WO2015088306A1 - Solutions based on plant extracts for disinfecting coriander (coriandrum sativum) - Google Patents

Abstract

The invention relates to compositions or formulations for the effective disinfection and/or preservation of fruit and vegetables contaminated with pathogenic and deteriorating micro-organisms. The formulations described contain compounds with an anti-microbial effect derived from plants, which can act independently or combined with other disinfecting agents, such as organic acids and compounds of chlorine, and surfactants such as polysorbate 80. The compositions of the invention can eliminate or inactivate microbial contamination, including that of pathogenic micro-organisms, of fruit and vegetables, especially coriander, without altering the nutritious and/or alimentary and or sensory properties thereof.

Description

 SOLUTIONS BASED ON PLANT EXTRACTS TO DISINFECT

 CYLINDER ICoriandrum sativum)

TECHNICAL FIELD OF THE INVENTION

The present invention relates to the development of formulations containing compounds with antimicrobial activity 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 extracts of Jamaican chalices {Hibiscus sabdariffa), the method by which it is obtained and its uses as an effective formulation to eliminate pathogenic bacteria from plant-based foods, such as fruits and vegetables, but with the most high effectiveness for coriander (Coriandrum sativum).

STATE OF THE TECHNIQUE

Coriander (Coriandrum sativum) is an agricultural product widely consumed throughout the world. In Mexico there is a little more than 6 thousand hectares dedicated to planting (Sagarpa, 2013). Coriander is an annual herb of the apiaceae family (formerly called umbellifers). It is the only species of the genus Coriandrum, which is also the only member of the Coriandreae tribe. All parts of the plant are edible, however, fresh leaves and dried seeds are the most common culinary. Fresh leaves are an essential ingredient in Mexican salsa verde and guacamole. Chopped leaves are also used as garnish, added at the end of cooking or just before serving, on soups and other dishes. Fresh cilantro is never cooked because heat totally destroys its aroma and flavor. It must be kept in the refrigerator in airtight containers, trying to consume it in a few days, as it quickly withers. It should not be dried or frozen because it loses the aroma.

In Mexico its use is very extensive, it is used in the preparation of various sauces and moles, as a flavoring in soups and broths, and fresh and chopped as a dressing of different types of tacos and snacks. Coriander is also known as Chinese parsley or Arabic parsley, in Spanish it is also identified by the following names: wild or hydrangea coriander, ceandro, celandria, cilandro, coantrillo, coendro, coentro, coentro das hortas, colentro, coriander, cuantrillo, coriander, jilantro, salandria and xendro.

Only 3 varieties of coriander have been reported, from which the cultivars of coriander types that exist worldwide are derived: sativum, microcarpum and Alef vulgare.

However, along with the increase in coriander consumption worldwide, disease outbreaks due to pathogenic bacteria and parasites have occurred due to the consumption of raw coriander (Campbell et al., 2001; CDC, 2013). For example, recently (November, 2013) there was a multi-state outbreak (that is, it affected several states) in the American union where the vehicle involved was coriander and the microorganism that was in the coriander was Ciclospora (CDC, 2005) . A little more 30 outbreaks of disease of microbial etiology have been reported since 1970 to date in the United States of America (USA) and in different parts of the world due to the consumption of raw coriander. This type of outbreaks has caused strict regulation inside and outside the USA for all coriander producers, and of course, also for all coriander producers that export to the USA or other parts of the world such as Mexico. All this has meant that shipments of coriander are frequently withheld at the borders, partial or total closure of the export of this product to the countries and economic losses by the producers when they do not comply with microbiological standards. However, in the case of Mexico, the largest production of coriander is consumed in the country.

It should be noted that although in Mexico there are no reports of disease outbreaks of microbial etiology associated with the consumption of coriander, due to the poor hygiene practices that generally occur during the cultivation, harvest, transport and commercialization of coriander, the participation of the coriander in disease outbreaks and as a vehicle for pathogenic bacteria. A fact that supports this observation is the frequency with which Salmonella strains have recently been isolated from coriander (Quiroz-Santiago et al., 2009), as well as Salmonella, pathogenic groups of Escheríchia cotí, and other pathogenic bacteria, from different raw products in Mexico (Castañeda-Ramírez et al., 2011; Castro-Rosas et al., 2012).

Recent outbreaks of food diseases associated with coriander created the need to determine the sources of coriander contamination and to understand the survival and / or growth of pathogenic microorganisms in it; These have led to the development of innovative control technologies. In general, pathogens in coriander could be controlled by preventing contamination during cultivation and harvesting of products, also by using disinfectants with antimicrobial power in the harvested product, and by storing coriander at low temperature. However, disinfection has been identified as the most important stage for the microbial safety of raw coriander.

Usually coriander is not consumed directly as it is 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 coriander 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 damage the coriander.

Coriander contamination prevention is also a control strategy because for several pathogenic microorganisms 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 the coriander is affected by the location of the pathogen in the product, product quality, storage temperature, type of packaging, and relative humidity. Coriander surface usually has few nutrients, which limits the growth of pathogens during storage at room temperature or refrigeration. However, it should be noted that pathogenic microorganisms such as Salmonella are able to survive for a long time on the coriander surface both in refrigeration and at room temperature (BrandI and Mandrell, 1998). Also once on the coriander, pathogens such as Salmonella could produce extracellular polymers on the coriander which leads to the formation of a biofilm (BrandI and Mandrell, 1998) that could protect them against disinfectants; This behavior of pathogenic microorganisms has been observed in different vegetables such as tomatoes (Iturriaga et al., 2007). A noteworthy fact is that pathogenic microorganisms such as Salmonella, for example, are able to multiply in chopped cilantro by significantly increasing their concentration and making the food much more dangerous (BrandI and Mandrell, 1998).

Pathogenic microorganisms on the surface of the coriander can contaminate the internal tissues and infiltrate (BrandI and Mandrell, 1998) and later during the cutting of the coriander they could contaminate the chopped product and grow in it. Several research findings indicate that bacterial pathogens can infiltrate plant products, such as coriander when there is a temperature differential between the plant product and the wash water (BrandI and Mandrell, 1998; Kroupitski et al., 2009; Bartz , 1982; Guo et al., 2002; Ibarra-Sanchez et al., 2004; Zhuang and Beuchat 1995) and by 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 coriander wash water is the control of the spread of pathogens, their inactivation and / or preventing their infiltration into The leaves and stem. However, currently available chemical disinfectants have limited benefits over plant products, such as coriander. 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), iodine (iodophore), chlorine dioxide or 100 mg / L of a commercial product based on grapefruit seed extract (citricidal) reduced alfalfa sprout content in 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 ™ 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 bacteria Pathogens 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 Logos 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 oxidants that act at the level of membranes and other cellular constituents (Harmon et al., 1987). However, the former has the disadvantage of reacting easily with organic matter, so it is inactivated faster. In the second, the interference is minimal (Castro-Rosas and Escartin, 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 coriander. 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 of 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 peduncle scar 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 of exposure) (Yuk et al., 2005). The use of 10 mg / liter of ozone completely inactivates 7 log CFU of Salmonella enterítidis 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 coriander producers 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 post-harvest diseases have obtained complete control of the brown rot of the peaches caused by Monilinia fructicola (Roller, 2003). Yucel and Karapinar (2005) evaluated the reduction of S. typhimuríum in onions by applying lemon juice, vinegar and their mixtures, observing a respective reduction of 0.87-293, 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, Extractos Vegetales SA) on the survival of L. monocytogenes, P. fluorescens, E. coli, Erwinia caratovora and S. typhimuríum in apple 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 the tomato surface. 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 apple. 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 mint (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 enteritidis was eliminated in snacks under all 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 £. 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, taramasalata or mayonnaise, reactions such as pH increase, 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 to the present application, the antimicrobial effect of about 60 different plants used in herbalism has been evaluated (Cruz-Gálvez et al., 2013); where some of these have shown a high antimicrobial power against different pathogenic microorganisms, such as Salmonella or Escheríchia coli 0157: H7, among others, as well as against food spoilage microorganisms (Pseudomonas aeruginosas, for example), and the plant that has the greatest antimicrobial effect It has been shown that the chalices of the Jamaican flower have been, in some cases the antimicrobial effect being greater than that of commercial disinfectants based on hypochlorite, iodine, colloidal silver.

