Herein, the term'plant matter'generally refers to an entire or whole plant, to a portion of an entire or whole plant, to a component thereof, or, to a combination thereof.

The form of the plant matter is selected from the group consisting of a raw form, a partly processed form (that is, substantially solid and visually recognizable as originating from a plant), a loose form, a bundled form, and combinations thereof.

Primary examples of plant matter particularly relevant to the field of the present invention are fruits, vegetables, and flowers, in a raw or partly processed, loose or bundled, form. Examples of plant matter as entire or whole plants, also relevant to the field of the present invention, are trees, shrubs, bushes, grass, and moss, in a raw or partly processed, loose or bundled, form. Examples of plant matter as a portion of an entire or whole plant, or, a component thereof, also relevant to the field of the present invention, are leaves,

blossoms, beans, seeds, grains, stems, stalks, fibers, and roots, in a raw or partly processed, loose or bundled, form.

Herein, 'harvested'plant matter generally refers to any of the above types of plant matter which has been harvested, that is, manually, mechanistically, and/or automatically, separated, detached, or removed, such as by pulling or cutting, from the point or location of cultivation, development, or growth, of the plant matter, such that the harvested plant matter is no longer cultivated, developed, or grown. Typically, the harvested plant matter is gathered or collected, and subjected to a variety of numerous processes and sequences, involving activities such as bundling, packaging, storing, transporting, and treatment, in a variety of forms before eventually being sold in commercial wholesale and retail (shelf) environments.

Active oxygen species (AOS) can cause oxidative damage to many cellular components, including membrane lipids, proteins, and nucleic acids (Shalata et al. , 2001; Halliwell and Gutteridge, 1989). In plants, both enzymatic and non-enzymatic processes participate in AOS detoxification. Low molecular mass antioxidants, either hydrophilic, such as ascorbic acid (ASC) or reduced glutathione (GSH), or, lipophilic, such as a- tocopherol or carotenoids, can quench various kinds of AOS. During development of plants, during storage of fruits, and when plants or fruits, such as harvested plant matter, are subjected to environmental stresses, oxidative damage may result due to the balance between production of AOS and their detoxification by alteration of the antioxidative system (Hernandez et al. , 1993,1995 ; Gomez et al. , 1999; Shalata et al. , 2001).

Ascorbic acid (vitamin C), schematically illustrated immediately below, is a relatively small, water-soluble antioxidant molecule, which acts as a primary substrate in the cyclic pathway for enzymatic detoxification of hydrogen peroxide (H ? 02)- In addition, it acts to directly neutralize superoxide radicals (O ?'), single oxygen, and as secondary anti-oxidant during reductive recycling of the oxidized form of a-tocopherol (vitamin E), another lipophilic anti-oxidant molecule (Noctor and Foyer, 1998; Shalata and Neumann, 2001). Ascorbic acid, identified as the antiscorbutic factor about 70 years ago (Davies et al. , 1991), is well known as an effective antioxidant which protects humans from diseases, primarily by functioning as a scavenger of AOS involved in most human diseases (Arrigoni and Tullio, 2000; Horemans et al., 1997).

AA, R=R1=R2=R3=H R, R1, R2, R3, alkyl or alkanoy AA = ASCORBIC ACID (ASC) This property and behavior of ascorbic acid are clearly evidenced by the resulting rapid changes in the concentration or redox status of ascorbic acid observed in response to air pollutants such as ozone (Castillo and Greppin, 1988 ; Polle et al. , 1990; Luwe et al., 1993) and NO2 (Ramage et al. , 1993). However, cell growth and development have also been attributed to the presence of ascorbic acid in the cell wall matrix. For example, the addition of extracellular ascorbic acid to onion (Allium Cepal) Italic, or pea (Pisum Sativum L. ) Italic, roots, enhance the elongation of cells (Hidalgo et al. , 1991), and regulate lignification processes in the cell wall. When ascorbic acid is used in different physiological processes, it becomes oxidized to dehydroascorbate (DHA). For this reason, plant cells have developed efficient ways to regenerate ascorbic acid in different cell compartments, such as according to the Halliwell-Asada cycle (Foyer and Halliwell, 1976; Rose, 1988). However, regeneration of ascorbic acid does not provide full protection because ascorbic acid is insufficiently stable for long periods of time.

Ascorbic acid, being vitamin C, is an important quality component of many types and varieties of fruits and vegetables. It is now generally accepted that vitamin C is one of the most important free radical scavengers in plants, animals, and humans (MacKersie and Lesham, 1994), which is the main reason that vitamin C is considered to play a significant role in reducing carcinogenesis and cardiovascular diseases. Protection against free oxygen and other metabolic radicals is also an important characteristic of vitamin C in the physiology of plants and fruits (Elstner, 1982). Vitamin C may also be involved in preventing certain physiological fruit disorders, such as superficial scald in apples, and preventing oxidative damage. Vitamin C is also involved in many reactions in basic plant

metabolism (Chinoy, 1984), and is widely used in the fruit processing industry. Because of this fundamental importance, there is a need to preserve the content of vitamin C in fruits during prolonged periods of storage and shelf life (Agar et al. , 1997). Vitamin C (ascorbic acid), as a natural antioxidant (Kohler et al., 1988), is added to foodstuffs to protect color, aroma, and/or nutrient content.

Ascorbic acid is very sensitive to heat, light, and the action of oxidizing agents and metal ions. Ascorbic acid is readily oxidized, especially in aqueous solutions, by reacting with atmospheric oxygen (Stryer, 1988). Since ascorbic acid is generally unstable, use of it as an antioxidant for foodstuffs is significantly limited, especially with respect to commercial applications. Additionally, the highly hydrophilic behavior of ascorbic acid either prevents or at best significantly limits use of it in the presence of fats and/or oils, in preservative types of formulations or compositions for agricultural applications, and in cosmetics applications.

There are, however, readily synthesized derivatives of ascorbic acid, in particular, at positions 6,2, or 3, which are relatively stable, and retain antioxidant properties and behavior (Tanaka et al. , 1966). For example, alkanoyl-L-ascorbic acid ester (saturated fatty acid ester) derivatives of ascorbic acid, characterized by an amphiliphic structure, have better solubility and miscibility properties in hydrophobic environments, and also appear to enhance radical scavenging, compared to ascorbic acid (Liu et al. , 1996).

A known method for synthesizing alkanoyl-L-ascorbic acid esters (saturated fatty acid esters) of ascorbic acid is based on enzymatic catalysis, involving the use of a lipase catalyst, of a trans-esterification reaction, during which there is trans-esterification of the saturated fatty acid ester and the ascorbic acid reactants (Humeau et al. , 1998-a, 1998-b).

For example, 6-octyl-ascorbate (6-octanoyl-L-ascorbate) and 6-decyl-ascorbate (6- decanoyl-L-ascorbate) were each synthesized and obtained with a yield of 65 % using this method (Youchun et al. , 1999). However, this type of enzymatic method is not suitable or cost effective for commercial large-scale manufacturing applications for the following main reasons: (1) the enzymatic reaction is relatively long, (2) the separation of products is not practical using standard extraction or recrystalization procedures due to the need for having excess quantities of saturated fatty acid ester reactant relative to the ascorbic acid, (3) limited stability of the ascorbic acid ester products, and (4) relatively high cost of a sufficiently active enzyme needed for obtaining a good yield of product.

