JP6966818B1 - Manufacturing method of aqueous solution for sterilization - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing an aqueous solution for sterilization having high bactericidal activity and less side effects on the human body or the like. A method for producing an aqueous solution for sterilization, which comprises a step of preparing a mixed solution of sulfuric acid (H2SO4) and heavy water (D2O) so that the heavy water component contains 1.5 to 20% by weight, and water (H2O). ) Is used to dilute and prepare the mixed solution so that the pH is 0.5 or more and less than 2.0. Further, a step of weighing and preparing sulfuric acid and heavy water so that the content of the heavy water component is 1.5 to 20% by weight, and when water is added to the prepared sulfuric acid and heavy water. It has a step of weighing and preparing water for adjusting the pH to 0.5 or more and less than 2.0, and a step of mixing the prepared weighed sulfuric acid, heavy water and water. [Selection diagram] Fig. 5

Description

The present invention relates to a method for producing an aqueous solution for sterilization.

Sterilization liquids are used in many situations such as sterilization of living environment, sterilization of medical equipment, and sterilization of food containers. Its usage is wide-ranging. For example, the operation of wiping with a cloth, the operation of pouring a sterilizing solution into a tank and immersing surgical instruments in it for ultrasonic cleaning, the operation of shower cleaning a food container, and the operation of atomizing indoors. It is an operation of spraying in the air.

The most important property of the bactericidal solution is its high bactericidal activity. The more toxic the bactericidal solution is, the more effective it is, so it can be said that the higher the toxicity, the higher the performance. On the other hand, when handling the sterilizing solution, if the side effects on the person handling it are high, it becomes difficult to handle. In general, the higher the bactericidal activity of the bactericidal solution, the higher the side effects in handling. Therefore, in reality, a bactericidal solution having as high a bactericidal activity as possible is used with the acceptable side effects as the limit.

As an acidic sterilizing solution, a sterilizing solution containing hypochlorous acid as a main component and a sterilizing solution containing peracetic acid and / or acetic acid as a main component are generally known. The sterilizing solution described in Patent Document 1 is a sterilizing solution containing chloric acid as a main component, and is either a chloric acid and an inorganic acid or an inorganic acid salt, or an organic acid or an organic acid salt, or two or more kinds thereof. It is a sterilizing solution containing a single substance or a combination thereof, and is used as a pretreatment bactericidal agent for food processing. This sterilizing solution is produced by adjusting and maintaining the pH of sodium chlorate to 2.3 to 3.4.

The sterilizing solution described in Patent Document 2 is a peracetic acid-based sterilizing composition solution containing peracetic acid, acetic acid, hydrogen peroxide, and catalase enzyme, and the sterilizing solution has a pH in the range of 2.6 to 5.0. It is characterized by being used for sterilizing food containers and the like.

Japanese Unexamined Patent Publication No. 2013-100436 International Publication WO2014 / 020961

A bactericidal solution containing sodium hypochlorite as the main component is commonly used, but it is said that if it gets on the skin or gets into the eyes, delayed treatment may lead to skin necrosis or blindness. ing. In addition, since it is corrosive to metals, it is necessary to avoid using it for metal products, or to thoroughly rinse and wipe it with water after use. In the bactericidal solution described in Patent Document 1, the pH is adjusted to 2.3 to 3.4 in order to compromise between bactericidal activity and side effects.

Peracetic acid and / or a bactericidal solution containing acetic acid as a main component is used as a high-level bactericidal agent for sterilizing surgical instruments such as endoscopes, but it adheres to the skin and is exposed to steam on the human body. It is said that the adverse effects of the product should be noted, and sufficient rinsing after use is required. In the bactericidal solution described in Patent Document 1, the pH is adjusted to 2.6 to 5.0 to compromise the bactericidal activity and side effects.

Acidic bactericidal solutions show high bactericidal activity when the pH is less than 2.0, but they have an adverse effect on the human body.
Since there is an adverse effect on surgical instruments and cleaning tanks, lowering the pH value makes side effects practically unacceptable and makes commercialization difficult.

An object of the present invention is to provide a method for producing an aqueous solution for sterilization, which has high bactericidal activity and has few side effects on the human body and the like.

