JP3210142B2 - Bacterial control confirmation test method, medium used for it - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for confirming bacterial control in a process of filling contents into containers by controlling bacterial contamination such as aseptic filling, and more particularly to a method for confirming bacterial control in a manufacturing plant upon delivery or start-up. or when bacterial control confirmation test methods to simplify the operation of confirming the bacterial control status performed after maintenance of repair and maintenance of the apparatus relates to a medium used for it.

[0002]

2. Description of the Related Art Filling of contents such as a potion pack for coffee which is not subjected to sterilization or sterilization after filling the container and subjected to refrigerated distribution at room temperature employs filling means which controls bacterial contamination such as aseptic filling. I have. For aseptic filling, usually
Asepticity must be achieved in all processes from sterilizers, lines, surge tanks, aseptic filling machines aseptic chambers and filling nozzles, so maintenance such as delivery or start-up of manufacturing plants or repair / repair of equipment, etc. It is necessary to check the sterility after the inspection to ensure the sterility of the manufacturing process.

[0003] In addition to the aseptic filling, it is necessary to control the bacterial contamination in the above-mentioned process also in the filling for eliminating contamination of specific bacteria such as Escherichia coli.

[0004] One of the means for confirming the bacterial control state in such a process before the actual production is to fill the container in the same manner as in the actual production by using a liquid content (actual liquid) actually filled in the container as a product. This is a sampling inspection (culture inspection). This method seems to have high inspection accuracy at first glance because the bacterial control of the process can be confirmed under the same conditions as this production.However, in this method, a large amount of real liquid sample must be collected in order to increase the inspection accuracy. In addition, in the bacterial test, the collected sample is opened, a certain sample is transferred to a separately prepared bacterial test medium, and subjected to a cultivation test. There is a possibility of secondary contamination, and in reality, the inspection accuracy and reliability are low. Therefore, even if the control of bacteria in the process is confirmed for the time being with the actual liquid filling, the operation time of this production cannot be extended (one operation time Is about one hour).

[0005] Not only in the case of aseptic filling, but in the step of controlling bacterial contamination, it is necessary to check the presence or absence of contamination by performing a bacterial test by filling with actual liquid in the process control, and has the same problem.

[0006]

SUMMARY OF THE INVENTION In view of the above-mentioned problems of the prior art, the present invention requires confirmation of non-contamination of bacteria at the time of delivery or start-up of a manufacturing plant or after maintenance and inspection such as repair and repair of equipment. To solve problems such as the complexity of sampling and bacteria non-contamination work involved in the actual solution test and the incompleteness of the test accuracy in the process, and to enable the use of a specific test medium instead of the actual solution test. This greatly reduces sampling work and improves inspection accuracy.
It is another object of the present invention to significantly speed up the process of confirming bacterial contamination in a process by using a specific inspection container.

Another object of the present invention is to provide a test medium and a test container for achieving the above object.

[0008]

According to the present invention to achieve the above object, in a test for confirming bacterial control in a step of filling contents into a container from a filling machine and sealing the contents, a fluid equivalent to the contents is used instead of the contents. Bacterial control confirmation test method characterized by introducing a culture medium for bacteria test having the process into a process, filling and sealing the container, and detecting bacterial contamination by a change in the clarity of the culture medium to verify the bacterial control state in the process. It is. According to this method, secondary contamination at the time of sampling can be prevented, and the bacterial contamination in the process can be inspected under the same conditions as the actual liquid filling, so that the sampling operation is greatly reduced and the inspection accuracy is improved.

Further, the present invention is the above-mentioned bacterial control confirmation test method, characterized in that at least a part of a sealed container filled with a culture medium for bacteria test is transparent. According to this method, after filling the medium, the change in the clarity of the medium can be easily determined visually or the like from the outside without opening the container, so that the bacterial contamination confirmation work can be greatly speeded up. Fluctuations and leaks in the liquid can be observed.

Further, the present invention relates to a culture medium for bacterial test having the same fluidity as the contents used in the above-mentioned bacterial control confirmation test method, and at least a part of the culture medium filled with the bacterial test medium and sealed is transparent. There is a container. Since such a medium has the same fluidity as the contents, it is possible to prevent the medium solution from scattering at the time of filling and making it impossible to seal the cup.

Hereinafter, the present invention will be described in detail.