There is thus a need for, and it would be highly advantageous to have a method for extending the shelf life of harvested plant matter using compositions of the alkanoyl-L- ascorbic acid esters: 6-octanoyl-L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6-decyl-ascorbate), wherein the alkanoyl-L-ascorbic acid esters function as active ingredients exhibiting antioxidation properties, characteristics, and behavior, such that oxidation, and therefore, degradation, of the harvested plant matter are effectively inhibited during a period of time, thereby extending the shelf life of the harvested plant matter. Additionally, there is a need for having the compositions of the alkanoyl-L-ascorbic acid esters. Moreover, there is a further need for having a new method for synthesizing the alkanoyl-L-ascorbic acid esters, which is suitable and cost effective for commercial large-scale manufacturing applications.

Furthermore, there is a need for such an invention which is environmentally safe, and non- hazardous to humans or animals during handling and/or consumption of the treated harvested plant matter.

SUMMARY OF THE INVENTION The present invention relates to a method for extending the shelf life of harvested plant matter using compositions of alkanoyl-L-ascorbic acid esters; the compositions of the alkanoyl-L-ascorbic acid esters; and a method for synthesizing the alkanoyl-L-ascorbic acid esters. Particular alkanoyl-L-ascorbic acid esters featured in the present invention are 6-octanoyl-L-ascorbate, also known as and herein equivalently referred to as 6-octyl- ascorbate; 6-nonanoyl-L-ascorbate, also known as and herein equivalently referred to as 6- nonyl-ascorbate; and 6-decanoyl-L-ascorbate, also known as and herein equivalently referred to as 6-decyl-ascorbate. These alkanoyl-L-ascorbic acid esters, functioning as active ingredients exhibiting antioxidation properties, characteristics, and behavior, in compositions of solutions or suspensions applied onto harvested plant matter, are effectively utilized for significantly extending the shelf life of the harvested plant matter.

The method for synthesizing these alkanoyl-L-ascorbic acid esters is suitable and cost effective for commercial large-scale manufacturing applications.

The method for extending the shelf life of harvested plant matter, of the present invention, is based on the main step of applying onto the harvested plant matter an effective amount of a composition of a solution or suspension including an alkanoyl-L-

ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate, 6- nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, as an antioxidation active ingredient, such that oxidation, and therefore, degradation, of the harvested plant matter are effectively inhibited during a period of time, thereby extending the shelf life of the harvested plant matter.

The composition for use in extending the shelf life of harvested plant matter, of the present invention, is based on a solution or suspension including an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate, 6-nonanoyl-L- ascorbate, and 6-decanoyl-L-ascorbate, as an antioxidation active ingredient.

In the solution or suspension of the composition, of the present invention, the selected alkanoyl-L-ascorbic acid ester (6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, or 6-decanoyl-L-ascorbate), preferably having a concentration in a range of between about 1 mM and about 75 mM, and more preferably, having a concentration in a range of between about 5 mM and about 50 mM, dissolved or suspended in water, an alcohol, and/or a glycol, is active and effective as a sole antioxidation active ingredient, such that oxidation, and therefore, degradation, of the harvested plant matter are effectively inhibited during a period of time, thereby extending the shelf life of the harvested plant matter.

The solution or suspension of the composition, of the present invention, preferably includes a single alkanoyl-L-ascorbic acid ester selected from the group consisting of 6- octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, which functions as the sole antioxidation active ingredient. However, it is clearly within the scope of the present invention for the solution or suspension of the composition to include a combination of at least two alkanoyl-L-ascorbic acid esters selected from the group consisting of 6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L- ascorbate, wherein each alkanoyl-L-ascorbic acid ester functions as an antioxidation active ingredient. Moreover, for the solution or suspension of the composition including a combination of at least two of the alkanoyl-L-ascorbic acid esters, wherein each alkanoyl- L-ascorbic acid ester functions as an antioxidation active ingredient, the alkanoyl-L- ascorbic acid esters can be active and effective as a synergistic combination of antioxidation active ingredients, such that oxidation of the harvested plant matter is even more effectively inhibited during a period of time, thereby significantly extending the shelf life of the harvested plant matter.

Applying the solution or suspension of the composition onto the harvested plant matter is preferably performed using a dipping or spraying procedure performed at room temperature, during which the harvested plant matter is subjected or exposed to the applied composition for a time period of between thirty minutes and two hours, or, between three and ten seconds, respectively. Alternatively, the solution or suspension of the composition is brushed on the harvested plant matter using a suitable brush for such an application.

The method for synthesizing an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L- ascorbate, of the present invention, is based on direct esterification of an equimolar mixture of the corresponding saturated fatty acid (octanoic acid, nonanoic acid, and decanoic acid, respectively) and L-ascorbic acid, in a large molar excess of concentrated sulfuric acid (at least about 96 %), wherein the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid is greater than 10/1, and preferably, about 25/1.

Thus, according to the present invention, there is provided a method for extending the shelf life of harvested plant matter, the method comprising applying onto the harvested plant matter an effective amount of a composition which comprises a solution or suspension including an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, as an antioxidation active ingredient, such that oxidation of the harvested plant matter is effectively inhibited during a period of time.

According to further characteristics in preferred embodiments of the method of the invention described below, the plant matter is selected from the group consisting of an entire or whole plant, a portion of an entire or whole plant, a component thereof, and combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, the plant matter is in a form selected from the group consisting of a raw form, a partly processed form, a loose form, a bundled form, and combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, the plant matter is selected from the group consisting of fruits, vegetables, and flowers.

According to further characteristics in preferred embodiments of the method of the invention described below, the plant matter is selected from the group consisting of trees, shrubs, bushes, grass, and moss.

According to further characteristics in preferred embodiments of the method of the invention described below, the plant matter is selected from the group consisting of leaves, blossoms, beans, seeds, grains, stems, stalks, fibers, roots, and spices.

According to further characteristics in preferred embodiments of the method of the invention described below, applying the composition onto the harvested plant matter is performed using a procedure selected from the group consisting of dipping, rolling, brushing, wiping, rubbing, dripping, spraying, atomizing, and combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, applying the composition onto the harvested plant matter is performed using a procedure performed at a temperature in a range of between about 2 °C and about 70 °C.

According to further characteristics in preferred embodiments of the method of the invention described below, applying the composition onto the harvested plant matter is performed using a dipping procedure performed at a temperature in a range of between about 2 °C and about 70 °C, for a period of time in a range of between about one second and about eight hours, such that the harvested plant matter is substantially, entirely, dipped or immersed in, and surrounded by, the composition for the period of time.

According to further characteristics in preferred embodiments of the method of the invention described below, applying the composition onto the harvested plant matter is performed using a spraying or brushing procedure performed at a temperature in a range of between about 2 °C and about 70 °C, for a period of time in a range of between about one second and about three minutes, such that the harvested plant matter is subjected to the composition by the spraying or brushing for. the period of time.

