WO2015088307A1 - Phytochemical compositions for disinfecting chillis (capsicum annuum) - Google Patents

Abstract

The invention relates to compositions for effectively disinfecting and/or preserving fruit and vegetables in relation to pathogenic and destructive micro-organisms. The aforementioned compositions contain plant-derived extracts with antimicrobial activity, which can act either alone or in combination with other disinfectant agents, such as, for example, organic acids and chlorine compounds. The compositions of the invention can eliminate or inactivate microbial contamination of chillis, including by pathogenic micro-organisms, without altering the nutritional and/or dietary and/or sensory properties thereof.

Description

 PHYTOCHEMICAL COMPOSITIONS TO DISINFECT CHILES (Caosicum annuum)

TECHNICAL FIELD OF THE INVENTION

The present invention relates to the development of formulations containing phytochemicals present in plants and which are used as disinfectants and preservatives for food, for example of plant and animal origin; more particularly to aqueous formulations based on Jamaican chalices (Hibiscus sabdariffa), the method by which it is obtained and its uses as an effective formulation to eliminate pathogenic bacteria from fruits and vegetables, but with the highest effectiveness for chili peppers (Capsicum annuum)

STATE OF THE TECHNIQUE

Chile is one of the main agricultural products in Mexico with a little more than 138 thousand hectares dedicated to planting. The most important types of chili that are sown, both in the open field and in protected agriculture, are: Jalapeño (the one that is most produced), followed by the Serrano, Poblano, Ancho, Mirasol, Puya, Pasilla, Chilaca, Campana and to a lesser extent but also sown: Habanero, Colorado, Costeño, Tree (rat tail), Mulato, Tobaquero, Anaheim, Bell pepper, Caloro, Cayene, Water, Guajillo, Shadow mesh, Apple tree, Organic, Perón, Regional, Soledad, X-cat-ik, Morrón, Pepper, Ciltepin, of Time (Sagarpa, 2013)

However, along with the increase in the consumption of chili peppers, there have been outbreaks of diseases caused by bacteria associated with the consumption of chili peppers (CDC, 2008). For example, jalapeno and serrano peppers have been implicated in Salmonella outbreaks at least once in the period from 2008 to 2013.

This type of outbreaks has caused strict regulation for producers that export peppers to the United States (USA); and has often meant retention of shipments of peppers in the frontier, partial or total closing of the export of this product to the USA and economic losses on the part of the producers when not fulfilling the microbioiotic standards. It should be noted that although in Mexico there are no reports of disease outbreaks of microbial etiology associated with the consumption of chili peppers, due to the poor hygiene practices that generally occur during the cultivation, harvest, transport and commercialization of chili peppers, participation is expected of these raw products in disease outbreaks. A fact that supports this observation is the high frequency with which Salmonella strains and Escherichia coli pathogenic groups have recently been isolated from chili peppers in Mexico (Castro-Rosas et al., 2011; Cerna-Cortes et al., 2012).

Recent outbreaks of food diseases associated with chili peppers created the need to determine the sources of chili contamination and understand the survival and / or growth of pathogenic microorganisms in chili peppers; These have led to the development of innovative control technologies. In general, the pathogens in the peppers could be controlled by preventing contamination during the cultivation and harvest of the products, also by using disinfectants with antimicrobial power in the harvested product, and by storing the peppers at low temperature. However, the disinfection of chiles has been identified as the most important stage for the microbial safety of raw chiles. In general, chiles are not consumed directly as they are harvested.

After harvesting either in the field or in the industry (and even at home) they receive various treatments that tend to favor their conservation and / or safety. The application of washing and disinfection of chili improves its microbial image. However, it is difficult to safely achieve the inactivation or removal of pathogenic microorganisms even under extreme conditions of treatments that do not sensibly damage the peppers. The prevention of contamination of peppers is also a control strategy because the growth of pathogens is not required to cause disease. Therefore, additional control measures may be of value. It should be noted that the behavior of pathogenic microorganisms in peppers is affected by the location of the pathogen in the product, the quality of peppers, storage temperature, type of packaging, and relative humidity. The peppers usually have few nutrients on their surface which limits the growth of pathogens during storage at refrigeration temperatures. However, the microorganisms that constitute native fungal populations and yeast in raw tomatoes could injure the pericarp tissues of chili at a level that would favor the growth of pathogenic bacteria during storage (Wade and Beuchat, 2003).

It has been reported Salmonella is able to survive for a long time on the surface of whole peppers both in refrigeration and at room temperature and is also able to multiply in chopped peppers (Castro-Rosas et al., 2011). Also, once on the peppers Salmonella could produce extracellular polymers in the cuticles of the peppers, as has been observed in other vegetables such as tomatoes, which leads to the formation of a biofilm that can protect it against disinfectants (Iturriaga et aL, 2007 ), On the other hand, pathogens on the surface of peppers can contaminate internal tissues during slicing and can then survive or grow on slices. Several research findings indicate that bacterial pathogens can infiltrate plant products, such as chili peppers (Bartz, 1982; 1999; Guo et al., 2002; Ibarra-Sanchez et al., 2004; Zhuang and Beuchat 1995) when there are a temperature differential between the vegetable product and the wash water and by the hydrostatic pressure when the vegetables are submerged in the receiving tank (Bartz, 1982; Bartz and Showalter, 1981).

Bacterial infiltration increases in raw vegetables in the presence of wounds and punctures. Infiltrated pathogens are not eliminated by normal washing practices. The main benefit of the addition of antimicrobial chemicals (such as hypochlorite-based or organic acid-based chemical disinfectants) to the wash water of peppers is the control of propagation of pathogens, their inactivation and / or their infiltration into chiles. However, currently available chemical disinfectants have limited benefits over plant products, such as chili peppers. The antimicrobial effect of solutions of hypochlorite, hydrogen peroxide, peracetic acid and electrolyzed water in its ability to reduce pathogens in plant products during the washing process has been studied. However, it has been concluded that these treatments have a limited effect on the pathogenic microorganisms, presumably because the active agents do not have sufficient contact with the pathogenic microorganisms on the raw plant products.

The disinfection process refers to the physical destruction of microorganisms whose activity compromises the safety or sensory characteristics of a food. The effect can be achieved through physical or chemical means, its effectiveness depending on the microorganisms (type and number), the substrate on which they are (presence of organic matter), the structure of the material (which allows direct access from germicide to microorganisms) and germicide (concentration, temperature and contact time) (Fernández, 2000). In the disinfection process, the germicidal substance participates in chemical reactions, so that the greater the number of microorganisms, the greater the demand of the agent to achieve a total inactivation of the population. The susceptibility to a specific germicide varies among microorganisms; some are inactivated from the first moment of contact, while at the other end there may be survivors. Finally, it should be borne in mind that among the microorganisms it is possible to select strains with increasing resistance to the effect of a specific germicidal agent. Consequently, over time, much higher concentrations of the disinfectant than the initial ones are required to reach the same level of inactivation (Fernández, 2000).

