JP2631208B2 - Method for producing dry powder medium - 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 producing a dry powder medium.

[0002]

[Prior art]

[0003] Generally, a dry powder medium is composed of peptone, sugar,
Phosphoric acid, organic acids, amino acids, etc. are composed of highly hygroscopic components, and the particle size of each component is not uniform.
In order to obtain a uniform mixture, all components were crushed and mixed using a crusher such as a ball mill. The dry powder media produced in this way have a very small particle size and consequently a large surface area of the particles. For this reason, the conventional dry powder medium increases the particle surface area,
The hygroscopic medium component became more hygroscopic, and the oxidizable medium component became more oxidizable.

That is, the conventional dry powder medium is liable to form a solid mass or decrease in pH due to moisture absorption or oxidation which leads to performance degradation, and if the storage state is poor, it will not easily withstand practical use. . Further, even if a major problem does not occur in quality, a change in weight due to moisture absorption is not preferable because it causes inaccurate weighing.

At present, as a means for avoiding such a problem, only a passive and troublesome method such as taking a strict moisture-proof measure or a sealing measure using a desiccator or the like is known. Further, the dry powder medium having a small particle size has drawbacks in use such as being easily scattered at the time of weighing and being hardly dissolved in water.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a new technique for suppressing the hygroscopicity and oxidizability of a dry powder medium.

[0007]

The present invention is a method for producing a dry powder medium comprising the following steps 1) and 2). 1) A step of spraying a liquid onto a mixed and fluidized powdery medium component to form fine particles 2) A finely divided medium component is selected from the group consisting of natural pastes, synthetic pastes, sugars, and fats and oils At least one
A process of spraying and drying a solution in which various materials are dissolved

[0008] As the natural paste in the present invention, agar,
Gelatin, collagen, tamarind gum, gum arabic, tragacanth gum, pectin and the like are used as synthetic pastes such as carboxymethyl cellulose (hereinafter abbreviated as CMC), sodium salt of carboxymethyl cellulose (hereinafter abbreviated as CMC-Na), methyl cellulose (hereinafter abbreviated as MC). ), Sodium salts of methylcellulose (hereinafter abbreviated as MC-Na), polyvinylpyrrolidone (hereinafter abbreviated as PVP), polyvinyl alcohol (hereinafter abbreviated as PVA) and the like, and disaccharides such as lactose as saccharides , Trisaccharides, sucrose and the like, and oils and fats include sucrose fatty acid ester, monoglyceride, Tween 80, lecithin and the like.

The above various materials can be used alone or in any combination. Particularly preferred materials are agar, gelatin, CMC, CMC-Na, collagen and the like. In addition, when a paste is used, problems such as adhesion of the paste to the inner wall of the container during the high-pressure sterilization of the culture medium and adhesion of the dye to the further attached paste may occur, so that PVP and Tween 80, It is preferable to use a combination of CMC and sucrose fatty acid ester, gum arabic and lecithin, gum tragacanth and monoglyceride, or collagen and Tween 80. When the medium component is a liquid medium (broth), it is preferable to use sugars and fats and oils that do not solidify at room temperature than agar or gelatin.

At this time, if the medium component contains peptone, it is advantageous to use a peptone solution as the spray solution in step 1) for the following reasons. That is, in general, peptone is produced through the steps of digesting, concentrating, and spray-drying the raw material. However, when spray drying, which is the final step, is also used as the step of spraying the medium material solution, the production process is simplified and peptone is used. This is because prevention of thermal deterioration can be expected.

Examples of the medium material used in the present invention include various conventionally used medium materials such as various peptones, sugars, phosphoric acids, organic acids, amino acids, selection agents, and indicators. Furthermore, the medium composition is not limited at all, and is applicable to various medium compositions currently provided as a dry powder medium, for example, SS agar medium, desoxycholate medium, BGLB medium, tryptic soy agar medium, heart infusion medium and the like. It is possible.

According to the method for producing a dry powder culture medium of the present invention, the medium components acquire sufficient moisture absorption resistance and oxidation resistance. However, when the surface area per unit weight is reduced by increasing the particle diameter, the performance is further improved. Can be enhanced.

[0013]

The step 2) according to the present invention gives the particles comprising the medium components an effect of absorbing moisture or preventing oxidation. In addition, when the size of the medium component is increased as an ancillary action, it also acts as an excipient and a binder for maintaining its shape.

