FR3049285A1 - DETERGENT ENZYMATIC COMPOSITIONS FOR THE AUTOMATIC CLEANING OF SURGICAL AND / OR MEDICAL INSTRUMENTS AND / OR DEVICES. - Google Patents

Abstract

The invention relates to the use of a concentrated aqueous detergent composition for cleaning surgical and / or medical instruments in a disinfector washer, said concentrated aqueous detergent composition comprising: - a subtilisin, - a lipase, - an amylase, - a biodegradable chelating agent selected from methylglycine diacetic acid and iminodisuccinate and water, said concentrated detergent composition being free of alkanolamine.

Description

The present invention relates to the use of enzymatic detergent compositions for the automatic cleaning of surgical and / or medical instruments and / or devices.

In hospital settings, surgical instruments are usually subjected to an automatic instrument washer-disinfector cleaning (LDI) using strongly alkaline detergents followed by thermal or chemical disinfection. These detergents generally have a pH greater than 12 which makes them very corrosive.

To avoid the damage caused by these alkaline detergents, enzymatic detergents have been proposed. For example, MEDICLEAN® forte by Dr. Weigert is known and is the subject of European patent EP 1 327 674 B1. This detergent contains triethanolamine, nitrilotriacetic acid (NIA) as chelating agent, phosphates and a protease. The simultaneous presence of an alkanolamine and an enzyme in the same composition poses stability problems for the enzyme, especially when the composition is very concentrated. That's why Dr. Weigert developed the NEodisher® SYSTEM ALPHA which consists of two highly concentrated components, namely an enzymatic component, the neodisher® SystemAct, and an alkaline component, the neodisher® SystemClean. The enzymatic component contains a subtilisin while the alkaline component contains triethanolamine and methylglycine diacetic acid (MGDA). This system is also the subject of the international patent application WO2015 / 043777. Even if this system makes it possible to solve the problems related to the instability of the enzyme in the presence of high concentrations of alkaline agents such as alkanolamines, it presents a major disadvantage in that it requires specific equipment to be able to supply the scrubber instrument disinfector with both components simultaneously. The disinfecting washers of instmments do indeed have only one supply of detergent composition.

There is therefore still a need for a means for effectively cleaning the surgical and medical instruments in the machine without presenting the stability problems of the solutions proposed in the state of the art and without the need for specific equipment to power the machines used for the cleaning.

It is the merit of the Applicant to have found in the fruit of much research that these disadvantages could be overcome by using a concentrated detergent composition which is based on the combination of three enzymes and a powerful chelating agent and which does not contain any alkanolamine. The invention therefore relates to the use of a concentrated aqueous detergent composition for cleaning surgical and / or medical instruments in a disinfector washer, said concentrated aqueous detergent composition comprising: - a subtilisin, - a lipase, - an amylase, - a biodegradable chelating agent selected from methylglycine diacetic acid (MGDA) and iminodisuccinate (IDS) and water, said concentrated detergent composition being alkanolamine-free.

Surprisingly, this specific combination of a subtilisin, a lipase and an amylase with MGDA or IDS makes it possible to overcome the need for the addition of alkanolamine while maintaining effective cleaning of surgical and / or medical instruments contaminated with a washer disinfector. Thus, the concentrated aqueous detergent composition of the invention does not exhibit the enzymatic stability problems encountered in the prior art enzyme compositions containing alkanolamines.

Moreover, since the main components of biological tissues that constitute major stains of surgical and medical instruments are proteins, lipids (triglycerides) and carbohydrate polymers (glycogen), the tri-enzymatic cocktail of the concentrated aqueous detergent composition of the The invention will simultaneously hydrolyze the three targets and thereby facilitate detergency.

In addition, the absence of alkanolamine makes it possible to formulate the detergent composition used according to the invention in a very concentrated manner. Thus, the water content of the concentrated aqueous detergent composition can be reduced up to 25% by weight relative to the total weight of composition without having to add significant amounts of third-solvent, impact the stability of the enzymes and harm cleaning efficiency.

