EA041635B1 - ANTIMICROBIAL COMPOSITION AND METHOD FOR TREATMENT OF FOOD PRODUCTS AND HARD SURFACES - Google Patents

Description

Cross-reference to related applications

This application claims the benefit of US Provisional Application No. 62/084,278, filed November 25, 2014, the contents of which are incorporated herein by reference.

Region

This is a description of the invention in the field of antimicrobial treatment of food and hard surfaces using compositions including inorganic acids, organic acids, or a combination thereof; copper(II) salt; and buffer salt, detergent, or combinations thereof.

Contamination of surfaces with toxic concentrations of bacteria, viruses and parasites (pathogens) is a significant problem. Antimicrobial formulations that are used in the food industry must be able to reduce the number of pathogens on the surface, and they must also be safe for humans. In addition, antimicrobial formulations should not adversely affect the quality of the products or the treated surface. Antimicrobial food and surface treatments also need to be easy to use and relatively inexpensive to be cost effective for the expected payoff.

Thus, there is a need for antimicrobial formulations that are safe, effective, easy to administer, and economically/commercially viable. This disclosure addresses these and other important needs.

This disclosure provides formulations containing inorganic acids, organic acids, or combinations thereof; a copper(II) salt; and a buffer salt, detergents, or combinations thereof. Methods for using these formulations to reduce the number of pathogens on surfaces are also described.

The general description and the following detailed description are exemplary and illustrative only and do not limit the invention as defined in the appended claims. Other aspects of the present invention will become apparent to those skilled in the art upon review of the detailed description of the invention that is provided herein.

Brief description of the drawings

In FIG. 1 presents the results of individual tests (214 data points) at a pH value of 2.2 of samples of the wash solution formed by the composition specified in the disclosure.

Fig. 2 shows the results of the accelerated shelf life test at 45°F. Approximately 30% reduction in active degradation was achieved with treatment with the antimicrobial solution formed by the disclosure formulation compared to standard treatment with hypochlorous acid.

Fig. 3 displays the results of measurements of organic pollution load (in DCO, approximate colony forming unit concentration) directly on the surfaces of lettuce leaves from 250 randomized samples taken over a 6-week period. The lettuce leaf samples were treated with either the disclosure formulation or the standard hypochlorous acid treatment, which is widely used in the manufacturing industry and includes a combination of citric acid and chlorine.

Detailed description of private embodiments of the invention

The present invention may be better understood by reference to the following detailed description accompanied by the drawings and examples which form part of it. It should be understood that this invention is not limited to the specific devices, methods, applications, conditions or parameters described and/or shown in this document, and that the terminology used here is for the purpose of describing particular ways of implementing the invention by way of example only and is not intended to limit the claimed inventions. And also, the use in the description, including the appended claims, of singular nouns includes the plural, and reference to a specific numerical value includes at least that value, unless the context clearly dictates otherwise. When a range of values is shown, another embodiment includes from one particular value and/or to another particular value. Similarly, when values are expressed as approximate values, using the terms about, about, about, it should be understood that a particular value constitutes another embodiment. All ranges are inclusive and allow combinations.

It should be appreciated that certain features of the invention, which are described herein for clarity in the context of separate embodiments, may also be represented by a combination in one embodiment. Conversely, various features of the invention, which for brevity are described in the context of one embodiment, may also be available individually or in any subcombination. Further, reference to values specified within a range includes each and every value within that range.

In the context of this document, the term food refers to solid and liquid foods that are suitable for human or pet consumption. Solid food products include, but are not limited to, meat products such as poultry products (for example, chicken, duck, turkey products), eggs, beef products, pork products, and seafood products (for example, fish, shrimp, crustaceans, mollusks, echinoderms, algae). Solid foods also include agricultural products such as fruits, vegetables, algae, seeds, grains, sprouts, legumes, soybeans, and nuts. Solid foods also include dairy products such as hard, soft and semi-soft cheeses. Liquid food products may include beverages (for example, juices, carbonated drinks), liquid dairy products (for example, milk and cream), fermented drinks (for example, beer and wine), and liquid food components and liquid foods used in animal nutrition (for example, fermented liquid feed as feed for farm animals and poultry).