[0014]

【Example】

Embodiment 1 FIG. Agar Spray SS Agar Medium An SS agar medium having the following composition was sprayed with an agar solution per liter of the medium. Meat extract (Eiken Chemical) 5 g Brilliant green (Merck) 0.00033 g Neutral red (Kishida Chemical) 0.025 g Agar (Eiken Chemical) 13.5 g Peptone (Eiken Chemical) 5 g Lactose 10 g Bile salt No. 2 (Eiken Chemical Co., Ltd.) 8.5 g Sodium citrate 8.5 g Iron citrate 1 g

A peptone solution in which peptone is dissolved in twice the amount of purified water, a pigment solution in which brilliant green and neutral red are dissolved in 30 times the amount of purified water, and a part of agar (about 1 g)
Was dissolved to prepare a 1 W / V% agar solution. While blowing at 90 ° C., the undissolved one of the above compositions is mixed and fluidized, and the peptone solution, the dye solution, and finally the agar solution are sprayed and then dried to obtain an agar-sprayed SS agar medium. Was. The water content immediately after preparation of the agar-sprayed SS agar medium produced in this example was 2.4% (4.5% for the conventional product).

Embodiment 2 FIG. Agar-sprayed desoxycholate medium A desoxycholate medium having the following composition per liter of the medium was sprayed with an agar solution. Peptone (Eiken Chemical) 10 g Lactose 10 g Sodium desoxycholate (Eiken Chemical) 1 g Sodium chloride 5 g Dipotassium phosphate 2 g Iron ammonium citrate 2 g Neutral red 0.033 g Agar (Eiken Chemical) 15 g

A peptone solution in which peptone was dissolved in twice the volume of purified water, a neutral red solution in which neutral red was dissolved in 30 times the volume of purified water, and a portion (about 1 g) of agar were dissolved in purified water. A 1 W / V% agar solution was prepared. Except that a neutral red solution was used instead of the dye solution. An agar-sprayed desoxycholate medium was obtained by the same conditions and operations as in the above. The water content immediately after preparation of the agar-sprayed desoxycholate medium produced according to this example was 2.4% (3.8% for the conventional product).

Embodiment 3 FIG. Lactose sprayed BGLB medium A BGLB medium having the following composition per liter of the medium was sprayed with a lactose solution. Peptone (Eiken Chemical) 10 g Lactose 10 g Beef bile powder (Mikuni Chemical 20 g Brilliant Green (Merck) 0.0133 g

A lactose solution in which a part (about 3 g) of lactose was dissolved in an equal amount of purified water, and a brilliant green solution in which brilliant green was dissolved in a 40-fold amount of purified water were prepared. While blowing at 90 ° C., the undissolved one of the above compositions was mixed and fluidized, and a brilliant green solution was sprayed first, and then a lactose solution was sprayed and dried to obtain a lactose sprayed BGLB medium. The water content immediately after the preparation of the lactose sprayed BGLB medium produced according to this example was 2.0.
% (2.9% for the conventional product).

Embodiment 4 FIG. 1. CMC-Na sprayed desoxycholate medium Under the same conditions as described above, a peptone solution and a neutral red solution were prepared, and 2,000 ml of a 1 W / V% solution of CMC-Na was prepared.

While blowing at 90 ° C., the undissolved one of the above compositions is mixed and fluidized, first spraying a peptone solution, then neutral red solution, and finally CMC-N
The solution a was sprayed and then dried to obtain a CMC-Na sprayed desoxycholate medium. CM manufactured according to the present embodiment
The water content immediately after preparation of the C-Na sprayed desoxycholate medium was 2.5% (3.8% for the conventional product).

Embodiment 5 FIG. 1. Desoxycholate medium sprayed with sucrose fatty acid ester A desoxycholate medium having the same composition as in Example 1 was sprayed with a sucrose fatty acid ester solution. 2. A peptone solution and a neutral red solution were prepared under the same conditions as described above, and 1000 ml of a 0.5% W / V solution of sucrose fatty acid ester was prepared.

While blowing at 80 ° C., the undissolved one of the above compositions is mixed and fluidized, first spraying a peptone solution, then spraying a neutral red solution, and finally spraying a sucrose fatty acid ester solution, followed by drying. In this manner, a desoxycholate medium sprayed with a sucrose fatty acid ester was obtained. The water content immediately after the preparation of the sucrose fatty acid ester-sprayed desoxycholate medium produced in this example was 2.3% (3.8% for the conventional product).