The extracts of the chalices of Jamaica have been separated by column chromatography to obtain fractions with greater antimicrobial power; With selected fractions, solutions have been developed that have been evaluated to determine their antimicrobial potential.

Different researchers have also reported that the chalices of the Jamaica flower (Hibiscus sabdariffa L) have substances with high antimicrobial power (Aziz et al., 1998; Fernández et al., 1996; Kang et al., 2007). A range of phytochemical compounds that could be responsible for the observed antimicrobial effect, such as polyphenols (Tajkarimi er al., 2010), including some phenolic acids (Tajkarimi et al. ., 2010), as well as flavonoids, catechins and epicatechin (Friedman et al. 2002). 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 US20100323048 describes the use of a crude extract of the calyces of Jamaica to produce a drug to treat urinary infections caused by Escherichia coliy Candida albicans.

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 antimicrobial protective compositions to prevent and / or combat microbial contamination of food, especially those consumed raw, such as coriander, in order to preserve and consume them without the risk of acquiring 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 such as those 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 the Coriander, whereby it is possible with the present invention to obtain harmless raw coriander from the microbial point of view.

BRIEF DESCRIPTION OF THE FIGURES

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

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

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

PROTÓN ( ¹ H) from the collection of fractions referred to as III which was obtained from an acetonic extract of the calyces of Jamaica and which was the collection that was used in the present invention.

 Figure 3. The nuclear magnetic resonance (NMR) spectrum of

PROTÓN ( ¹ H) of the collection of fractions denominated as IV that was obtained from an acetonic extract of the calyces of Jamaica and that was the collection 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 coriander (Coriandrum sativum) even under extreme conditions of treatment, but that do not sensory damage to the food.

The present invention relates to compositions containing phytochemicals present in plant extracts that are used as food disinfectants of vegetable and animal origin, for example aimed at the disinfection and preservation of fruits and vegetables, particularly the disinfection and / or preservation of coriander (Coriandrum sativum).

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 specific chromatographic fractions obtained from! acetonic and methanolic extract of the calyces of Jamaica, which are useful for eliminating pathogens present in food (disinfectant effect) and for delaying food spoilage or preserving 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 , those that 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 L) 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 Jamaica plant {Hibiscus sabdariffa L.) that have a disinfectant or conservative effect when applied to foods of plant origin, preferably coriander. 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 relates to the method for obtaining specific chromatographic fractions with antimicrobial effect obtained from the acetonic or methanolic extract of the calyxes of Jamaica, which are useful for eliminating pathogens present in food (disinfectant effect) and for delaying food spoilage or preserve its safety (conservative effect).

Another embodiment of the present invention is a method of treating and / or preserving 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 development of an efficient disinfectant to eliminate the pathogenic bacteria present in raw vegetables, such as coriander. In the present invention, an extract of Jamaican chalices and specific bernia fractions is described by column chromatography from an acetonic and / or methanolic extract of Jamaican calyces, which comprises phytochemicals, which can be used as a disinfectant and / or food preservative due to its efficiency in the elimination of pathogenic bacteria from raw vegetables such as coriander.

Unlike other compositions known so far for the same purpose, the compositions of the present invention are capable of eliminating the pathogenic bacteria present in raw vegetables, such as coriander per se, without altering their nutritional properties as well as quality characteristics. of the product. Consequently, the application of the compositions of the present invention in vegetables raw, allows its preservation, as well as its 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, for example, methanolic extracts of Jamaica and specific chromatographic fractions obtained from acetonic and methanolic extracts of Jamaican calyces, either alone or in combination with other components with proven activity disinfectant, such as for example organic acids that include acetic acid and chlorine compounds that include sodium hypochlorite. In the case of the disinfection of raw vegetables such as coriander, the compositions of the invention that include a mixture of the methanolic extract of plants with antimicrobial activity as well as acetic acid and sodium hypochlorite and polysorbate, and a mixture of specific chromatographic fractions obtained from the acetonic and / or methanolic extract of the calyces of Jamaica as well as acetic acid and sodium hypochlorite and polysorbate 80, are usually very effective in eliminating the microorganisms resident in the plant, achieving at the same time that their organoleptic and / or nutritional properties they are not affected and without being altered, for example the commercial quality of coriander.

For purposes of the present invention, the compositions described herein comprise:

 a) Extracts derived from plants, which exhibit antimicrobial properties, such as extracts derived from Jamaican chalices (Hibiscus sabdariffa),

 b) Chromatographic fractions obtained from acetonic and methanolic extracts derived from plants, which exhibit antimicrobial properties, such as chromatographic fractions obtained from chalices of Jamaica {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. themselves, in a w / w concentration of 0.01% to 10%, preferably 0.1% to 1%,

 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%, and e) A surfactant with emulsifying activity of natural fats or waxes found on the surface of! Coriander, such as polysorbates, Polysorbate 80, Polysorbate 20, C12-C18 alkyl dimethyl betaine (cocobetaine, C10-C16 alkyl dimethylbetaine (lauryl betaine), Sulphobetaine acyl (C10-C14 fatty) amidopropylene (hydroxypropylene B, cyclodextrin, C-cyclodextrin, C-cyclodextrin cyclodextrins and β-Cyclodextrin and mixtures thereof in a w / w concentration of 0.1% to 5%, preferably 0.5% to 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 the plant in disinfectant solutions or through devices that allow its 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 herein may contain only plant extracts with antimicrobial activity, such as extracts derived from Jamaican chalices, or chromatographic fractions obtained from acetonic and / or methanolic extracts of Jamaican calyces, which are put in contact with food, for example raw plant-based foods such as coriander, in order to disinfect and / or preserve them. In the present invention, the disinfectant activity of extracts derived from Jamaica and specific chromatographic fractions obtained from the extracts of the chalices of Jamaica, in the disinfection and / or preservation of foods, for example raw fruits and vegetables, is described by which can be used directly or as part of compositions that contain them. In this sense, chromatographic extracts or fractions derived from the chalices of Jamaica can be added or contacted with the 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 tai, which 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 foods such as coriander, 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 and the chromatographic fractions can be obtained by the following method: a) Place the dried plant in a container under aseptic conditions, add solvent ratio 1: 9; preferably 100 g of the dried plant are placed in a container (flask) under aseptic conditions, 900 ml of solvent are added and allowed to stand for 7 days; b) Remove the calyces and recover the solvent extract; preferably the resulting extract is recovered after pressing the plant on the walls of the flask 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) Completely remove the solvent 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; f) Obtain fractions of the extracts by column chromatography using solvent eluents of different polarity;

g) Remove the solvent from the fractions obtained by rotary evaporation at a temperature of 40 ° C, a rotation of 80 rpm and a vacuum pressure of 72 mbar; and h) Recover the dry fractions; preferably in a container.