The present invention (1) is a method for producing an aqueous solution for sterilization, which comprises sulfuric acid (H ₂ SO ₄ ) and heavy water (D _2).
A step of preparing a mixed solution containing 1.5 to 20% by weight of a heavy water (D ₂ O) component with O), and a pH of 0.5 or more of the mixed solution using _{water (H 2 O).} It is a method for producing an aqueous solution for sterilization, which comprises a step of diluting and preparing so as to be less than 2.0.

Sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O), heavy water (D ₂ O) component is 1.5 to 20% by weight
Make a mixed solution so that it contains, and dilute this mixed solution with water to a pH of 0.5 or more and less than 2.0 _{to alleviate the side effects of sulfuric acid (H 2} SO ₄ ) on the human body, etc. It can be a bactericidal solution having high bactericidal activity due to low pH.

Sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O), heavy water (D ₂ O) component is 1.5 to 20% by weight
The mixture was prepared to contain, but pH is not necessarily clear side effect of diluting the prepared dilute sulfuric acid decreases mechanisms to be less than 0.5 to 2.0, in heavy water (D 2 _O) It is presumed to be due to the so-called deuterium effect that slows down the reaction rate due to the contained deuterium component.

Since the heavy water (D ₂ O) component is expensive, _{the upper limit of the heavy water (D 2} O) component contained in the diluent is preferably less than 20% by weight, and 19% by weight or less, 18% by weight or less, and 17% by weight. Less than,
16% by weight or less, 15% by weight or less, 14% by weight or less, 13% by weight or less, 12% by weight or less,
11% by weight or less, 10% by weight or less, 9% by weight or less, 8% by weight or less, 7% by weight or less, and 6% by weight or less are more preferable in this order. The lower limit of heavy water _(D 2 O) component is preferably not less than 1.5 wt%, 2 wt% or more, 3% by weight or more, 4% by weight or more, in turn, is more preferable. This is to ensure the side effect reduction effect of the deuterium component.

The upper limit of the diluted pH is preferably 2.0 or less, 1.9 or less, 1.8 or less, 1.7 or less, 1.6 or less, 1.5 or less, 1.4 or less, 1.3 or less, 1.2 or less, 1.1 or less, 1.
0 or less, 0.9 or less, 0.8 or less, 0.7 or less, and 0.6 or less are more preferable in this order. This is because the sterilizing water of the present application uses dilute sulfuric acid as a basic substance, and therefore, the smaller the pH, the higher the attack power against bacteria and the like. If the lower limit of pH is 0.5 or more, it is preferable because it can effectively sterilize bacteria and the like that require strong bactericidal activity. However, depending on the type of bacteria to be sterilized, the pH can be increased to more than 0.5 if the sterilizing effect is sufficient even if the pH is higher than 0.5.

The present invention (1) is also effective in terms of pH stability. Sulfuric acid is basically non-volatile and its concentration does not change easily, so its pH is stable. Further, it is presumed that since the sterilizing water of the present application contains a deuterium component, the water of the diluted solution is also difficult to evaporate. Diluted water (H ₂₎
O) is replaced partially with deuterium (D) of heavy water _(D 2 O), it is considered to be because the HDO.

The present invention (2) is a method for producing an aqueous solution for sterilization, which comprises sulfuric acid (H ₂ SO ₄ ) and heavy water (D _2).
O), a step of weighing and preparing _{heavy water (D 2 O) with sulfuric acid (H 2} SO ₄ ) and heavy water (D 2 O) so as to have a predetermined content containing 1.5 to 20% by weight of the heavy water (D _{2 O) component. 2. When} water (H ₂ O) is added to the prepared sulfuric acid (H ₂ SO ₄ ) and heavy water (D 2 O) weighed, the pH is 0.5 or more.
_{The step of weighing and preparing water (H 2} O) which is less than 0 and for making a predetermined pH, and the weighed prepared sulfuric acid (H ₂ SO ₄ ), heavy water (D ₂ O) and water (H _2). It is a method for producing an aqueous solution for sterilization having a step of mixing O).