In the present invention, the step of filling the contents into the container from the filling machine and sealing the container is such that the viable cell count of the filling in the closed container is adjusted to a certain level or less, preferably to zero. The process of controlling the bacterial contamination of the contents at the stage of filling the contents from the filling machine into the container, and filling and sealing the contents. The maintenance is performed at the time of delivery or start-up of the manufacturing plant or at the time of repair / repair of the equipment. This refers to the process that requires confirmation of bacterial contamination after inspection. The control of bacterial contamination includes not only control of sterilizing the contents but also control of making Escherichia coli negative and reducing the contamination of general bacteria to a certain level or less. The treatment after filling and sealing of the container is not particularly limited, but usually, a process in which sterilization and sterilization treatment is not performed after filling and sealing is targeted. The type of bacteria to be controlled does not matter. Means for controlling bacteria are not particularly limited, such as heat sterilization / sterilization, centrifugation / filter removal of bacteria, use of bactericides / bacteriostatic agents, and the like. In addition, the above-mentioned steps include all steps related to bacterial contamination control, and in general, lines from the sterilization / sterilization / bacteriostatic steps of the contents to the filling machine to the filling machine and the filling from the filling machine to the container. And the entire process up to sealing. For example, in the case of aseptic filling, the process includes steps from the inlet to the outlet of a sterile chamber of a surge tank, a buffer tank, a small filling tank, and an aseptic filling machine. The contents are fluid at the time of filling regardless of the properties of the contents after filling, and the distribution process after filling is refrigerated or normal temperature, and food or functional foods in which bacterial growth during distribution or storage becomes a problem, Drugs and the like are not particularly limited. Foods, pharmaceuticals, and the like in the form of solutions, suspensions, emulsions, foams, gels, aggregates thereof, thread-like, band-like gel aggregates, and the like are included. The contents may be solidified after filling and may be solid at the time of distribution. For example, foods that require aseptic filling such as potion pack sterilization means, milk,
Examples include foods, pharmaceutical drinks, medical ampules, medical infusions and the like, such as milk drinks and fruit juices, which are required to be filled with the coliform group negative.

The fluidity of the contents is such that the contents are usually filled into the container by a nozzle, so that the nozzle can be filled. The fluidity of the contents is not particularly limited and is appropriately adjusted by a filling machine or the like used. Is what is done.

The bacteria control confirmation test of the present invention includes, for example, the time of delivery or start-up of a manufacturing plant, the time of installation of equipment, the repair or modification of equipment, the replacement of parts, the change of manufacturing conditions, or the occurrence of microbial troubles. This is performed before (not limited to immediately before) or after (not limited to immediately after) the actual production using actual liquid, but it is necessary to verify that bacterial control in this production is ensured. This can be done at any time.

In the present invention, the fluidity of the culture medium for bacterial test is adjusted to the same level as that of the contents. Since the medium has the same fluidity as the contents, the medium can be filled into the container under the same conditions as when filling the contents, so that the amount of the test sample that accurately reflects the bacterial contamination control state varies. It can be collected without. The fact that the bacterial test medium has the same fluidity as the contents does not mean that it is necessary to have completely the same fluidity, but the medium may be scattered when the medium is filled instead of the contents. The fluidity is identical to such an extent that a bacterial control state can be ensured without causing a cup seal failure, and means a fluidity in which such functions and actions are exhibited. The fluidity can be defined rheologically by viscosity and elasticity, but these properties are not particularly limited as long as the function and action at the time of filling are exhibited.
As one guide, a fluidity corresponding to a viscosity of about 70 to 250 cps is generally preferred for exhibiting the above functions and effects. In addition, in general, it is preferable that the spinnability is small in terms of breakage at the time of filling, but when the content to be manufactured has a high spinnability, it has a similar spinnability. Good.