Obtained extracts and fractions, these are stored at room temperature until use. Once the extracts and chromatographic fractions are 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 they are formed to form solutions and / or suspensions capable of being subsequently applied 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 Cilantro example. As can be seen later, the compositions of the invention are capable of disinfecting and / or eliminating microorganisms present in coriander very efficiently, so that it is possible to have microbiologically safe coriander safe 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 calyces (Hibiscus sabdariffa) of the Creole variety of Oaxaca were used, while in the case of coriander (Coriandrum sativum), this was obtained from a local producer. I work with the leaves; sheets of an approximate size of 3 cm ^{2 were selected} ; sheets of a uniform size and no visible damage were used.

1.2. Bacterial strains

 Strains of E. coli 0157 were used: H7 (P1C6, isolated from an outbreak of disease), enteroinvasive E. coli (4VC81-5, isolated from clinical case) E. Enterotoxigenic coli (1620 TL, isolated from clinical case), E Enteropathogenic coli (52 GM 291, isolated from clinical case), Salmonella typhimuríum (ATCC 14028), Salmonella choleraesuis (ATCC 10708), Listeria monocytogenes (ATCC 19115), Listeria monocytogenes Scott A, Staphylococcus epidermis (ATCC 1222c), Staphyloco (Staphyloco (ATCC 1222c)) ATCC 25923), Pseudomonas aeruginosa (ATCC 27853), Bordetella (ATCC 12741) Sri / ge // a sonnei (ATCC 25931) and Shigella flexnerí (ATCC 12022), V. cholerae (87151, Inaba serotype isolated from the environment) and Pseudomonas aeruginosa (ATCC 27853). The strains of £. 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 chalices were placed in an Erlenmeyer flask, to which 900 mL of distilled water was added, leading to boil 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 bar. The dried extract was recovered in a sterile bottle and stored at room temperature until use.

1.4. Obtaining methanolic and acetonic extract from Jamaica.

 Under aseptic conditions 100 g of Jamaican chalices were placed in a 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. Obtaining fractions from the methanolic extract by column chromatography

Once the dry extract (solvent-free) was obtained, it was mixed with silica (in order to make the extract manageable, since it still had moisture), it was added to the packed column. Cotton was placed at the bottom of the column with the help of a rod to prevent the silica gel from detaching when the key was opened, the column was held with two tweezers and made sure it was straight. The silica gel was mixed with hexane approximately 8: 1g. (silica gel: extract), this amount was mixed with hexane until a fluid paste was obtained, the paste was poured into the column, the amount of hexane added should be sufficient to prevent the silica from drying out or air entering the paste. The extract is added little by little, a small layer of sodium sulfate (this serves as a drying agent) was added, on top of this a layer of cotton was placed to cushion the solvent drop when added and thus prevent the dispersion of calcium sulfate and the extract, after this procedure, the column was filled with the solvent (hexane) and the key was opened to begin lowering the fractions with the different solvent mixtures, recovering them in amounts of 50 ml each, which were subsequently evaporated with help of the rotary evaporator, and these were placed in vials, considering each of these as a fraction.

Table 1. Solvents and mixtures used to obtain fractions from methanolic extract

To change the solvent mixture, thin plate chromatography was performed, and finding clearly visible differences (due to the appearance of different bands in size and shape) between fractions the mixture was changed from lower to higher polarity (hexane, ethyl acetate and methanol ) Table 1 shows the solvent mixtures used and with which it was eluted. From a total of 28.5 g of methanolic extract, 193 fractions were obtained, after determining the similarity of the fractions by thin layer chromatography, 7 fraction collections were obtained (Table 2).

Table 2. Number of fractions collected in each collection obtained from the methanolic extract

 1.6. Obtaining fractions from the acetonic extract by column chromatography.

Once the dry extract (solvent-free) was obtained, it was mixed with silica (in order to make the extract manageable, since it still had moisture), it was added to the packed column. Cotton was placed at the bottom of the column with the help of a rod to prevent the release of the silica gel when the key was opened, the column was fastened with two tweezers and secured in such a way that it was straight. The silica gel was mixed with chloroform approximately 8: 1g. (silica gel: extract), this amount was mixed with chloroform until a fluid paste was obtained, the paste was poured into the column, the amount of chloroform added should be sufficient to prevent the silica from drying out or air entering the paste, subsequently the extract was added little by little, a small layer of sodium sulfate (this serves as a drying agent) was added, a cotton layer was placed on top of it to cushion the solvent drop when added and thus avoid dispersion of the calcium sulfate and the extract, after this procedure the column was filled with the solvent (chloroform) and the key was opened to begin lowering the fractions with the different solvent mixtures, recovering them in amounts of 50 ml each, which subsequently evaporated with the help of the rotary evaporator, and these were placed in vials, considering each of these as a fraction. To change the solvent mixture, thin plate chromatography was performed, and finding differences between fractions the mixture was changed from lower to higher polarity (chloroform-acetone) in table 3 the solvent mixtures used and with which it was eluted are shown.

Table 3. Solvents and mixtures used to obtain fractions from acetonic extract

From a total of 20.5 g of acetonic extract, 1,117 fractions were obtained, after determining the similarity of the fractions by thin layer chromatography, 7 fraction collections were obtained (Table 4).

Table 4. Number of fractions collected in each collection obtained from the acetonic extract

1.7. Determination of the antimicrobial activity of aqueous, methanolic and acetonic extracts, acetic acid, hypochlorite and the corresponding fractions from Jamaican calyces in culture medium (in vitro studies).

1.7.1. 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.

1.7.2. Preparation of solutions of extracts or fractions.

From the dry extracts or dry fractions (collections), 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 and acetonic fractions (fraction III) and methanolic fractions (fraction IV) and were solubilized in distilled water while acetonic extracts and non-polar or low polarity fractions were solubilized in the solution of Polysorbate 80: water. To water or polysorbate 80: water, dry extracts or fractions were added in a ratio of 1: 10 and 1: 100 (water: extract or water: fraction) separately and will be deposited in sterile bottles. ^!