Different studies show that disinfection treatments of raw agricultural products often have a weak or limited effect. For example washing and disinfection with 200 mg / L of active chlorine (hypochlorite), of iodine (iodophore), chlorine dioxide or 100 mg / L of a commercial product based on grapefruit seed extract (citricidal) reduced the content of alfalfa sprouts by only 1-2 logi ₀ ; the decrease in S. typhi or V. cholerae 01 inoculated in the laboratory was not greater than 1.5 log ₁₀ CFU / g (Castro-Rosas and Escartín, 1999),

 The food industry has a variety of germicidal agents. Its virtues and limitations require carefully selecting those that best fit each particular need (Fernández, 2000). The inactivation of pathogenic bacteria in food processing plants is a basic requirement to control them and prevent their access to the finished product (Álvarez, 1998).

It is common for a germicide to be considered effective when it demonstrates the ability to inactivate at least 3 Log ₁₀ of a microorganism suspension in 30s (Fernández, 2000).

Chlorine-based solutions are a cheap and available disinfectant as hypochlorite or in its slow-release forms (chloramines, for example) (Lelieveld et al., 2013). Hypochlorites have a broad spectrum of antibacterial activity, although they are less effective against spores than against non-spore-forming bacteria and have a low effect against mycobacteria (Russell et al., 2004). Chlorine solutions, such as sodium hypochlorite or chlorine dioxide, are widely used by the food industry as a disinfectant. Both are strong oxidizers that acféam at the level of membranes and other cellular constituents (Harmon et al., - ^ 87) 7 However, the former has the disadvantage of reacting easily with organic matter, so it is inactivated faster . In the second, the interfeteieia is minimal (Castro-Rosas and Escartín, 1999). The main disadvantage of? Sodium hypochlorite is that humidity, heat, light and especially the presence of organic matter, increase the loss rate of free chlorine. The germicidal activity of chlorine has generally been attributed to hypochlorous acid (HOCI), which is generated in aqueous solutions of sodium hypochlorite and other chlorine-containing compounds. Disinfectants can be incorporated into the wash water and thus contribute to the reduction of the microbial load. The effectiveness of hypochlorite is not only affected by the exposure time and concentration of free chlorine, but also by other factors such as temperature, pH, type of strain, as well as presence and type of organic matter (Álvarez, 1998). However, some authors point out that the efficiency of hypochlorite in reducing pathogenic microorganisms present in vegetables is limited (Adams et al., 1997). Chemical compounds derived from chlorine, iodine and silver have typically been used as vegetable disinfectants, such as chili peppers. However, recently several studies show that disinfection treatments with these compounds are inefficient in eliminating or reducing levels of microbial pathogens. For this reason, many countries have abandoned the use of hypochlorite or iodine solutions for the disinfection of raw vegetables.

Organic acids have traditionally been used as food preservatives or in solutions to disinfect raw vegetables. Its antimicrobial effect is exerted through the undissociated form causing a decrease in pH.

Acetic acid is a harmless substance; There are no official limits for daily intake in man. When acetic acid is incorporated into a food, two effects are expressed, one acidifying and the other preservative. At a concentration of 1-2% it inhibits almost all the total flora within reasonably high initial load limits. At 0.1% it acts on the majority of pathogens and sporulates; 0.5% has an effect on toxigenic fungi. The efficacy of acetic acid against some specific pathogens has been evaluated using some foods as a medium. Published reports are often difficult to compare because acid concentrations have been variables expressed as percentage, molarity or final pH of the acidified test medium. The antimicrobial activity depends on the exposure time, temperature, type of acid, acid concentration, dissociation level and pH (Harmon et al., 1987). However, the general results show that the effectiveness of acetic acid increases as the concentration increases, the pH decreases, the temperature increases and the microbial load decreases (Harmon et al., 1987). Among bacteria, Gram positive are usually more resistant than Gram negative bacteria (Rameshkumar et al., 2007). Bacterial spores and viruses are more resistant than vegetative cells. However, organic acids have also shown little effectiveness in disinfecting raw vegetables (Fernández, 2000).

Recent research indicates that antimicrobial chemicals in the vapor phase can significantly reduce plant surface pathogen populations. The use of 5 mg / liter of chlorine dioxide gas for 1 h was significantly more effective against Salmonella in the scar of the peduncle of tomatoes that were aqueous solutions of 200 ppm of sodium hypochlorite (2 min of exposure) and 1200 ppm of acidified sodium hypochlorite (2 min exposure) (Yuk et al., 2005). The use of 10 mg / liter of ozone completely inactivates 7 log CFU of Salmonella enteritidis from the surface of cherry tomatoes after 15 min, however, the color of tomatoes is affected (Das et al., 2003).

Because the vapor phase antimicrobial agents can be effective against bacteria adhering to locations of raw agricultural products not reached by active agents in aqueous solution, their use in packaged products (in plastic bags) or during product processing (in the company) could provide an extra benefit in pathogen control. However, this type of steam treatment would not be an optional or practical treatment for primary producers of chili peppers in the field, as producers generally sell their product packed in cardboard boxes or wood among other things for ease and to avoid accumulation and humidity what would happen if plastic bags were used. In addition, this would not be a practical treatment to apply in restaurants or homes. The use of chemical substances as raw vegetable disinfectants to improve or preserve their safety is a procedure universally used by producers. However, some of these antimicrobials may be toxic to consumers; This is the case of hypochlorite solutions. Recent reports indicate that hypochlorite in solution can form cancer precursors. In addition, many of the chemical disinfectants, such as solutions based on iodine or colloidal silver, show limited or varied antimicrobial effects in products such as raw vegetables; A similar situation occurs with food preservatives.

Due to this, disinfectants and preservatives obtained from plants have recently emerged as a viable alternative, since these could have equal or greater antimicrobial potential and with minimal risk to consumers.

The application of garlic extracts in fresh fruit against postharvest diseases has obtained complete control of the brown rot of peaches caused by Monilinia fructicola (Roller, 2003). Yucel and Karapinar (2005) evaluated the reduction of S. typhimurium in onions by applying lemon juice, vinegar and their mixtures, observing a respective reduction of 0.87-2.93, 0.66-2.92 and 0.86-3.24 Log CFU / g.

Essential oils from plants are capable of inactivating the pathogens of interest in fresh products. Of 96 different types of essential oils examined, only 3 were effective against E. coli 0157: H7 and enteric Salmonella which were of oregano, thyme, and cinnamon. In another study, 16 individual compounds of the most effective oils against E. coli 0157: H7 and Salmonella were tested and the most effective compounds were found to be thymol, cinnamaldehyde, and carvacrol (Friedman et al., 2002). This information was obtained using the oil in the liquid phase. There is limited information available on the effectiveness of essential oils in the form of steam. In another study, Muñoz (2003) evaluated the effect of two concentrations of carvacrol and the commercial disinfectant Boradantix © (EVESA, Vegetable Extracts SA) in the survival of L. monocytogenes, P. fíuorescens, E. coli, Erwinia caratovora and S. typhimurium in lettuce and carrot juice. All study microorganisms were inhibited at both concentrations of carvacrol. The bacteria studied showed greater sensitivity towards carvacrol than Boradantix ©. Lin et al., (2000) evaluated the effect of allyl and methyl isocyanate (AITC / MITC) (key components of green mustard) on L. monocytogenes, E. coli 0157: H7 and S. montevideo, inoculated on lettuce surface and tomato AITC was more effective against Salmonella and E. coli, achieving 8 Log reduction with a generated steam treatment of 400 μΙ of AITC after 4 and 2 days, respectively on lettuce. 8 Logs of reduction of S. Montevideo on tomato cuticle with 500 μΙ of AITC were also reached.

