bacteria in the fermenter and the pipes around the fermentor. However, if miscellaneous bacteria such as lactic acid bacteria are mixed into the collected sample when collecting the sample, it may be that the bacteria were originally mixed in the fermenter or pipes, or the miscellaneous bacteria mixed only in the collected sample from the outside at the time of sampling. It became impossible to distinguish whether or not there was, which caused inconvenience in quality control of the product.

For these reasons, it is extremely important to prevent contamination of bacteria in the fermentation tank and the pipes around it, especially when collecting samples in quality control of products that undergo fermentation processes.

Conventionally, for sampling from a fermenter, a sample conduit is provided in the fermentor, the sample conduit and the drain pipe are connected by a sample valve, steam is introduced into the drain pipe, and the drain pipe itself is 121 ° C. After sterilizing by letting steam circulate for about 30 minutes in the above-retained state, it was left to cool, and then sterilized air was passed through the drain pipe after being left to cool, drain water was discharged and dried to sterilize the drain pipe at high temperature. After that, it has been carried out by a method of transferring a sample solution from a fermenter through a drain tube to a sample tube connected to the drain tube.

However, high temperature sterilization with steam is not suitable for some fermented products. For example, in beer production, it is necessary to control the fermenter at a temperature of 0 to 10 ° C., and since the sample valve connected to the drain pipe, the sample conduit, and the fermenter are also heated by high-temperature heat sterilization with steam, Temperature control of the fermenter becomes difficult. for that reason,
Sterilization with ethanol instead of steam has been proposed for products in which it is not desirable to heat the fermenter at high temperatures.

FIG. 5 shows a typical one of the conventional sampling methods.
It is a flowchart which shows an example. In the figure, 501 is a fermenter, 502 is a sample conduit, 503 is a sample valve, 504 is a drain pipe, 505 is a sample pipe, 506 is a sample liquid sampled, 5
Reference numeral 07 is a steam introducing pipe, 508 is an ethanol or ethanol aqueous solution introducing pipe, and 509 is a sterilizing air introducing pipe. However, in this method, it is necessary for a human to replace the sample tube, so the replacement process is complicated, and
In order to avoid this complication, there is a problem that a large-scale facility such as a clean room is required in order to automatically sterilize the sample tubes and perform complete sterilization.

On the other hand, there has been proposed a method in which a sample valve provided in a fermenter is sterilized by flame to prevent contamination of various bacteria, and then a sample is directly collected from a sample valve in an open system into a previously sterilized sample tube. . In this method, the complicated sample tube exchange found in a closed system is unnecessary, but since the sample collection is performed in an open system, it is necessary to collect the sample while preventing the sample from scattering and mixing of various bacteria. , Required highly skilled humans. Further, even when a sample is collected by a highly skilled person, contamination of the product, which is considered to be caused by contamination with other bacteria during the sample collection, that is, deterioration of product quality, occurs several times a year.

[0009]

SUMMARY OF THE INVENTION In view of the above-mentioned prior art, it is an object of the present invention to provide a method for collecting a sample in a closed system in a completely sterilized state by simplifying sample tube exchange in the closed system. And

Another object of the present invention is to automate sample collection and sample tube replacement in place of manual work with human error without requiring large-scale equipment such as a clean room.

[0011]

The present invention for achieving the above object has a sterilizing material inlet for introducing a sterilizing material, a drain outlet for discharging unnecessary sterilizing material and a sample, and a sample inlet for introducing a sample. A drain pipe, a sterilizing cylinder having a sterilizing material introducing port for introducing a sterilizing material, and one end open, a sampling tube that is detachably attachable to the sterilizing cylinder and sealed under reduced pressure sterilization in advance, A sample collecting device that is mounted and has a hollow needle that connects a drain tube, a sterilizing cylinder, and a sample collecting tube to transfer the sample in the drain tube to the sample collecting tube when the sample is transferred.

Also, the device of the present invention comprises one end of the drain pipe,
One end of the sterilization cylinder and one end of the sample collection tube are made of silicone rubber, and a hollow needle is passed through the silicone rubber portion to connect the drain tube, the sterilization cylinder and the sampling tube to transfer the sample.