In the context of this document, the term hard surfaces refers to surfaces including, but not limited to, wood (hardwood and parquet), travertine, MDF (medium density fibreboard), plywood, ceramics, concrete, porcelain, linoleum, laminate, granite, marble, quartz, soapstone, stainless steel, copper, metal alloys, ferrous metal, plastic, brick, other masonry materials, gypsum board, gypsum/plaster, glass, textile board, corrugated board, cardboard, rubber, latex, plastic, composite materials, which may include two or more of the above materials, and the like.

In the context of this document, the term pathogens refers to bacteria, viruses and parasites. An example of bacteria that can be reduced with formulations described in the disclosure are Gram-positive and Gram-negative bacteria, such as Salmonella enterica, Listeria monocytogenes, Escherichia coli, Clostridium botulinum, Clostridium difficile, Campylobacter, Bacillus cereus, Vibrio parahaemolyticus, Vibrio cholerae, Vibrio vulnificus, Staphylococcus aureus, Yersinia enterocolitica, Shigella, and combinations thereof. Examples of Salmonella enterica serovars that can be reduced by the formulations of this disclosure include, for example, Salmonella Enteriditis, Salmonella Typhimurium, Salmonella Poona, Salmonella Heidelberg, and Salmonella Anatum. Examples of viruses that may be reduced by the composition described in the Disclosure include viruses of the genera Enterovirus, Norovirus, Influenza, Rotavirus, or combinations thereof. Examples of parasites include Cryptosporidium, Toxoplasma gondii, Giardia duodenalis, Cyclospora cayetanensis, Trichinella spiralis, Taenia saginata, Taenia solium, or combinations thereof.

In one aspect, the present disclosure provides compositions containing an inorganic acid, organic acids, or combinations thereof; a copper(II) salt; and a buffer salt, a detergent, or combinations thereof. In preferred embodiments, these compositions do not include chlorine and sources of chlorine. For example, preferred compositions in the invention exclude hypochlorous acid salts, such as sodium hypochlorite and calcium hypochlorite, as their basis.

Preferred inorganic acids are known in the art. Inorganic acids effective in disclosure formulations and methods have a pKa value less than or equal to 1. Preferred inorganic acids include, for example, sulfuric acid, phosphoric acid, hydrochloric acid, nitric acid, boric acid, hydrobromic acid, perchloric acid, hydroiodic acid, and their combinations. In some cases, the inorganic acid is sulfuric acid. In other cases, the inorganic acid is phosphoric acid. In other cases, the inorganic acid is hydrochloric acid. The inorganic acid may be present in such an amount to provide a predetermined pH value, as described herein. In some embodiments, the inorganic acid is present at a concentration of from about 0.1 to about 3 wt.%, preferably from about 0.1 to about 1 wt.% based on the weight of the composition.

Preferred organic acids are known in the art. Organic acids suitable for the formulations and methods described in the disclosure will have a pKa between 1 and 7. Organic acids include citric acid, ascorbic acid, lactic acid, acetic acid, peracetic acid, formic acid, propionic acid, butyric acid, valeric acid , caproic acid, oxalic acid, malic acid, benzoic acid, carbonic acid, phenol, uric acid, taurine, p-toluenesulfonic acid, trifluoromethanesulfonic acid, aminomethylphosphonic acid, and combinations thereof. In some cases, the organic acid is citric acid. In other cases, the organic acid is ascorbic acid. In other cases, the organic acid is lactic acid. In some cases, the organic acid is acetic acid. In some cases, the organic acid is peracetic acid. The organic acid may be present in any amount, so as to achieve a predetermined pH, as described here. In some embodiments, the organic acid is present at a concentration of from about 0.1% to about 3% by weight. Or from about 0.1 to about 2 wt.%, Preferably from about 0.1 to about 1 wt.%, based on the weight of the composition.

Disclosure formulations may include an inorganic acid or combinations of inorganic acids. Disclosure formulations may include an organic acid or combinations of organic acids. Disclosure formulations may include a mixture of inorganic acid(s) and organic acid(s). In those embodiments consisting of a mixture of inorganic acids, a mixture of organic acids, or a mixture of inorganic acid(s) and organic acid(s), the total amount of acid is sufficient to achieve the desired pH value, as described in this document. In some embodiments, the combination of acids at a concentration of from about 0.1 to about 3 wt.%, from about 0.1 to about 2 wt.%, preferably from about 0.1 to about 1 wt.% based on the weight of the composition.