According to further characteristics in preferred embodiments of the method of the invention described below, the solvent of the composition is a low molecular weight polar solvent selected from the group consisting of water, a low molecular weight alcohol, a low molecular weight polar glycol, and miscible combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, the low molecular weight alcohol is selected from the group consisting of ethanol, propanol, and miscible combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, the low molecular weight glycol is selected from the group consisting of propylene glycol, ethylene glycol, and miscible combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, the molar concentration of the alkanoyl-L-ascorbic acid ester in the composition is in a range of between about 1 mM and about 75 mM.

According to further characteristics in preferred embodiments of the method of the invention described below, the composition further includes at least one additional component or ingredient for improving physicochemical properties, characteristics, behavior, and activity, of the composition.

According to further characteristics in preferred embodiments of the method of the invention described below, the at least one additional component or ingredient is selected from the group consisting of a dispersing agent, an emulsifier, an acidulant, a plasticizer, a complexing agent, a preservative, a firming and/or sequestering agent, a protein, an antioxidation agent, a coating, and combinations thereof.

According to further characteristics in preferred embodiments of the method of the invention described below, the composition includes a single the alkanoyl-L-ascorbic acid ester, wherein the alkanoyl-L-ascorbic acid ester functions as a sole antioxidation active ingredient.

According to further characteristics in preferred embodiments of the method of the invention described below, the composition includes at least two alkanoyl-L-ascorbic acid esters, wherein each alkanoyl-L-ascorbic acid ester functions as an antioxidation active ingredient.

According to further characteristics in preferred embodiments of the method of the invention described below, the at least two alkanoyl-L-ascorbic acid esters are active and effective as a synergistic combination of the antioxidation active ingredients.

According to further characteristics in preferred embodiments of the method of the invention described below, the alkanoyl-L-ascorbic acid ester is active and effective as a fungicide.

According to further characteristics in preferred embodiments of the method of the invention described below, the alkanoyl-L-ascorbic acid ester is 6-octanoyl-L-ascorbate.

According to further characteristics in preferred embodiments of the method of the invention described below, the alkanoyl-L-ascorbic acid ester is 6-nonanoyl-L-ascorbate.

According to further characteristics in preferred embodiments of the method of the invention described below, the alkanoyl-L-ascorbic acid ester is 6-decanoyl-L-ascorbate.

According to another aspect of the present invention, there is provided a composition for use in extending the shelf life of harvested plant matter, the composition comprising a solution or suspension including an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6- decanoyl-L-ascorbate, as an antioxidation active ingredient.

According to further characteristics in preferred embodiments of the composition of the invention described below, the molar concentration of the alkanoyl-L-ascorbic acid ester in the solution or suspension is in a range of between about 1 mM and about 75 mM.

According to further characteristics in preferred embodiments of the composition of the invention described below, the solution or suspension further includes at least one additional component or ingredient for improving physicochemical properties, characteristics, behavior, and activity, of the solution or suspension.

According to further characteristics in preferred embodiments of the composition of the invention described below, the at least one additional component or ingredient is selected from the group consisting of a dispersing agent, an emulsifier, an acidulant, a plasticizer, a complexing agent, a preservative, a firming and/or sequestering agent, a protein, an antioxidation agent, a coating, and combinations thereof.

According to further characteristics in preferred embodiments of the composition of the invention described below, the solution or suspension includes a single alkanoyl-L- ascorbic acid ester, wherein the alkanoyl-L-ascorbic acid ester functions as a sole antioxidation active ingredient.

According to further characteristics in preferred embodiments of the composition of the invention described below, the solution or suspension includes at least two alkanoyl- L-ascorbic acid esters, wherein each alkanoyl-L-ascorbic acid ester functions as a antioxidation active ingredient.

According to further characteristics in preferred embodiments of the composition of the invention described below, the at least two alkanoyl-L-ascorbic acid esters are active and effective as a synergistic combination of the antioxidation active ingredients.

According to further characteristics in preferred embodiments of the composition of the invention described below, the alkanoyl-L-ascorbic acid ester is active and effective as a fungicide.

According to further characteristics in preferred embodiments of the composition of the invention described below, the alkanoyl-L-ascorbic acid ester is 6-octanoyl-L- ascorbate.

According to further characteristics in preferred embodiments of the composition of the invention described below, the alkanoyl-L-ascorbic acid ester is 6-nonanoyl-L- ascorbate.

According to further characteristics in preferred embodiments of the composition of the invention described below, the alkanoyl-L-ascorbic acid ester is 6-decanoyl-L- ascorbate.

According to another aspect of the present invention, there is provided a method for synthesizing an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6- octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, comprising the steps of : (a) preparing an equimolar mixture of a saturated fatty acid selected from the group consisting of octanoic acid, nonanoic acid, and decanoic acid, respectively, and, L- ascorbic acid, such that molar ratio of the saturated fatty acid to the L-ascorbic acid equals 1 ; (b) adding an excess quantity of sulfuric acid having a concentration of at least about 96 % to the equimolar mixture, such that molar ratio of the sulfuric acid to sum of the saturated fatty acid and the L-ascorbic acid is greater than 10/l, for forming a reaction mixture; (c) stirring the reaction mixture, such that reaction takes place for forming the corresponding alkanoyl-L-ascorbic acid ester, and (d) separating the formed alkanoyl-L- ascorbic acid ester from the reaction mixture.

According to further characteristics in preferred embodiments of the synthesis method of the invention described below, the saturated fatty acid is the octanoic acid for forming 6-octanoyl-L-ascorbate as the alkanoyl-L-ascorbic acid ester.

According to further characteristics in preferred embodiments of the synthesis method of the invention described below, the saturated fatty acid is nonanoic acid for forming 6-nonanoyl-L-ascorbate as the alkanoyl-L-ascorbic acid ester.

According to further characteristics in preferred embodiments of the synthesis method of the invention described below, the saturated fatty acid is decanoic acid for forming 6-decanoyl-L-ascorbate as the alkanoyl-L-ascorbic acid ester.

According to further characteristics in preferred embodiments of the synthesis method of the invention described below, the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid is 25/'l.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention is herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative description of the preferred embodiments of the present invention only, and are presented in the cause of providing what is believed to be the most useful and readily understood description of the principles and conceptual aspects of the present invention. In this regard, no attempt is made to show structural details of the present invention in more detail than is necessary for a fundamental understanding of the invention, the description taken with the drawings making apparent to those skilled in the art how the several forms of the invention may be embodied in practice. In the drawings: FIG. 1 is a schematic diagram illustrating the reaction schemes of the reactions for synthesizing the alkanoyl-L-ascorbic acid esters: 6-octanoyl-L-ascorbate, 6-nonanoyl-L- ascorbate, and 6-decanoyl-L-ascorbate, in accordance with the present invention; FIGS. 2 and 3 are complementary bar graphs illustrating the effect on % firm melons and % soft melons, respectively, after a treatment of spraying the melons with different compositions at room temperature, then cold storage at 5 °C for 14 days, 3 days, and 7 days, followed by shelf life at 20 °C, in accordance with Example 4 of the present invention; FIG. 4 is a bar graph illustrating the effect of different treatments on brown spots (%) of melons after spraying with different compositions at room temperature, then cold