There have been relatively few studies of the antimicrobial action of essential oils in model food systems and in real foods. However, the effectiveness of essential oils in vitro is often much better than in vivo or in situ, that is in food. Generally, when applying a plant antimicrobial to a food or in vitro, 10 to 100 times the concentration of antimicrobial is needed than what is observed in vivo. For example, the essential oil of peppermint (Mentha piperita) has been shown to inhibit the growth of Salmonella enteritidis and L monocytogenes in culture media for 2 days at 30 ° C. However, the effect of peppermint essential oil on Greek snacks tzatziki (pH 4.5) and taramasalata (pH 5.0) and in pate (pH 6.8) at 4 ° C and 10 ° C was variable (Roller, 2003). Salmonella enterítidis was eliminated in snacks under all the conditions examined but not when it was inoculated in pate and kept at 10 ° C. In this same study, L monocytogenes behaved similarly, since the microbial count decreased in snacks but increased in pate (Roller, 2003). The growth of E. coli, Salmonella sp., L. monocytogenes and Staphylococcus aureus were inhibited by the essential oil of oregano in culture broths. However, when these oils were tested in foods such as eggplant, taramasalate or mayonnaise, reactions such as an increase in pH, temperature increase were observed and in the case of emulsions separation of used oil (Roller, 2003). In another study L monocytogenes and S. typhimurium were inhibited in meat treated with essential oil of clove and oregano, respectively. A marked reduction in Aeromonas hydrophila has also been reported in cooked pork that was treated with clove or coriander oils, vacuum packed or air-packed and stored at 2 ° C and 10 ° C. (Roller, 2003).

The differences observed between studies of antimicrobial effect when directly applied oils extracted from plants on microorganisms (microorganisms in aqueous suspension) and those in which there is a food or organic matter in between, it is possible that it occurs by interference with the components of food or organic matter (proteins, fats, sugars, salts). Therefore, it is very possible that only a proportion of the essential oil added to the food has antibacterial activity. On the other hand, the spatial distribution of the different phases (solid / liquid) in a food and the lack of homogeneity of factors such as pH, at _w among others, can play a role in efficacy. Due to all of the above, studies aimed at the search for alternative antimicrobials are under way in various parts of the world (Jongen, 2005). Among the new disinfectant alternatives, natural compounds with a broad antimicrobial capacity have been chosen.

It should be noted that the extracts obtained from some plants have shown antimicrobial effect against strains of antibiotic multiresistant pathogens, which opens up a whole new field for the development of new antimicrobials for use in humans and animals.

As an antecedent of the present application, the antimicrobial effect of about 60 different plants used in herbalism has been evaluated (Cruz-Galvez et al., 2013); where some of these have shown a high antimicrobial power against different pathogenic microorganisms, such as Salmonella or Escherichia coli 0157: H7, among others, as well as against food spoilage microorganisms (Pseudomonas aeruginosas, por example), and the plant that has shown the greatest antimicrobial effect has been the chalices of the Jamaican flower, in some cases the antimicrobial effect being greater than that of commercial disinfectants based on hypochlorite, iodine, colloidal silver or that of antibiotics such as Penicillin

Different researchers have also reported that the chalices of the flower of Jamaica {Hibiscus sabdariffa L) have substances with high antimicrobial power (Aziz er a /., 1998; Fernández er a /., 1996; Kang et al., 2007).

 Jamaica is one of the plants in which the presence of antimicrobial compounds in dehydrated chalices has recently been reported (Aziz et al., 1998; Fernández et al., 1996; Kang et al., 2007). In the chalices of the flower of Jamaica a range of phytochemical compounds have been detected that could be responsible for the observed antimicrobial effect, such as for example polyphenols (Tajkarimi eí al., 2010), among them some phenolic acids (Tajkarimi et al ., 2010), as well as flavonoids, catechins and epicatechin (Friedman, 2007). However, there are no specific studies that show which are the chemical molecules or compounds responsible for the antimicrobial effect observed in the chalices of Jamaica. Different researchers agree that further studies are necessary to identify the specific molecules responsible for the antimicrobial effect caused by the calyxes of Jamaica in solution.

There are few patent documents that describe extracts of the chalices of the flower of Jamaica (Hibiscus sabdariffa) and its use as a material with antimicrobial properties.

For example, patent application JP2002128602 describes its use in an agrochemical composition to protect plants in fields of crops, while application US20100323046 describes the use of a crude extract of the calyces of Jamaica to produce a drug to treat urinary infections caused by Escheríchia coliy Candida albícans. Patent application KR20080092186 describes an extract from Jamaica that is used to improve the quality of beef, pork and chicken and to increase its storage stability. The extract is prepared by ethanol extraction and subjected to a cold drying process. The concentration of the extract in the composition is 500 mg / ml and the meat is treated with a preparation of 0.5 to 3.0% (by weight).

On the other hand, in the application US20120015062, compositions comprising extract of the Agapanthus africanus plant and compositions comprising this extract plus other extracts of other different plants are described, such as for example plants of the Rosa or alfalfa family for use as agents in the Biological protection of other plants including their seeds. Although this patent application document refers to the article published by Leksomboon et al. (Kasetsart, Journal Natural Science 35: 392-396, 2001.) where it is mentioned that extracts obtained from various plants (Hibiscus sabdariffa, Psidium guctjava, Punica granatum, Spondias pinnata and Tamarindus indica) have an antimicrobial function, this document does not provide no experimental evidence involving the extracts of Hibiscus sabdariffa for the same use that is given to the extracts of Agapanthus africanus.

Therefore, it is necessary to have effective protective antimicrobial compositions to prevent and / or combat microbial contamination of food, mainly those that are consumed raw, such as chili peppers, in order to preserve and consume them without the risk of acquire diseases caused by their contamination with pathogenic microorganisms.

Until before the present invention, it had not been possible to develop effective compositions to disinfect efficiently and without damage to the product as described herein, and which at the same time allowed to preserve the nutritional properties of fruits and vegetables and not affect, for example the quality of chili peppers, whereby it is possible with the present invention to obtain mirobiologically harmless raw chiles. BRIEF DESCRIPTION OF THE FIGURES

Figure 1. The nuclear magnetic resonance (NMR) spectrum of

PROTON ( ¹ H) of the dry ACETONIC extract obtained from the chalices of Jamaica that was used in the present invention.

Figure 2. The nuclear magnetic resonance (NMR) spectrum of

PROTON ( ¹ H) of the dry METHANOLIC extract obtained from the Jamaican calyces that was used in the present invention.

BRIEF DESCRIPTION OF THE INVENTION

In accordance with the aforementioned problems, there is a need to provide a more effective formulation to inactivate or remove pathogenic microorganisms from chili peppers (Capsicum annuum) even under extreme conditions of treatment, but which do not sensorially damage the food.