Preferred copper(P) salts include copper(P) sulfate, copper(P) chloride, copper(P) bromide, and the like. A preferred copper(II) salt is copper(II) sulfate, together with copper(II) sulfate pentahydrate, which is particularly preferred. In some embodiments, the copper(II) salt is copper(II) chloride. In other embodiments, the copper(II) salt is copper(II) bromide. Combinations of copper(II) salts are permitted under disclosure.

The concentrations of copper salts used in the compositions described in the disclosure can be determined by a person skilled in the art, and are needed at a value that is effective to reduce the number of pathogens on a particular treated surface. For example, the copper salt concentration may be up to 1000 ppm, between 500 and 1000 ppm, between 100 and 500 ppm, or 1 to 100 ppm. According to the disclosure, the concentration of copper salts used in the composition of the described invention is from about 1 ppm to about 80 ppm, from about 1 ppm to about 70 ppm, from about 1 ppm to about 60 ppm, from about 1 ppm and to about 50 ppm, from about 1 ppm to about 40 ppm, from about 1 ppm to about 30 ppm, from about 1 ppm to about 20 ppm, from about 1 ppm to about 15 ppm, from about 1 ppm to about 1 ppm and to about 10 ppm, from about 3 ppm to about 10 ppm, or from about 35 ppm to about 40 ppm. Other preferred compositions described in the application include about 0.1; 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9; 1; 2; 3; 4; 5; 6; 7; 8; 9; 10; eleven; 12; 13; 14; 15; 16; 17; 18; 19; 20; 21; 22;

23; 24; 25; 26; 27; 28; 29; thirty; 31; 32; 33; 34; 35; 36; 37; 38; 39; 40; 41; 42; 43; 44; 45; 46; 47; 48; 49; 50; 51;

52; 53; 54; 55; 56; 57; 58; 59; 60; 61; 62; 63; 64; 65; 66; 67; 68; 69; 70; 71; 72; 73; 74; 75; 76; 77; 78; 79; 80;

81; 82; 83; 84; 85; 86; 87; 88; 89; 90; 91; 92; 93; 94; 95; 96; 97; 98; 99 or 100 ppm copper(II) salts. In some embodiments, the copper(II) salt concentration is from about 1 ppm to about 19 ppm for fruit and vegetable processing, or from about 1 ppm to less than 60 ppm for meat, seafood, and/or poultry products.

Preferred buffer salts for use as described in the disclosure include ammonium sulfate, sodium sulfate, sodium chloride, calcium sulfate, and combinations thereof. Sodium sulfate, ammonium sulfate and calcium sulfate are particularly preferred buffer salts. In some cases, the buffer salt is ammonium sulfate. In other cases, the buffer salt is sodium sulfate. In some cases, the buffer salt is sodium chloride. In other cases, the buffer salt is calcium sulfate.

Preferred detergents include formulations at concentrations that are recognized as safe by the US Food and Drug Administration. For the purposes of this disclosure, the term detergent refers to surfactants or mixtures of surfactants. Particularly preferred detergents for use in the formulations of this invention include dodecyl sulfate, n-alkylbenzene dodecyl sulfate, alkyl polyglycoside, sodium lauryl sulfate, polysorbates (eg, polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80), and combinations thereof. Particularly preferred alkyl polyglycosides include alkyl polyglucoside, decyl polyglucose, lauryl polyglucose, and combinations thereof.

The compositions described in the disclosure may include an inorganic acid, an organic acid, or combinations thereof, a copper(II) salt, and a buffer salt, while excluding the detergent. Other disclosure formulations may include an inorganic acid, an organic acid, or a combination thereof; copper(II) salt, detergent, excluding buffer salt. Other compositions of the invention may include an inorganic acid, an organic acid, or a combination thereof; copper(II) salt; buffer salt, and detergent.

According to the disclosure, buffer salt and/or detergent are present in the composition, each in concentrations ranging from 0.01 to 0.5 wt.%, from 0.01 to 0.4 wt.%, from 0, 01 to 0.3 wt.%, from 0.01 to 0.25 wt.%, from 0.01 to 0.20 wt.%, from 0.01 to 0.15 wt.%, or from 0.01 to 0.10 wt.% based on the mass of the composition. Preferred amounts of buffer salts and/or detergents maintain the pH of the aqueous solutions of this disclosure within a predetermined range. Preferred amounts of buffer salts and/or detergents can also control surface voltage changes, which helps to reduce the number of pathogens on the surface. Other preferred compositions of this disclosure include a buffer salt and/or detergent, each at a concentration of about 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.20; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.30; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.40; 0.5; 0.6; 0.7; 0.8; 0.9 or 1 wt.% based on the weight of the composition. Preferred amounts of buffer salts and/or detergents can provide a reduction in organoleptic damage at low pH values and can provide a reduction in alkalinity, a reduction in toxicity, as well as a safer use by processors, which was not observed using previously developed formulations in this area.