storage at 5 °C for 14 days, followed by shelf life at 20 °C for 3 days and for 7 days, in accordance with Example 4 of the present invention; FIGS. 5-10 are photographs illustrating the effect of different treatments on initiation and development of brown spots and discoloration of melons after spraying with different compositions at room temperature, then cold storage at 5 °C for 14 days, followed by shelf life at 20 °C for 3 days (FIGS. 5-7) and for 7 days (FIGS. 8-10); in accordance with Example 4 of the present invention; FIGS. 11,12, and 13 are photographs illustrating the effect on preventing decay and microbial growth of three different species (golden apples, green apple, and red apple, respectively), of cut apples, after dipping the cut applies in a 6-octanoyl-L-ascorbate solution at room temperature, following shelf life storage of 0 days at 20 ° C, in accordance with Example 5 of the present invention; FIGS. 14,15, and 16 are photographs illustrating the effect on preventing decay and microbial growth of three different species (golden apples, green apple, and red apple, respectively), of cut apples, after dipping the cut applies in a 6-octanoyl-L-ascorbate solution at room temperature, following shelf life storage of 1 day at 20 ° C, in accordance with Example 5 of the present invention; FIGS. 17,18, and 19 are photographs illustrating the effect on preventing decay and microbial growth of three different species (golden apples, green apple, and red apple, respectively), of cut apples, after dipping the cut applies in a 6-octanoyl-L-ascorbate solution at room temperature, following shelf life storage of 8 days at 20 ° C, in accordance with Example 5 of the present invention ; FIGS. 20,21, and 22 are photographs illustrating the effect on preventing decay and microbial growth of three different species (golden apples, green apple, and red apple, respectively), of cut apples, after dipping the cut applies in a 6-octanoyl-L-ascorbate solution at room temperature, following shelf life storage of 7 days at 4 ° C, in accordance with Example 5 of the present invention; FIGS. 23,24, and 25 are photographs illustrating the effect on preventing decay and microbial growth of three different species (golden apples, green apple, and red apple, respectively), of cut apples, after dipping the cut applies in a 6-octanoyl-L-ascorbate solution at room temperature, following shelf life storage of 7 days at 4 ° C and 21 days at 20 ° C, in accordance with Example 5 of the present invention;

FIGS. 26 and 27,28 and 29, and, 30 and 31, are photographs illustrating the effect on preventing decay and browning of petiole grapes, after spraying the grapes with different compositions at room temperature, followed by shelf life at 20 °C for 2 days, 4 days, and 6 days, respectively, in accordance with Example 6 of the present invention; FIGS. 32 and 33 are photographs illustrating the effect of various treatments of compositions of the present invention on the visual and organoleptic quality of cut lettuce, corresponding to data in Tables 12-15, in accordance with Example 7 of the present invention; FIG. 34 is a set of photographs illustrating the effect on preventing decay and browning of lettuce heads, after brushing the lettuce heads with a 6-octanoyl-L-ascorbate composition at room temperature, followed by storage at 6 °C for periods of 0, 18,48, 72, 96,120, and 216 hours, in accordance with Example 8 of the present invention; FIGS. 35 and 36 are photographs illustrating the effect on preventing decay, browning, discoloration, and spoilage, of bananas, after dipping the bananas into different compositions at room temperature, followed by storage at 20 °C for periods of 0 days, and 8 days, respectively, in accordance with Example 9 of the present invention; and FIG. 37 are photographs illustrating the effect on preventing decay and browning of lettuce heads, after the lettuce heads were brushed with different compositions at room temperature, directly covered with nylon pieces and stored at 2 °C in a cold room for a period of 7 days, in accordance with Example 10 of the present invention.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention relates to a method for extending the shelf life of harvested plant matter using compositions of alkanoyl-L-ascorbic acid esters; the compositions of the alkanoyl-L-ascorbic acid esters; and a method for synthesizing the alkanoyl-L-ascorbic acid esters. Particular alkanoyl-L-ascorbic acid esters featured in the present invention are 6-octanoyl-L-ascorbate, also known as and herein equivalently referred to as 6-octyl- ascorbate; 6-nonanoyl-L-ascorbate, also known as and herein equivalently referred to as 6- nonyl-ascorbate; and 6-decanoyl-L-ascorbate, also known as and herein equivalently referred to as 6-decyl-ascorbate. These alkanoyl-L-ascorbic acid esters, functioning as active ingredients exhibiting antioxidation properties, characteristics, and behavior, in compositions of solutions or suspensions applied onto harvested plant matter, are

effectively utilized for significantly extending the shelf life of the harvested plant matter.

The method for synthesizing these alkanoyl-L-ascorbic acid esters is suitable and cost effective for commercial large-scale manufacturing applications.

The method for extending the shelf life of harvested plant matter, of the present invention, is based on the main step of applying onto the harvested plant matter an effective amount of a composition of a solution or suspension including an alkanoyl-L- ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate (6-octyl- ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6- decyl-ascorbate), as an antioxidation active ingredient, such that oxidation, and therefore, degradation, of the harvested plant matter are effectively inhibited during a period of time, thereby extending the shelf life of the harvested plant matter.

The composition for use in extending the shelf life of harvested plant matter, of the present invention, is based on a solution or suspension including an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate (6-octyl- ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6- decyl-ascorbate), as an antioxidation active ingredient.

In the solution or suspension of the composition, of the present invention, the selected alkanoyl-L-ascorbic acid ester (6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, or 6-decanoyl-L-ascorbate), preferably having a concentration in a range of between about 1 mM and about 75 mM, and more preferably, having a concentration in a range of between about 5 mM and about 50 mM, dissolved or suspended in water, an alcohol, and/or a glycol, is active and effective as a sole antioxidation active ingredient, such that oxidation, and therefore, degradation, of the harvested plant matter are effectively inhibited during a period of time, thereby extending the shelf life of the harvested plant matter.

Applying the solution or suspension of the composition onto the harvested plant matter is preferably performed using a dipping or spraying procedure performed at room temperature, during which the harvested plant matter is subjected or exposed to the applied composition for a time period of between thirty minutes and two hours, or, between three and ten seconds, respectively. Alternatively, the solution or suspension of the composition is brushed on the harvested plant matter using a suitable brush for such an application.

The method for synthesizing an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate

(6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6-decyl-ascorbate), of the present invention, is based on direct esterification of an equimolar mixture of the corresponding saturated fatty acid (octanoic acid, nonanoic acid, and decanoic acid, respectively) and L- ascorbic acid, in a large molar excess of concentrated sulfuric acid (at least about 96 %), wherein the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L- ascorbic acid is greater than 10/1, and preferably, about 25/1. Reaction schemes of the alkanoyl-L-ascorbic acid ester synthesis reactions are illustrated in FIG. 1.