The composition used according to the invention advantageously has a subtilisin content of 0.5 to 15% by weight relative to the total weight of the composition, preferably from 1 to 10% by weight relative to the total weight of the composition, preferably from 1 to 5% by weight relative to the total weight of the composition, more preferably from approximately 3% by weight relative to the total weight of the composition. The lipase content is advantageously from 0.05 to 2% by weight relative to the total weight of the composition, preferably from 0.5 to 1.5% by weight relative to the total weight of the composition, more preferably from about 1% by weight relative to the total weight of the composition. The content of amylase is advantageously from 0.05 to 2% by weight relative to the total weight of the composition, preferably from 0.5 to 1.5% by weight relative to the total weight of the composition, more preferably from about 1% by weight relative to the total weight of the composition. The biodegradable chelating agent is chosen from methylglycine diacetic acid (MGDA) and an iminodisuccinate (IDS), in particular tetrasodium iminodisuccinate. Preferably, the biodegradable chelating agent is methylglycine diacetic acid (MGDA). Without wishing to be bound by any theory, the inventors believe that the chelating agent used in the present invention not only serves to soften water but also to complex particularly efficiently bi-metal ions and trivalents, especially Ca ^^, Fe ^^, Fe ^^ and Cu ^^, contained in the protein residues typically found on surgical and / or medical instruments. It seems that these chelating agents sequester these metal ions which destabilizes the tertiary structure of proteins and makes them more easily hydrolyzable.

The composition used according to the invention advantageously has a content of biodegradable chelating agent of 10 to 50% by weight relative to the total weight of the composition, preferably from 20 to 40% by weight relative to the total weight of the composition, more preferably 25 to 35% by weight relative to the total weight of the composition, more preferably still approximately 30% by weight relative to the total weight of the composition.

The concentrated aqueous detergent composition is advantageously free from any non-bio-degradable chelating agent, especially ethylene-diamine-tetraacetic acid (EDTA), diethylene triamine pentaacetic acid (DTPA), phosphates and polyphosphates. Advantageously, the concentrated aqueous detergent composition is also free of any chelating agent classified as reprotoxic such as nitrilotriacetic acid and its sodium salt (NTA).

In one embodiment, the concentrated aqueous detergent composition is also free of alkali and alkaline earth hydroxides.

The water content of the concentrated detergent composition used according to the invention is advantageously from 10 to 50% by weight relative to the total weight of the composition, preferably from 15 to 45% by weight relative to the total weight of the composition. more preferably from 20 to 40% by weight relative to the total weight of the composition and more preferably still from 25 to 40% by weight relative to the total weight of the composition. In addition to water, the concentrated detergent composition may also contain a third-solvent such as a polyol. Examples of suitable polyols include alkylenes glycols such as ethylene glycol, propylene glycol and glycerine (1,2,3-propanetriol). Preferably, the third-solvent is glycerin. Glycerine has the advantage of not only serving as a third-solvent but also stabilizing the enzymes.

The content of the third-solvent is advantageously from 5 to 50% by weight relative to the total weight of the composition, preferably from 10 to 40% by weight relative to the total weight of the composition, more preferably from 13 to 25% by weight. weight relative to the total weight of the composition.

The concentrated detergent composition used in accordance with the invention may further comprise one or more nonionic surfactants. The nonionic surfactant (s) may, in particular, be chosen from alkoxylated nonionic surfactants and alkylpolyglucosides. The alkoxylated nonionic surfactants are advantageously chosen from alkoxylated nonionic surfactants, especially from alkoxylated fatty alcohols, especially ethoxylated and / or propoxylated, and block copolymers EO-PO. In a preferred embodiment, the nonionic surfactant or surfactants are chosen from alkylpolyglucosides, preferably it is a C1-alkylpolyglucoside. This alkylpolyglucoside has the triple advantage of acting as a stabilizer, provide a favorable wetting effect for detergency and a non-foaming character compatible with a washer-disinfector application.

The content of nonionic surfactant in the detergent composition used according to the invention is advantageously from 1 to 10%, preferably from 2 to 5%, more preferably from approximately 3% by weight relative to the total weight of the composition.