In the present invention (2), similarly to the present invention, the heavy water (D ₂ O) component is a predetermined content of 1.5 to 20% by weight with _{respect to sulfuric acid (H 2} SO ₄ ) and heavy water (D _{2 O).} When water (H ₂ O) is added, the pH is 0.5 or more and less than 2.0, and the pH becomes a predetermined pH, so that the same effect as that of the present invention (1) can be obtained.

Provided is a method for producing a safe aqueous solution for sterilization, which is highly effective as an aqueous solution for sterilizing bacteria and the like, and is easy to handle.

In the test of side effects using the leaves of Pachira grabula, the state at the start of the test is shown. In the test of side effects using the leaves of Pachira grabula, the state after 1 hour has passed is shown. In the side effect test using the leaves of Pachira grabula, the state after 2 hours has passed. In the side effect test using the leaves of Pachira grabula, the state after 19 hours has passed. In the side effect test using the leaves of Pachira grabula, the state after 43 hours has passed. The pH that changes with the passage of time of the sterilized water of the present application and the comparative electrolyzed water is shown. Comparison with the sterilized water of the present application The pH that changes with the passage of time when animal tissue is mixed with electrolyzed water is shown. Comparison with the sterilized water of the present application The pH that changes with the passage of time when a metal substance is mixed in the electrolyzed water is shown. Comparison with the sterilized water of the present application The pH that changes with the passage of time when a resin substance is mixed in electrolyzed water is shown.

Hereinafter, the best embodiment of the present invention will be described with reference to the figures and tables.
The sterilizing aqueous solution of the present invention _{comprises a step of preparing a mixed solution of sulfuric acid (H 2} SO ₄ ) and heavy water (D ₂ O) so that the heavy water (D ₂ O) component contains 1.5 to 20% by weight. , It is made in a manufacturing process comprising the step of preparing the mixed solution using water (H _{2 O) so that the pH is 0.5 or more and less than 2.0.} Sulfuric acid (H ₂ SO ₄ ) used was manufactured by Fuji Film Wako Pure Chemical Industries, Ltd. and had a concentration of 10% by weight. Heavy water _(D 2 O) was used as Fuji Film Wako Pure Chemical purity 99.8% Co., Ltd.. Water _(H 2 O) was used tap water. The method for producing an aqueous solution for sterilization of the present invention is composed of these three substances, but a trace component that does not affect the pH value or has an extremely small effect may be added. The pH was measured using a model METRO-51 manufactured by HORIBA, Ltd.

(Preparation of the first test solution group)
In the first test solution group, sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O) are used, and the heavy water (D ₂ O) component is 1.
5 created to contain by weight%, in which the pH of the disinfecting solution diluted prepared in water (H 2 _O) was a predetermined value.

(Preparation of the second test solution group)
In the second test solution group, sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O) are used, and the heavy water (D ₂ O) component is 3.
Prepared as containing 0 wt%, in which the pH of the disinfecting solution diluted prepared in water (H 2 _O) was a predetermined value.

(Preparation of the third test solution group)
The third test solution group contains sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O), and the heavy water (D ₂ O) component is 5.
Prepared as containing by weight%, in which the pH of the disinfecting solution diluted prepared in water (H 2 _O) was a predetermined value.

(Preparation of 4th test solution group)
In the fourth test solution group, sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O) are used, and the heavy water (D ₂ O) component is 1.
.. 5 created to contain by weight%, in which the pH of the disinfecting solution diluted prepared in water (H 2 _O) was a predetermined value.

(Test method for sterilization test)
Using SCDLP agar medium (manufactured by Nihon Pharmaceutical Co., Ltd.) as a medium for measuring the number of bacteria, the method is carried out by the pour-mixed plate cultivation method, the temperature is 36 ° C. ± 1 ° C., and the temperature is 48 ± 3 hours. As the preparation of the test bacterial solution, B
One platinum loop inoculates the test strain into HI liquid medium (Brain Heart Infusion) and inoculates at 36 ° C. for 18 to 24 hours. The bacterial solution after culturing is smeared on a Scriptic Soy plate medium and cultured at 36 ° C. for 18 to 24 hours. Scrape the cells from the plate medium after culturing, and 0.
Stir with glass beads for 3 minutes in a 1% tryptone 0.85% NaCl solution and stir to make a test bacterial solution.