It is to be understood that the present invention is not limited in its application to the details of the order or sequence, and number, of steps and sub-steps of implementation of the method for extending the shelf life of harvested plant matter using compositions of alkanoyl-L-ascorbic acid esters; or of implementation of the method for synthesizing the alkanoyl-L-ascorbic acid esters, set forth in the following description, drawings, or examples. Additionally, the present invention is not limited in its application to the components and ingredients, or solvents, included in the compositions of the alkanoyl-L- ascorbic acid esters for use in extending the shelf life of harvested plant matter, set forth in the following description, drawings, or examples. For example, the following description refers to application of the alkanoyl-L-ascorbic acid ester compositions in the form of either purely aqueous solutions or suspensions, or, aqueous solutions or suspensions including an additional component or ingredient such as gum arabic and/or potassium sorbate, onto the harvested plant matter, in order to illustrate implementation of the present invention. The present invention is capable of other embodiments or of being practiced or carried out in various ways. Although methods and materials similar or equivalent to those described herein can be used for practicing or testing the present invention, suitable methods and materials are described herein.

It is also to be understood that unless otherwise defined, all technical and scientific words, terms, and/or phrases, used herein have either the identical or similar meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Phraseology, terminology, and, notation, employed herein are for the purpose of description and should not be regarded as limiting. As used herein, the term'about'refers to 10 % of the associated value. Additionally, as used herein, indication of room temperature corresponds to a temperature of about 20 +/-2 °C.

For example, description of the present invention relates to the alkanoyl-L-ascorbic acid esters: 6-octanoyl-L-ascorbate (6-octyl-ascorbate); 6-nonanoyl-L-ascorbate (6-nonyl- ascorbate); and 6-decanoyl-L-ascorbate (6-decyl-ascorbate). These alkanoyl-L-ascorbic acid esters are also known as saturated fatty acid esters. These compounds are each well known by one of ordinary skill in the art to which this invention belongs, thereby precluding the need for providing and/or referring to additional synonyms, alternative names, abbreviations, and/or acronyms, of these chemical compounds.

As previously stated above, herein, the term'plant matter'generally refers to an entire or whole plant, to a portion of an entire or whole plant, to a component thereof or, to a combination thereof. The form of the plant matter is selected from the group consisting of a raw form, a partly processed form (that is, substantially solid and visually recognizable as originating from a plant), a loose form, a bundled form, and combinations thereof.

Primary examples of plant matter particularly suitable for implementation of the present invention are fruits, vegetables, and flowers, in a raw or partly processed, loose or bundled, form. Examples of plant matter as entire or whole plants, also suitable for implementation of the present invention, are trees, shrubs, bushes, grass, and moss, in a raw or partly processed, loose or bundled, form. Examples of plant matter as a portion of an entire or whole plant, or, a component thereof, also suitable for implementation of the present invention, are leaves, blossoms, beans, seeds, grains, stems, stalks, fibers, roots, and spices, in a raw or partly processed, loose or bundled, form.

As previously stated above, herein, 'harvested'plant matter generally refers to any of the above types of plant matter which has been harvested, that is, manually, mechanistically, or automatically, separated, detached, or removed, such as by pulling or cutting, from the point or location of cultivation, development, or growth, of the plant matter, such that the harvested plant matter is no longer cultivated, developed, or grown.

Typically, the harvested plant matter is gathered or collected, and subjected to a variety of numerous processes and sequences, involving activities such as packaging, storing, transporting, and treatment, in a variety of forms before eventually being sold in commercial wholesale and retail (shelf) environments.

Steps, sub-steps, components, ingredients, and implementation, of exemplary preferred embodiments, alternative preferred embodiments, specific configurations, and,

additional and optional aspects, characteristics, or features, thereof, of a method for extending the shelf life of harvested plant matter using compositions of alkanoyl-L- ascorbic acid esters; of the compositions of the alkanoyl-L-ascorbic acid esters; and of a method for synthesizing the alkanoyl-L-ascorbic acid esters, according to the present invention, are better understood with reference to the following description, accompanying drawings, and examples. Throughout the following description, accompanying drawings, and examples, same names used for identifying the various different alkanoyl-L-ascorbic acid ester compositions, controls, blank solvents, and/or'comparative or reference' compositions of solutions, refer to the same types of compositions, controls, blank solvents, and/or'comparative'compositions of solutions, respectively.

Immediately following, there is first provided description of the method for extending the shelf life of harvested plant matter using compositions of alkanoyl-L- ascorbic acid esters. Thereafter, is provided description of the alkanoyl-L-ascorbic acid ester compositions for use in extending the shelf life of harvested plant matter. Thereafter, is provided description of the method for synthesizing the alkanoyl-L-ascorbic acid esters.

Following thereafter, are provided illustrative descriptions of several examples relating to actual experimental support and implementation of the present invention.

In the following description of the present invention, included are main or principal steps and sub-steps needed for sufficiently understanding proper'enabling'utilization and implementation of the disclosed invention. Accordingly, description of various possible required and/or optional preliminary, intermediate, minor, and/or sub-steps, of secondary importance with respect to enabling implementation of the invention, which are readily known by one of ordinary skill in the art, and/or which are available in the prior art and technical literature relating to methods for extending the shelf life of harvested plant matter using ascorbic acid esters, and syntheses thereof, are at most only briefly indicated herein.

The preferred embodiment of the method for extending the shelf life of harvested plant matter, according to the present invention, includes the main step of applying onto the harvested plant matter an effective amount of a composition of a solution or suspension including an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6- octanoyl-L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6-decyl-ascorbate), as an antioxidation active ingredient, such that oxidation, and therefore, degradation, of the harvested plant matter are effectively

inhibited during a period of time, thereby extending the shelf life of the harvested plant matter.

The solution or suspension of the composition, of the present invention, preferably includes a single alkanoyl-L-ascorbic acid ester selected from the group consisting of 6- octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, which functions as the sole antioxidation active ingredient. However, it is clearly within the scope of the present invention for the solution or suspension of the composition to include a combination of at least two alkanoyl-L-ascorbic acid esters selected from the group consisting of 6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L- ascorbate, wherein each alkanoyl-L-ascorbic acid ester functions as an antioxidation active ingredient. Moreover, for the solution or suspension of the composition including a combination of at least two of the alkanoyl-L-ascorbic acid esters, wherein each alkanoyl- L-ascorbic acid ester functions as an antioxidation active ingredient, the alkanoyl-L- ascorbic acid esters can be active and effective as a synergistic combination of antioxidation active ingredients, such that oxidation of the harvested plant matter is even more effectively inhibited during a period of time, thereby significantly extending the shelf life of the harvested plant matter.

The alkanoyl-L-ascorbic acid ester solution or suspension compositions of the present invention are applied onto the harvested plant matter by using any of a variety of application or treatment techniques, procedures, along with corresponding equipment and conditions thereof, known and employed in the art. Well known and employed application or treatment techniques and procedures of dipping, rolling, brushing, wiping, rubbing, dripping, spraying, atomizing, and combinations thereof, along with corresponding equipment and conditions thereof, are suitable for applying onto the harvested plant matter an effective amount of the solution or suspension compositions. The specific application or treatment technique, procedure, along with corresponding equipment and conditions thereof, used for implementing the method of the present invention, depend upon the particular harvested plant matter, as indicated in the specific Examples illustratively described herein below. Preferably, application or treatment techniques and procedures of dipping, spraying, and brushing, along with corresponding equipment and conditions thereof, are used for implementing the present invention.