Further, according to the present invention, the sterilizing material is introduced into the drain pipe through the sterilizing material introducing port by using the above-mentioned sampling device.
The process of discharging the sterilizing material from the drain discharge port, washing and sterilizing the inside of the drain pipe, and drying, and introducing the sterilizing material into the drain pipe through the sterilizing material introducing port, and introducing the sterilizing material into the sterilization tube. Discharging from the tip, washing and sterilizing the inside of the hollow needle, drying, inserting the sample collection tube into the sterilization cylinder to a position where it does not contact the hollow needle, and sterilizing material into the sterilization cylinder from the sterilization material inlet. Introduce the atmosphere in the sterilization cylinder and the washing needle sterilization of the hollow needle in the sterilization cylinder and the external surface of the sampling tube, the step of introducing the sample into the drain tube from the sample introduction port, and the sterilized gas, A step of transferring the sample into the sample collecting tube by inserting the hollow needle into the sample collecting tube whose inside is sterilized in advance under reduced pressure while being introduced into the sterile tube from the sterilizing material introduction port of the sterilizing cylinder. This is a sampling method.

Further, the method of the present invention includes that the sterilizing material is an aqueous ethanol solution and sterilized gas.

Hereinafter, the present invention will be described in detail with reference to the drawings.

FIG. 1 is a flow chart showing one embodiment of the sampling device of the present invention.

In the figure, reference numeral 1 is a test tank such as a fermenter in which a sample is collected by the device of the present invention (hereinafter simply referred to as a fermenter), and a sample conduit 2 is usually installed at the tip of the sample conduit 2. Is provided with a sample valve 3.

In the present invention, the sterilizing material for sterilizing and drying the inside of the drain pipe 4, the hollow needle 5 and the sterilizing cylinder 8 includes a sterilizing liquid 12 having a washing and sterilizing action and a gas for evaporating and drying the sterilizing liquid 12. It is preferable that

The sterilizing solution 12 has (1) a concentration of 60
% Or more ethanol aqueous solution, or (2) concentration 1 pp
An aqueous solution of m or more and 4000 ppm or less of hypochlorous acid, sodium hypochlorite or potassium hypochlorite, or (3) an aqueous solution of hydrogen peroxide having a concentration of 3% or more and 35% or less can be used alone or in combination. . When the sterilizing liquids of (2) and (3) above are used alone or in combination, they are washed and sterilized with these sterilizing liquids, and finally washed and sterilized with the aqueous ethanol solution of (1) at least once. Preferably.

As the gas for evaporating and drying the sterilizing liquid 12, air sterilized through the sterilizing filters 21 and 38, nitrogen, helium, argon or the like can be used. It is preferable to use sterilized air when the bacteria are aerobic bacteria such as lactic acid bacteria and nitrogen, helium, argon or the like when the bacteria are anaerobic bacteria such as methane bacteria.

For example, when a solution using ethanol is used as the sterilizing solution 12, its sterilizing action works when ethanol is evaporated, as is well known. Therefore,
Drying gas supplier 17, sterilizing filter 21, disperser 2
By supplying a sterilized gas via 8, sterilization and drying are performed simultaneously. At this time, the concentration of the aqueous ethanol solution is 90 to 100% when the main purpose is drying.
The drying time is shortened, and when the main purpose is sterilization, it is 60 to 80%, and more preferably 65 to 75% where the sterilization effect is remarkable.

Further, as described above, in the case where at least one of hypochlorous acid, sodium hypochlorite, potassium hypochlorite and an aqueous solution of hydrogen peroxide is used as the sterilizing liquid 12 in combination with an aqueous solution of ethanol. Is preferably washed with 100% ethanol and dried from the viewpoint of removing salts from the sterilized solution.

The pressure for supplying the sterilized gas can be set according to the size of the drain pipe, the length and the inner diameter of the hollow needle, and is generally 10 mm to 20 mm in a water column.
If it is adjusted so that it can be supplied at 0 mm, there will be no particular inconvenience. Although it depends on the length and inner diameter of the hollow needle 5, if the water column is less than 10 mm, sterilization and drying may take too long. If the water column exceeds 200 mm, it is possible to use sterilized gas more than necessary. Because there is.