The detergent composition used in accordance with the invention may further comprise an acid or buffer system for adjusting the pH. Preferably, an organic acid is added, chosen from the C 2 -C 6 mono-, di- and tricarboxylic acids, chosen in particular from citric acid, tartaric acid, malic acid, lactic acid, glutaric acid, gluconic acid, glycolic acid, succinic acid and adipic acid. Citric acid is particularly preferred. It has the advantage of serving at the same time as complexing agent as well as tartaric acid and gluconic acid.

Advantageously, the composition will contain no antimicrobial agent and will therefore have no claim for antimicrobial activity at the dose of use. This is to be distinguished from a bacteriostatic effect that can be sought. For this purpose, the detergent composition used according to the invention may also comprise a preservative agent. The preserving agent may be chosen from the usual preserving agents, such as, for example, 5-chloro-5-methyl-4-isothiazolin-3-one, 2-methyl-4-isothiazolin-3-one, p-acid and the like. hydroxybenzoic acid and its methyl esters, 5-bromo-5-nitro-1,3-dioxane and salicylic acid, benzyl alcohol and sodium benzoate. These preservatives make it possible to avoid microbial and fungal contamination by providing a bacteriostatic and static fungi effect.

The detergent composition used according to the invention may also comprise a hydrotropic agent. This hydrotrope agent may for example be selected from xylene sulfonates, toluenesulfonates, cumenesulfonates, urea, methylacetamide, polyethylene glycol and polypropylene glycol.

The concentrated detergent composition used according to the invention may further comprise other adjuvants such as, for example, dispersing agents and / or antifoaming agents. The dispersing agents may for example be acids of homo-, co- or terpolymers based on acrylic acid or alkali metal salts thereof (polyacrylates).

In the use according to the invention, the detergent composition is implemented in a washer disinfector (LD) such as an instrument washer-disinfector (LDI) or an endoscope washer-disinfector (LDE). The term "washer-disinfector" is to be understood as defined in ISO 15883-1: 2006, namely a machine for cleaning and disinfecting medical devices and all other articles used in the medical, dental, pharmaceutical and veterinary. Also, the term "surgical and / or medical instruments" as used in the present application is to be broadly understood as referring to medical devices and all other articles used in the medical, dental, pharmaceutical and veterinary environments requiring cleaning. with aqueous detergents before reuse.

In the present invention, this cleaning is advantageously carried out at a temperature of 20 to 60 ° C, preferably 45 to 55 ° C, more preferably 50 to 55 ° C. The cleaning time is advantageously between 20 s and 10 min, preferably between 30 s and 5 min.

The present invention also relates to the concentrated aqueous detergent composition implemented in the use according to the invention as described above. This composition is to be diluted with water, preferably softened, demineralized or osmosis, before use. The dilution is generally from 0.03% to 1% (v / v), preferably from 0.1% to 0.6% (v / v), more preferably from 0.2 to 0.4% (v / v). . The pH of the diluted ready-to-use composition is advantageously from 8 to 11, preferably from 8.5 to 9.5, more preferably from 8.8 to 9.2, and even more preferably from about 9. determination of the pH of the diluted composition, softened water is used (degree of hardness about 5 ° f). The invention will now be described using non-limiting examples of embodiment of the invention.

FIGURES

Figure 1: Photos of the detergency tests embedded for compositions Cl, C2etC3.

Figure 2: Evaluation of the activity of subtilisin by gel electrophoresis.

Figure 3: Evaluation of the activity of the lipase by thin layer chromatography.

Figure 4: Evaluation of the activity of amylase by thin layer chromatography.

EXAMPLES

The following three concentrated detergent compositions (Table 1) were evaluated for their ability to effectively clean up residues typically found on surgical and medical instruments.