The culture medium for bacterial tests used in the present invention includes:
In addition to having the predetermined fluidity as described above, those having excellent transparency and non-antibacterial properties are preferred. This is because if the culture medium is transparent, detection of a bacterially contaminated sample becomes extremely easy, and if the culture medium has antibacterial properties, it may not be possible to accurately detect even if bacterial contamination has occurred. The medium is a liquid that exhibits fluidity at the time of filling, but does not hinder solidification after filling. However, if the liquid state remains after filling, it is possible to visually confirm the state of liquid swaying and liquid leakage not only at the time of filling but also in the container being conveyed, which is useful for investigating the cause of troubles such as leakage. be able to. Incidentally, unlike the solid medium, the liquid medium does not contain a solidifying agent such as agar, so that turbidity or the like does not usually matter.However, in the present invention, a liquid medium having an increased viscosity is used. Agents must be used, and the issues of transparency and antibacterial properties must be solved. Conventionally, there is a technique of using xanthan gum and locust bean gum in combination in place of agar to prevent clouding of the agar medium (Japanese Patent Application Laid-Open No. 62-186784), but almost no liquid medium having a relatively high viscosity is known. Not been.

The culture medium for bacterial test used in the present invention basically comprises a basal medium and a stabilizer. The basal medium can be appropriately selected as appropriate for the culture of the bacteria to be controlled,
For example, a broth medium, which is a basal medium for bacteria, a lactose broth medium for testing coliform bacteria, a BGLB medium, peptone,
A broth medium or the like can be used, and peptone or the like is preferable from the above viewpoint. As a stabilizer, it is at least soluble in warm water, the solution is transparent, it is heat stable,
Those exhibiting a stable viscosity in a small amount are good, for example, locust bean gum, guar gum, tragacanth gum, karaya gum and the like can be used. It is purified guar gum. Further, agar, gelatin or the like may be used depending on the fluidity of the actual solution.

The composition of the medium for testing general bacteria is, for example, as follows. (Blending ratio) ・ Purified salt 0.5% ・ Yeast extract 0.3% ・ Peptone 1.0% ・ Glucose 0.5% ・ Stabilizer 0.5-0.6% ・ Purified water 97.1-97. 2% (pH 6.5 to 6.8) (viscosity 70 to 170 cps) The preparation of the culture medium may be performed by mixing the raw materials into powder, adding heated water thereto, sufficiently dispersing and dissolving, and then sterilizing. This preparation step may be carried out separately from the content production step, but preferably the equipment can be simplified by using a mixing line, a tank, a sterilizer or the like in the actual liquid filling step.

Next, as a container used for a bacterial test in the present invention, in principle, a container used for filling contents, that is, a container filled with actual liquid can be applied as it is. By using a container filled with actual liquid, the production line of the filling process can be used as it is, so that inspection materials and equipment can be simplified and labor can be reduced, and the control of bacteria can be accurately confirmed. It can be carried out. Further, the state of the liquid sway in the container can be accurately grasped as a simulation in the case of the actual liquid. The container may be composed of one member, or may be composed of two members, a lid member and a main body (bottom member), or more members.

The test container is preferably at least partially transparent. When it is transparent, the properties of the medium inside can be easily inspected by visual observation from outside without opening the container, so that the inspection efficiency can be greatly improved. For example, in the case of a real liquid container in which a transparent bottom portion is formed and a film-shaped opaque lid material is thermally welded thereto,
By directly using this as a test container, the inside medium can be easily observed through the transparent bottom portion, and the state of the inside liquid during filling and transportation can also be observed. In the case of an aseptic potion pack having a volume of about 5 to 10 ml, the bottom portion is generally made of PET / OPS.
The lid portion is made of an opaque material such as PET / Al foil, so that when used as it is as an inspection container, the inside of the container cannot be observed from the outside. You can open the lid at the time of inspection and inspect the inner clarity,
In order to save the trouble of opening, it is preferable that a part of the container is transparent. In the case of aseptic filling, the container is usually sterilized and molded in a sterile room, so replacing the bottom part with an inspection part would be cumbersome.In that case, it is better to use a transparent material for the lid part. It is preferable because there is a wide selection range and there are few restrictions on the work process. The transparent lid material is preferably a material capable of being sterilized by means of sterilization means, for example, hydrogen peroxide (H ₂ O ₂ ) in a sterile room, and capable of sealing to the bottom portion and easily punching after welding to the bottom portion. is there. It is convenient in terms of equipment that the device used for the actual liquid filling can be used as it is for punching after welding. As the transparent lid member,
PET / OPS or the like can be used.

The size of the test container is the same as that of the actual liquid container.
Generally, the thickness is about 50 to 600 μm (the thickness of the lid is 25 to 30 μm).

[0022]

The present invention operates, for example, as follows (in the case of aseptic filling).