By using an application or treatment technique and procedure of dipping, the harvested plant matter, for example, fruit, vegetable, or flower, is dipped into one of the embodiments of the alkanoyl-L-ascorbic acid ester compositions, being a solution or suspension, at a pre-determined temperature, and for a pre-determined period of time.

During the dipping procedure, the harvested plant matter is at a temperature different from, or the same as, the temperature of the composition applied by dipping. Preferably, the pre- determined temperature refers to the same temperature of both the harvested plant matter and the composition applied by dipping. The pre-determined period of time refers to the time during which the harvested plant matter is substantially, entirely, dipped inside of, immersed within, and surrounded by, the composition applied by dipping.

The dipping procedure is performed at a pre-determined temperature in a range of between about 2 °C and about 70 °C. Preferably, the dipping procedure is performed at room temperature, that is, about 20 +/-2 °C, whereby the harvested plant matter and the applied composition are each at room temperature. The dipping procedure is performed for a pre-determined period of time in a range of between about one second and about eight hours, whereby the harvested plant matter is substantially, entirely, dipped or immersed in, and surrounded by, the applied composition, for the particular period of time.

Preferably, the dipping procedure is performed for a period of time in a range of between about ten minutes and about four hours, and more preferably, for a period of time in a range of between about thirty minutes and about two hours.

By using an application or treatment technique and procedure of spraying or brushing, the harvested plant matter, for example, fruit, vegetable, or flower, is sprayed or brushed with one of the embodiments of the alkanoyl-L-ascorbic acid ester composition, being a solution or suspension, at a pre-determined temperature, and for a pre-determined period of time. During the spraying or brushing procedure, the harvested plant matter is at a temperature different from, or the same as, the temperature of the composition applied by spraying. Preferably, the pre-determined temperature refers to the same temperature of both the harvested plant matter and the composition applied by spraying or brushing. The pre-determined period of time refers to the time during which the harvested plant matter is exposed or subjected to the composition applied by spraying or brushing.

The spraying or brushing procedure is performed at a pre-determined temperature in a range of between about 2 °C and about 70 °C. Preferably, the spraying or brushing

procedure is performed at room temperature, that is, about 20 +/-2 °C, whereby the harvested plant matter and the applied composition are each at room temperature. The spraying or brushing procedure is performed for a pre-determined period of time in a range of between about one second and about three minutes, whereby the harvested plant matter is exposed or subjected to the composition applied by spraying or brushing, for the particular period of time. Preferably, the spraying or brushing procedure is performed for a period of time in a range of between about one second and about thirty seconds, and more preferably, for a period of time in a range of between about three seconds and about ten seconds.

A wide variety of different procedures, such as drying, room temperature storage, low temperature or cold storage, handling, processing, and/or, eventual shelf display and sale, of the treated harvested plant matter, are used following the dipping or spraying treatment of the present invention. Specific procedures and conditions thereof depend upon the particular harvested plant matter.

For example, in some cases of implementing the present invention, following the dipping, spraying, or brushing, treatment, the treated harvested plant matter is first allowed to dry at room temperature (about 20 +/-2 °C) for a period of time of up to about two to three hours, then stored at room temperature (about 20 +/-2 °C) for a period of time, determined according to possible further use, processing, and/or, eventual shelf display and sale, of the treated harvested plant matter.

Alternatively, for example, in other cases of implementing the present invention, following the dipping, spraying, or brushing, treatment, the treated harvested plant matter is first allowed to dry at room temperature (about 20 +/-2 °C) for a period of time of up to about two to three hours, followed by low temperature or cold storage, for example, at 2 °C or 6 °C, in a cold room for a period of time, determined according to possible further use, processing, and/or, eventual shelf display and sale, of the treated harvested plant matter.

The preferred embodiment of the composition for use in extending the shelf life of harvested plant matter, of the present invention, is a solution or suspension including an alkanoyl-L-ascorbic acid ester selected, from the group consisting of 6-octanoyl-L- ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6- decanoyl-L-ascorbate (6-decyl-ascorbate), as an antioxidation active ingredient.

The solvent of the alkanoyl-L-ascorbic acid ester composition of the present invention, is a solvent which sufficiently dissolves or disperses the alkanoyl-L-ascorbic acid ester to the extent that a solution or suspension, respectively, is formed. Preferably, the solvent is a low molecular weight polar solvent selected from the group consisting of water, a low molecular weight alcohol, a low molecular weight polar glycol, and miscible combinations thereof. An exemplary preferred low molecular weight alcohol is selected from the group consisting of ethanol, propanol, and miscible combinations thereof. An exemplary preferred low molecular weight glycol is selected from the group consisting of propylene glycol, ethylene glycol, and miscible combinations thereof. More preferably, the solvent is water.

The molar concentration of the alkanoyl-L-ascorbic acid ester in the solution or suspension of the composition of the of the present invention, is in a range of between about 1 mM and about 75 mM, and more preferably, in a range of between about 5 mM and about 50 mM.

The alkanoyl-L-ascorbic acid ester composition of the present invention, is prepared, at room temperature, according to the main steps of : (a) weighing a pre- determined quantity of the alkanoyl-L-ascorbic acid ester, in dry powdered form; (b) placing the weighed alkanoyl-L-ascorbic acid ester into an appropriate beaker or container; (c) adding an appropriate quantity of the previously described solvent, preferably, water, to the beaker or container; and (d) mixing or dispersing the alkanoyl-L-ascorbic acid ester throughout the solvent until a solution or suspension is formed. The solution or suspension composition is then stored at room temperature, and is ready for applying, preferably, by a dipping, spraying, or brushing, procedure, onto the harvested plant matter.

In alternative, specific embodiments of the preferred embodiment of the alkanoyl- L-ascorbic acid ester composition, of the present invention, the composition, being a solution or suspension including the alkanoyl-L-ascorbic acid ester as the antioxidation active ingredient, further includes at least one additional component or ingredient, for improving the physicochemical properties, characteristics, behavior, and activity, of the composition, and therefore, for further improving the effectiveness of the composition for use in extending the shelf life of the harvested plant matter. Preferably, in a non-limiting manner, at least one additional component or ingredient selected from the group consisting of a dispersing agent, an emulsifier, an acidulant, a plasticizer, a complexing agent, a

preservative, a firming and/or sequestering agent, a protein, an antioxidation agent, a coating, and combinations thereof.

Exemplary dispersing agents are selected from the group consisting of carboxymethylcellulose (CMC), hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), methyl cellulose (MC), guar gum, locust bean gum, pectin, xanthan gum, modified starch, carrageenan, gum arabic, and combinations thereof.

Exemplary emulsifiers are selected from the group consisting of monoglycerides, diglycerides, glycol esters of fatty acids, polyglycol esters of fatty acids, polyoxyethylene sorbitan oleates, polyoxyethylene sorbitan stearates, polyoxyethylene sorbitan laurates, lecithins, and combinations thereof.