1.7.3 Antimicrobial effect of extracts and fractions in culture medium.

 Separately, 100 µL of the first dilution of the cultures of the pathogens were inoculated on boxes of supplementary AST with 10 mg / L of the antibiotic rifampin, the inoculum was distributed over the entire surface of the agar by the technique extensiónte extension by surface . On the inoculated boxes, separately; 10 µL aliquots of the solution of the extracts (aqueous, methanolic or acetonic), or of the chromatographic fractions were cotocated. 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.

1.8. Evaluation of the antimicrobial effect of aqueous, methanolic, acetonic extracts, chromatographic fractions and specific formulations in the reduction of Salmonella and E. coli 0157.Ή7 in contaminated coriander. 1.8.1. Preparation of disinfectant solutions.

 The solutions of Jamaican calyx extracts, mixtures based on extracts and chromatographic fractions as well as mixtures containing acetic acid, hypochlorite and / or polysorbate 80% were prepared at the concentrations, proportions or mixtures described in Table 5. By For example, to prepare 100 ml of a solution containing 1% Jamaican calyces methanolic extract, 0.1% acetic acid and 100 mg / L hypochlorite: to 100 mL of distilled water was added 1 g of dry methanolic extract of calyces of Jamaica, in addition 1 ml of a 10% acetic acid solution and 0.2 ml of a 5% hypochlorite solution.

1.8.2. Strains

For these studies we worked with 7 Salmonella serotypes: (3 typhimurium [ATCC 14028, one isolated from tomato, J1, and another from alfalfa seed, GA1], Salmonella choleraesuis [ATCC 10708], typhi, gaminara, and montes / ideo ) and £ 3. co // ^" 0157: H7 (two isolated in our laboratory from raw ground beef [P1C6 and M5C8] and another isolated from an outbreak caused by meat consumption in the United States of America [E09]), this strain It was donated by Dr. Eduardo Fernández Escartin of the Autonomous University of the State of Hidalgo, from the native strains mutant strains resistance to the antibiotic rifampin (R +) were obtained, this to be used in the studies. Cultivation to monitor the behavior of the mutant strains would eliminate the interference of the native microbial flora of the extracts, fractions and plant material of study (Castro-Rosas and Escartin, 2000).

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

1.8.4. Coriander Inoculation

Coriander leaves of a uniform or similar size (approx. 3 cm ² ) were used and which did not show visible damage. Separately, individual leaves were inoculated by placing in the central part of each leaf 5 drops or 10 L aliquots of a suspension of each type of pathogenic bacteria mixture (Salmonella or E. coli 0157: H7) containing approximately 1 x 10 ⁷ CFU , the 5 inoculums were close without coming together the inoculated leaves 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 conditions, in other words, to have a model that resembled as err - as possible what happens when the Coriander is contaminated by natural or common sources of contamination with pathogenic bacteria.

1.8.5. Cilantro leaves disinfection treatment.

 After two hours in the bioclimatic chamber, each leaf was separated by law to remove microorganisms that did not adhere, the washing consisted of submerging and stirring each coriander leaf inoculated in distilled water for 10 s, the part was allowed to drain washed at room temperature until Jgftál dryness and subsequently separately the leaves were immersed for 10 min in Jas different disinfectant solutions indicated in Table 5. A treatment only with distilled water served as a positive control.

1.8.6. Counting of microorganisms survivors to the tr amientos

After the treatment, the coriander leaves 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 box 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 shaken manually by pressing and rubbing the inoculated part and the entire coriander sheet 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.

Table 5. Treatments to which coriander leaves contaminated with mixtures of Salmonella or E. coli strains 0157: H7 were subjected separately

 No Treatments

 1 No treatment (control)

 2 EA 1%

 3 EAc 1%

 4 MS 1%

 5 Fac 1%

 6 Fm 1%

 7 HS 100 ppm

 8 Here 0.1%

 9 Here 0.5%

 10 EA 1% + Here 0.1% + HS 100 ppm

 11 EA 1% + Here 0.5% + HS 100 ppm

 12 EAc 1% + Here 0.1% + HS 100 ppm

 13 EAc 1% + Here 0.5% + HS 100 ppm

 14 MS 1% + Here 0.1% + HS 100 ppm

 15 MS 1% + Here 0.5% + HS 100 ppm

 16 Fac 1% + Here 0.1% + HS 100 ppm

 17 Fac 1% + Here 0.5% + HS 100 ppm

 18 Fm 1% + Here 0.1% + HS 100 ppm

 19 Fm 1% + Here 0.5% + HS 100 ppm

 20 EA 1% + Here 0.1% + HS 100 ppm + Po80 2%

 21 EA 1% + Here 0.5% + HS 100 ppm + Po80 2%

 22 EAc 1% + Here 0.1% + HS 100 ppm + Po80 2%

 23 EAc 1% + Here 0.5% + HS 100 ppm + Po80 2%

 24 MS 1% + Here 0.1% + HS 100 ppm + Po802%

 25 MS 1% + Here 0.5% + HS 100 ppm + Po802%

 26 Fac 1% + Here 0.1% + HS 100 ppm + Po802%

 27 Fac 1% + Here 0.5% + HS 100 ppm + Po80 2%

 28 Fm 1% + Here 0.1% + HS 100 ppm + Po802%

 29 Fm 1% + Here 0.5% + HS 100 ppm + Po802%

 30 MS 1% + Fac 1% + Here 0.1% + HS 100 ppm + Po80 2%

 31 MS 1% + Fac 1% + Here 0.5% + HS 100 ppm + Po80 2%

 32 MS 1% + Fm 1% + Here 0.1% + HS 100 ppm + Po80 2%

33 MS 1% + Fm 1% + Here 0.5% + HS 100 ppm + Po80 2% 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) Polyoxyethylene Sorbitan monoleate or polysorbate 80 (Polysorbate 80), d) Sterile distilled water at pH 6

1.8.7. 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.

1.9. Nuclear Magnetic Resonance (NMR) of extracts and collections.

The NMR spectrum of the proton ( ¹ H) of the dry methanolic extract obtained from the chalices of Jamaica and also of the collections of fractions obtained from the acetonic extract (collection III) and the methanolic extract (collection IV) was determined. These extracts and collections of fractions analyzed with NMR were those that were used in the formulations.

The dried extracts and the collections were solubilized in deuterated water. NMR spectra were obtained using a nuclear magnetic resonance spectrometer (They vary 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 ¹ H, ¹³ C, ¹ 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 movement of rotation 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 6). All the microorganisms tested were inhibited from the first moments of contact. 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.

In order to separate, isolate and / or concentrate the antimicrobial substances present in the extracts of the chalices of -Jamaica, the extracts were separated into different compounds or groups of compounds based on their polarity; for this, the compounds were separated by column chromatography, in this way different groups of compounds or groups of fractions were obtained (collections of fractions, see methodology). Subsequently, the antimicrobial effect of the collections of fractions obtained was tested. Because the acetonic and methanolic extract had a greater antimicrobial effect than the aqueous extract, the components of the aqueous extract were not chromatographed.