The raw materials for the test medium are all weighed and mixed with powder, and a part (about 30%) of hot water (50 to 60%) is prepared.
), The raw material is dissolved in, for example, a desper mill, and then directly introduced into, for example, a TK tank, while the remaining mixed water (about 7
0 ° C.) from the desper mill into the TK tank, and uniformly mix the whole by a TK homomixer high-speed rotation (about 25 minutes). pH adjusted, direct sterilizer (about 150 seconds at 150 ° C)
And then cooled by, for example, a plate cooling unit (about 10
° C).

This product (the amount required for one test) is introduced into an aseptic filling chamber through a real liquid filling line, for example, a path from a surge tank to a buffer tank to a small filling tank.
The sterile room is sterilized with hydrogen peroxide, and the inspection container (lid member and bottom member) is similarly sterilized. The continuous filling is started, and the medium is filled into the bottom member forming the container using the air pressure or the like in the cushion tank (the contents can be filled using the air pressure because of the same fluidity. Become). The number of samples to be used for filling is a number necessary for performing sterility determination with high accuracy. For example, a sample with 70 nozzles has 100 to 300 shots (7000 to 21000 shots).
) Sampling (the number of samples, which was impractical in the conventional method, can be easily processed). The sample number can be identified by marking the nozzle number or the like on the bottom surface or the like of the bottom member. After the filling, the transparent lid member is welded to the upper surface of the bottom member on which the container is formed by heat sealing, and the container is sealed. The collected sample was cultured in an incubator at 30 ° C. for 5 days (may be 1 to 2 days depending on the degree of contamination), and the change in the clarity of the medium was determined by visual observation without opening the container lid. I do. As a result of the determination, when it is recognized that there is a problem in sterility, it can be estimated that a problem in quality occurs in the actual liquid product such as a lot and a nozzle corresponding to the sample. In this way, it is not necessary to open the container when confirming sterility or to inoculate a separately prepared medium with the actual liquid product.

Here, the change in the clarity of the medium includes not only the change in the average clarity of the whole medium but also the change in the local clarity such as the occurrence of one colony. The change in clarity can be determined not only visually but also by automatic detection using various optical measuring devices.

After collecting the sample filled with the test medium, purified sterilized water is used to remove the test medium from the sterilizer, line, aseptic surge tank (surge tank to buffer tank to small filling tank), nozzle and the like. It is important to load the liquid into the line to drain the liquid, and to perform further sufficient washing, whereby the test medium is completely removed from the process. After confirming the determination of the sterility of the cultured sample, the actual production using the actual solution is performed. Further, the bacterial control confirmation test of the present production can be carried out after completion of the filling with the actual solution for changing production conditions and the like. In this case, it is important to fill the line with purified sterilized water to draw out the actual liquid, and then to perform sufficient washing to completely shut off the actual liquid from the test medium in order to enhance the inspection accuracy.
After the actual solution is removed from the process, a test medium is introduced into the actual solution line, filled in a test container, and the sample is cultured to determine sterility.

In the conventional inspection by extracting with actual liquid filling, etc., the reliability of the determination of sterility was low, although much labor was required. For this reason, for example, when a new manufacturing plant is installed, it is necessary to spend several days to several tens of days to perform a bacterial control confirmation test and confirm the sterility of the plant before starting full-scale production. . According to the method of the present invention,
Although the sterility of the process can be confirmed with extremely high accuracy despite the simple method, the time required for the bacterial control confirmation test can be significantly reduced, and the reliability can be improved.

It is also possible to conduct a sterility confirmation test by using a paper with bacteria and the like in combination with a culture medium for examination to make the judgment of sterility more complete.

Further, even in cases other than the aseptic filling, the above-described processing can be applied to the step of controlling bacteria.

[0030]

EXAMPLES The present invention will be further described below with reference to examples.

(Filling Line and Packaging Material) A sterility confirmation test was carried out on a continuous aseptic filling line for a potion cream for coffee (5 ml). 70 filling nozzles are set, and the production capacity of the line is 102,000 pieces /
h. The bottom material is opaque HIPS and is pressed in a sterile chamber, while the lid material is a transparent PET / OPS film (PET 16 μm / OPS 25 μm). After the contents are filled into the bottom member (container) in the chamber, the bottom member Is heat-sealed to the upper surface. Each component is sterilized with hydrogen peroxide in a sterile chamber before being used. The lid material at the time of this production was opaque PET /
It is an Al film. The filling amount of the medium for each portion was 5 ml. (Preparation of medium) The mixing ratio of the medium was as follows.