Embodiment 6 FIG. 1. Gum arabic sprayed desoxycholate medium A desoxycholate medium having the same composition as was sprayed with a gum arabic solution. 2. A peptone solution and a neutral red solution were prepared under the same conditions as described above, and 1000 ml of a 3 W / V% gum arabic solution was prepared. While blowing at 80 ° C., the undissolved one of the above compositions is mixed and fluidized, and the peptone solution is sprayed first, the neutral red solution is sprayed, and finally the gum arabic solution is sprayed and dried to spray gum arabic. Dessociate medium was obtained. The water content immediately after the preparation of the gum arabic sprayed desoxycholate medium manufactured according to this example is 2.5% (the conventional product is 3.8%).
%)Met.

Comparative Example 1. Comparison of hygroscopicity Example 1 ~ 6. The hygroscopicity of the dry powder medium according to the present invention obtained in the above was compared with the dry powder medium of the corresponding conventional dosage form as a control. The method is as follows. 10 g of each dry powder medium is placed in a Petri dish, and the humidity is 60% RH at room temperature.
And the weight after 10, 30, and 60 minutes was measured. The results are as shown in FIGS.

Comparative Example 2 Comparison of pH drop rate Example 1 ~ 6. P of the dry powder medium according to the present invention obtained in
The H reduction rate was compared with the dry powder medium of the corresponding conventional dosage form as a control. The method is as follows.

In order to prevent quality deterioration due to light, 100 g of each dry powdered medium is sealed in an aluminum-evaporated bag, stored at room temperature, dissolved in purified water after 1, 2 and 3 years, and then dissolved in purified water.
H was measured. The results are as shown in FIGS.

Comparative Example 3 Comparison of changes in supportive power for microbial growth ~ 6. The changes in the microbial growth supporting capacity of the dry powder medium according to the present invention obtained in the above were compared with the dry powder medium of the corresponding conventional dosage form as a control. The method is as follows.

Comparative Example 2 Each medium stored under the same conditions as described above was opened after 1, 2 and 3 years, dissolved and sterilized, and the growth degree was compared by pour culture using several kinds of standard strains. The inoculum amount was 10 ⁻³ , 10 ⁻⁴ , 1
The dilution was set to four serial dilutions of 0 ^-5 and 10 ^-4 , and the culture conditions were 37 ° C. and 24 hours. 37 for BGLB medium
After culturing at 24 ° C. for 24 hours, the turbidity of the medium was visually observed and determined. The results are shown in Tables 1 to 6. The standard strains tested are as follows.

SS agar medium: Shigella flexner 2C1 (Shigella flexneri) Shigella flexner SHGL-2 Shigella sonnei 131 Shigella sonnei 133 Salmonella typhi Salmonella typhi desoxycholate medium Escherichia coli NI Esherichia coli ・ Escherichia coli FSCH-263 ・ Salmonella entrididis ・ Citrobacter freundii BGLB medium ・ Escherichia coli NIHJ ・ Escherichia coli ES-63 ・ Escherichia coli ES-2 ・ Escherichia coli ES-2 4

[0031]

[Table 1]

[0032]

[Table 2]

[0033]

[Table 3]

[0034]

[Table 4]

[0035]

[Table 5]

[0036]

[Table 6]

[0037]

The dry powder medium produced according to the present invention has a low hygroscopicity and a low rate of decrease in pH. And Comparative Example 2. It is clear from all the results. In particular, Comparative Example 1. Of agar-sprayed BGLB medium (FIG. 3) and gum arabic sprayed desoxycholate medium (FIG. 6), and Comparative Example 2. (FIGS. 7 to 12), the improvement of the pH reduction rate is remarkable. Therefore, according to the present invention, the storage stability of the dry powder medium can be dramatically improved.

This is shown in Comparative Example 3. Flexner SHGL-2 and Shigella sonnei 133 on SS agar medium (Table 1) and Escherichia coli NIHJ and ES-63 on desoxycholate medium (Table 2).
This is also evidenced by the difference in one-order growth support in the third year data.

Further, the dry powder medium obtained by the production method of the present invention can maintain uniform quality even if the particles are coarser than conventional products, so that the particles are less likely to be scattered and easily dissolved at the time of weighing. It becomes possible. Further, the dry powder culture medium produced by the production method of the present invention has a small weight change due to moisture absorption, and thus enables accurate weighing.

[Brief description of the drawings]

FIG. 1 is a graph showing the results of comparing the hygroscopicity of an agar-sprayed SS agar medium manufactured according to the present invention with a conventional dry powder medium. The vertical axis indicates the water increase rate, and the horizontal axis indicates the standing time.

FIG. 2 is a graph showing the results of comparing the hygroscopicity of an agar-sprayed desoxycholate medium manufactured according to the present invention with a conventional dry powder medium. The vertical axis indicates the water increase rate, and the horizontal axis indicates the standing time.