Exemplary acidulants are selected from the group consisting of citric acid, propionic acid, lactic acid, gluconic acid, succinic acid, tartaric acid, fumaric acid, ascorbic acid, and combinations thereof.

Exemplary plasticizers are selected from the group consisting of polyethylene glycol, glycerol, propylene glycol, carnauba wax, candellila wax, stearic acid, oleic acid, sorbitol, soybean oil, beeswax, mannitol, and combinations thereof.

Exemplary complexing agents are selected from the group consisting of citric acid, cyclodextrins, acidic polyphosphates, and combinations thereof.

Exemplary preservatives are selected from the group consisting of calcium chloride, sodium propionate, calcium propionate, benzoic acid, sodium benzoate, potassium sorbate, sodium bisulfite, and combinations thereof.

Exemplary firming and/or sequestering agents are selected from the group consisting of calcium chloride, calcium gluconate, calcium lactate, citric acid and a salt thereof, ethylenediamine tetraacetic acid and a salt thereof, and combinations thereof.

Exemplary proteins are selected from the group consisting of soy protein, whey, casein, gelatin, zein, and combinations thereof.

Exemplary antioxidation agents are selected from the group consisting of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tertiary-butylated hydroquinone (TBHQ), propyl gallate, ascorbic acid, ascorbyl-6-palmitate, tocopherols, spice extracts which have antioxidant properties, and combinations thereof.

Exemplary coatings are selected from the group consisting of gum arabic, chitosan, and combinations thereof. Chitosan, a polysaccharide polymer obtained from skeletal matter of an invertebrate, is originally derived, for example, from crabs.

In the above described alternative, specific embodiments of the preferred embodiment of the alkanoyl-L-ascorbic acid ester composition, of the present invention, each of the at least one additional component or ingredient is of a concentration, preferably less than about 10 %, wt/vol, in the solution or suspension which does not interfere with, inhibit, and/or decrease, the desirable activity and effectiveness of the alkanoyl-L-ascorbic acid ester in the solution or suspension, for use in extending the shelf life of the harvested plant matter.

As described above, the alkanoyl-L-ascorbic acid ester compositions of the present invention, are each in the form of a solution or a suspension, immediately ready for treating and applying to the harvested plant matter. In alternative embodiments, the alkanoyl-L-ascorbic acid ester compositions are in a variety of different forms. In particular, in a form selected from the group consisting of a dry or essentially dry form, a partly solution or partly suspension form and a partly dry or partly essentially dry form, and a concentrated solution or concentrated suspension form.

In a dry or essentially dry form, the main component, alkanoyl-L-ascorbic acid ester, of the composition of the present invention, is in a loose powdered form, a pelleted powder form, or a tableted powder form. In these embodiments, the composition is prepared by dissolving or dispersing the alkanoyl-L-ascorbic acid ester in the appropriate solvent, for forming the desired solution or suspension, respectively, which is to be applied onto the harvested plant matter, as previously described hereinabove.

In a partly solution or partly suspension form, and, in a partly dry or partly essentially dry form, each alkanoyl-L-ascorbic acid ester composition is prepared by mixing the dry or essentially dry component or components with the partly dissolved or partly suspended component or components, along with adding the appropriate solvent, for forming the desired solution or suspension which is to be applied onto the harvested plant matter, as previously described hereinabove.

In a concentrated solution or concentrated suspension form, each alkanoyl-L- ascorbic acid ester composition is prepared by adding the appropriate solvent to the concentrated solution or concentrated suspension form, for forming the desired solution or

suspension which is to be applied onto the harvested plant matter, as previously described above.

The preferred embodiment of the method for synthesizing an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L-ascorbate (6-octyl- ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6- decyl-ascorbate), according to the present invention, includes the main steps of : (a) preparing an equimolar mixture of a saturated fatty acid selected from the group consisting of octanoic acid, nonanoic acid, and decanoic acid, respectively, and, L-ascorbic acid, such that the molar ratio of the saturated fatty acid to the L-ascorbic acid equals 1, (b) adding an excess quantity of sulfuric acid having a concentration of at least about 96 % to the equimolar mixture, such that the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid is greater than 10/1, for forming a reaction mixture, (c) stirring the reaction mixture, such that reaction takes place for forming the corresponding alkanoyl-L-ascorbic acid ester, and (d) separating the formed alkanoyl-L-ascorbic acid ester from the reaction mixture, for obtaining the alkanoyl-L-ascorbic acid ester.

Preferably, the synthesis method further includes main step (e), purifying the formed alkanoyl-L-ascorbic acid ester, for obtaining a pure form of the alkanoyl-L- ascorbic acid ester, for example, at least about 98 % pure. Preferably, the synthesis method further includes steps for performing appropriate analytical techniques, for the purposes of identifying and confirming the chemical structure, and for determining the purity, of the purified alkanoyl-L-ascorbic acid ester.

Reaction schemes of the alkanoyl-L-ascorbic acid ester synthesis reactions are illustrated in FIG. 1. It is noted that the method for synthesizing the alkanoyl-L-ascorbic acid ester, according to the present invention, is based on direct esterification of an equimolar mixture of the corresponding saturated fatty acid (octanoic acid, nonanoic acid, and decanoic acid, respectively) and L-ascorbic acid, in large molar excess of concentrated sulfuric acid (at least about 96 %), wherein the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid is greater than 10/1, and preferably, about 25/1.

In Step (a), of the preferred embodiment of the method for synthesizing an alkanoyl-L-ascorbic acid ester selected from the group consisting of 6-octanoyl-L- ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, according to the present

invention, there is preparing an equimolar mixture of a saturated fatty acid selected from the group consisting of octanoic acid, nonanoic acid, and decanoic acid, respectively, and, L-ascorbic acid, such that the molar ratio of the saturated fatty acid to the L-ascorbic acid equals 1.

In Step (a), according to the present invention, clearly, for synthesizing each individual alkanoyl-L-ascorbic acid ester, 6-octanoyl-L-ascorbate, 6-nonanoyl-L- ascorbate, or 6-decanoyl-L-ascorbate, there is using only the corresponding saturated fatty acid, octanoic acid, nonanoic acid, or decanoic acid, respectively, and mixing a molar quantity of the corresponding saturated fatty acid with the same molar quantity of L- ascorbic acid.

In Step (b), there is adding an excess quantity of sulfuric acid having a concentration of at least about 96 % to the equimolar mixture, such that the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid is greater than 10/1, for forming a reaction mixture.

In Step (b), the concentration of the sulfuric acid is at least 96 %, and in general, in a range of between at least about 96 % and about 98 %. By adding the excess quantity of sulfuric acid to the equimolar mixture of the saturated fatty acid and the corresponding saturated fatty acid, the sulfuric acid functions as a main reactant and as the solvent of the reaction mixture. This procedure is in strong contrast to, most organic reactions which include the use of concentrated sulfuric acid, wherein only relatively very small quantities (typically, on the order of a few drops, amounting to less than 1 mL, for an approximately 1 liter reaction volume) of concentrated sulfuric are added to the reactant mixture, and wherein the sulfuric acid simply functions as a catalyst for catalyzing the chemical reaction, and not as a main reactant in the form of the solvent participating in the reaction for forming the desired product. In the reaction mixture of the present invention, the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid is greater than 10/1, and preferably, about 25/1.