The present invention relates to solutions containing phytochemicals present in plant extracts that are used as disinfectants of plant and animal foods, for example directed to the disinfection and preservation of plant foods, particularly disinfection and / or preservation of chili peppers (Capsicum annuum).

One embodiment of the present invention relates to obtaining a vegetable preparation comprising a methanolic extract of the chalices of the flower of Jamaica {Hibiscus sabdariffa) or an acetonic extract of the calyces of the flower of Jamaica, which are useful for eliminate pathogens present in food (disinfectant effect) and to delay the deterioration of food or preserve its safety (conservative effect).

Another embodiment of the present invention relates to obtaining plant-derived extracts that are used as disinfectants against pathogenic microorganisms present in food and to delay the deterioration of food and / or preserve its safety, that is, as food preservatives, which constitute an alternative to the use of traditional disinfectants that can become toxic to humans, animals or the environment.

Another embodiment of the present invention relates to the preparation of compositions containing the extract of Jamaican calyces (Hibiscus sabdariffa) that have a food disinfectant and preservative function together with other compounds that have disinfectant properties such as acetic acid, hypochlorite, etc.

Another embodiment of the present invention relates to obtaining extracts obtained from Jamaican chalices that have a disinfectant or preservative effect when applied to food. One aspect of this modality refers to the application of extracts obtained from chalices of the Jamaican plant {Hibiscus sabdariffa) that have a disinfectant or conservative effect when applied to foods of plant origin, preferably chili peppers. Another embodiment of the present invention is the development of a method for obtaining acetonic extract from Jamaican calyces, an extract that proves to be useful as a food disinfectant and preservative.

Another embodiment of the present invention is the development of a method for obtaining the methanolic extract from calyxes of Jamaica, an extract that turns out to be useful as a disinfectant and food preservative.

Another embodiment of the present invention is a method of treatment and / or preservation of foods of animal and / or vegetable origin by applying compositions containing extracts of Jamaican chalices that allow disinfection and preservation thereof. The use of Jamaican calyx extracts as a disinfectant and / or food preservative is another modality described in the present invention. Compounds from the calyces of Jamaica can be useful in the preparation of an efficient disinfectant to eliminate pathogenic bacteria present in raw plant-based foods, such as chili peppers. The present invention describes two types of extracts of Jamaican chalices, one methanolic and another acetonic, which can be used as a disinfectant and / or food preservatives due to their efficiency in the elimination of pathogenic bacteria from raw plant-based foods such as chili peppers.

Unlike other compositions known so far for the same purpose, the compositions of the present invention are capable of eliminating pathogenic bacteria present in raw plant-based foods, such as chiles per se, without altering their nutritional properties as well as Product quality features. Consequently, the application of the compositions of the present invention in raw plant-based foods allows their preservation, as well as their effective disinfection, which makes them safe food for consumption.

DETAILED DESCRIPTION OF THE INVENTION The compositions of the present invention comprise plant extracts with known antimicrobial activity, such as aqueous and acetonic extracts of chalices of the flower of Jamaica, either alone or in combination with other components with proven disinfectant activity, such such as organic acids that include acetic acid and chlorine compounds that include sodium hypochlorite. In the case of the disinfection of raw plant-based foods such as chili peppers, the compositions of the invention that include a mixture of calyx extracts from the Jamaican flower as well as acetic acid and sodium hypochlorite, which are usually very effective to eliminate the microorganisms resident in the plant, while achieving that their organoleptic and / or nutritional properties are not affected and without being altered, for example the commercial quality of the peppers

 For the purposes of the present invention, the compositions described herein comprise: a) A methanolic extract derived from plants, which exhibit antimicrobial properties, such as extracts derived from chalices of the Jamaican flower (Hibiscus sabdariffa),

 b) An acetonic extract derived from plants, which exhibit antimicrobial properties, such as extracts derived from chalices of the Jamaican flower (Hibiscus sabdariffa),

 c) An organic acid with disinfectant activity, such as acetic acid, lactic acid, citric acid, peracetic acid, octanoic acid, peroxyethanoic acid and 1-hydroxyethylidene-1,1-diphosphonic acid, and mixtures thereof, in a concentration p / p from 0.01% to 10%, preferably from 0.1% to 1%, and

 d) A chlorine compound with disinfectant activity, such as sodium hypochlorite, calcium hypochlorite, chlorine dioxide and mixtures thereof in a w / w concentration of 0.001% to 10%, preferably 0.001% to 0.1%,

For purposes of the invention, the compositions are added to food to be disinfected and / or preserved through methods known in the art, such as direct application, through aerosols, the complete immersion of fruits and vegetables in disinfectant solutions. or by means of devices that allow their adequate dispersion in the food to be treated. The compositions of the invention can be added or contacted with food in an amount of 0.1 mL per 1000g of food, preferably 0.1 to 1mL per 100g of food, or added in larger volumes according to the needs of disinfection. of food. After applied, the compositions can remain the necessary time until obtaining the desired disinfectant and / or preservation effect on fruits and vegetables. Prior to consumption, the fruits and vegetables treated with the compositions described herein are simply washed with drinking water to remove said compositions. The compositions described herein can be obtained by mixing their components in the desired concentrations, to then store them at room temperature, so they are ready to be applied to food when deemed necessary. For the purposes of the invention, the compositions described must be in admixture with plant extracts with antimicrobial activity, such as extracts derived from Jamaican chalices, which are brought into contact with food, for example, raw plant-based foods such as the peppers, in order to disinfect and / or preserve them. In the present invention, the disinfectant activity of extracts derived from Jamaican chalices is described, in the disinfection and / or preservation of foods, for example to raw plant-based foods, so they can be used directly or as part of compositions that contain them. In this sense, extracts derived from the chalices of Jamaica can be added or contacted with food to be disinfected and / or preserved in a w / w concentration of 0.001% to 10%, preferably 0.1% to 1%.

The disinfectant and / or preservation effectiveness of the compositions described herein is such that it inactivates or eliminates human pathogenic bacteria or food spoilage that may be present in them, while at the same time not affecting the organoleptic properties. and / or nutritious food. In the case of fresh-food such as chili peppers, the compositions of the invention adequately disinfect the food without affecting its nutritional properties, while at the same time not affecting the organoleptic or quality properties.

The plant extracts of the present invention can be obtained by the following method: a) Place the dried plant in a container under aseptic conditions, add methanol, or acetone in a 1: 9 ratio; preferably 100 g of the dried plant are placed in a container (flask) under aseptic conditions, 900 ml of water or acetone are added, and in the case of acetone it is allowed to stand for 7 days;

b) Remove the calyces and recover the methanolic and / or acetonic extract; preferably the resulting extract is recovered after pressure on the flask walls to remove excess liquid;

c) Pass the extract through a sieve and recover the filtered extract; preferably the extract is passed through a No. 200 sieve;

d) Remove the methanol or acetone from the extract by rotary evaporation at a temperature of 40 ° C, a rotation of 80 rpm and a vacuum pressure of 72 mbar;

e) Recover the dry extract; preferably in a previously sterile contain.