The sterilizing material inlet 48 provided in the drain pipe 4 for introducing the sterilizing material as described above into the drain pipe 4 is supplied from the inlet of the sterilizing liquid 12 supplied by the pump 18 and the drying gas supplier 17. The inlet for the drying gas may be the same or may be provided separately. Further, a disperser 28 for efficiently dispersing the sterilizing material inside the drain pipe is preferably provided at the tip of the sterilizing material introducing port 48.

FIG. 2 is a schematic diagram showing an example of the disperser 28. As the disperser 28, a pipe having a plurality of holes opened on the surface of a pipe whose both ends are closed can be used, but it is not particularly limited as long as the surface is provided with holes for dispersing a sterilizing material. The inner diameter of the hole depends on the size of the disperser 28, but may be about 0.5 to 2 mm. The material of the disperser 28 is not particularly limited as long as it is a material that is inert to the sample and the sterilizing material, and, for example, stainless steel, pressure resistant glass, polytetrafluoroethylene or the like can be used.

The drain tube 4 may have any shape as long as it has a small amount of sample retention and is easy to clean, sterilize and dry. In FIG. 1, a rectangular parallelepiped is connected to the upper portion of the horizontally oriented cylindrical tube, and the cylindrical tube bottom portion on the side to which the rectangular parallelepiped is connected is cut out for easy cleaning. In this example, the rectangular parallelepiped portion is provided with a disperser 28 or the like for dispersing the sterilizing material introduced from the sterilizing material introducing port 48, and the sideways cylindrical tube portion serves as a sample retaining portion. It should be noted that the volume of the portion of the drain pipe 4 where the sample is retained depends on the amount of the sample to be collected, but may be selected to be approximately 2 times or more and 3 times or less the required amount of the analysis sample.

As the material of the drain pipe 4, any material can be used without particular limitation as long as it is an inert material with respect to the sample and the sterilizing material. For example, stainless steel or the like can be used.

The drain discharge port 43 is preferably provided in the lower portion of the drain pipe 4 where the liquid stays.

The sample introduction port is connected to the sample valve 3 attached to the fermenter 1, and the connection method can use a normal connecting means such as a bolt nut, a sanitary joint, etc. Is polytetrafluoroethylene,
A packing such as silicone rubber may be used.

It is preferable that the sterilization cylinder 8 has a cylindrical shape with one end open and is provided with one or more spray ports 7 for spraying a sterilizing material.

FIG. 3 is a plan view of the sterilization cylinder 8 seen from the open end. In the figure, 5 is a hollow needle, 8 is a sterilization cylinder, and 7 is a spray port provided in the sterilization cylinder 8.

At the closed end of the sterilization cylinder 8, a plug 6 through which the hollow needle 5 can pass is provided. FIG. 4 is a schematic cross-sectional view in which the periphery of the stopper 6 through which the hollow needle 5 of the sterilization cylinder 8 penetrates is enlarged. The sterilization cylinder 8 and the hollow needle 5 are connected and sealed by a stopper 6 attached to the sterilization cylinder 8. The stopper 6 may be a silicone rubber stopper or the like, and is fixed by the presser 45 and the cap 46. The material of the retainer and the cap is not particularly limited, but a metal material such as stainless steel or a polymer material such as polytetrafluoroethylene, polystyrene, or polyethylene may be used. The fixation of the cap 46 and the sterilization cylinder 8 is achieved by threading the sterilization cylinder 8 and the cap 46.

The sterilization cylinder 8 sends the sterilization liquid 12 and sterilized gas into the sterilization cylinder 8 to sterilize the inside of the sterilization cylinder 8 and the outside of the hollow needle 5 and the sampling tube 10 inserted into the sterilization cylinder 8. And it is installed for the purpose of drying. On the other hand, it also has the purpose of cleaning the drain discharged from the tip of the hollow needle 5 with a sterilizing material supplied through the spray port 7.

The material of the sterilization cylinder 8 can be used without any problem as long as it is stable with respect to the analysis sample 13 and the sterilization material. Further, when visually observing the analytical sample 13, it is preferable to use transparent glasses such as hard glass and quartz glass, and transparent polymer products such as polyester and polyethylene.