Table 6. Antimicrobial effect of the aqueous extract of Jamaica diluted 1: 10 and that of a penicillin solution (control) on different microorganisms

Diameter Diameter Diameter Diameter

Type of inhibition inhibition inhibition inhibition microorganism Extract Extract Methanolic acetonic aqueous penicillin extract (control)

E. coli 11 * 13 * 14 25

£ co // 0157: H7 11 14 14 22

E. coli enteroinvasive 11 11 13 25

E. coli 10 12 11 20 enteropathoqena

 E. coli 11 12 12 20 enterotoxigenic

 S. aureus 11 13 14 26

V. choteras OI 11 14 14 22

S. Typhimurium 12 14 14 24

S. Choleraesuis 11 12 12 20

P. aeruginosas 11 13 14 21

S. flexneri 10 12 14 22

S. sonnei 12 14 13 24

* (mm) L monocytogenes 10 13 14 23 Example 3. Antimicrobial effect of different chromatographic fractions.

 Table 7 shows the inhibitory effect expressed in length of the inhibition halo in millimeters (mm), which was observed in petri dishes seeded with different microorganisms, due to the effect of different collections of chromatographic fractions grouped by polarity obtained from acetonic extract of the calyces of Jamaica. It is observed that only collections II and III from acetonic extract show antimicrobial or inhibitory effect. Collection III is the one that showed the greatest antimicrobial effect (Table 7). This fraction III was used to make the mixtures or formulations that were used in the cilantro disinfection experiments.

Table 8 shows the inhibitory effect in mm of different collections of chromatographic fractions grouped by polarity obtained from the methanoic extract of the calyxes of Jamaica. It is observed that all the collections caused halos of inhibition which is interpreted as the antimicrobial effect of the collections. However, the IV collection is the one that showed the greatest antimicrobial effect (Table 8). This fraction IV was used to make the mixtures or formulations that were used in the cilantro disinfection experiments.

Table 7. Antimicrobial effect of acetonic collections

 Collection Collection Collection Collection Collection Collection Collection

Type of bacteria

 I II III IV V VI VII

 S. Choleraesuis 0 0 14 0 0 0 0

6. co / (0157: H7 0 9 * 18 0 0 0 0

S. flexneri 0 19 25 0 0 0 0

S. aureus 0 13 12 0 0 0 0

S. Typhimurium 0 22 18 0 0 0 0

L. monocytogenes 0 18 22 0 0 0 0

S. epidermis 0 13 21 0 0 0 0

P. aeruginosas 0 14 19 0 0 0 0

Bordetella 0 0 12 0 0 0 0

S. sonnei 0 11 18 0 0 0 0

* Halo of inhibition expressed in millimeters (mm); Zero (0) means that no inhibitory effect was observed. Table 8. Antimicrobial effect of methanolic collections

Type of Colecció Colecció Collection Collection Colecció Collection Collection bacteria n i n ll III IV n V VI VII

L

 6 10 15 24

 monocytogenes 12 11 15

S. sonnei 8 11 10 30 10 12 6

P. aeruginosas 6 10 11 22 8 8 7

S. Choleraesuis 10 12 13 36 9 13 6

S. fíexneri 7 14 13 29 10 12 8

S. Typhimurium 7 11 10 32 12 10 10

Bordetella 8 10 8 23 9 13 6

E. C0ft ^' O157: H7 6 17 20 26 14 11 6

S. aureus 13 22 17 30 11 10 7

S. epidermal 6 15 12 28 10 12 6

* Halo of inhibition expressed in millimeters (mm); Zero (0) means that no inhibitory effect was observed

Example 4. Disinfectant potential of extracts and fractions alone or in mixtures with acetic acid, sodium hypochlorite and / or Polysorbate 80.

It was found that all treatments had antimicrobial effect with respect to the control. The data of this study are reported in Table 9. It is observed that although all treatments show antimicrobial effect only 4 combinations managed to eliminate the concentration of mixtures of each pathogen at undetectable levels: treatments 30, 31, 32 and 33 reduced the concentration of both pathogens by 5 log ™ (Table 9).

In the present invention, with 4 specific combinations of three antimicrobials and one surfactant (polysorbate) the total elimination of the pathogenic microorganisms inoculated in the coriander was achieved; 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 should be noted the potential role of polysorbate 80 in the observed antimicrobial effect, since being a surfactant it is possible that it has favored the emulsification of natural coriander wax which could increase the effect of the disinfectant solution by eliminating or diminishing the effect protector that the wax would be providing to the microorganisms inoculated on the coriander.

Therefore, the 4 compositions of the present invention are an excellent alternative for the disinfection and / or preservation of foods, for example fresh foods, without altering their nutritional properties. In this sense, the compositions described here allow the effective disinfection of pathogenic microorganisms of fruits and vegetables, preferably coriander allowing the safe consumption of such products.

Table 9:

Table 9. Concentration of £ coli 0157: H7 and S. Typhimurium in Coriander at the beginning and after different treatments

