JPH048298A - Environment retaining composition and transporting and preserving tool using the same composition - Google Patents

Abstract

PURPOSE:To inexpensively and simply operate an environment retaining composition, by blending a deoxidizer with a high water absorbent and a hygroscopic immobilizing agent. CONSTITUTION:0.3-2.0 pts.wt. deoxidizer such as active iron oxide or ascorbic acid is blended with 0.05-0.1 pt.wt. high water absorbent such as acrylic acid derivative (salt), etc., and 0.02-0.5 pt.wt. hygroscopic immobilizing agent such as polyethylene glycol having 1,000 20,000 molecular weight to give an environment retaining composition in the dosage form selected from blended agent, multi-layer agent and tablet. Then the composition 1 is put in a holder 4 of a container 2, a medium 5 for anaerobic microorganisms is packed into the container and the container is sealed with a rubber stopper 3 to prepare a transporting and preserving tool.

Description

[Detailed description of the invention] [Industrial application field]

The present invention relates to a composition for maintaining an environment for transporting and preserving specimens collected for testing for anaerobic bacteria, or for transporting and preserving the anaerobic bacteria themselves, as well as transport and storage using the same.
This relates to preservation equipment. More specifically, the present invention relates to an environmental preservation composition characterized by comprising an oxygen scavenger, a high moisture absorbent, and a hygroscopic fixing agent, and a transportation/storage device using the environment preservation composition. .

[Conventional technology]

Culture media for transporting and preserving collected specimens (body fluids such as urine, blood saliva, stool, pus, tissue pieces, etc.) during clinical bacteriological tests include Stuart's transport medium and Carey Blair. (Cary Blair) transport medium and the like are known. Conventionally, when transporting and storing specimens collected for testing for anaerobic bacteria, or the anaerobic bacterial specimen itself,
The above medium is usually sterilized using high-pressure steam at 121°C and then dispensed aseptically into non-ventilated containers, but this aseptic operation after sterilization requires special equipment and is complicated. It was also time consuming. Furthermore, it is difficult to maintain an anaerobic state after dispensing, and it is not necessarily suitable for transporting and preserving specimens or anaerobic bacteria.

[Problem to be solved by the invention]

Many attempts to maintain an anaerobic environment have been reported. These methods include replacing the air in the container with nitrogen gas, adding a reducing agent as a medium component, or using special devices and equipment. For example, Kenki Porter (registered trademark: ■Clinical Supply) is a transparent glass container filled with carbon dioxide gas.
products are known. Furthermore, gas bags (manufactured by Roche) and Culturet (registered trademark: manufactured by Marion Laboratories) are known as products that require special devices and instruments. However, Kenkiporter cannot be expected to preserve bacteria for more than 2 to 3 days when taking samples in and out, and the anaerobic environment deteriorates significantly when taking samples in and out.In addition, Culturette is a disposable product that cannot be used repeatedly. Previous attempts required special culture media, instruments, special equipment, and expensive equipment, and were complicated to operate and were not common. Recently, an oxygen absorber that can be placed in a sealed culture medium container for bacterial culture has been reported (Japanese Patent Laid-Open No. 59-78682
(see publication). This provides an oxygen scavenger that easily creates an anaerobic environment with a constant carbon dioxide concentration. By placing the oxygen scavenger together with a small synthetic resin bag, it is possible to obtain an anaerobic state extremely easily. However, these techniques were only devised when culturing bacteria, and were not applicable to specimens collected for testing for anaerobic bacteria, or to the transportation and preservation of anaerobic bacteria. Further, it still required sterilization of the culture medium and subsequent dispensing into containers, which was a troublesome aseptic operation. If a container containing a culture medium containing an oxygen scavenger is subjected to aseptic processing such as high-pressure steam sterilization, moisture from the container and the culture medium will be generated as condensation, and the water-soluble components of the oxygen scavenger will elute, causing the oxygen scavenger to sterilize. As a result, the preservation effect of specimens or anaerobic bacteria becomes extremely low as a result of lowering the oxygen absorption capacity of the anaerobic bacteria and further affecting it by flowing back into the culture medium. Furthermore, when the conventional container described above is opened, the anaerobic environment is released, making it impossible to maintain the anaerobic state. Therefore, simply enclosing an oxygen absorber in specimens collected for testing for anaerobic bacteria, or in medium-containing utensils for storing and transporting anaerobic bacteria, will not provide sufficient anaerobic and appropriate moisture environment. Retention was impossible. In contrast, the anaerobic environment preservation composition of the present invention can be used for specimens collected for testing anaerobic bacteria, for which conventional oxygen scavengers alone were insufficient, or for transporting and preserving anaerobic bacteria. The present invention makes it possible to provide transport and storage equipment that can maintain a safe environment without requiring special culture media, equipment, or special or expensive equipment, and which can be operated very easily.