Some of the formulations described in the disclosure consist primarily of an inorganic acid, a copper(II) salt, and a buffer salt and/or detergent, as described herein. They are disclosed formulations that include an inorganic acid, a copper(II) salt, and a buffer salt and/or detergent as described herein, and [include] only those additional materials, such as water, that do not significantly affect the main and innovative features of the invention described here. Preferably, these formulations exclude chlorine and chlorine sources such as hypochlorite salts.

Some formulations of this disclosure consist primarily of an organic acid, a copper(II) salt, and a buffer salt and/or detergent, as described herein. They are the contemplated formulations of this disclosure, including an organic acid, a copper(II) salt, a buffer salt, and/or a detergent, as described herein, and [comprising] only those additional materials, such as water, that do not significantly affect the basic and innovative characteristics of the invention described here. Preferably, these compositions exclude, or contain in negligible amounts, chlorine and chlorine sources such as hypochlorite salts.

In some cases, the compositions of this application, which include an organic acid, such as ascorbic acid, have antioxidant properties. In other cases, the compositions may additionally contain an inorganic acid antioxidant. Antioxidants that are recognized as safe by the Food and Drug Administration are known in the art and include, for example, herbal extracts. Antioxidants can be included in concentration ranges from about 0.01 to about 1.0 wt.%, from about 0.01 to about 0.2 wt.%, from about 0.01 to about 0.15 wt. %, from about 0.01 to about 0.10 wt.%, or from about 0.01 to about 0.05 wt.%.

In some embodiments, the preferred formulations of the invention are non-oxidizing agents and may contain no or negligible amounts of ozone, peracetic acid, chlorine dioxide, and hypochlorite salts. The non-oxidizing formulations of this disclosure are beneficial when treated with recirculating water and can result in significant water savings.

In particularly preferred embodiments, the compositions of the invention are aqueous compositions. The concentrations of mineral and/or organic acid, copper salt, and buffer salt and/or detergent in aqueous solutions may vary depending on the particular treatment in which the aqueous solution is used. In some embodiments, the ratio of water and other components of the composition is 1:1. In other embodiments, the ratio of water and other components is from 1:1 to 2:1. In other embodiments, the ratio of water to the rest of the formulation is from about 1:1 to about 20:1, or from about 1:1 to about 500:1. In some embodiments, the ratio of water to the rest of the composition is from about 20:1 to about 500:1. The concentration suitable for final use in aqueous solutions may be lower than the concentrations in the original solid form, which can be diluted by about 500, about 450, about 400, about 350, about 300, about 250, about 200, about 150, about 100, about 90, about 80, about 70, about 60, about 50, about 40, about 30, about 20, about 10, about 5 , about 4, about 3 or even about 2 times. An appropriate dilution may be selected based on the effectiveness of the concentration, and the expected pathogenic contamination of the treated object.

In aqueous solutions of this disclosure, the pH may be neutral (pH about 7) or acidic (pH less than 7) or slightly alkaline (pH up to and including 9). In preferred embodiments, the pH is between about 1 and 7. In some cases, the pH is between about 2 and 7. In other embodiments, the pH is between about 3 and 7. In yet other embodiments, the pH is between about 4 and 7. Preferably, the pH value of the aqueous solutions of this invention is about 1; 1.1; 1.2; 1.3; 1.4; 1.5; 1.6,; 1.7; 1.8; 1.9; 2.0; 2.1; 2.2; 2.3; 2.4; 2.5; 2.6; 2.7; 2.8; 2.9; 3; 3.1; 3.2; 3.3; 3.4; 3.5; 4; 4.5; 5; 5.5; 6; 6.5; 7; 7.5; 8; 8.5 or about 9. In still other embodiments, the pH of the aqueous solutions of the invention is between 1.6 and 6.5, between 2.1 and 6.5, between 1.6 and 4.5, between 1.6 and 3.5 , or between 2.0 and 3.0. According to the disclosure, the pH of aqueous solutions is tested at about 5° C. or about 22° C., depending on the treatment chosen, using methods known in the art. One skilled in the art will recognize that the pH of aqueous solutions of this disclosure can be adjusted by adjusting the amount of acid in the solution.