As a test operation, 9 mL of a sample is inoculated with 1 mL of the test bacterial solution to prepare a test solution. The test solution is stored at room temperature, and after 2 minutes and 10 minutes of storage, the test solution is immediately diluted 10-fold with SCDLP liquid medium (manufactured by Nippon Pharmaceutical Co., Ltd.), and the viable cell count in the test solution is used as a medium for measuring the bacterial count. Measure using. Also, as a control, the same test is performed using sterile physiological saline, and the viable cell count is measured.

To confirm the inactivation of the sample, 9 mL of SCDLP liquid medium (inactivating agent) and lmL of the sample
Add lmL of the test bacterial solution to the mixture that has been shaken and stirred, store at room temperature for 10 minutes, and after storage, measure the viable cell count and confirm that there is no difference in the viable cell count. Depending on the test, the bactericidal activity was evaluated by counting the number of colonies. Also, depending on the test, Log infectious titer / 0.1 ml
Evaluation was made by law. Furthermore, the parts of the individual tests that differ from the above test methods will be described separately in the explanation of each test result. As for the evaluation, if it is recognized that the bactericidal power is killed within a few minutes, it is marked with 〇, if it is recognized that the bactericidal effect appears after 10 minutes or more, it is marked with △, and if it is judged to be insufficient as sterilizing water, ×. Distinguish by mark. Among the items corresponding to 〇, the ones that are extremely good are marked as ◎.

(Test method for side effect test)
The above-mentioned various test solutions and control solutions are dropped onto the leaves of Pachira grabula to wet them, and follow-up is performed. Follow-up is performed focusing on the color and range of discoloration.

(Test method for pH stability test)
A test solution prepared to a predetermined pH, a test solution obtained by immersing an animal tissue, a metal substance, or a resin substance in this test solution, and a control solution are allowed to stand at room temperature, and a change in pH is examined. The volume of the test solution and the control solution is 100 ml, and the test solution and the control solution are opened and allowed to stand without a lid. The animal tissue to be soaked was 5 grams of pork, the metal substance was one iron nail (4 grams), and the resin substance was a PET bottle cap (2 grams). As the control solution, hypochlorous acid water (strong acid hypochlorous acid water) purified by First Maintenance Co., Ltd. is used.

(Result of bactericidal test)
(Test 1)
Four test solutions having pH = 1.5, 2.0, 2.5, and 3.0 were used as the test solutions of the first test solution group. Escherichia coli and Staphylococcus aureus were used as sterilization targets. The results are shown in Table 1 below. In addition, in this specification, the "control test body" means the thing which was left untreated without using the test solution.
[Table 1]

(Test 2)
Four test solutions having pH = 2.0, 2.5, 3.5, and 4.4 were used as the test solutions of the first test solution group. Escherichia coli and Staphylococcus aureus were used as sterilization targets. The results are shown in Table 2 below.
[Table 2]

(Test 3)
Four test solutions having pH = 0.5, 1.2, and 2.5 were used as the test solutions of the first test solution group. Feline calicivirus was used as the sterilization target. Evaluation was performed by the Log infectious titer / 0.1 ml method. The results are shown in Table 3 below.

[Table 3]

(Test 4)
Two test solutions having a pH of 1.2 and 4.0 were used as the test solutions of the first test solution group. The subjects to be sterilized were Escherichia coli and general bacteria, and the number of colonies per 1 ml was counted. Escherichia coli was tested by JIS K 0102-1993 72.3 (deoxycholic acid salt medium method), and general bacteria were tested by JIS K 0102-1993 72.2 (standard agar medium method). The test results are shown in Table 4 below.
[Table 4]

(Test 5)
Three test solutions having a pH of 2.1, 3.2, and 4.2 were used as the test solutions of the first test solution group. The target of sterilization was Escherichia coli, and the test target bacteria were inoculated into a BHI liquid medium, cultured at 36 ° C. overnight, and then the bacterial solution was diluted 100-fold to obtain a test bacterial solution. 20 ml of the test solution was placed in an Erlenmeyer flask, and 0.1 ml of the test bacterial solution was further inoculated to prepare a test solution. After inoculation with the bacterial solution, leave it at room temperature for 2 minutes and 10 minutes, sample 1 ml of the test solution, and pour it on a standard agar medium by a 10-fold dilution method at 36 ° C.
-After culturing for 1 day, the viable cell count was measured. As a control, a similar test was conducted using sterile purified water. For the subjects, the viable cell count was measured even at the start of the test. The results are shown in Table 5 below.