 Eco // 0157: H7 Salmonella

 Treatment

 Number Number Number Number

Initial Final Initial Final

 1 No treatment (control) 5.00 ± 0.30 '* 4.80 ± 0.30 4.90 ± 0.30 4.70 ± 0.30

2 EA1% 5.00 ± 0.20 3.80 ± 0.30 4.90 ± 0.30 3.70 ± 0.20

3 EAc 1% 5.00 ± 0.20 3.50 ± 0.30 4.90 ± 0.30 3.50 ± 0.30

4 EM1% 5.00 ± 0.20 3.60 ± 0.30 4.90 ± 0.30 3.70 ± 0.40

5 Fac1% 5.00 ± 0.20 3.30 ± 0.40 4.90 ± 0.30 3.60 ± 0.20

6 Fm 1% 5.00 ± 0.20 3.40 ± 0.30 4.90 ± 0.30 3.50 ± 0.30

7 HS 100 ppm 5.00 ± 0.20 3.90 ± 0.40 4.90 ± 0.30 3.70 ± 0.20

8 Here 0.1% 5.00 ± 0.20 4.10 ± 0.30 4.90 ± 0.30 3.90 ± 0.30

9 Here 0.5% 5.00 ± 0.20 4.10 ± 0.30 4.90 ± 0.30 3.70 ± 0.30

10 EA 1% + Here 0.1% + HS 100 ppm 5.00 ± 0.20 3.10 ± 0.20 4.90 ± 0.30 3.40 ± 0.30

11 EA 1% + Here 0.5% + HS 100 ppm 5.00 ± 0.20 2.70 ± 0.20 4.90 ± 0.30 3.10 ± 0.30

12 EAc 1% + Here 0.1% + HS 100 ppm 5.00 ± 0.20 2.60 ± 0.30 4.90 ± 0.30 2.80 ± 0.30

13 EAc 1% + Here 0.5% + HS 100 ppm 5.00 ± 0.20 2.30 ± 0.30 4.90 ± 0.30 2.60 ± 0.30

14 MS 1% + Here 0.1% + HS 100 ppm 5.00 ± 0.20 2.80 ± 0.30 4.90 ± 0.30 2.80 ± 0.40

15 MS 1% + Here 0.5% + HS 100 ppm 5.00 ± 0.20 2.50 ± 0.30 4.90 ± 0.30 2.50 ± 0.40

16 Fac 1% + Here 0.1% + HS 100 ppm 5.00 ± 0.20 2.10 ± 0.20 4.90 ± 0.30 2.60 ± 0.30

17 Fac 1% + Here 0.5% + HS 100 ppm 5.00 ± 0.20 1.80 ± 0.30 4.90 ± 0.30 2.10 ± 0.30

18 Fm 1% + Here 0.1% + HS 100 ppm 5.00 ± 0.20 2.50 ± 0.20 4.90 ± 0.30 2.60 ± 0.30

19 Fm 1% + Here 0.5% + HS 100 ppm 5.00 ± 0.20 2.20 ± 0.20 4.90 ± 0.30 2.50 ± 0.30

20 EA 1% + Here 0.1% + HS 100 ppm + Po802% 5.00 ± 0.20 2.00 ± 0.20 4.90 ± 0.30 2.40 ± 0.20

21 EA 1% + Here 0.5% + HS 100 ppm + Po802% 5.00 ± 0.20 1.90 ± 0.30 4.90 ± 0.30 2.30 ± 0.20

22 EAc 1% + Here 0.1% + HS 100 ppm + Po802% 5.00 ± 0.20 1.90 ± 0.20 4.90 ± 0.30 2.10 ± 0.30

23 EAc 1% + Here 0.5% + HS 100 ppm + Po802% 5.00 ± 0.20 1.60 ± 0.20 4.90 ± 0.30 1.90 ± 0.20

24 MS 1% + Here 0.1% + HS 100 ppm + Po802% 5.00 ± 0.20 1.80 ± 0.20 4.90 ± 0.30 2.00 ± 0.30

25 MS 1% + Here 0.5% + HS 100 ppm + Po802% 5.00 ± 0.20 1.60 ± 0.20 4.90 ± 0.30 1.70 ± 0.30

26 Fac 1% + Here 0.1% + HS 100 ppm + Po802% 5.00 ± 0.20 1.60 ± 0.30 4.90 ± 0.30 1.90 ± 0.30

27 Fac 1% + Here 0.5% + HS 100 ppm + Po802% 5.00 ± 0.20 1.40 ± 0.30 4.90 ± 0.30 1.10 ± 0.30

28 Fm 1% + Here 0.1% + HS 100 ppm + Po802% 5.00 ± 0.20 1.70 ± 0.30 4.90 ± 0.30 1.80 ± 0.30

29 Fm 1% + Here 0.5% + HS 100 ppm + Po802% 5.00 ± 0.20 1.40 ± 0.30 4.90 ± 0.30 1.60 ± 0.30

 30, 1% MS + 1% Fac + Here 0.1% + HS 100 ppm + Po802% 5.0010.20 0.00 4.9010.30 0.00

 31 MS 1% + Fac 1% + Here 0.5% + HS 100 ppm + Po802% 5.0010.20 0.00 4.90 0.30 0.00

 32 MS 1% + Fm 1% + Here 0.1% + HS 100 ppm + Po802% 5.00 0.20 0.00 4.90 0.30 0.00

 33 MS 1% + Fm 1% + Here 0.5% + HS 100 ppm + Po802% 5.00 and 0.20 0.00 4.9010.30 0.00

Acetonic extract, MS: Methanolic extract, Fac: Acetonic fraction, Fm: Methanolic fraction, Here: Acetic acid, HS: Sodium hypochlorite, Po80: Polysorbate 80

Example 5. NMR spectrum obtained from the dried matanolic extract.

The NMR spectrum obtained from the dry methanolic extract of the chalices 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 acetonic extract used in the formulations in Table 9.

Example 6. NMR spectrum obtained from chromatographic collection III from acetonic extract.

From the chromatographic collection III obtained from the acetonic extract, the NMR spectrum was obtained. Figure 2. Several and different characteristic peaks of the chromatographic collection that we use in the formulations are observed. This spectrum characterizes the chromatographic collection III used in the antimicrobial formulations in Table 9.

Example 7. NMR spectrum obtained from the IV chromatographic collection from the methanolic extract.

From the chromatographic collection IV obtained from the methanolic extract, the NMR spectrum was obtained. Figure 3. Several and different characteristic peaks of the chromatographic collection that we use in the formulations are observed. This spectrum characterizes the IV chromatographic collection used in the antimicrobial formulations in Table 9.

References.

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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 coriander (Coriandrum sativum), characterized in that it comprises: a) Jamaican chalices methanolic extract {Hibiscus sabdariffa);

 b) A collection of otteaide chromatographic fractions of an acetonic extract of Jamaican chalices (Hibiscus sabdariffa);

 c) Acetic acid;

 d) Sodium hypochlorite;

 e) Sorbitan Polyoxyethylene Monooleate, or Polysorbate 80 (Polysorbate 80)

2. The solution with antimicrobial activity according to claim 1, characterized in that the a) metariótic extract of Jamaican chalices (Hibiscus sabdariffa) is present in a concentration between 0 ^% to 10%.

3. The solution with antimicrobial activity according to co-aPlication 1, characterized in that the b) collection of chromatographic fractions obtained from an acetonic extract of Jamaican chalices (Hibiscus. Sabdariffa) is present in a concentration between 0.01% to 10%.

4. The solution with antimicrobial activity in accordance with * claim 1, characterized in that the c) acetic acid is present in a concentration between 0.01 to 10%.

5. The solution with antimicrobial activity according to claim 1, characterized in that the d) sodium hypochlorite is present in a concentration between 10 to 1000 ppm.

6. The solution with antimicrobial activity according to claim 1, characterized in that the polyoxyethylene sorbitan e) monooleate, or polysorbate 80 is present in a concentration between 0.1 to 10%.

7. The solution with antimicrobial activity according to claim 1, characterized in that the f) methanolic extract of Jamaican chalices {Hibiscus sabdariffa) has a nuclear magnetic resonance (NMR) spectrum as seen in Figure 1.

8. The solution with antimicrobial activity according to claim i, characterized in that the g) chromatographic fraction collection obtained from an acetonic extract of Jamaican chalices (Hibiscus sabdariffa) has a nuclear magnetic resonance (NMR) spectrum as observed in Figure 2.

9. The solution of claim 1, characterized in that it has a presentation as an aqueous formulation.

10. The solution of claim 1 according to the preceding claims, characterized in that where one or more parts of Jamaica (Hibiscus sabdariffa) can be used to obtain the methanolic extract.

11. The solution of claim 1 according to the preceding claims, characterized in that where one or several parts of Jamaica (Hibiscus sabdariffa) can be used to obtain the chromatographic fraction collections of an acetonic extract;

12. The solution of claim 1 according to the preceding claims, wherein the part of the Jamaica plant that is used to obtain the extracts and the fraction collections are the calyces.

13. The solution of claim 1 according to the preceding claims, characterized by a nuclear magnetic resonance (NMR) spectrum of the methanolic extract obtained from the calyces of Jamaica (Hibiscus sabdariffa) (Figure 1).