FIG. 3 is a graph showing the results of comparing the hygroscopicity of a lactose sprayed BGLB medium produced according to the present invention and a conventional dry powder medium. The vertical axis indicates the water increase rate, and the horizontal axis indicates the standing time.

FIG. 4 is a graph showing a comparison result of hygroscopicity between a CMC-Na sprayed desoxycholate medium manufactured according to the present invention and a conventional dry powder medium. The vertical axis indicates the water increase rate, and the horizontal axis indicates the standing time.

FIG. 5 is a graph showing the results of comparing the hygroscopicity of a desoxycholate medium sprayed with a sucrose fatty acid ester prepared according to the present invention and a conventional dry powder medium. The vertical axis indicates the water increase rate, and the horizontal axis indicates the standing time.

FIG. 6 is a graph showing the results of comparing the hygroscopicity of a gum arabic sprayed desoxycholate medium manufactured according to the present invention with a conventional dry powder medium. The vertical axis indicates the water increase rate, and the horizontal axis indicates the standing time.

FIG. 7 is a graph showing a comparison result of a pH reduction ratio between an agar-sprayed SS agar medium manufactured according to the present invention and a conventional dry powder medium. The vertical axis indicates the change in pH value, and the horizontal axis indicates the storage period.

FIG. 8 is a graph showing a comparison result of a pH reduction ratio between an agar-sprayed desoxycholate medium manufactured according to the present invention and a conventional dry powder medium. The vertical axis indicates the change in pH value, and the horizontal axis indicates the storage period.

FIG. 9 is a graph showing a comparison result of a pH decrease rate between a lactose sprayed BGLB medium manufactured according to the present invention and a conventional dry powder medium. The vertical axis indicates the change in pH value, and the horizontal axis indicates the storage period.

FIG. 10 is a graph showing a comparison result of a pH decrease ratio between a CMC-Na sprayed desoxycholate medium manufactured according to the present invention and a conventional dry powder medium. The vertical axis indicates the change in pH value, and the horizontal axis indicates the storage period.

FIG. 11 is a graph showing a comparison result of a pH decrease rate between a desoxycholate medium sprayed with a sucrose fatty acid ester manufactured according to the present invention and a conventional dry powder medium. The vertical axis is p
The horizontal axis indicates the change in the H value and the storage period.

FIG. 12 is a graph showing a comparison result of a pH decrease rate between a gum arabic sprayed desoxycholate medium manufactured according to the present invention and a conventional dry powder medium. The vertical axis indicates the change in pH value, and the horizontal axis indicates the storage period.

Continued on the front page (51) Int.Cl. ⁶ Identification number Reference number in the agency FI Technical display location C12R 1:42) (C12N 1/20 C12R 1:19)

Claims (8)

(57) [Claims] 1. A method for producing a dry powder medium comprising the following steps 1) -2) 1) A step of spraying a liquid onto a mixed and fluidized powdery medium component to form fine particles 2) Fine particles At least one selected from the group consisting of natural pastes, synthetic pastes, sugars, and fats and oils.
A process of spraying and drying a solution in which various materials are dissolved 2. A natural paste comprising agar, gelatin, collagen, tamarind gum, gum arabic, gum tragacanth,
2. The method for producing a dry powder medium according to claim 1, wherein the method is selected from the group consisting of pectin and pectin. 3. The dry powder of claim 1, wherein the synthetic paste is selected from the group consisting of carboxymethylcellulose, carboxymethylcellulose sodium salt, methylcellulose, methylcellulose sodium salt, polyvinylpyrrolidone, and polyvinylalcohol. Medium production method 4. The method according to claim 1, wherein the saccharide is selected from the group consisting of a disaccharide, a trisaccharide, and a sucrose. 5. The method according to claim 4, wherein the disaccharide is lactose. 6. The method according to claim 1, wherein the fats and oils are selected from the group consisting of sucrose fatty acid esters, monoglycerides, Tween 80, and lecithin. 7. A method for stabilizing a dry powder medium comprising the following steps 1) -2) 1) A step of spraying a liquid onto a mixed and fluidized powdery medium component to form fine particles 2) Fine particles At least one selected from the group consisting of natural pastes, synthetic pastes, sugars, and fats and oils.
A process of spraying and drying a solution in which various materials are dissolved 8. A method for preventing moisture in a dry powder medium comprising the following steps 1) -2) 1) A step of spraying a liquid to a mixed and fluidized powdery medium component to form fine particles 2) Fine particle formation At least one selected from the group consisting of natural pastes, synthetic pastes, sugars, and fats and oils.
A process of spraying and drying a solution in which various materials are dissolved