[0032]

[Table 1] Based on the above blending ratio, the amount charged in the preparation is 1,000 kg
The raw materials are all weighed and mixed with powder and powder so as to obtain, and then about 30% of hot water (50 to 60 ° C.) is used in the prepared water,
The raw material was melted in a desper mill and put directly into the TK tank, while the rest of the formulated water (about 70 ° C.) was put into the TK tank from the desper mill. After dissolving with TK homomixer at high speed for 25 minutes and adjusting the pH to 6.8 with 4% sodium hydroxide, the medium was sterilized directly with a sterilizer (150 ° C, 4 seconds), and then the plate cooling unit At 10 ° C
And cooled. The viscosity of this product is 126 cps (B-type viscometer), and the viscosity of coffee potion cream is 150 ~
It was about the same as 250 cps, and the state of filling from the nozzle was similar to that of the cream. (Sterility test) Extraction of product using actual solution (conventional method) For comparison, products filled and manufactured using actual solution are collected periodically (once / two hours) (70 pieces x 3 times) Then, this was opened, and a certain amount was inoculated on a separately prepared solid agar medium and subjected to a culture test, and then sterility was determined. Medium filling (the present invention) Before the present production, the medium in the tank was introduced into a sterile room, and the container was filled under the same conditions as in the present production. The collected sample was then subjected to a culture test to determine the presence or absence of turbidity. 1
The number of samples collected per production was 11,130 (1
59 shots x 70 filling nozzles). (Results of Test for Confirming Sterility) The following table shows the persons and time required for confirming the complete sterility of the samples collected above.

[0033]

[Table 2] As is clear from the table, in the present invention in which the medium was filled into the container using the transparent lid material, unlike the extraction test, the lid material was not required to be opened and the inoculation to another culture medium was not required, so that a large amount was visually observed. Since the complete sterility of the sample can be easily confirmed, the labor can be significantly reduced.

[0034]

As described above, in a process requiring control of bacterial contamination, a culture medium for bacterial test having the same fluidity as that in place of the contents is introduced into the process, and the container is filled and sealed, Bacterial contamination is detected based on the change in the clarity of the culture medium, thereby verifying the bacterial control status of the process, thereby preventing secondary contamination at the time of sample collection and enabling bacterial contamination testing of the process under the same conditions as the actual liquid filling. As a result, the sampling work has been greatly reduced and the inspection accuracy has been improved.
In addition, since the test container is at least partially transparent, after the medium is filled, the change in the clarity of the medium can be easily determined visually from the outside without opening the container,
Bacterial contamination confirmation work has been greatly speeded up, and it has become possible to ascertain the liquid state during filling and transport. Since the test medium had the same fluidity as the contents, it was possible to prevent the medium liquid from scattering at the time of filling and making cup sealing impossible. These effects are also effective when applied to a process in which E. coli is negative and the number of general bacteria is reduced to a certain level or less, but is particularly remarkable in the case of aseptic packing in which sterility must be ensured in all processes. As a result, the work of checking the sterility, such as filling a portion pack, was ensured and simplified.

Continuation of the front page (72) Inventor Kenji Sakai 2-19-8 Chuo, Miyashiro-machi, Minamisaitama-gun, Saitama (56) References JP-A-1-317383 (JP, A) (58) Fields investigated (Int. . ^7, DB name) C12M 1/34 C12Q 1/00 - 1/24

Claims (3)

(57) [Claims] 1. In a bacterial control confirmation test in a step of filling contents into a container from a filling machine and sealing the container, a medium for bacterial test having the same fluidity as the contents is introduced into the process in place of the contents, and the container is introduced into the container. A test method for confirming the control of bacteria, characterized by verifying the bacterial control state in the step by filling and sealing, and detecting bacterial contamination by a change in the clarity of the medium. 2. The method according to claim 1, wherein at least a part of the sealed container filled with the culture medium for bacteria test is transparent.
Bacterial control confirmation test method described in 1. 3. A culture medium for bacterial test having fluidity equivalent to the contents used in the bacterial control confirmation test method of claim 1.