As the sample collecting tube 10, a commercially available blood collecting tube made of polyester, hard glass or the like which has been decompressed and sterilized in advance can be used. When a non-commercially available product is used, the following method can be used as the vacuum sterilization and sealing method. (1) A rubber stopper is attached to a cylindrical tube made of polyester, hard glass or the like prepared in advance, and (2) a glass inlet / outlet is provided through the stopper, or
The entrance and exit is provided by a separately prepared hollow needle whose tip is sealed and has a hole in the side. (3) 65 to 7 where the sterilization effect is remarkable from the entrance provided
After filling with a 5% ethanol aqueous solution, a pre-sterilized gas for drying is flowed to sterilize and dry the inside. (4) After sterilization and drying, 200 mmHg-
The pressure is reduced to about 300 mmHg. (5) If a glass door is provided through the stopper,
The inlet / outlet may be burnt out and sealed. On the other hand, if the inlet / outlet is provided by a hollow needle having a sealed tip and a lateral hole, the hollow needle may be removed.

The stopper 9 is preferably made of silicone rubber or butyl rubber so that the hollow needle 5 can penetrate therethrough.

The diameter of the sampling tube 10 and the length in the long axis direction are defined by the amount of the analysis sample 13 to be collected, but in the case of normal sampling, it is usually used in a commercially available blood collection tube. Any size is acceptable. The sampling tube 10
The length in the long axis direction of is preferably at least longer than the length of the sterilization cylinder 8 in the long axis direction in terms of operability.

The sterilizing cylinder 8 and the sample collecting tube 10 are preferably cylindrical in consideration of the ease of attachment and detachment of the sample collecting tube 10. When inserting the sample collection tube 10 into the sterilization cylinder 8, the gap between them is set to be equal at any position within a range of 1 to 4 mm. At this time, a guide plate or the like for inserting the sample collection tube 10 into a correct position may be provided as needed. By making the gap between the sterilization cylinder 8 and the sample collection tube 10 uniform, the mixed fluid of the sterilization liquid vapor and the sterilization gas or the sterilization gas is uniformly discharged out of the system of the sterilization cylinder 8. When the gap is less than 1 mm, the sampling tube 10
When inserting into the sterilization cylinder 8 or withdrawing it from the sterilization cylinder 8, the operability deteriorates, and when the gap exceeds 4 mm, the uniformity of the fluid discharged to the outside of the system deteriorates.

The hollow needle 5 for transferring the sample 13 for analysis from the drain pipe 4 to the sampling pipe 10 may be attached or fixed by providing a hollow needle connection port in the drain pipe 4, but it may be washed or replaced. For convenience, it is preferable to penetrate the rubber plug 14 or the like as shown in FIG. 1 and to connect the rubber plug 14 to the drain pipe 4 with a cap 16 over the presser 15 so that the rubber plug 14 does not come off from the drain pipe 4. The material of the presser foot 15 and the cap 16 is not particularly limited, but a metal material such as stainless steel or a polymer material such as polytetrafluoroethylene, polystyrene or polyethylene is preferable. Also, presser foot 15
Is preferably provided with a valve function for opening and closing the hollow needle 5.

The material of the hollow needle 5 is a metal such as stainless steel, iron or aluminum.

Next, a method of sampling with the above-described sampling device will be described.

Usually, when collecting a sample for analysis, at least the sample staying in the sample conduit 2 is repeatedly discharged several times as a drain, but this is also the case in the present invention.

First, the drain pipe 4 is connected to the sample valve 3 by the above method, the valve of the presser foot 15 is opened, the valve 44 is partially opened, and when the sample valve 3 is opened, it stays in the sample conduit 2. The existing sample is introduced into the drain pipe 4. If the valve 44 is closed at the time when all the sample staying in the sample conduit 2 is transferred into the drain pipe, no more sample is transferred into the drain pipe 4. Then, the sample valve 3 is closed, the valve 44 is opened, and the sterilized drying gas is supplied through the drying gas supplier 17, the sterilizing filter 21 and the disperser 28, and the sample in the drain pipe 4 is discharged. . At this time, the valve of the presser foot 15 is in a closed state. When the presser 15 does not have a valve function, the tip of the hollow needle 5 may be left in the middle of the stopper 9 of the sampling tube 10, or a small silicone rubber stopper or the like prepared separately may be sterilized in advance. It is also possible to cover the tip of the hollow needle 5 and seal it.