[Table 5]

(Test 6)
Two test solutions having a pH of 0.5 and 2.3 were used as the test solutions of the second test solution group. Escherichia coli was used as the sterilization target. For the subjects, the viable cell count was measured even at the start of the test. The average value of 3 trials was calculated. The results are shown in Table 6 below.
[Table 6]

(Test 7)
Two test solutions having a pH of 0.5 and 2.3 were used as the test solutions of the third test solution group. Escherichia coli was used as the sterilization target. For the subjects, the viable cell count was measured even at the start of the test. The average value of 3 trials was calculated. The results are shown in Table 7 below.
[Table 7]

(Test 8)
Two test solutions having a pH of 0.5 and 2.3 were used as the test solutions of the 4th test solution group. Escherichia coli was used as the sterilization target. For the subjects, the viable cell count was measured even at the start of the test. The average value of 3 trials was calculated. The results are shown in Table 6 below.
[Table 8]

Looking at the results of tests 1 to 8, the first test solution group (15% by weight of heavy water), the second test solution group (30% by weight of heavy water), the third test solution group (5% by weight of heavy water) and the first test solution group. In any of the four test liquid groups (heavy water is 1.5% by weight), the bactericidal activity of the sterilizing aqueous solution having a pH of 0.5 or more and less than 2.0 shows good results.

(Results of side effect test)
Prepare three leaves pachira glabra was examined whether dropwise disinfecting aqueous sulfuric acid (H 2 _{SO 4),} leaves pachira glabra over time how changes. FIG. 1 shows a state at the start of the test. Of the three columns, the leftmost column is for H ₂ SO ₄ that does not contain heavy water.
Comparative examples with pH of 0.5, 2.0, and 2.3 are shown in order from the top. The middle row shows examples with 1.5% by weight of heavy water and 0.5, 2.0, and 2.3 pH in order from the top. The rightmost column is an example in which heavy water is 5% by weight, and the pH is 0.5, 2.0 in order from the top.
An embodiment of 2.3 is shown.

FIG. 2 shows the state after one hour has passed. The _{pH of the comparative solution was H 2} SO ₄ and the pH was 0.5, and the inside of the white dotted line turned brown. No noticeable changes were observed in the other 8 samples.

FIG. 3 shows a state after 2 hours have passed. In _{the comparative solution having H 2} SO ₄ and a pH of 0.5, the region that had turned brown was widened as shown by the white dotted line. The white dotted line in the center of the top of the middle row with 1.5% by weight of heavy water and 0.5 of pH was slightly discolored to brown.

FIG. 4 shows the state after 19 hours have passed. pH is of the three top of 0.5, damage is greatest those that do not contain the left end of the D _{2 O,} damage as go right had been reduced. The same was true for the pH of 2.0. The discoloration seen in the white dotted line area on the far right was very small, and there was almost no discoloration. The same was true for those having a pH of 2.3.

FIG. 5 shows the state after the lapse of 43 hours. The tendency was the same as that after 19 hours in FIG. Looking at FIGS. 1 to 5, it can be seen that by impregnating sulfuric acid (H ₂ SO ₄ ) with heavy water, side effects (damage) to cell tissues are reduced. Even if the pH is 0.5 and the acidity is very strong, side effects do not appear after 1 hour and only slightly appear after 2 hours. This shows that it is very easy to handle. For example, even if it gets on the skin, it can be handled safely because the reaction is slow and there are no immediate side effects. Normally, when cleaning and sterilizing surgical instruments and the like, it is common to immerse the instruments and the like in sterilizing water and cleaning water contained in a tank and perform ultrasonic cleaning for several to 10 minutes. When sterilizing water is used for sterilization, damage to instruments and the like does not occur in a short time, and side effects can be completely removed by rinsing. Therefore, using the sterilizing water of the present invention facilitates handling.

(Test result of pH stability test)
The test liquids of the first test liquid group, which have pH of 0.5, 1.0, 2.0, and pH of 2.
FIG. 6 shows the results of examining the change in pH with the lapse of one week from immediately after the test solution in which the animal tissue, the metallic substance, and the resin substance were immersed in the weakly acidic electrolyzed water (comparative example) of No. 5 by allowing them to stand still. ~ Figure 9
Shown in.

FIG. 6 shows changes in the elapsed time (days) and pH of each test solution. PH with sterilized water of the present invention
At 0.5, 1.0 and 2.0, the pH did not change at all. In comparative electrolyzed water, the initial pH
Was 2.5, but increased by 0.3 after 1 day, and increased by 0.4 after 7 days, 2.9.
It became.

FIG. 7 shows changes in the elapsed time (days) and pH of each test solution when the animal tissue was mixed. When the pH of the sterilized water of the present invention was 0.5 and 1.0, the pH did not change at all. pH
The sterilized water with a pH of 2.0 gradually increased after one day, and the pH was initially 2.5.
After 7 days, it increased by 1.2 to 3,2. In comparative electrolyzed water, the pH is 1.0 after one day.
After 7 days, the pH was initially 2.5, but increased by 1.7 to 4.2.

FIG. 8 shows changes in the elapsed time (days) and pH of each test solution when a metallic substance is mixed. When the pH of the sterilized water of the present invention was 0.5 and 1.0, the pH did not change at all. pH
The sterilized water with a value of 2.0 changed slightly between 5 days and 6 days, but on the 7th day it was the original 2
.. It became 0 and there was no change. In the comparative electrolyzed water, the pH gradually increased after 2 days, and after 7 days, the pH was initially 2.5, but increased by 0.4 to 2.9.

FIG. 9 shows changes in the elapsed time (days) and pH of each test solution when the resin substance is mixed. When the pH of the sterilized water of the present invention was 0.5, 1.0 and 2.0, the pH did not change at all. In the comparative electrolyzed water, the pH was initially 2.5, but after 7 days, it increased by 0.4 2
.. It became 9. In the comparative electrolyzed water, the pH gradually increased after 2 days, and initially p after 7 days.
What H was 2.5 increased by 0.4 to 2.9.

In the sterilized water of the present invention, the pH is set to 0.5 or more and less than 2.0. When the pH was 1.0 or less, there was no change even in the case of contamination with animal tissue. PH 2 mixed with animal tissue
.. Even if it was 0, there was no change until the lapse of 1 day, and the pH increased by 0.4 after the lapse of 2 days. In the case of contamination with a metallic substance or a resin substance, there was almost no change if the pH was 2 or less. _{Sulfuric acid (H 2} SO ₄ ), which does not contain heavy water components, showed changes even when mixed with metallic substances or resin substances, and those mixed with animal tissues were significantly changed at an early stage.

Since the bactericidal aqueous solution of the present invention has both strong bactericidal activity and low side effects, it can be suitably used in various places.

Claims (2)

It is a method for producing an aqueous solution for sterilization.
Sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O), heavy water (D ₂ O) component is 1.5 to 20% by weight
Steps to create a mixture to include,
Water (H 2 _O) production method of disinfecting solution and a step of diluting preparing the mixture to pH 0.5 to 1.2 with. It is a method for producing an aqueous solution for sterilization.
Sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O), heavy water (D ₂ O) component is 1.5 to 20% by weight
A step of weighing and preparing _{sulfuric acid (H 2} SO ₄ ) and heavy water (D ₂ O) so as to contain a predetermined content, and
When water (H ₂ O) is added to the weighed prepared sulfuric acid (H ₂ SO ₄ ) and heavy water (D ₂ O), the pH is 0.5 or more and 1.2 or less , and the pH is set to a predetermined pH. Steps to weigh and prepare water (H _{2 O) for}
A method for producing an aqueous solution for sterilization, which comprises a step of mixing the prepared weighed sulfuric acid (H ₂ SO ₄ ), heavy water (D ₂ O) and water (H _{2 O).}

Images