Obtained extracts these are stored at room temperature until use. Once the extracts have been obtained, they can be used alone or in combination with other disinfectants to obtain the compositions of the invention, which can be obtained by methods known in the art where it involves the combination of the various elements that form them to form solutions. and / or suspensions to be applied subsequently to food to be disinfected and / or preserved, by methods known in the art.

The present invention constitutes the first report of the use and effectiveness of compositions containing plant extracts with microbial activity, either alone or in combination with other disinfectants, for the disinfection and / or preservation of food, particularly fruits and vegetables, as per Example the chiles. As can be seen later, the compositions of the invention are capable of disinfecting and / or eliminating microorganisms present in peppers in a very efficient way, which is It is possible to have microbiologically safe and safe chili peppers for consumption.

The following examples are included below for the sole purpose of illustrating the present invention, without implying any limitation on its scope.

Example 1. Materials and methods.

 1.1. Vegetal material.

 Dry Jamaican chalices (Hibiscus sabdariffa) of the Creole variety of Oaxaca were used, while in the case of chiles (Capsicum annuum) the jalapeno and serrano varieties were used. The peppers of each variety had a uniform size.

1.2. Bacterial strains

Strains of £ were used. coli 0157: H7 (P1C6, isolated from an outbreak of disease), enteroinvasive E. coli (4VC81-5, isolated from clinical case) Enterotoxigenic coli (1620 TL, isolated from clinical case), enteropathogenic E. coli (52 GM 291, isolated from clinical case), Salmonella typhimurium (ATCC 14028), Salmonella choleraesuis (ATCC 10708), Listeria monocytogenes (ATCC 19115), Listeria monocytogenes Scott A, Staphylococcus epidermis (ATCC 12228), Staphylococcus aureus (ATCC 25923onas) (ATCC 27853), Bordetella (ATCC 12741) Shigella sonnei (ATCC 25931) and Shigella flexneri (ATCC 12022), V. cholerae (87151, Inaba serotype isolated from the environment) and Pseudomonas aeruginosa (ATCC 27853). The strains of E. coli 0157: H7 and that of V. cholerae 01 were donated by Dr. Fernández Escartin of the Autonomous University of Querétaro. All strains were marked with resistance to the antibiotic rifampicin (R +) to eliminate interference from the native microbial flora of the extract (Castro-Rosas and Escartin, 2000). This resistance to the antibiotic was maintained throughout the study. The strains were maintained at 4-7 ° C in blood-based agar (ABS, Merck®, Germany) with biweekly transfers, activating in tryptosesein soy broth (CST, Bioxon®, Mexico) with incubation at 35 ° C / 24h. 1.3. Obtaining aqueous extract from the chalices of Jamaica.

 Under aseptic conditions, 100 g of Jamaican calyces were placed in an Erlenmeyer flask, to which 900 mL of distilled water was added, boiling the mixture for 20 minutes. Once the treatment was finished, it was allowed to cool to room temperature. The calyces were removed from the extract (after pressure on the flask walls to remove excess liquid from it) and subsequently the extract was passed through a No. 200 sieve (MONTIMAX) to remove particles. Finally, all the water was removed from the extract by broken evaporation using a broken evaporator (Buchi R-205) using the following conditions: temperature of 40 ° C of the tub, rotation of 80 rpm and a vacuum pressure of 72 mbar. The dried extract was recovered in a sterile bottle and stored at room temperature until use.

1.4. Obtaining methanolic and acetonic extract from the chalices of Jamaica.

 Under aseptic conditions 100 g of Jamaican chalices were placed in an Erlenmeyer flask, to which 900 mL of methanol or acetone was added and stored for 7 days at room temperature. Once the treatment was finished, the calyces were removed from the extract (after pressure on the flask walls to remove excess liquid from it) and subsequently the extract was passed through a No. 200 sieve (MONTIMAX) to remove particles. Finally, all methanol or acetone was removed from the extract by broken evaporation using a broken evaporator (Buchi R-205) using the following conditions: temperature of 40 ° C of the tub, rotation of 80 rpm and a vacuum pressure of 72 mbar. The dried extracts (methanolic or acetonic) were recovered separately in a sterile bottle and stored at room temperature until use.

1.5. Determination of the antimicrobial activity of aqueous, methanolic and acetonic extracts of Jamaican calyces in culture medium (in vitro studies).

1.5.1. Preparation of inoculum of strains. Test tubes with 24 h cultures in CST of each R + strain were centrifuged at 3500 rpm for 20 min. Subsequently, the supernatant was discarded; The cell packet was resuspended by adding 3 mL of sterile isotonic saline and stirred in vortex for 10 s. The above procedure was repeated two more times. Subsequently, the concentration of each strain was approximately 1x10 ⁹ CFU / mL. Finally, each strain was diluted decimally in isotonic saline solution only once.

1.5.2. Preparation of the solutions of the extracts.

 From the dried extracts, aqueous solutions were prepared using sterile distilled water or a solution of Polysorbate 80: water in a 20:80 ratio. The aqueous and methanolic extracts were solubilized in distilled water while the acetonic extracts were solubilized in the solution of Polysorbate 80: water. To water or polysorbate 80: water, dried extracts were added in a 1: 10 ratio (water, extract) separately and deposited in sterile bottles.

1.5.3 Antimicrobial effect of extracts in culture medium.

Separately, 100 pL of the first dilution of the cultures of the pathogens were inoculated on boxes of AST supplemented with 10 mg / L of the antibiotic rifampin, the inoculum was distributed over the entire surface of the agar using the surface extension technique. On the inoculated boxes, separately, 10 pL aliquots of the extract solution (aqueous, methanolic or acetonic) were placed. Four repetitions were performed for each treatment. After the extract or fractions were absorbed by the agar, the culture boxes were incubated at 35 ± 1 ° C, for 24 h. Finally, the diameter of each of the inhibition halos formed on the surface of the inoculated medium was measured. 1.6. Evaluation of the antimicrobial effect of aqueous, methanolic, acetonic, acetic acid, hypochlorite and specific formulations in the reduction of Salmonella and £ coli 0157.Ή7 in contaminated peppers. 1.6.1. Preparation of disinfectant solutions.

 Solutions of Jamaican calyx extracts as well as mixtures containing extracts, acetic acid and hypochlorite were prepared at the concentrations, proportions or mixtures described in Table 1. For example, to prepare 100 ml of a solution containing methanolic extract of 1% Jamaica calyces, 0.1% acetic acid and 100 mg / L hypochlorite: 100 g of dried methanolic extract of Jamaican calyces was added to 100 mL of distilled water, in addition to 1 ml of a 10% acetic acid solution % and 0.2 ml of a 5% hypochlorite solution.

1.6.2. Strains

 For these studies we worked with 7 Salmonella serotypes: (3 typhimuríum [ATCC 14028, one isolated from tomato, J1, and another from alfalfa seed, GA1], Salmonella choleraesuis [ATCC 10708], typhi, gaminara, and montevideo) and 3 of E. coli 0157: H7 (two isolated in our laboratory from raw meat [P1C6 and M5C8] and another isolated from an outbreak caused by meat consumption in the United States of America [E09]). All strains were marked with resistance to the antibiotic rifampicin (R +) to eliminate interference from the native microbial flora of the extract (Castro and Escartín, 2000).

Table 1. Treatments to which chiles contaminated with mixtures of Salmonella strains or strains of E. co / 0157: H7 were subjected separately

No Treatments

 1 No treatment (control)

 2 1% aqueous extract

 3 1% acetonic extract

 4 1% methanolic extract

 5 Sodium hypochlorite 50 ppm

 6 Sodium hypochlorite 100 ppm

 7 Acetic acid 0.1%

 8 Acetic acid 0.5%

 9 1% aqueous extract + Sodium hypochlorite 50 ppm

 10 1% aqueous extract + 100 ppm sodium hypochlorite

 11 Acetonic extract 1% + Sodium hypochlorite 50 ppm

 12 Acetonic extract 1% + Sodium hypochlorite 100 ppm

13 1% Methanolic Extract + Sodium Hypochlorite 50 ppm 14 1% Methanolic Extract + Sodium Hypochlorite 100 ppm

 15 1% aqueous extract + 0.1% acetic acid

 16 1% aqueous extract + 0.5% acetic acid

 17 Acetonic extract 1% + acetic acid 0.1%

18 1% acetonic extract + 0.5% acetic acid

 19 1% methanolic extract + 0.1% acetic acid

 20 1% methanolic extract + 0.5% acetic acid

 21 1% aqueous extract + 0.1% acetic acid + sodium hypochlorite 50 ppm

22 1% aqueous extract + 0.5% acetic acid + sodium hypochlorite 50 ppm 23 1% aqueous extract + 0.1% acetic acid + sodium hypochlorite 100 ppm

24 1% aqueous extract + 0.5% acetic acid + sodium hypochlorite 100 ppm

25 1% acetonic extract + 0.1% acetic acid + sodium hypochlorite 50 ppm

26 Acetonic extract 1% + acetic acid 0.5% + sodium hypochlorite 50 ppm

27 Acetonic extract 1% + acetic acid 0.1% + sodium hypochlorite 100 ppm

 28 Acetonic extract 1% + acetic acid 0.5% + sodium hypochlorite 100 ppm

 29 1% methanolic extract + 0.1% acetic acid + sodium hypochlorite 50 ppm

30 1% methanolic extract + 0.5% acetic acid + sodium hypochlorite 50 ppm

 31 1% methanolic extract + 0.1% acetic acid + 100 ppm sodium hypochlorite

 32 1% methanolic extract + 0.5% acetic acid + 100 ppm sodium hypochlorite

 For the elaboration of the solutions the base was used:

 A) the dry extract of the calyces of Jamaica in the previous section, B) Sodium hypochlorite solution with 5% free hypochlorite, C) 10% glacial acetic acid, D) Sterile distilled water at pH 6

1.6.3. Preparation of inoculum of strains.

Test tubes with 24 h cultures in CST of each strain +, were centrifuged at 3500 rpm for 20 min. Subsequently, the supernatant was discarded; The cell packet was resuspended by adding 3 mL of sterile isotonic saline and stirred in vortex for 10 s. The above procedure was repeated two more times. The resulting concentration of each strain was approximately 1x10 ⁹ CFU / mL. One milliliter of each Salmonella strain was mixed in an empty test tube to have a mixture of the 7 strains of

Salmonella examined. The same was done with strains of E. coli 0157: 1-17, to have a mixture of the three strains of E. coli 0157: H7.

1.6.4. Chilli Inoculation

Chili peppers of the jalapeño and serrano varieties were used; The peppers were obtained from a local producer. Prior to inoculation, the peppers were cleaned with a clean cloth to remove dust particles. In each variety, peppers of a uniform or similar size were used and did not show visible damage. Separately, individual chiles were inoculated by placing in the central part (not in peduncle) of the 10 L chili of a suspension of each type of pathogenic bacteria mixture (Salmonella or E. coli 0157.Ή7) microorganisms containing approximately 1 x 10 ⁷ CFU Inoculated chiles were placed in a tray and placed in a bioclimatic hood for two hours at a relative humidity of 90 ± 1% and 26.5 ± 1 ° C. The purpose of this treatment was to cause the adhesion or infiltration of the cells of the pathogenic bacteria under study to simulate the natural or common conditions of contamination. 6.5. Chili disinfection treatment.

 After two hours in the bioclimatic chamber, each chili was washed separately to remove microorganisms that did not adhere, the washing consisted of submerging and stirring the inoculated part of the chili in distilled water for 10 s, the washed part was drained at room temperature until total dryness and subsequently separately the inoculated part of different peppers was immersed for 10 min in the different disinfectant solutions indicated in Table 1. A treatment with only distilled water served as a positive control.

1.6.6. Counting of microorganisms surviving the treatments

After treatment, the peppers were removed from the disinfectant solution and to remove the remaining disinfectant, the inoculated part was immersed in distilled water for 10 s, then the inoculated part was cut (a frame of approximately 2 x 2 cm and with a depth of approx. 2 cm) with the help of a sterile scalpel, each portion was placed independently in a plastic bag and 10 ml of peptone diluent was added. Subsequently, the materials were manually agitated by pressing and rubbing the inoculated part and the entire portion of chili from the outside of the bag for a minute. After this time, each bag was counted using the plate pouring technique using agar for standard methods (Bioxon, Mexico) added 100 mg / L of Rifampicin (Sigma, Mexico), the boxes were incubated at 35 ° C / 24-48 h. This procedure was performed in duplicate for each replica. Each treatment was carried out in quintuplicate. 6.7. Statistic analysis

 The results obtained were statistically analyzed with a one-way analysis of variance (ANOVA) comparing the means with the Tukey test, with a significance level of 0.05.

1.7. Nuclear Magnetic Resonance (NMR) of extracts

The NMR spectrum of the proton ( ¹ H) of both the dried methanolic and acetonic extract obtained from the chalices of Jamaica was determined. The extracts were solubilized in deuterated water. NMR spectra were obtained using a nuclear magnetic resonance spectrometer (Varian NMR, 400 MHz).

NMR spectroscopy studies atomic nuclei. This spectroscopic technique can only be used to study atomic nuclei with an odd number of protons or neutrons (or both), to determine the structures of organic compounds. This situation occurs in the atoms of H, ¹³ C, ⁹ F and ³¹ P. These types of nuclei are magnetically active, that is, they have spin, just like electrons, since the nuclei have a positive charge and have a rotational movement. on an axis that makes them behave as if they were small magnets. The NMR spectrometer detects these signals and records them as a graph of frequencies versus intensity, which is called the NMR spectrum

Example 2. Antimicrobial effect of extracts from Jamaican chalices.

The three types of extracts (aqueous, methanolic and acetonic) showed a marked antimicrobial effect (Table 2). All the microorganisms tested were inhibited from the first moments of contact. The inhibitory effect observed suggests the presence of antimicrobial substances in the extracts. This effect can cause lethal damage to the cell or only cause a sublethal effect or cellular stress (Busta, 1976). Different components of the vegetable could be responsible for this antimicrobial effect. It is possible that the chalices of Jamaica contain different antimicrobials; that is, the antimicrobial effect may be due to the effect of different molecules with antimicrobial activity, this observation is based on the fact that the extracts are they obtained with three solvents of different polarity, and the three types of extracts showed antimicrobial activity.

Table 2. Antimicrobial effect of the aqueous extract of Jamaica diluted 1:10 and that of a solution of penicillin (control) on different microorganisms

 * (mm) Example 3. Disinfectant potential of extracts alone or in mixtures with acetic acid and sodium hypochlorite.

It was found that all treatments had an antimicrobial effect with respect to the control (Tables 3 and 4). However, only 4 combinations were 100 percent effective (Tables 3 and 4). In Tables 3 and 4, it is observed that only 4 combinations managed to eliminate the concentration of the mixtures of each pathogen at undetectable levels: treatments 27, 28, 31 and 32 reduced the concentration of both pathogens by 5 log (Tables 3 and 4 ). In the present invention, 4 specific combinations (treatment 29, 30, 33 and 34; Tables 3 and 4) of three antimicrobials and one surfactant (polysorbate), the total elimination of the pathogenic microorganisms inoculated on both types of peppers (jalapeno) was achieved and mountain); This is an example of what is currently known as multiple barrier treatment. Multiple barriers are the combination of antimicrobial treatments that enhance the overall antimicrobial effect, which results in stable, safe and safe food.

It is worth noting the potential role of polysorbate 80 in the observed antimicrobial effect, since being an active tense it is possible that it has favored the emulsification of the natural wax of ios chiles which could increase the effect of the disinfectant solution by eliminating or decreasing The effect protector that the wax would be providing the microorganisms inoculated on the peppers. It is important to highlight the disinfectant effect that was observed with miasmas 4 more active solutions in the 2 types of peppers; This data strongly suggests the high possibility that such an effect can be observed when these four solutions are applied to other types of peppers (Poblano, for example) contaminated with pathogenic bacteria. It is possible that in other types of chili peppers, a disinfectant effect greater than or equal to that observed in jalapeno and serrano peppers is observed. Therefore, the results in Table 3 and 4 support the application of these 4 formulations to efficiently disinfect other types of peppers.

 Therefore, the compositions of the present invention are an excellent alternative for the disinfection and / or preservation of foods, for example raw plant-based foods, without altering their nutritional properties. In this sense, the compositions described here allow the effective disinfection or inactivation of pathogenic microorganisms of raw plant-based foods, preferably chiles, allowing the safe consumption of such products.

Example 4. Nuclear magnetic resonance (NMR) spectrum obtained from the acetonic extract.

 The NMR spectrum obtained from the dry acetonic extract of the calyxes of Jamaica is presented in Figure 1. In the spectrum there are several characteristic peaks of the extract that we use in the formulations. This spectrum characterizes the aqueous extract used in the antimicrobial formulations of Tables 3 and 4.

Example 5. Nuclear magnetic resonance (NMR) spectra obtained from the methanolic extract.

The NMR spectrum obtained from the dry methanolic extract of the chalices of Jamaica is presented in Figure 2. In the spectrum, several characteristic peaks of the extract of the formulations are observed. This spectrum characterizes the acetonic extract used in the antimicrobial formulations in Tables 3 and 4. References. 1. Adams MR 1997. Food microbiology. Editorial Acribia SA

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Campylobacter jejuni, Escherichia coli, Listería monocytogenes, and

Enteric Salmonella J. Food Prot. 65: 545-1560.

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Publication: Cambridge Woodhead Publishing. 21. Kang PS, Seok JH, Kim YH, Eun JS, and Oh SH. 2007. Antimicrobial and antioxidative effects of reselle {Hibiscus sabdariffa L) flower extract and its fractions on skin microorganisms and oxidation. Food Sci Biotechnol. 16: 409-414.

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Claims

CLAIMS Having sufficiently described my invention, I consider as a novelty and therefore claim as my exclusive property, what is contained in the following clauses:

1. A solution with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized in that it comprises: a) Methanolic extract of Jamaican calyxes (Hibiscus sabdariffa);

b) Acetic acid;

c) Sodium Hypochlorite;

2. The solution with antimicrobial activity according to claim 1, characterized in that the a) methanolic extract of Jamaican calyxes

(Hibiscus sabdariffa) is present in a concentration between 0.01% to 10%.

3. The solution with antimicrobial activity according to claim 1, characterized in that b) acetic acid is present in a concentration between 0.01 to 10%.

4. The solution with antimicrobial activity according to claim 1, characterized in that c) Sodium hypochlorite is present in a concentration between 10 to 1000 ppm.

5. The solution with antimicrobial activity according to claim 1, characterized in that d) methanolic extract of Jamaican calyxes (Hibiscus sabdariffa) presents a nuclear magnetic resonance (NMR) spectrum as seen in Figure 1.

6. The solution of claim 1, characterized in that it is presented as an aqueous formulation.

7. The solution of claim 1 in accordance with the previous claims, characterized in that where one or several parts of Jamaica (Hibiscus sabdariffá) can be used to obtain the extract.

8. The solution of claim 1 in accordance with the previous claims, wherein the part of the Jamaica plant that is used is the calyxes.

9. The solution defined in the previous claims, characterized by a nuclear magnetic resonance (NMR) spectrum of the methanolic extract obtained from the calyxes of Jamaica (Hibiscus sabdariffá) (Figure 1).

10. The solution of the previous claims, characterized in that it is useful as a disinfectant and preservative for foods of plant and animal origin.

11. The solution of the previous claims, characterized in that the aqueous formulation is useful as a disinfectant and preservative for fruits and vegetables, especially chili peppers.

12. The solution with antimicrobial activity in accordance with the previous claims, characterized in that the main varieties of chili peppers (Capsicum annuum) on which it acts as a disinfectant are selected from Serrano, Poblano, Ancho, Mirasol, Puya, Pasilla, Chilaca, Campana and to a lesser extent but which are also planted: Habanero, Colorado, Costeño, Tree (rat tail), Mulato, Tabaquero, Anaheim, Bell pepper, Caloro, Cayene, Agua, Guajillo, Shadow mesh, Manzano, Organic, Perón, Regional, Soledad, X-cat-ik, Morrón, Pimiento, Ciltepin, de Tiempo.

13. A method for the disinfection and/or preservation of chili peppers (Capsicum annuum), characterized in that it comprises applying the solution defined in the previous claims to the chili pepper.

14. A plant extract with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized in that it is obtained through the following steps: a) Place the dry plant in a container under aseptic conditions, add methanol and store at 22° ± T C for 7 days,

b) Pass the extract through a sieve and remove the methanol from the extract, and

c) Recover the dry extract.

15. The extract of claim 14, characterized in that it is obtained with methanol.

16. The extract according to claim 14, wherein one or more parts of the plant can be used to obtain the extract.

17. The extract of claim 14, characterized in that the plant is the Jamaican plant (Hibiscus sabdariffa).

18. The extract of claim 14 in accordance with the previous claims, characterized in that the extract is obtained from the calyxes of the

Jamaica.

19. The extract of claim 14 in accordance with the previous claims, characterized in that it presents a nuclear magnetic resonance (NMR) spectrum as seen in Figure.

20. The extract of claim 14 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative for foods of animal and plant origin.

21. The extract of claim 14 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative for fruits and vegetables, preferably chili peppers.

22. The extract of claim 14 in accordance with the previous claims, characterized in that the main varieties of chili (Capsicum annuum) on which it acts as a disinfectant are selected from Serrano, Poblano, Ancho, Mirasol, Puya, Pasilla, Chilaca, Campana and to a lesser extent but which are also planted: Habanero, Colorado, Costeño, Tree (rat tail), Mulato, Tabaquero, Anaheim, Bell pepper, Caloro, Cayene, Agua, Guajillo, Shadow mesh, Manzano, Organic, Perón , Regional, Soledad, X-cat-ik, Morrón, Pimiento, Ciltepin, de Tiempo.

23. A method for the disinfection and/or preservation of chili peppers (Capsicum annuum), characterized in that it comprises applying the extract defined in claims 14 to 22 to the chili.

24. A method to obtain a plant extract with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized in that it comprises the following steps: a) Place the dry plant in a container under aseptic conditions, add methanol and store at 22 ° ± 2 ^o C for 7 days,

b) Pass the extract through a sieve and remove the methanol from the extract, and

c) Recover the dry extract.

25. The method for obtaining the plant extract of claim 24, characterized in that they can have a solid or liquid presentation.

26. A method for the preparation of a solution with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized by comprising the steps of: a) Place dried Jamaica (Hibiscus sabdaríffa) in a container under aseptic conditions, add methanol and store at 22° ± 2 ^o C for 7 days,

b) Pass the extract through a sieve and remove the methanol from the extract and, c) Recover the dry extract,

d) Prepare the aqueous solution in a container containing: water, dry methanolic extract of the plant, acetic acid and sodium hypochlorite,

27. The method according to claim 26, wherein the part of the Jamaica plant used is the calyxes.

28. A solution with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized in that it comprises: a) Acetonic extract of Jamaican calyxes (Hibiscus sabdariffa);

b) Acetic acid;

c) Sodium Hypochlorite;

29. The solution with antimicrobial activity according to claim 28, characterized in that a) acetone extract of Jamaican calyxes {Hibiscus sabdariffa) is present in a concentration between 0.01% to 10%.

30. The solution with antimicrobial activity according to claim 28, characterized in that b) acetic acid is present in a concentration between 0.01 to 10%.

31. The solution with antimicrobial activity according to claim 28, characterized in that c) Sodium hypochlorite is present in a concentration between 10 to 1000 ppm.

32. The solution with antimicrobial activity according to claim 28, characterized in that d) acetone extract of Jamaican calyxes (Hibiscus sabdariffa) presents a nuclear magnetic resonance (NMR) spectrum as seen in Figure 2.

33. The solution of claim 28, characterized in that it is presented as an aqueous formulation.

34. The solution of claim 28 in accordance with the previous claims, characterized in that where one or several parts of Jamaica (Hibiscus sabdaríffa) can be used to obtain the extract.

35. The solution of claim 28 in accordance with the previous claims, wherein the part of the Jamaica plant that is used is the calyxes.

36. The solution of claim 28 in accordance with the preceding claims, characterized by a nuclear magnetic resonance (NMR) spectrum of the acetone extract obtained from the calyxes of Jamaica (Hibiscus sabdaríffa) (Figure 2).

37. The solution of claim 28 in accordance with the previous claims, characterized in that it is useful as a disinfectant and preservative for foods of plant and animal origin.

38. The solution of claims 28 in accordance with the preceding claims, characterized in that the aqueous formulation is useful as a disinfectant and preservative for fruits and vegetables, especially chili peppers.

39. The solution with antimicrobial activity according to claim 28, characterized in that the main varieties of chili peppers (Capsicum annuum) on which it acts as a disinfectant are selected from Serrano, Poblano, Ancho, Mirasol, Puya, Pasilla, Chilaca, Campana and to a lesser extent but which are also planted: Habanero, Colorado, Costeño, Tree (rat tail), Mulato, Tabaquero, Anaheim, Bell pepper, Caloro, Cayene, Agua, Guajillo, Shadow mesh, Manzano, Organic, Perón, Regional, Soledad, X-cat-ik, Morrón, Pimiento, Ciltepin, de Tiempo.

40. A method for disinfecting and/or preserving chili peppers (Capsicum annuum), characterized in that it comprises applying the solution defined in claims 28 to 39 to the chili.

41. A plant extract with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized in that it is obtained through the following steps: a) Place the dry plant in a container under aseptic conditions, add acetone and store at 22° ± 2° C for 7 days,

b) Pass the extract through a sieve and remove the acetone from the extract, and

c) Recover the dry extract.

42. The extract of claim 41, characterized in that it is obtained with acetone

43. The extract of claim 41, because where one or more parts of the plant can be used to obtain the extract.

44. The extract of claim 41, characterized in that the plant is the Jamaican plant (Hibiscus sabdariffa).

45. The extract of claim 41 in accordance with the preceding claims, characterized in that the extract is obtained from the calyxes of Jamaica.

46. The extract of claim 41 in accordance with the preceding claims, characterized in that it presents a nuclear magnetic resonance (NMR) spectrum as seen in Figure 2.

47. The extract of claim 41 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative for foods of animal and plant origin.

48. The extract of claim 41 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative for fruits and vegetables, preferably chili peppers.

49. The extract of claim 41 in accordance with the previous claims, characterized in that the main varieties of chili {Capsicum annuum) on which it acts as a disinfectant are selected from Serrano, Poblano, Ancho, Mirasol, Puya, Pasilla, Chilaca, Campana and to a lesser extent but which are also planted: Habanero, Colorado, Costeño, Tree (rat tail), Mulato, Tabaquero, Anaheim, Bell pepper, Caloro, Cayene, Agua, Guajillo, Shadow mesh, Manzano, Organic, Perón , Regional, Soledad, X-cat-ik, Morrón, Pimiento, Ciltepin, de Tiempo.

50. A method for the disinfection and/or preservation of chili peppers (Capsicum annuum), characterized in that it comprises applying the extract defined in claims 41 to 49 to the chili.

51. A method to obtain a plant extract with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized in that it comprises the following steps: a) Place the dry plant in a container under aseptic conditions, add acetone and store at 22 ° ± 2° C for 7 days,

b) Pass the extract through a sieve and remove the acetone from the extract, and

c) Recover the dry extract.

52. The method for obtaining the plant extract of claim 51, characterized in that the extract can have a solid or liquid presentation.

53. A method for the preparation of a solution with antimicrobial activity to disinfect and/or preserve chili peppers (Capsicum annuum), characterized by comprising the steps of: a) Place dried Jamaica (Hibiscus sabdanffa) in a container under aseptic conditions, add acetone and store at 22° ± 2 ^o C for 7 days,

b) Pass the extract through a sieve and remove the acetone from the extract and,

c) Recover the dry extract,

d) Prepare the aqueous solution in a container containing: water, dry acetone extract of the plant, acetic acid and sodium hypochlorite.

54. The method according to claim 53, wherein the part of the Jamaica plant used is the calyxes.