Table 1

EXAMPLE 1: On-board detergent tests in a MIELE G7836CD washer disinfector 1) Cycle sequence - Detergent injection at 35 ° C, dilution of 0.3% (v / v) in softened water

- Wash with softened water for 3 minutes at 55 ° C - Rinse with cold water system Drying 2) The different onboard tests used

Simicon (SIMICON PET-K-10 from SIMICON GmbH, Munich, Germany) - Wash check H (Getinge, Sweden) - Load Check Browne (Steris, USA)

The different tests were used according to the instructions provided by the respective manufacturer. 3) Test results

The performance tests showed that the three compositions Cl, C2 and C3 give quite satisfactory results under the test conditions. The best results are however obtained with the composition Cl. With this composition, no trace is indeed detected on the three tests used (FIG. 1).

The tests were repeated at a cleaning temperature of 40 ° C. and a dilution of the compositions at 0.5% (v / v). The results obtained are comparable to those obtained at 40 ° C and thus confirm the effectiveness of the compositions according to the invention. EXAMPLE 2 Comparative Detergent Tests in MIELE PG 8528 Washer Disinfector

The performance of the composition Cl was compared to that of Anisoyme® DLM Maxi (Anios Laboratories, Hellemmes, France), a tri-enzyme detergent containing a protease, a lipase, an amylase and the EDTA chelating agent. The embedded test used was the TOSI AMCOR (reference 35002577).

For each test, the treatment cycle is as follows: - Prewash: cold mains water, 1 min, - Wash: softened water (5 ° f), 5 min, temperature and dosage according to Table 2 below, - Rinsing : cold network water, 1 min, - Final rinse: water osmosis at 90 ° C, 1 min with Anios RDA at 0.05% (v / v).

Table 2 below summarizes the test conditions and results.

Table 2

These tests show that the composition according to the invention allows a more efficient cleaning at low temperatures and dosage. This is all the more surprising since the chelating activity of EDTA is higher than that of MGDA, which would allow a higher cleaning efficiency for the Aniosyme® DLM Maxi. EXAMPLE 3 Evaluation of the Activity of Enzymes in the Cl Composition

The respective stabilities of subtilisin, lipase and amylase in the composition C1 were evaluated by the following tests. 1) Activity of subtilisin The activity of the subtilisin contained in the detergent is followed by the technique of electrophoresis on polyacrylamide gel, the substrate used is a solution of human serum albumin at 0.3% (representative soil of an application in a health facility).

After migration, the proteins and peptides are revealed by exposure in a Coomassie blue staining solution followed by decolorization. A revelation with Coomassie blue reveals the degradation products resulting from the hydrolysis of human serum albumin.

FIG. 2 shows four electrophoresis tracks: lane 1 corresponds to the control (human serum albumin (0.3%), non-degraded), lanes 2, 3 and 4 to the composition Cl after manufacture, 3 weeks of storage in a container tightly closed at 30 ° C and 7 weeks storage in a tightly closed container at 30 ° C respectively. It is found that the enzymatic activity of subtilisin does not degrade for at least 7 weeks. 2) Lipase Activity The lipase activity was followed by the Thin Layer Chromatography (TLC) technique, the substrate used was a triglyceride and the eluent a mixture of 50 mL of heptane, 20 mL of diethyl ether, 0.5 mL of acetic acid and 1.5 mL of ethanol.

Triglyceride-degrading lipase, the absence of triglycerides, and the presence of diglycerides and monglycerides demonstrate the enzymatic activity of lipase.

Figure 3 shows six TLC tracks: lanes 1, 2, 3 correspond to controls (1: triglycerides, 2: diglycerides, 3: monoglycerides), lanes 4, 5 and 6 to composition Cl after manufacture, after 3 weeks storage in a tightly closed container at 30 ° C and after 7 weeks of storage in a tightly closed container at 30 ° C respectively. It is found that the enzymatic activity of the lipase does not degrade for at least 7 weeks. 3) Activity of amylase The activity of amylase was followed by the Thin Layer Chromatography (TLC) technique, the substrate used is glycogen and the eluent a mixture of 70 ml of ethanol, 1 mL of acetic acid and 29 mL of demineralized water. The glycogen degrading amylase, the absence of glycogen and the presence of glucose demonstrate the enzymatic activity of the amylase.