The drying gas supplier 17 is not particularly limited and can be a cylinder or a compressor, and the sterilizing filters 21 and 38 are not particularly limited and can be commercially available products.

After the sample accumulated in the sample conduit 2 is discharged as a drain, the inside of the drain pipe 4 and the inside of the hollow needle are washed and sterilized and dried.

First, the sterilizing solution 12 in the sterilizing solution tank 11 is supplied to the drain pipe 4 via the pump 18 and the disperser 28,
Drain through valve 44. When the drain discharged from the line 47 becomes colorless and transparent, the valve 44 is closed and the valve of the presser 15 is opened to discharge the sterilizing liquid from the tip of the hollow needle 5. After repeating the above operation several times, the sample 13 for analysis is again received in the drain tube 4 by the same procedure.

Then, a sterilizing material is introduced from the spray port 7 to wash and sterilize the inside of the sterilizing cylinder and dry it.

The sterilization conditions are common to the inside of the drain pipe 4, the inside of the hollow needle 5, the atmosphere inside the sterilization cylinder 8 and the outside of the hollow needle 5 and the sampling pipe 10 inside the sterilization cylinder 8. is there.

After washing and sterilizing each part of the apparatus, the sampling tube 10 is further inserted into the sterilization cylinder 8 while sterilizing gas is passed through the sterilization cylinder 8, and the hollow needle 5 is penetrated into the stopper 9.
Since the inside of the sample collecting tube 10 has been depressurized in advance, when the hollow needle 5 is inserted into the sample collecting tube 10, the analysis sample 13 in the drain tube 4 is transferred into the sample collecting tube 10. After the transfer, a sample that may remain in the hollow needle 5 is sent to the sampling tube 10 and the pressure inside the sampling tube 10 is returned to normal pressure. Is sent to the sampling tube 10. Then, the sampling tube 10 is pulled out.

In the present invention, since the sample-collecting tube 10 is pulled out from the hollow needle 5 in a sterilized atmosphere, it is possible to prevent contamination of live bacteria such as lactic acid bacteria in the air through the stopper 9 of the sample-collecting tube.

In the above description, the procedure in the case of manual analysis has been described, but automation including semi-automation by forming a sequence can be easily performed by a combination of conventional techniques.

[0052]

The present invention will be described in more detail with reference to the following examples, but these specific examples do not limit the present invention.

Example 1 Using the apparatus shown in FIG. 1, 50 ml of an analytical sample 13 was sampled from the beer fermenter 1 having a volume of 50 kl by the following procedure.

Outline of sampling device: A stainless drain tube 4 having a volume of 500 ml was prepared. The shape of the liquid retention portion of the drain pipe 4 is cylindrical, and the volume of that portion is 120
ml. The hollow needle 5 has an inner diameter of 1 mm and an axial length of 50.
mm. The presser foot 15 is made of polytetrafluoroethylene and has a valve function. The disperser 28 had a shape in which both ends of a stainless steel pipe having an inner diameter of 10 mm and a length of 50 mm were closed, and a total of 100 holes having a diameter of 2 mm were provided on the cylindrical side surface.

As the sample collecting tube 10, a commercially available product having an inner diameter of 30 mm, an outer diameter of 33 mm and a total volume of 100 ml was used.

The sterilizing cylinder 8 has a cylindrical bottom having an inner diameter of 35 mm and a length of 100 mm in the long axis direction, one of the bottom surfaces of which is open, and the other end which is closed has a hole of 6 mm in diameter, and the hole is made of silicon as shown in FIG. Attach the rubber stopper 6, presser foot 45 and cap 4
Fixed at 6.

Preparation for sample collection: Fermenter 1 with the above sampling device
The sample valve 3 was attached. Silicon rubber was used for the packing.