[Means for Solving the Problems] The present invention provides an environmental preservation composition characterized by comprising an oxygen scavenger, a high moisture absorbent, and a hygroscopic fixing agent, and a holder filled with the environment preservation composition. A container for transporting and sealing a medium-filled container for anaerobic bacterial culture with a rubber stopper equipped with
It provides storage equipment. The oxygen scavenger of the present invention includes iron, iron powder such as iron carbide, inorganic salts such as ferric salts, polyphenols such as hydroquinone and pyrogallol, reducing sugars such as glucose, or reducing sugars such as ascorbic acid. Any material containing as a main component a reducing agent selected from the group consisting of polyhydric alcohols can be used, but activated iron oxide or ascorbic acid has particularly good oxygen absorption ability. Therefore, it is preferable. As the high moisture absorbent of the present invention, starch derivatives such as acrylic acid graft polymers, or acrylic acid derivatives such as crosslinked sodium polyacrylates can be used. As the hygroscopic fixing agent of the present invention, higher alcohols, aliphatic hydrocarbons, etc. can be used, preferably with a molecular weight of 1000.
~20,000 polyethylene glycols can be used. The anaerobic environment preservation composition of the present invention can be made into various dosage forms convenient for use by mixing or layering an oxygen scavenger, a high moisture absorbent, and a hygroscopic fixing agent in appropriate proportions. be. For example, 0.3 to 2.0 parts by weight of an oxygen scavenger, 0.05 to 0.1 parts by weight of a high moisture absorbent, and 0.02 to 0 parts by weight of a hygroscopic fixing agent.
．． Mix at a ratio of 5 parts by weight and knead until uniform,
Use as a mixture. Alternatively, the mixture is made into tablets by a known tableting method. Furthermore, various dosage forms can be made, such as by layering a high moisture absorbent and a hygroscopic fixing agent on top of an oxygen scavenger to create a dosage form of a layered agent. The culture medium that can be used in the present invention is for specimens collected for testing for anaerobic bacteria, or for transporting and preserving anaerobic bacteria, and it is ensured that the number of bacteria will not change during the transportation or storage period. preferable. An example of a suitable medium composition is listed below, but other commercially available media such as Carey-Blair medium (manufactured by Eiken Chemical Co., Ltd.) may also be used. Medium formulation: Sodium thioglycolate 1.0 g Potassium chloride 0.2 g Sodium chloride 3.0 g Sodium phosphate-hydrogen
1.15 g potassium dihydrogen phosphate
0.2gB-Agar 8.0
g Sodium glycerophosphate 0.5 g L-cysteine hydrochloride 0.4 g Methylene blue
0.0006g magnesium chloride
0.1g purified water total amount 1
,000 ml (pH 7.05) Next, a storage/transport device obtained using the above-mentioned environmental preservation composition of the present invention will be explained. The device according to the present invention is characterized by comprising a container containing a culture medium suitable for anaerobic bacteria and having a sealable opening, and a rubber stopper equipped with a holder containing the environmental preservation composition of the present invention. It is something to do. The holder is a cylindrical container with an open top, and a cylindrical opening with a diameter approximately half that of the holder passes through the holder. Further, the holder is attached to the inner part of the rubber stopper in a specific direction by a convex protrusion partially provided at the bottom for positioning, and the holder holding part formed in a ring shape at the bottom of the rubber stopper allows It is held so as not to fall into the container, and can be fitted and sealed with the rubber stopper into the mouth of the container. or,
There is a slight gap between the upper surface of the holder and the rubber stopper, and the holder is air permeable to the inside of the container through the cylindrical opening of the holder. When the device according to the present invention having the above configuration is used, the condensed water eluted during sterilization such as high-pressure steam sterilization is removed from the high moisture absorbent and hygroscopic fixative contained in the environmental preservation composition of the present invention. This makes it possible to prevent a decrease in oxygen absorption capacity, prevent accumulated condensed water from coming into contact with the culture medium, and enable sterilization without affecting culture performance. In addition, when the anaerobic environment preservation composition of the present invention is in the form of a multilayer agent, excess water is absorbed by the high water absorbent, and the large amount of water causes oxidation to proceed at once, resulting in deoxidation. It also has the effect of preventing the oxygen scavenging ability of the agent from decreasing significantly. Generally, when an oxygen absorber is immersed in water, oxygen is blocked by the water and its oxygen absorption capacity is reduced. To prevent this, it is necessary to isolate the oxygen absorber from direct contact with water. For this purpose, water is absorbed by an absorbent to isolate the oxygen scavenger and make it work better.゛・Furthermore, through the pierced cylindrical opening provided in the holder inside the rubber stopper, the liquid specimen is inoculated into the culture medium from the specified prick hole on the rubber stopper with a syringe needle through the opening. This is possible without compromising the anaerobic environment.