FIG. 4 shows five TLC tracks: lanes 1 and 2 correspond to the controls (1: glycogen, 2: glucose), lanes 3, 4 and 5 to the Cl composition after manufacture, after 3 weeks of storage in a hermetically sealed container closed at 30 ° C and after 7 weeks of storage in a tightly closed container at 30 ° C respectively. It is found that the enzymatic activity of the amylase does not degrade for at least 7 weeks. EXAMPLE 4 Influence of the Enzymes on the Degreasing Cleansing Power The influence of the presence of the three subtilisin, lipase and amylase enzymes in the detergent composition was evaluated by comparing the composition Cl with compositions without enzymes (C4), with subtilisin (C5 ) and with lipase and amylase (C6).

For these tests, the degreasing cleansing power of the various compositions was evaluated with respect to English soil according to the ISO 15883 standard which was applied by brush on stainless steel blades (one blade per composition tested) and allowed to dry for one hour at ambient temperature. Then each slide was quenched with stirring (three-dimensional stirrer, 50 rpm) for 15 min at room temperature in one of the four detergent compositions diluted to 0.3% (v / v) in softened water (5 ° F). ).

The best results were obtained for the CL composition The slides treated with compositions C4, C5 and C6 indeed had significant residues of English soil visible to the naked eye. Residual protein assay using the MicroBCA technique (INTERCHBVI UPTIMA MicroBC Assay Kit, UP75860A) confirms these observations.

Claims (18)

Use of a concentrated aqueous detergent composition for cleaning surgical and / or medical instruments in a disinfector washer, said concentrated aqueous detergent composition comprising: a subtilisin, a lipase, an amylase, a biodegradable chelating agent selected from diacetic methylglycine and iminodisuccinate and water, said concentrated detergent composition being free of alkanolamine. 2. Use according to claim 1, characterized in that the detergent composition has a subtilisin content of 0.5 to 15% by weight relative to the total weight of the composition. 3. Use according to claim 1 or 2, characterized in that the detergent composition has a lipase content of 0.05 to 2% by weight relative to the total weight of the composition. 4. Use according to any one of the preceding claims, characterized in that the detergent composition has an amylase content of 0.05 to 2% by weight relative to the total weight of the composition. 5. Use according to any one of the preceding claims, characterized in that the biodegradable chelating agent is methylglycine diacetic acid. 6. Use according to any one of the preceding claims, characterized in that the detergent composition has a content of biodegradable chelating agent of 10 to 50% by weight relative to the total weight of the composition. 7. Use according to any one of the preceding claims, characterized in that the detergent composition also comprises glycerine. 8. Use according to any one of the preceding claims, characterized in that the detergent composition also comprises one or more nonionic surfactants. 9. Use according to any one of the preceding claims, characterized in that the cleaning is carried out at a temperature of 20 to 60 ° C. 10. Use according to any one of the preceding claims, characterized in that the cleaning time between 20 s and 10 min. A concentrated aqueous detergent composition comprising: - a subtilisin, a lipase, an amylase, - a biodegradable chelating agent selected from methylglycine diacetic acid and an iminodisuccinate and - water, said concentrated detergent composition being free of alkanolamine. 12. Detergent composition according to claim 11, characterized in that it has a subtilisin content of 0.5 to 15% by weight relative to the total weight of the composition. 13. Detergent composition according to claim 11 or 12, characterized in that it has a lipase content of 0.05 to 2% by weight relative to the total weight of the composition. 14. Detergent composition according to any one of claims 11 to 13, characterized in that it has an amylase content of 0.05 to 2% by weight relative to the total weight of the composition. 15. Detergent composition according to any one of claims 11 to 14, characterized in that the biodegradable chelating agent is methylglycine diacetic acid. 16. Detergent composition according to any one of claims 11 to 15, characterized in that it has a content of biodegradable chelating agent of 10 to 50% by weight relative to the total weight of the composition. 17. Detergent composition according to any one of claims 11 to 16, characterized in that it also comprises glycerin. 18. Detergent composition according to any one of claims 11 to 17, characterized in that it also comprises one or more nonionic surfactants.