14. The solution of claim 1 according to the preceding claims, characterized by a nuclear magnetic resonance (NMR) spectrum of the chromatographic fraction collection obtained from an acetonic extract of Jamaican calyces (Hibiscus sabdariffa) (Figure 2).

15. The solution of claim 1 according to the preceding claims, characterized in that it is useful as a disinfectant and preservative of foods of plant and animal origin.

16. The solution of claim 1 according to the preceding claims, characterized in that the aqueous formulation is useful as a disinfectant and preservative of fruits and vegetables, especially coriander.

17. The solution of claim 1 according to the preceding claims, characterized in that the main coriander varieties (Coriandrum sativum) on which it acts as a disinfectant are selected from sativum, microcarpum and Alef vulgare.

18. A method for the disinfection and / or preservation of coriander (Coriandrum sativum), characterized in that it comprises applying the solution defined in the preceding claims to the coriander.

19. A plant extract with antimicrobial activity to disinfect and / or preserve coriander (Coríandrum sativum), characterized in that it is obtained through the following stages: a) Place the dried 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 methanolic extract.

20. The extract of claim 19, characterized in that it is obtained with methanol.

21. The extract according to claim 19, wherein where one or more parts of the plant can be used to obtain the extract.

22. The extract of claim 19, characterized in that the plant is the Jamaica plant (Hibiscus sabdariffa).

23. The extract of claim 19 according to the preceding claims, characterized in that the extract is obtained from the calyces of Jamaica.

24. The extract of claim 19 according to the preceding claims, characterized in that it has a nuclear magnetic resonance (NMR) spectrum as seen in the Figure.

25. The extract of claim 19 according to the preceding claims, characterized in that it is useful as a disinfectant and preservative of foods of animal and plant origin.

26. The extract of claim 19 according to the preceding claims, characterized in that it is useful as a disinfectant and preservative of fruits and vegetables, preferably coriander.

27. The extract of claim 19 according to the preceding claims, characterized in that the main coriander varieties {Coriandrum sativum) on which it acts as a disinfectant are selected from sativum, microcarpum and Alef vulgare.

28. A method for the disinfection and / or preservation of coriander (Coriandrum sativum), characterized in that it comprises applying to the coriander the extract defined in claims 19 to 27.

29. A method for obtaining a plant extract with antimicrobial activity to disinfect and / or preserve coriander (Coriandrum sativum), characterized in that it comprises the following stages: a) Place the dried 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.

30. The method of obtaining the plant extract of claim 29, characterized in that the extract can have a solid or liquid presentation.

31. A collection of chromatographic fractions with antimicrobial activity to disinfect and / or preserve coriander (Coriandrum sativum), characterized in that it is obtained through the following stages: a) Place the dried plant 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,

 c) Recover the dry extract,

 d) Carry out chromatography to separate the dry acetonic extract into chromatographic fractions using solvents and solvent mixtures of different polarities,

 e) Recover chromatographic fractions of different polarities in containers, f) Remove solvent from the fractions,

 g) Group or collect the fractions in containers to form groups (collections) of equal or similar polarities,

 h) Perform microbiological tests with the collections,

i) Recover the collections of fractions with antimicrobial activity

32. The collection of chromatographic fractions according to claim 31, wherein where one or more parts of the plant can be used to obtain the collection of chromatographic fractions.

33. The collection of chromatographic fractions of claim 31 in accordance with the preceding claims, characterized in that the plant is Jamaica {Hibiscus sabdariffa).

34. The collection of chromatographic fractions of claim 31 in accordance with the preceding claims, characterized in that the fractions are obtained from the chalices of Jamaica.

35. The collection of chromatographic fractions with antimicrobial activity of claim 31 in accordance with the preceding claims, characterized in that the collection of fractions has a nuclear magnetic resonance spectrum (R N) as seen in Figure 2.

36. The collection of chromatographic fractions of claim 31 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative of foods of animal and plant origin.

37. The collection of chromatographic fractions of claim 31 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative of fruits and vegetables, preferably coriander.

38. The collection of chromatographic fractions with antimicrobial activity according to claim 31 in accordance with the preceding claims, characterized in that the main varieties of coriander (Coriandrum sativum) on which it acts as a disinfectant are selected from sativum, microcarpum and Alef vulgare.

39. The method to disinfect and / or preserve fruits and vegetables, preferably coriander with the collection of chromatographic fractions with activity antimicrobial extract of the chalices of Jamaica defined in claims 31 to 38.

40. A method to obtain collections of chromatographic fractions with antimicrobial activity from a plant extract to disinfect and / or preserve coriander (Coriandrum sativum), characterized in that it comprises the following stages: a) Place the dried plant in a container in 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,

 c) Recover the dry extract,

 d) Carry out chromatography to separate the dry acetonic extract into chromatographic fractions using solvents and solvent mixtures of different polarities,

 e) Recover chromatographic fractions of different polarities in containers, f) Remove solvent from the fractions,

 g) Group or collect the fractions in containers to form groups (collections) of equal or similar polarities,

 h) Perform microbiological tests with the collections,

 i) Recover the collections of fractions with antimicrobial activity.

41. The method according to claim 40, characterized in that the fraction collections can have a solid or liquid presentation.

42. A method for the preparation of a solution with antimicrobial activity to disinfect and / or preserve coriander {Coriandrum sativum), characterized by understanding the steps of: a) Place the dried Jamaica plant in a container under aseptic conditions, add methanol and storing at 22 ° C ± 2 for 7 days,

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

c) Recover the dry methanolic extract, d) Place dried Jamaica plant in a container under aseptic conditions, add acetone and store at 22 ° ± 2 ^o C for 7 days,

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

 f) Recover the dry acetonic extract,

 g) Carry out chromatography to separate the dry acetonic extract into chromatographic fractions using solvents and solvent mixtures of different polarities,

 h) Recover chromatographic fractions of different polarities in containers, i) Remove solvent from the fractions,

 j) Group or collect the fractions in containers to form groups (collections) of equal or similar polarities,

 k) Perform microbiological tests with the collections,

 I) Recover the collections of fractions with antimicrobial activity, m) Prepare the aqueous solution in a container containing: water, dry methanolic extract of Jamaica, a collection of chromatographic fractions with antimicrobial activity from the dry acetonic extract of Jamaica, acetic acid, hypochlorite of sodium and polyoxyethylene sorbitan monooleate or polysorbate 80.

43. The method according to claim 44, wherein the part of the Jamaica plant that is used is the calyces.

44. A solution with antimophobic activity to disinfect and / or preserve coriander (Coriandrum sativum), characterized in that it comprises a) Methanolic extract of Jamaican chalices (Hibiscus sabdariffa);

 b) A collection of chromatographic fractions obtained from a methanolic extract of Jamaican chalices (Hibiscus sabdariffa);

 c) Acetic acid;

 d) Sodium hypochlorite;

e) Sorbitart Polyoxyethylene Monooleate, or Polysorbate 80 (Polysorbate 80)

45. The solution with antimicrobial activity according to claim 44, characterized in that the a) metanolic extract of Jamaican chalices (Hibiscus sabdariffa) is present in a concentration between 0.01% to 10%.