First, the inside of the drain pipe, the inside of the sterilization cylinder, the outside of the sampling pipe, and the inside and outside of the hollow needle were washed, sterilized and dried as follows.

For cleaning the inside of the drain pipe and the inside of the hollow needle,
From the ethanol tank 11 storing the 0% ethanol aqueous solution 12, 150 ml of 70% ethanol aqueous solution is pumped by the lines 35, 33, 32, 31, 29, 25,
It carried out by injecting into the drain pipe 4 through 27 and the disperser 28. The valve of the presser foot 15 is repeatedly opened and closed several times, and it is confirmed that the 70% ethanol aqueous solution is flowing out from the tip of the hollow needle 5, and the valve 44 is opened by keeping the valve closed and the 70% ethanol aqueous solution is discharged from the drain pipe 4. Released.

On the other hand, for sterilization inside the sterilization cylinder, outside the sampling tube and outside the hollow needle, 50 ml of 70% ethanol aqueous solution 1
2 by the pump 18 through lines 35, 33, 42,
It was sprayed from the spray port 7 into the sterile tube 8.

The inside of the drain pipe 4 and the inside of the sterilization cylinder 8 were simultaneously cleaned. Then, a drying gas supplier 17
Through sterilizing filters 21 and 38 for 10
Air sterilized at a rate of 00 ml / min was supplied to the drain pipe 4 and the sterilization cylinder 8 to carry out sterilization and drying.

The supply of sterile air to the drain pipe 4 was continued for 5 minutes and then stopped, and the transfer of the sample to the drain pipe 4 was started. First, the sample valve 3 is opened and the valve 4 is delayed or at the same time.
4 was opened, and a sample of about 3 times the internal volume of the sample conduit 2 whose internal volume had been measured beforehand was taken out as a drain and discharged from the drain discharge port 43.

Then, in order to receive the sample for analysis 13, the inside of the drain tube 4 was washed, sterilized and dried according to the above procedure. After washing three times, sterilization and drying were performed.

100 ml of the sample 13 for analysis is placed in the drain tube 4.
The hollow needle 5 is received, the silicone rubber stopper 9 is penetrated, the valve of the presser 15 is opened, and the sample 13 for analysis is placed in the sampling tube 10.
Received 50 ml. The ethanol concentration and sugar concentration of the obtained analytical sample were measured by high performance liquid chromatography. 45 ml of analytical sample has a grain size of 0.2μ
m pre-sterilized nitrocellulose filter paper,
This filter paper was cultured in an agar medium for lactic acid bacteria for 72 hours, and the number of living lactic acid bacteria was measured. The analysis results are shown in Table 1.

Example 2 The beer fermenter 1 of Example 1 having a volume of 50 kl was replaced with a fermenter containing 50 liters of sterilized broth medium, and the same operation as in Example 1 was carried out to obtain 50 ml of a sample for analysis. After holding the obtained 50 ml sample for analysis in a constant temperature bath at 37 ° C. for 48 hours, the presence or absence of contamination by various bacteria was examined. The results are shown in Table 2.

Reference Example 1 Using the fermenter 1 of Example 1, a sample solution was sampled by the conventional method shown in FIG. As a sterilizing material, 121 ° C
Of water vapor was used. The inside and outside of the sample collection tube were sterilized and stored in a clean room in advance. The collected analysis sample was analyzed in the same manner as in Example 1. The results are also shown in Table 1.

Reference Example 2 Using the fermenter 1 of Example 2, a sample for analysis was sampled in the same manner as in Reference Example 1 and examined in the same manner as in Example 2. The results are also shown in Table 2.

Reference Example 3 In an open system, a sample was sampled directly from the sample valve 3 of the fermenter 1 of Example 1 into a sample collection tube. The inside and outside of the sample collection tube were sterilized and stored in a clean room in advance. The collected analysis sample was analyzed in the same manner as in Example 1. The results are also shown in Table 1.

Reference Example 4 In an open system, a sample was sampled directly from the sample valve of the fermentor of Example 2 into a sample collection tube. The inside and outside of the sample collection tube were sterilized and stored in a clean room in advance. The collected analytical sample was examined in the same manner as in Example 2.
The results are also shown in Table 2.