【Example】

The present invention will be explained below based on Examples and Experimental Examples, but the present invention is not limited thereto. Example 1 Production of the environmental preservation composition (mixture) of the present invention 30 g of sodium polyacrylate (product name: Aqualic) was used as a high moisture absorbent, and polyethylene glycol 6000 (Wako Pure Chemical Industries, Ltd.) was used as a hygroscopic fixing agent. Place 50g of the product in a mortar, grind it into a homogeneous powder, and mix. Thereafter, 50 g of activated iron oxide powder (manufactured by Mitsubishi Gas Chemical Co., Ltd., trade name "Ageless") was added as an oxygen scavenger, and quickly kneaded to obtain a mixture. Example 2 Production of the environmental preservation composition (tablet) of the present invention Example 1. 1.3 g of a mixture consisting of the environmental preservation composition of the present invention obtained in
is directly compressed into tablets using a tablet machine. Example 3 Production of the environmental preservation composition (multilayer) of the present invention In the holder,
On top of 0.5 g of active iron oxide powder (Ageless) as an oxygen scavenger, 0.3 g of sodium polyacrylate as a high moisture absorbent,
and the hygroscopic fixative polyethylene glycol 6000
Layer 0.5 g of powder thoroughly mixed in a mortar. Experimental Example 1゜In order to confirm the oxygen absorption and water absorption effects of the anaerobic environment preservation composition of the present invention, manufacture of an anaerobic bacteria transport/storage device filled with the anaerobic environment preservation composition of the present invention time, 1 month, 3 months, 6 months, and 12 months from the time of manufacture.
After several months, the amount of oxygen absorbed by the oxygen scavenger and the amount of water absorbed by the high moisture absorbent were measured. The amount of oxygen absorbed was determined by measuring the degree of negative pressure in the container using a manometer, and the amount of water absorbed was determined by measuring the increased weight using an electronic balance (manufactured by Mettler). The results are shown in Table 1. As a control, a composition obtained by removing the high moisture absorbent from the environment-preserving composition of the present invention was used, and the oxygen absorption amount of the oxygen scavenger was also measured as a control.The results are also shown. From the results shown in Table 1, it was confirmed that the environmental preservation composition of the present invention has excellent oxygen absorption and moisture absorption effects. Experimental Example 2 The following anaerobic bacteria species were inoculated into an anaerobic bacteria transportation/storage device using the environmental preservation composition of the present invention. And the number of viable bacteria on day 18 was measured by pour culture. The results are shown in Table 2. Inoculum Bacteroides fragilis (B, fragilis)
sl, Hubbacterium nucleatum (F, n
ucleaturn) Pebtococcus magnus (P
, fragilis), Staphylococcus aureus (S, aureus), Streptococcus aureus
According to the results in Table 2, with the anaerobic bacteria transport and storage device using the environmental preservation composition of the present invention, no change in the number of viable bacteria was observed even after storage for 18 days. It was confirmed that it maintained a good state of preservation. Margin below

【Effect of the invention】

As mentioned above, the environmental preservation composition of the present invention is capable of absorbing condensed water generated during sterilization such as high-pressure steam sterilization using the high moisture absorbent and hygroscopic fixing agent that are the constituent components of the environmental preservation composition of the present invention. The anaerobic bacteria transport/storage device according to the present invention using this composition can absorb and fix the bacteria, and prevents a decrease in oxygen absorption capacity, as well as preventing accumulated condensed water from contacting the culture medium. It is possible to perform sterilization treatment without affecting oxygen absorption capacity and culture ability. Specimens collected for testing for anaerobic bacteria, for which oxygen scavengers alone were insufficient in the past, or transportation and storage of anaerobic bacteria, require special culture media, equipment, special equipment, and expensive equipment. without,
It has excellent effects such as being extremely easy to operate.

[Brief explanation of drawings]

FIG. 1 shows a cross-sectional view of one embodiment of the transportation/storage device of the present invention, FIG. 2 shows the same (a front view thereof, and FIG. 3 shows a back view of the same without the container).

Claims (1)

[Scope of Claims] 1) An environmental preservation composition characterized by comprising an oxygen scavenger, a high moisture absorbent, and a hygroscopic fixing agent. 2) The oxygen scavenger contains iron oxide or ascorbic acid as a main component. 3) The environment-preserving composition according to claim 1, characterized in that the high moisture absorbent is an acrylic acid derivative or a salt thereof 4) The environment-preserving composition according to claim 1, characterized in that 5) The environment-preserving composition according to claim 1, wherein the hygroscopic fixing agent is polyethylene glycol. 6) The environment-preserving composition according to claim 1, characterized in that anaerobic bacteria Transport/storage device formed by sealing a container filled with culture medium 7) The holder has a cylindrical shape with an opening at the top, and consists of a accommodating portion and a cylindrical penetrating opening portion having a diameter approximately half that of the holder. The transportation/storage device according to claim 6, characterized in that: