JP5230549B2 - Cleaning method for medical equipment - Google Patents

Description

  The present invention relates to a method for cleaning a medical instrument.

  Blood instruments, body fluids, and the like adhere to steel instruments such as scissors, forceps, and insulators and medical instruments such as rigid and flexible endoscopes after being used for examination, treatment, surgery, and the like. These stains may contain bacteria, viruses, pathogenic proteins such as abnormal prions, etc., and must be reused after being cleaned, disinfected and sterilized. At that time, if the cleaning is insufficient and dirt remains, it is said that the effect as much as disinfection and sterilization cannot be expected cannot be achieved and complete disinfection and sterilization may not be achieved. In addition, the protein remaining in the washing process is very strong and difficult to remove due to protein denaturation caused by sterilization with a disinfectant such as glutaraldehyde or peracetic acid used in the next process or high-pressure steam or ethylene oxide. Become. Therefore, the use of a neutral enzyme detergent is recommended in order to surely clean the protein soil before the disinfection or sterilization process (Patent Document 1). In recent years, it has been found that alkaline detergents have higher detergency than neutral enzyme detergents, and therefore, alkaline detergents are also used in washing using a washing machine.

  The detergency using these cleaning agents at the medical site is usually judged visually, and if dirt that can be visually confirmed remains, re-washing is performed. However, in recent research, when instruments used in the medical field are washed with a commercially available neutral enzyme cleaner or alkaline cleaner, most of the instruments are visually free of residual stains. When it is observed in detail using a fluorescence microscope after staining, it is revealed that the adhered protein stain remains (Non-Patent Document 1).

JP 2001-31999 A

Journal of Hospital Infection (2008) 68, 52-58

  The blood-derived protein soil adhering to the substrate surface is very difficult to remove, and the amount seems to be about several micrograms per square centimeter mail. Such blood-derived protein stains adhering to the substrate surface cannot be detected (remaining confirmation) by, for example, the visual determination method employed in the example of Patent Document 1 or the determination method using a commercially available measurement kit. is there. Moreover, in the Example of patent document 1, the serum which is the blood component from which the factor related to the coagulation of blood was excluded is used, and the blood dirt which coagulates and adheres does not generate | occur | produce. Therefore, Patent Document 1 does not teach a higher-level cleaning that can remove blood-derived protein soil adhered to the substrate surface. In addition, the detergent composition as described in Patent Document 1 cannot remove blood-derived protein stains adhered to the substrate surface.

  In order to prevent nosocomial infections caused by medical devices, it is important to sufficiently remove such a minute and strong protein stain, but no cleaning method that can achieve this has been found so far.

  The subject of this invention is providing the washing | cleaning method which can remove efficiently the protein dirt which adhered to the medical device.

  The present invention comprises (a) monoethanolamine [hereinafter referred to as component (a)], (b) alkaline protease [hereinafter referred to as component (b)], (c) nonionic surfactant [hereinafter referred to as component (c). And a treatment liquid containing water and having a pH of 9 or more is brought into contact with a medical instrument on which protein soil adheres.

  ADVANTAGE OF THE INVENTION According to this invention, the washing | cleaning method which can remove effectively the protein stain | pollution | contamination which cannot be visually observed adhered to the base material which adhered to the medical device can be provided.

  The component (a) is monoethanolamine. Further, an alkali agent other than the component (a) [hereinafter referred to as the component (a ′)] can also be used. As the component (a ′), one or more selected from alkanolamines other than monoethanolamine, alkali metal hydroxides, carbonates, phosphates, and silicates can be blended. Examples of alkanolamines other than monoethanolanolamine include diethanolamine, triethanolamine, N-methylpropanolamine, and 2-amino-2-methyl-1-propanol. Alkali metal hydroxides, carbonates, phosphates and silicates include potassium hydroxide, sodium hydroxide, potassium carbonate, sodium carbonate, potassium phosphate, sodium phosphate, No. 1 potassium silicate, No. 1 sodium silicate No. 2, potassium silicate, No. 2, sodium silicate, ortho orthosilicate, ortho orthosilicate, and the like.

  The ratio of the component (a) in the total of the component (a) and the component (a ′) is preferably 50% by mass or more, more preferably 60% by mass or more, and 70% by mass or more from the viewpoint of the effect of removing protein stains. Is more preferable, 80% by mass or more is even more preferable, and 90% by mass or more is particularly preferable.

  In the treatment liquid used in the present invention, the content of the component (a) is preferably 0.004 to 1% by mass, from the viewpoint of the effect of removing protein stains and cost and influence on the substrate, and is preferably 0.005 to 0.5 mass% is more preferable, 0.008-0.2% mass% is further more preferable, and 0.01-0.1% mass% is still more preferable.

  Further, when the component (a ′) is used, the content of the component (a ′) in the treatment liquid used in the present invention is preferably 0.05% by mass or less from the viewpoint of further enhancing the effect of removing protein stains. 0.02% by mass or less is more preferable, 0.01% by mass or less is more preferable, and 0.001% by mass or less is even more preferable.

  The alkaline protease as component (b) may be any enzyme as long as it has an optimum pH from neutral to alkaline, and combinations thereof are also possible. The component (b) is preferably a subtilisin protease derived from Bacillus SP, and among them, a subtilisin protease derived from Bacillus Halodurans or Bacillus clausii is preferred. Commercially available alkaline proteases include, for example, KAP available from Kao Corporation, Alcalase, Sabinase, Evalase, Esperase, Cannase, Obozyme, Genefector International, Plafect, Properase available from Novozymes Japan. and so on.

  In the treatment liquid used in the present invention, the content of the component (b) is preferably 0.002 to 1% by mass, more preferably 0.005 to 0.5% by mass, from the viewpoint of the protein stain removal effect and cost. Preferably, 0.008 to 0.3% by mass is more preferable, and 0.01 to 0.1% by mass is even more preferable.

  In addition, the content of the component (b) (proteolytic activity) in the treatment liquid of the present invention is preferably 0.01 to 200 PU per 1 kg of the treatment liquid from the viewpoint of the effect of removing the fixed protein and cost. 100PU is more preferable and 0.05-20PU is further more preferable.

The proteolytic activity (PU / g) is measured by the following method.
1 mL of 50 mmol / L borate buffer solution (pH 10.5) containing casein (Hammerstein: Merck) at a concentration of 1 w / v% was kept at 30 ° C. for 5 minutes, and then mixed with 0.1 g of enzyme solution. The reaction is performed at 30 ° C. for 15 minutes. Add 2 mL of the reaction stop solution (0.11 mol / L trichloroacetic acid-0.22 mol / L sodium acetate-0.33 mol / L acetic acid) and let stand at room temperature for 10 minutes. Next, the acid-denatured protein was filtered (No. 2 filter paper; manufactured by Whatman), and an alkaline copper solution [1 w / v% potassium tartrate / sodium tartrate aqueous solution: 1 w / v% copper sulfate aqueous solution: sodium carbonate was added to 0.5 mL of the filtrate. 0.1 mol / L aqueous sodium hydroxide solution (sodium carbonate concentration 2 w / v%) = 1: 1: 100 (V / V)] 2.5 mL is added, and the mixture is kept at 30 ° C. for 10 minutes. Further, 0.25 mL of a diluted phenol reagent [phenol reagent (manufactured by Kanto Chemical Co., Ltd.) diluted twice with ion-exchanged water] is added, and the mixture is incubated at 30 ° C. for 30 minutes, and then the absorbance at 660 nm of this sample is measured. Moreover, after mixing a reaction stop liquid with said enzyme reaction system, what added an enzyme solution is measured similarly as a blank. Next, the amount of acid proteolytic product that has been liberated (the amount converted to tyrosine) is obtained from the difference in absorbance between the sample and the blank, and this is the reaction time (in this case: 15 minutes) and the amount of enzyme solution. Dividing by (in the case of this condition: 0.1 g), the proteolytic activity value can be determined. Note that 1 PU is the amount of enzyme that liberates an acid-soluble proteolytic product corresponding to 1 mmol of tyrosine per minute under the above reaction conditions. The above blending amount is determined based on the proteolytic activity obtained by this method.

Component (c) is a nonionic surfactant, and one or more nonionic surfactants selected from the group consisting of the following (1) to (3) are preferable from the viewpoint of the effect of removing protein stains.
(1) a nonionic surfactant represented by the following general formula _{(c1) RO- (EO) l} - (PO) m - (EO) n -H (c1)
(R is a hydrogen atom or a hydrocarbon group having 6 to 18 carbon atoms, and when R is a hydrogen atom, l, m and n are each independently a number of 1 to 350, and R is a carbon number of 8 to 18) 1 is a number of 1 to 10, and m and n are independently a number of 0 to 15).
(2) Amine oxide having a hydrocarbon group having 6 to 16 carbon atoms (preferably an alkyl group or alkenyl group) (3) Glyceryl ether having a hydrocarbon group having 6 to 12 carbon atoms (preferably an alkyl group or alkenyl group)

  Regarding the nonionic surfactant of (1), those in which R in the general formula (c1) is a hydrogen atom preferably have a weight average molecular weight of 1,000 to 15,000, and in the general formula (c1) l, m, and n are preferably numbers corresponding to the weight average molecular weight. This nonionic surfactant is available from BASF, for example, under the trade names Pluronic and Pluronic R.

  The weight average molecular weight is a weight average molecular weight converted using standard polyethylene glycol by gel permeation chromatography (GPC).

  As for the nonionic surfactant (1), R in the general formula (c1) is a hydrocarbon group having 6 to 18 carbon atoms, and the carbon number of R is preferably 8 to 16, and more preferably 8 to 14. The hydrocarbon group for R is preferably an alkyl group or an alkenyl group, more preferably a linear or branched alkyl group, and still more preferably a linear alkyl group. L is preferably a number of 2 to 10, and m and n are preferably a number of 0 to 10. This nonionic surfactant can be obtained, for example, from Kao Corporation under the trade name Emulgen or from BASF under the trade name Pullurafac.

  The nonionic surfactant (2) is an alkyl group or alkenyl group (preferably linear or branched alkyl) having 6 to 16 carbon atoms (preferably 6 to 14 carbon atoms, more preferably 8 to 12 carbon atoms). Amine oxide having a group, more preferably a linear alkyl group) is preferred.

  The nonionic surfactant (3) is an alkyl group or alkenyl group (preferably linear or branched alkyl) having 6 to 12 carbon atoms (preferably 6 to 10 carbon atoms, more preferably 8 to 10 carbon atoms). Group, more preferably a glyceryl ether having a branched alkyl group).

  Among the nonionic surfactants (1) to (3), one or more nonionic surfactants selected from the group consisting of (1) and (2) are preferable from the viewpoint of the effect of removing protein stains. 1 or more types of nonionic surfactant chosen from (1) are more preferable. Among these, among the nonionic surfactants (1), nonionic surfactants in which R in the general formula (c1) is a hydrocarbon group having 6 to 18 carbon atoms are preferable.

  In the treatment liquid used in the present invention, the content of the component (c) is preferably 0.002 to 1% by mass, more preferably 0.005 to 0.5% by mass, from the viewpoint of the protein stain removal effect and cost. Preferably, 0.008 to 0.3% by mass is more preferable, and 0.01 to 0.1% by mass is even more preferable.

  In the present invention, by using a treatment solution having a predetermined pH in which the components (a), (b), and (c) coexist, among the stains such as blood, the stains adhered to the hard surface of the medical device. In addition to the visible portion covering the top, the protein stains that are in direct contact with and adhered to the substrate surface can be sufficiently removed.

  The pH of the treatment liquid used in the present invention is 9 or more, and preferably from 9.5 to 13, more preferably from 10 to 12, and more preferably from the viewpoint of the effect of removing protein stains and the influence on the substrate. More preferred is ˜11.5.

  In addition, pH can be calculated | required by measuring the process liquid used for this invention in 25 degreeC. Further, it may be the pH at the temperature of the treatment liquid when contacting the medical instrument.

  The treatment liquid used in the present invention is prepared by a method of preparing a high-concentration product in advance and diluting it to fall within the set concentration range at the time of dilution, in addition to the method of adjusting the concentration by adding each component separately at the time of use. be able to. As high-concentration products, those containing all three components (I) (a), (b) and (c), (II) those containing (a) and (b), (III) The component containing (b) component and (c) component is mentioned, (II) and (III) are preferable from a viewpoint of enzyme stability. Moreover, in (II) and (III), the other component finally mix | blended is added separately. A high-concentration product can be used, for example, diluted 10 to 200 times with water.

  As a high concentration product containing all three components (a), (b) and (c), (a) monoethanolamine 0.4 to 10% by mass, (b) alkaline protease 0.5 -10% by mass, (c) 1-10% by mass of a nonionic surfactant and water are included, and the high-concentration product is further, from the viewpoint of preventing deterioration of enzyme activity over time, (d) 10-70 mass% of 1 or more types chosen from ethylene glycol, propylene glycol, and glycerol can be contained.

  The high-concentration product may contain 0.01 to 5% of a water-soluble calcium salt, boric acid or a salt thereof, a boron compound such as borax, formic acid or a salt thereof as an enzyme stabilizer. it can.

  The treatment liquid used in the present invention is a surfactant other than the component (c), an alkali buffer, a hydrotrope, a pH adjuster, a thickener, a viscosity adjuster, and a solvent as long as the object of the present invention is not impaired. , Fragrances, colorants, antioxidants, preservatives, bleaches, bleach activators and the like can be blended. You may mix | blend these components in a high concentration goods.

  Examples of medical instruments that are the subject of the present invention include steel instruments such as scissors, forceps, and insulators, resin instruments such as catheters, tubes, and bite blocks, and rigid or flexible endoscopes. The contact of the treatment liquid is performed so as to come into contact with a site where protein stains derived from blood or the like of these medical instruments are attached, and can be applied to the site by a method such as coating, dipping, or spraying. The temperature of the treatment liquid is preferably 5 to 50 ° C, more preferably 10 to 40 ° C. The contact time is preferably 30 seconds to 30 minutes, more preferably 1 minute to 15 minutes.

Example 1 and Comparative Example 1
Using the treatment liquid shown in Table 1, the cleaning effect on protein stains derived from blood was evaluated by three methods: a visual judgment method, a protein staining method, and a fluorescent staining method. The results are shown in Table 1.

[I] Visual Judgment Method and Protein Staining Method 50 μL of 0.1% calcium chloride was added to 1 mL of horse-preserved blood and then immediately stirred. This was uniformly applied to a circle having a diameter of 15 mm on a Teflon (polytetrafluoroethylene, registered trademark) plate of 20 μL and 30 mm × 80 mm, and dried at room temperature for 2 hours and 30 minutes to obtain a test piece.

In a 100 mL glass beaker, 100 mL of the treatment solution shown in Table 1 was placed at 30 ° C. The test piece was immersed for 20 minutes and then gently rinsed with ion exchange water. The effect of the cleaning effect was determined by visually checking for residual blood, then immersed in Coomassie Protein Assay Reagent (reagent supplied with a protein quantification kit, manufactured by Thermo Scientific) for 3 minutes, and then thoroughly rinsed with ion-exchanged water. It was determined according to the following criteria in the subsequent staining state. The test was conducted five times, and the average value is shown in Table 1. In addition, pH of the processing liquid was adjusted with lactic acid as needed.
* Visual criteria ○: No residual blood is observed.
Δ: Slight blood residue is observed. ×: Many blood residues are observed.

* Criteria after staining 5: Almost unstained.
4: Almost half or less of the blood application surface is lightly stained.
3: Almost half or more of the blood application surface is lightly stained.
2: Almost half or less of the blood application surface is deeply stained.
1: Almost half or more of the blood application surface is darkly stained.

[II] Fluorescent staining method The test piece washed in the same manner as in [I] is stained with SYPRO Ruby Protein Gel Stain (manufactured by SIGMA) for 10 minutes, rinsed thoroughly with distilled water, dried, and then fluorescence microscope (Keyence Corporation) Using a 20 × objective lens manufactured by Biozero, the exposure time was changed, irradiation with excitation light of 470 nm was performed, and reflected light of 510 nm or more was detected and observed on a monitor, and judged according to the following criteria. It means that the amount of protein increases as color develops in a shorter exposure time.
Criteria 5: Color development is hardly recognized at an exposure time of 3 seconds 4: Partial color development occurs at an exposure time of 3 seconds 3: Almost the entire surface is colored at an exposure time of 0.4 to 1 second 2: The entire surface is colored with an exposure time of 0.05 to 0.3 seconds.
1: The entire surface is colored with an exposure time of 0.03 seconds.

The components in the table are as follows.
-Alkaline protease: KAP [manufactured by Kao Corporation, 2.2 (PU / g)]
Nonionic surfactant A: Emulgen 106 [polyoxyethylene (average number of moles of added ethylene oxide 6) lauryl ether, manufactured by Kao Corporation]
Nonionic surfactant B: Emulgen LS106 [in general formula (c1), l = 3.0, m = 1.5, n = 3.0, R is an alkyl group having 12 carbon atoms, manufactured by Kao Corporation]
・ Nonionic surfactant C: Plurafac LF901 (manufactured by BASF)
Nonionic surfactant D: Penetol GE-EH (2-ethylhexyl glyceryl ether, manufactured by Kao Corporation)
Nonionic surfactant E: Amphital 20N (Lauryldimethylamine oxide, manufactured by Kao Corporation)

  As shown in Table 1, in Examples 1-1 to 1-7 using the treatment liquid containing the component (a), the component (b), and the component (c) and having a pH of 9 or more, Protein stains adhered to the material surface can be effectively removed. In particular, the fluorescent staining method employed in this example can confirm the presence of a small amount of protein stains that cannot be distinguished by conventional determination methods (such as amide black staining or orthotoluidine method). Even with a highly accurate evaluation method, the presence of protein contamination is hardly confirmed, indicating that an excellent cleaning effect is obtained.

  On the other hand, protein stains are generally considered to have higher solubility and higher detergency as the pH is increased and alkalinity. However, as in Comparative Examples 1-1 to 1-3, an alkaline agent that is not component (a) is used. It can be seen that in the system in which the pH is in the alkaline region, the protein soil adhered to the substrate surface cannot be completely removed.

  Further, in the treatment liquid that does not contain the alkaline protease (b) component, the cleaning agent can remove stains at the visual level, but almost removes protein stains adhering to the substrate surface as in Comparative Example 1-4. You can see that it was not done.

  Moreover, when (c) component is not contained, it turns out that the stain | pollution | contamination fixed to the base-material surface is not able to be removed completely like Comparative Example 1-5.

  Moreover, even if it is an alkanolamine, when diethanolamine or triethanolamine is used, as in Comparative Examples 1-6 and 1-7, even if it looks as if the dirt is removed by the visual judgment method, the fluorescent staining method It can be seen that the protein soil adhering to the substrate surface was not sufficiently removed because of poor evaluation at. That is, even if it is a compound which belongs to alkanolamine, the effect of this invention will not be acquired unless it is the monoethanolamine of (a) component of this invention.

  Moreover, even when it contains (a) component, (b) component, and (c) component, when pH is made into neutral vicinity, it turns out that a washability falls remarkably like Comparative Example 1-8. In addition, pH of Example 1-7 and Comparative Example 1-8 was adjusted by adding citric acid.

Although the mechanism of action of the cleaning method of the present invention is not clear, the soil fixed by the action of monoethanolamine and a nonionic surfactant is easily affected by the alkaline protease, and the protein soil that has been decomposed and removed is not a nonionic interface. It is thought that it was effectively dispersed by the activator.

Claims (5)

(A) monoethanolamine (hereinafter referred to as component (a)), (b) alkaline protease (hereinafter referred to as component (b)), (c) nonionic surfactant (hereinafter referred to as component (c)) and water A method for cleaning a medical device, comprising the step of bringing a treatment liquid containing 9 and a pH of 9 or more into contact with a medical device to which protein soil has adhered ,
In the treatment liquid, the content of the component (a) is 0.004 to 1% by mass, the content of the component (b) is 0.002 to 1% by mass, and the content of the component (c) is 0.00. 002 to 1% by mass,
In the treatment liquid, the content of one or more alkali agents selected from (a ′) alkanolamines other than monoethanolamine, alkali metal hydroxides, carbonates, phosphates, and silicates is 0.05% by mass. Is
How to clean medical equipment. The cleaning method according to claim 1 , wherein the component (c) is at least one nonionic surfactant selected from the group consisting of the following (1) to (3).
(1) a nonionic surfactant represented by the following general formula _{(c1) RO- (EO) l} - (PO) m - (EO) n -H (c1)
(In the formula, R is a hydrogen atom or a hydrocarbon group having 6 to 18 carbon atoms, and when R is a hydrogen atom, l, m, and n are each independently a number of 1 to 350, and R is a carbon number. In the case of 6 to 18 hydrocarbon groups, l is a number from 1 to 10, and m and n are independently a number from 0 to 15.)
(2) Amine oxide having a hydrocarbon group having 6 to 16 carbon atoms (3) Glyceryl ether having a hydrocarbon group having 6 to 12 carbon atoms The method for cleaning a medical device according to claim 2 , wherein the component (c) is a nonionic surfactant represented by the following general formula (c1).
_{RO- (EO) l - (PO} ) m - (EO) n -H (c1)
(In the formula, R is a hydrocarbon group having 6 to 18 carbon atoms, l is a number of 1 to 10, and m and n are independently a number of 0 to 15). The method for cleaning a medical instrument according to any one of claims 1 to 3 , wherein the protein soil is blood-derived protein soil. (A) 0.4 to 10% by mass of component, 0.5 to 10% by mass of component (b), 1 to 10% by mass of component (c), and highly concentrated product containing water diluted with water The method for cleaning a medical instrument according to any one of claims 1 to 4 , wherein a treatment liquid is prepared.