In some disclosures, the formulations may alternatively be in solid form and presented as pellets, granules, powders, tablets, other soluble/water soluble films or packs, or pills. These formulations include 10% or less by weight of water/moisture. The compositions can be placed in a water-soluble film. Water soluble films can be made from, for example, polyvinyl alcohol film, aliphatic polyester film, or polyethylene glycol film. Such films are known in the art. Suitable films will be completely soluble or dispersible in water at temperatures above about 5°C. The films have a thickness of approx.

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0.0125 to about 0.125 mm, preferably from about 0.025 to about 0.075 mm. Water-soluble films can be sealed using, for example, thermal or ultrasonic welding, methods known in this field.

In other aspects, the present disclosure provides methods for reducing pathogens on surfaces, including applying any of the disclosure formulations to surfaces. The surface can be the surface of products or a solid surface. Preferably, the compositions used in the described methods are aqueous solutions. Surfaces can be treated in a variety of ways known in the art. For example, in preferred embodiments, any composition of this disclosure may be used to treat food surfaces by spraying, spraying, rinsing, soaking, dipping, washing, and the like. Deep immersion treatment can be used, and includes total immersion, either through mechanical or manual insertion, into single or multi-component tank designs. Spray treatments may be used, and be implemented with one or more nozzles or drip applicators. In some embodiments, ultrasonic nozzles may be used during spraying, or this may be combined with mechanical grinding to improve the surface of the contact zone.

Aqueous solutions used in the methods described may be at any commercially reasonable temperature that will not damage the surfaces treated with the solution, for example, from about 32°F (0°C) to 212°F (100°C) or from about 32°F (0°C) and up to 135°F (57°C). The solutions may be cold, such as about 32°F or about 38°F (about 3°C), or warm, such as about 135°F. In other variations, these solutions may be about 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52 , 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77 , 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102 , 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127 , 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139 or 140°F. In other cases, the aqueous solutions used in the described methods may be from about 140 to about 212°F. Higher temperatures may be preferred for hard surface treatments.

In preferred embodiments, the treatment comprises immersing the surface for a time sufficient to provide adequate coverage and/or penetration of the compositions described onto and/or within the surface. The immersion time may be, for example, up to 5 minutes, up to about 4 minutes, up to about 3 minutes, up to about 2 minutes, or up to about 1 minute. Immersion times of less than 1 minute are also provided and can be as short as two seconds at certain lower pH values. Preferred immersion times are from about 2 to about 180 seconds, from about 2 to about 120 seconds, from about 2 to about 90 seconds, from about 2 to about 60 seconds, from about 2 to about 45 seconds, or from about 2 to about 30 seconds.

Disclosure compounds are effective in reducing the number of pathogens, ie microbes, viruses, or parasites on surfaces. That is, surface treatment with disclosure formulations reduces the growth and/or multiplication of bacteria, viruses, and/or parasites (e.g., by killing or significantly slowing the growth of such bacteria, viruses, and/or parasites) on the surface compared to an untreated surface. composition of information disclosure. For example, the compositions of the invention are effective in reducing pathogens by about 10%, compared to a surface not treated with the disclosure composition. In other variations, disclosure formulations are more effective by about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300 , 400, 500%, or even more, in the reduction of pathogens on the surface, compared to surfaces that were not treated with the disclosure compound. Disclosure formulations are more effective in reducing pathogens by about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, or about 90% compared to a surface that has been treated with standard chlorine and citric acid. acid.

Hazy/smoky/high humidity and similar conditions increase the risks associated with disease causing pathogens spread through the digestive tract. Disclosure formulations are effective in reducing pathogens, even in foggy/smoky/high humidity and similar environments such as food processing environments.

The following examples are provided to illustrate the compositions, processes, and properties described herein. These examples are illustrative only and are not intended to limit disclosures regarding the materials, conditions, or process parameters set forth in this document.

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Examples

Example 1 General bacterial test.