[0070]

[Table 1] Note) * 1… The number of samples (numerator) in which the occurrence of lactic acid bacteria was confirmed with respect to the total number of samples (denominator) examined.

[0071]

[Table 2] Note) * 2… The number of samples (denominator) confirmed to be contaminated with various bacteria against the total number of samples (denominator) tested.

[0072]

According to the sampling device and the sampling method of the present invention, it is possible to collect a sample in a completely sterilized state in a closed system by simplifying the exchange of the sampling tube in the closed system.

Further, it becomes possible to automate the collection of the sample for analysis and the exchange of the sample collection tube in place of manual work with human error without requiring a large facility such as a clean room.

[Brief description of drawings]

FIG. 1 is a schematic view showing an example of one embodiment of a sampling device of the present invention.

FIG. 2 is a schematic view showing an example of one mode of a disperser provided in a drain pipe.

FIG. 3 is a plan view of the sterilization cylinder shown in FIG. 1 as viewed from the open side thereof.

FIG. 4 is an enlarged schematic view of a silicone rubber stopper portion of the sterilization cylinder shown in FIG.

FIG. 5 is a schematic view showing an example of a conventional sampling device.

[Explanation of symbols]

 1 Fermenter 2 Sample conduit 3 Sample valve 4 Drain pipe 5 Hollow needle 6, 9 Stopper 7 Spray port 8 Sterilization cylinder 10 Sample collection pipe 11 Sterilization liquid tank 12 Sterile liquid 13 Analytical sample 14 Rubber stopper 15, 45 Presser 16, 46 Cap 17 Drying gas supplier 21, 38 Sterile filter 28 Disperser 43 Drain outlet 48 Sterile material inlet 501 Test tank 503 Sample valve 504 Drain pipe 505 Sample sampling pipe 506 Sample liquid 507 Steam inlet pipe 508 Ethanol aqueous solution inlet pipe 509 Sterile air introduction tube

Claims (4)

[Claims] 1. A sterilizing material introducing port for introducing a sterilizing material, a drain discharging port for discharging unnecessary sterilizing material and a sample, and a sample introducing port for introducing a sample, and a sterilizing material introducing port for introducing a sterilizing material. A sterilization tube having one end opened, a sampling tube that is detachably attachable to the sterilization tube, and is sealed in advance under reduced pressure sterilization, and is attached to a drain tube in advance. The drain tube, the sterilization tube, and the sample when transferring the sample A sampling device having a hollow needle that is connected to the sampling pipe and transfers the sample in the drain pipe to the sampling pipe. 2. One end of the drain pipe, one end of the sterilization cylinder and one end of the sampling pipe are made of silicone rubber, and a hollow needle is penetrated through the silicone rubber portion to connect the drain pipe, the sterilization pipe and the sampling pipe. The sample collecting device according to claim 1, wherein the sample is transferred by means of a method. 3. A sterilizing material is introduced from the sterilizing material introducing port into the drain pipe by using the sampling device according to claim 1, the sterilizing material is discharged from the drain outlet, and the inside of the drain pipe is washed and sterilized and dried. And the step of introducing the sterilizing material into the drain pipe through the sterilizing material introducing port, discharging the introduced sterilizing material from the tip of the hollow needle inserted in the sterilization cylinder, washing and sterilizing the inside of the hollow needle, and drying. The step of inserting the sampling tube into the sterilization cylinder to the position where it does not come into contact with the hollow needle, and the sterilizing material is introduced into the sterilization cylinder through the sterilization material inlet, and the hollow needle inside the sterilization cylinder and the sampling needle Washing and sterilizing the outer surface of the, and drying the sample, introducing the sample into the drain tube from the sample inlet, while introducing the sterilized gas from the sterilizing material inlet of the sterilization tube into the sterilization tube, Insert inside the sample collection tube that has been sterilized under reduced pressure in advance Sampling method characterized by a step of transferring the sample into the sampling tube by Rukoto. 4. The sampling method according to claim 3, wherein the sterilizing material is an aqueous ethanol solution and a sterilized gas.