46. The solution with antimicrobial activity according to claim 44, characterized in that the b) collection of chromatographic fractions obtained from a methanolic extract of Jamaican chalices (Hibiscus sabdariffa) is present in a concentration between 0.01% to 10%.

47. The solution with antimicrobial activity according to claim 44, characterized in that the c) acetic acid is present in a concentration between 0.01 to 10%.

48. The solution with antimicrobial activity according to claim 44, characterized in that the d) sodium hypochlorite is present in a concentration between 10 to 1000 ppm.

49. The solution with antimicrobial activity according to claim 44, characterized in that the polyoxyethylene sorbitan e) monooleate, or polysorbate 80 is present in a concentration between 0.1 to 10%.

50. The solution with antimicrobial activity according to claim 44, characterized in that the f) methanolic extract of Jamaican chalices (Hibiscus sabdariffa) has a nuclear magnetic resonance (NMR) spectrum as seen in Figure 1.

51. The solution with antimicrobial activity according to claim 44, characterized in that the g) chromatographic fraction collection obtained from a methanolic extract of Jamaican chalices (Hibiscus sabdariffa) has a nuclear magnetic resonance (NMR) spectrum as observed in Figure 3.

52. The solution of claim 44, characterized in that it has a presentation as an aqueous formulation.

53. The solution of claim 44 according to the preceding claims, characterized in that where one or more parts of Jamaica (Hibiscus sabdariffá) can be used to obtain the methanolic extract.

54. The solution of claim 44 according to the preceding claims, characterized in that where one or more parts of Jamaica (Hibiscus sabdariffá) can be used to obtain chromatographic fraction collections of a methanolic extract.

55. The solution of claim 44 according to the preceding claims, wherein the part of the Jamaica plant that is used to obtain the extracts and the fraction collections are the chalices.

56. The solution of claim 44 according to the preceding claims, characterized by a nuclear magnetic resonance (NMR) spectrum of the methanolic extract obtained from the calyces of Jamaica (Hibiscus sabdariffá) (Figure 1).

57. The solution of claim 44 according to the preceding claims, characterized by a nuclear magnetic resonance (NMR) spectrum from the collection of chromatographic fractions obtained from methanolic urrextractcr of Jamaican calyces (Hibiscus sabdariffá) (Figure 3);

58. The solution of claim 44 according to the preceding claims, characterized in that it is useful as a disinfectant and coa3¾wtótof Ue food of plant and animal origin.

59. The solution of claim 44 according to the preceding claims, characterized in that the aqueous formulation is useful as a disinfectant and preservative of fruits and vegetables, especially coriander.

60. The solution of claim 44 according to the preceding claims, characterized in that the main coriander varieties (Coriandrum sativum) on which it acts as a disinfectant are selected from sativum, microcarpum and Alef vulgare.

61. A method for the disinfection and / or preservation of coriander (Coriandrum sativum), characterized in that it comprises applying the solution defined in claims 44 to 60 to the cilantro.

62. A plant extract with antimicrobial activity to disinfect and / or preserve coriander (Coriandrum sativum), characterized by claims from 19 to 30.

63. A collection of chromatographic fractions with antimicrobial activity to disinfect and / or preserve coriander (Coriandrum sativum), characterized in that it is obtained through the following stages: a) Place the dried 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 acetone from the extract,

 c) Recover the dry extract,

 d) Perform chromatography to separate the dry acetonic extract into chromatographic fractions using solvents-- and- solvent mixtures of different polarities,

 e) Recover chromatographic fractions of different polarities in containers, f) Remove the solvent from the fraaisnes,

 g) Grouping or collecting containers into sections to form groups (collections) of equal or similar polarities,

 h) Perform microbiological tests ^ eafrlas collections,

 i) Recover the collections of fractions with antimicrobial activity

64. The collection of chromatographic fractions according to claim 63, wherein where one or more parts of the plant can be used to obtain the collection of chromatographic fractions.

65. The collection of chromatographic fractions of claim 63 in accordance with the preceding claims, characterized in that the plant is Jamaica (Hibiscus sabdariffa).

66. The collection of chromatographic fractions of claim 63 in accordance with the preceding claims, characterized in that the fractions are obtained from the chalices of Jamaica.

67. The collection of chromatographic fractions with antimicrobial activity of claim 63 in accordance with the preceding claims, characterized in that the fraction collection has a nuclear magnetic resonance (NMR) spectrum as seen in Figure 3.

68. The collection of chromatographic fractions of claim 63 in accordance with the preceding claims, characterized in that it is useful as a disinfectant and preservative of foods of animal and plant origin.

69. The collection of chromatographic fractions of claim 63 according to the preceding claims, characterized in that it is useful as a disinfectant and preservative of fruits and vegetables, preferably coriander.

70. The collection of chromatographic fractions with arifimicrobial activity according to claim 63 in accordance with the preceding claims, characterized in that the main varieties of coriander (Coriandrum sativum) on which it acts as a disinfectant are selected from sativum, microcarpum and Alef vulgare.

71. The method for disinfecting and / or preserving fruits and vegetables, preferably coriander with the collection of chromatographic fractions with antimicrobial activity of the extract of the Jamaica chalices defined in claims 63 to 70.

72. A method to obtain collections of chromatographic fractions with antimicrobial activity from a plant extract to disinfect and / or preserve coriander (Coriandrum sativum), characterized in that it comprises the following stages: a) Place the dried plant in a container in 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,

 c) Recover the dry extract,

 d) Carry out chromatography to separate the dry acetonic extract into chromatographic fractions using solvents and solvent mixtures of different polarities,

 e) Recover chromatographic fractions of different polarities in containers, f) Remove solvent from the fractions,

 g) Group or collect the fractions in containers to form groups (collections) of equal or similar polarities,

 h) Perform microbiological tests with the collections,

 i) Recover the collections of fractions with antimicrobial activity.

73. The method according to claim 72, characterized in that the fraction collections can have a solid or liquid presentation.

74. A method for the preparation of a solution with antimicrobial activity to disinfect and / or preserve coriander (Coriandrum sativum), characterized by comprising the steps of: a) Place the dried Jamaica plant (Hibiscus sabdariffa) 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 methanolic extract.

 d) Place dried Jamaica plant in a container under aseptic conditions, add acetone and store at 22 ° ± 2 ° C for 7 days,

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

f) Recover the dry acetonic extract, g) Carry out chromatography to separate the dry acetonic extract into chromatographic fractions using solvents and solvent mixtures of different polarities,

h) Recover chromatographic fractions of different polarities in containers, i) Remove solvent from the fractions,

j) Group or collect the fractions in containers to form groups (collections) of equal or similar polarities,

k) Perform microbiological tests with the collections,

I) Recover the collections of fractions with antimicrobial activity, m) Prepare the aqueous solution in a container containing: water, dry metanóüco extract of Jamaica, a collection of chromatographic fractions with antimicrobial activity from the dry acetonic extract of Jamaica, acetic acid, hypochlorite of sodium and polyoxyethylene sorbitan monooleate or polysorbate 80. The method according to claim 74, wherein the part of the Jamaica plant used is calyces.