The Vivione RAPID-B instrument was run using the manufacturer's instructions (Vivione Biosciences, LLC, Pine Bluff, AR). Performance testing was performed in accordance with the manufacturer's instructions to verify performance and identify any potential problems. The Rapid-B system was then prepared to perform a total bacteria count test on Petri dishes. Treatment solutions were then prepared based on the protocol tested. Solutions were inoculated 99:1; which means 99 ml of treatment solution was mixed with 1 ml of bacterial wash from poultry, which forms a dilution of 10' ² . Serial dilutions (a 10' ³ dilution was commonly used) were made using sterile buffered peptone water to increase the dilution factor. Samples were prepared by combining 570 µl of sterile buffered peptone water, 330 µl of TPC reagent (TPC=total bacteria count), and 100 µl, from the dilution tube, was tested in the reaction tube. This process dilutes the sample an additional 10' ¹ (so a total dilution of 10' ⁴ was tested). The sample in the reaction tube was left for 15 min, periodically mixing on a vortex. A generic bacterial protocol was opened on the Vivione instrument and performance parameters were validated according to Vivione Biosciences guidelines. After 15 minutes, the reaction tube is placed in the sample holder and moved to the working position to start the analysis. After starting the sample, the holder returns to the flushing position to ensure flushing is completed.

Example 2 Processing green leafy vegetables slows down spoilage.

The antimicrobial formulation of this disclosure, referred to here as formulation A or formulation PS+, has been added to the wash water and is used in a deep tank to treat green leafy vegetables after harvest. Wash water treated with chlorine and citric acid as standard was also used for comparative testing. Wash water samples were used in deep tanks at approximately 38°F and were used to process leafy vegetables for thirty (30) seconds in turbulent water with over 250 samples taken directly from the surface of leafy vegetables for results.

Composition A, comprising an inorganic acid with a low pKa value (sulfuric acid) in combination with copper sulfate pentahydrate at 35-40 ppm (copper ions in an amount of 9-10 ppm), a buffer salt (ammonium sulfate 0.5%) in water at 38°F. The pH value of the wash water solution was 2.2.

The PS+ composition, which includes an inorganic acid with a low pKa value (sulfuric acid) in combination with copper sulfate pentahydrate at 35-40 ppm (copper ions in an amount of 9-10 ppm), a buffer salt (ammonium sulfate 0.05%) and surfactant (alkyl polyglycoside) in water at 38°F. The pH value of the wash water solution was 2.2.

Green leafy vegetables were treated with either Formulation A or standard chlorine and citric acid treatments. After processing, green leafy vegetables were checked for organoleptic characteristics and visible spoilage, microbiological contamination was measured by counting the total number of bacteria on Petri dishes. The results are presented in table. 1.

Table 1

Day Total number of bacteria: treatment with composition A Treatment composition A: organoleptic characteristics of the product Total bacteria count: standard chlorine treatment Standard chlorine treatment: organoleptic characteristics of the product 0 7265 Texture: crispy/brittle/fresh Color: bright, fresh look Smell: no unusual smell 208 217 Texture: crispy/ brittle/fresh Color: bright, fresh look Smell: no unusual smell 7 60 833 Texture: crispy/brittle/fresh 558 100 Texture: less crispy/brittle/fresh, less

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Color: bright, fresh look Smell: no unusual smell springy Color: some appearance deterioration (fading) Smell: slightly stale 14 652 500 Texture: crispy / brittle / fresh Color: bright, fresh look Smell: no unusual smell Visible degradation: < 1% General shelf life: acceptable, some increase in visual defects 1 064 652 Texture: starts wilting, texture less crispy/ brittle/fresh, less elastic Colour: less bright, duller appearance Odour: enhanced staleness Visible decomposition: 4 - 6% General shelf life: unacceptable, common deterioration, increase external defects, secondary spoilage 18 4,200,000 Texture: crisp/brittle/fresh Colour: bright Odour: slightly stale Visible decomposition: less than 3% Total suitability: unacceptable, general deterioration, increase in external defects 4 333 333 Texture: beginners wither leaves flabby, less elastic. Colour: less bright, even duller appearance Smell: amplification stale, musty Visible decomposition: 6- 8+% increase humidity General suitability: unacceptable, common deterioration, increase external defects, secondary spoilage

For Formulation A treated products, there was a significantly lower total bacteria count on day 0 [count using petri dishes] (7.268 vs 208.217). At day 7, the total bacteria count was still significantly lower than the standard treatment, while still providing a relatively improved texture, color and odor. At day 15, the total bacteria count is still well below standard processing values with a significant improvement in texture, color, odour, signs of spoilage, and commercial acceptability. After 14 to 16 days, there was an increase in visible wilting, primarily on the leaf sections, where the sections were jagged and dented, but still to a low degree and retaining the best texture, color, smell and signs of decay. There is a clear reduction in oxidative discoloration in intact leaves. Leaves treated with standard chlorine began to show general desiccation from day 10 to day 14 associated with tissue damage. Secondary spoilage foci developed rapidly in samples treated with standard chlorine, while spoilage in samples treated with composition A (even on the 20th day) was observed only on individual leaves.

Wash water samples with the addition of the PS+ composition were used at a pH value of 2.2. Subsequent tests showed a significant reduction in the number of microorganisms when quantified by plating on petri dishes [general bacterial test]. Individual test results (214 data points) at pH 2.2 are shown in FIG. 1. The results showed a 92% reduction in the number of micro-organisms in cultures on Petri dishes during all tests compared to optimized chlorine treatments using 20 ppm chlorine at pH 6.0. Decreases in [microorganism] numbers were generally highest under the most foggy/smoky conditions, in which the risks of foodborne illness are highest.

In FIG. 2 shows the results of accelerated shelf life tests at 45°C. Approximately 30% reduction in active spoilage was achieved with PS+ antimicrobial treatment compared to standard hypochlorous acid treatment. While not wishing to be bound by any particular theory, it is believed that this reduction can be attributed to a significant reduction in pathogen load when the treatment is put in place.

Example 3 Absence of Organisms in Wash Water Supplemented with an Antimicrobial Compound of this Disclosure.

Wash waters with the PS+ composition from Example 2 were used on yeasts and molds and for

- 7 041635 total count of microorganisms on Petri dishes after processing green leafy vegetables after harvest. For yeasts and molds, the number of colony forming units (cfu)/ml was measured. The limit of detection was 1 cfu/mL. Out of 30 PS+ wash water samples, 29 were found to be < 1 cfu/mL and 1 was reported as 1 cfu/mL, the limit of detection. Standard treatment with chlorinated wash water revealed <10 cfu/mL for yeast, an average of 55 cfu/mL for molds, and an average of 3,172 on a total plate count of bacteria.

Example 4 Significant Reduction of Pathogenic Organisms with Antimicrobial Compounds Disclosure.

Test cultures were treated with Formulation A and Formulation PS+ used in Example 2 at specified concentrations and exposure times, and then counted. The number of test cultures before treatment was compared with their number after treatment to determine the bactericidal ability of each formulation.

The following test organisms were evaluated:

Staphylococcus aureus (ATCC #6538),

Listeria monocytogenes (ATCC #7646),

Escherichia coli O157:H7 (ATCC #35150)

Salmonella enterica serovar Anatum (ATCC #9270), methicillin-resistant Staphylococcus aureus (MRSA) (ATCC #33592).

Cultures were prepared from lyophilized preparations according to the manufacturer's instructions (American Type Culture Collection (ATCC), Manassas, Virginia) or taken from petri dish stocks. Each culture was transferred to tryptic soy broth (TSB, Neogene, Lansing, MI) and incubated at 35±2°C for 24±2 h. water and resuspended to its original volume. The culture was sown on tryptic soy agar (TSA, Neogene) in appropriate dilutions that determine the actual final concentration.

The culture was exposed to each of the disinfectant solutions - composition A and composition PS+, diluted with sterile deionized water to pH 2.2 or 1.3. A separate 250 ml Erlenmeyer conical flask containing 99 ml of each disinfectant was prepared along with a duplicate flask containing 99 ml of Butterfield's phosphate buffer as a control. The disinfectant flasks were gently spun to create residual fluid movement, and then a 1 ml aliquot of test culture was added to the center of each flask, avoiding contact with the neck and walls of the flask during inoculation. Each flask was agitated by rotating for 1 min to thoroughly mix the contents, and then 1 ml of the mixture was added to a 9 ml test tube with a lecithin neutralizing solution prepared in accordance with the International Official Method 960.09 (Bactericidal and Detergent Sanitary Effects of Disinfectants) of the Association of Chemists -Analysts employed by US government organizations (EPA, Rockville, Maryland), which is incorporated herein by reference for all intents and purposes. This procedure was repeated as a control, using Butterfield's phosphate buffer and using the same test inoculum.

After processing and neutralization, the samples were poured into Petri dishes in serial dilutions up to 10 ^-6 (considering the neutralization tube as 10 ^-1 ) using Trypton Glucose Agar (TGEA, Neogene). Trypton Glucose Agar plates were incubated at 36±1° C. for 27±3 hours. After incubation, the dishes were counted using a Quebec colony counter (Model No. 3325, Reichert Technologies, Depew, NY).

The number of observed colonies on treated and untreated samples was recorded.

Raw observed data for each sample were converted to log10 cfu/mL. The number of test organisms present in the treated samples was compared with the number present in the control samples to determine the log reduction for each test organism.

The test results are given in table. 2.

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Claims (8)

table 2 Estimated (log reduction) by AOAS using disclosure formulations Treatment Composition A Composition PS+ pH 1.3 2.2 1.3 2.2 S. aureus 3.42 3.16 >5, 91 >5, 84 E. coll 3, 08 3, 60 >5, 56 >5, 67 Salmonella enterica serovar Anatum 3, 1 3, 02 >5, 7 >5.78 Listeria monocytogenes 3.79 3, 08 >5, 7 >5, 82 MRSA 3, 93 3, 93 >5, 73 >5, 73 Tests showed a significant reduction in microbial counts, with a synergistic effect being pronounced when copper ions were combined with detergent addition. An inorganic or organic acid buffered with a conjugated salt produces significant antimicrobial effects without organoleptic damage when combined with copper and detergent. EXAMPLE 5 Peak Organic Load Control Using Disclosure Compound Over Traditional Compositional Techniques. Disclosure formulations have shown significant suppression of peak organic load compared to conventional treatments. Fig. 3 shows the results of measurements of organic load (in DCO, the estimated concentration of colony forming units) on the actual surfaces of lettuce leaves from 250 randomized samples taken over a period of six weeks. Lettuce leaf samples were treated with the PS+ formulation of Example 2 or standard hypochlorous acid treatment. This is especially important to reduce the likelihood of unexpected peaks in organic load, which can lead to high levels of pathogens that can lead to foodborne illness. When ranges are used herein to describe physical properties, such as molecular weight, or chemical properties, such as chemical formulas, then all combinations and subcombinations in the ranges for specific embodiments are intended to be included. The disclosures of each patent, patent application, and publication cited or described herein are incorporated herein by reference in their entirety. Those skilled in the art will appreciate that numerous changes and modifications may be made to the preferred embodiments of the invention and that such changes and modifications may be made without departing from the spirit of the invention. For this reason, the appended claims are intended to cover all such equivalent variations as fall within the true spirit and scope of the invention. CLAIM 1. An antimicrobial composition containing an inorganic acid buffered with a conjugated salt; copper(II) salt, where the concentration of copper(II) salt is from 35 to 100 ppm; and an alkyl polyglycoside which is present in a concentration ranging from 0.01 to 0.3% by weight, based on the total weight of the composition, wherein the composition is an aqueous solution and has a pH of 1-4. 2. The composition of claim 1, wherein the copper(II) salt is copper(II) sulfate. 3. A composition according to any one of the preceding claims, wherein the copper(II) salt concentration is between 35 and 80 ppm, between 35 and 60 ppm, between 35 and about 50 ppm, or between 35 and 45 ppm. 4. A composition according to any one of the preceding claims, wherein the inorganic acid is sulfuric acid, phosphoric acid, hydrochloric acid, or a combination thereof. 5. The composition according to any one of the preceding claims, further comprising an organic acid, where the organic acid is citric acid, ascorbic acid, lactic acid, acetic acid, peracetic acid, or combinations thereof. 6. Composition according to any one of the preceding claims, wherein the conjugated salt is ammonium sulfate, sodium sulfate, sodium chloride, calcium sulfate, or combinations thereof. 7. The composition according to claim 6, in which the concentration of the conjugated salt is from 0.01 to 0.5 wt.%, from 0.01 to 0.4 wt.%, from 0.01 to 0.3 wt. .% or from 0.01 to 0.25 wt.%. 8. Composition according to any one of the preceding paragraphs, additionally containing an antioxidant.