In Step (c), there is stirring the reaction mixture, such that reaction takes place for forming the corresponding alkanoyl-L-ascorbic acid ester, 6-octanoyl-L-ascorbate, 6- nonanoyl-L-ascorbate, or 6-decanoyl-L-ascorbate.

In Step (c), the reaction mixture is stirred for a period of time in a range of between 15 and 30 hours, preferably, in a range of between 18 and 24 hours, and more preferably,

for 24 hours. The reaction mixture is stirred for the indicated time period, preferably, at room temperature, corresponding to a temperature of about 20 +/-2 °C. Stirring of the reaction mixture is intentionally performed at room temperature in order to avoid potential occurrence of undesired reactions, especially, of polymerization and/or oxidation of the L- ascorbic acid in the presence of the concentrated sulfuric acid solvent, which more readily take place at reaction temperatures higher than room temperature. During the time period of stirring, the desired alkanoyl-L-ascorbic acid ester, 6-octanoyl-L-ascorbate, 6-nonanoyl- L-ascorbate, or 6-decanoyl-L-ascorbate, is formed.

In Step (d), there is separating the formed alkanoyl-L-ascorbic acid ester from the reaction mixture, for obtaining the alkanoyl-L-ascorbic acid ester.

For separating the formed alkanoyl-L-ascorbic acid ester from the reaction mixture, the reaction mixture obtained at completion of Step (c) is subjected to the following sub- steps (i) through (vi): In sub-step (i), of Step (d), the reaction mixture of Step (c) is carefully poured into a quantity of ice water which is sufficient for diluting the concentrated sulfuric acid, for obtaining a diluted reaction mixture including the formed alkanoyl-L-ascorbic acid ester.

Ice water, instead of water at room temperature, is used for this purpose in order to absorb the exothermic heat released due to mixing of the water with the sulfuric acid. For example, for performing the synthesis method using a reaction mixture volume of about 500 mL, for obtaining an amount of pure alkanoyl-L-ascorbic acid ester product in the range of between about 10 and 100 grams, at a yield of about 60-70 %, typically, about 2 - 5 kg, and preferably, about 3 kg, of ice water is used for diluting the reaction mixture.

In sub-step (ii), of Step (d), the diluted reaction mixture of sub-step (i) is stirred at room temperature for a period of time.

For example, for performing the synthesis method using a reaction mixture volume of about 500 mL, for obtaining an amount of pure alkanoyl-L-ascorbic acid ester product in the range of between about 10 and 100 grams, the diluted reaction mixture is stirred at room temperature for a period of time in a range of between about 5 and 10 minutes, and preferably, for about 5 minutes.

In sub-step (iii), of Step (d), there is extracting the formed alkanoyl-L-ascorbic acid ester from the diluted reaction mixture of sub-step (iii), for obtaining an organic phase including the formed alkanoyl-L-ascorbic acid ester.

Preferably, the formed alkanoyl-L-ascorbic acid ester is extracted at least two times from the diluted reaction mixture, in order to sufficiently separate the formed alkanoyl-L- ascorbic acid ester from the water and the sulfuric acid of the diluted reaction mixture.

Extraction of the formed alkanoyl-L-ascorbic acid ester from the diluted reaction mixture is performed using an appropriate organic extraction solvent, such as ethyl acetate. For example, for performing the synthesis method using a reaction mixture volume of about 500 mL, for obtaining an amount of pure alkanoyl-L-ascorbic acid ester product in the range of between about 10 and 100 grams, at a yield of about 60-70 %, the formed alkanoyl-L-ascorbic acid ester is twice extracted from the diluted reaction mixture using 1.5 L of ethyl acetate.

In sub-step (iv), of Step (d), the organic phase including the formed alkanoyl-L- ascorbic acid ester, of sub-step (iii), is washed with water, for obtaining a washed organic phase including the formed alkanoyl-L-ascorbic acid ester.

Preferably, the organic phase is washed two times with water, in order to sufficiently remove the remaining sulfuric acid from the ethyl acetate organic phase. For example, for performing the synthesis method using a reaction mixture volume of about 500 mL, for obtaining an amount of pure alkanoyl-L-ascorbic acid ester product in the range of between about 10 and 100 grams, at a yield of about 60-70 %, the organic phase is washed two times with a volume of water in a range of between about 500 mL and about 1L, and preferably, with about 1 L of water.

In sub-step (v), of Step (d), the washed organic phase including the formed alkanoyl-L-ascorbic acid ester, of sub-step (iv), is dried with a drying agent, for example, anhydrous magnesium sulfate, for removing the water used for washing, for obtaining a dried organic phase including the formed alkanoyl-L-ascorbic acid ester.

In sub-step (vi), of Step (d), the organic extraction solvent, for example, ethyl acetate, is evaporated from the dried organic phase which includes the formed alkanoyl-L- ascorbic acid ester, of sub-step (v), for example, using a rotary evaporator, for obtaining a dry residue including the formed alkanoyl-L-ascorbic acid ester.

As stated hereinabove, preferably, the synthesis method of the present invention further includes the following purification step, Step (e).

In Step (e), there is purifying the formed alkanoyl-L-ascorbic acid ester, for obtaining a pure form of the alkanoyl-L-ascorbic acid ester.

For purifying the formed alkanoyl-L-ascorbic acid ester, for example, to an extent of at least about 98 % pure, the dry residue including the alkanoyl-L-ascorbic acid ester obtained at completion of Step (d) is subjected to a purification procedure.

For example, for performing the synthesis method using a reaction mixture volume of about 500 mL, for obtaining an amount of pure alkanoyl-L-ascorbic acid ester product in the range of between about 10 and 100 grams, at a yield of about 60-70 %, the dry residue including the alkanoyl-L-ascorbic acid ester is subjected to a re-crystallization procedure, followed by vacuum drying, for obtaining dry and pure alkanoyl-L-ascorbic acid ester product in the form of white crystals. In this case, an exemplary re- crystallization procedure includes dissolving the dry residue in about 140 mL of ether, and precipitating the pure alkanoyl-L-ascorbic acid ester product using about 440 mL of n- hexane, followed by vacuum drying for at least several hours, for obtaining the dry and pure alkanoyl-L-ascorbic acid ester product in the form of white crystals, having a purity of at least about 98 %.

As stated hereinabove, preferably, the synthesis method of the present invention further includes steps for performing appropriate analytical techniques, such as NMR spectroscopy and HPLC, for the purposes of identifying and confirming the chemical structure, and for determining the purity, respectively, of the purified alkanoyl-L-ascorbic acid ester, being 6-octanoyl-L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6- nonyl-ascorbate), or 6-decanoyl-L-ascorbate (6-decyl-ascorbate).

An exemplary NMR spectroscopy technique suitable for identifying and confirming chemical structure of each of the purified alkanoyl-L-ascorbic acid esters is based on using HI-NMR (200 MHz, DMSO-d6), with pure standards, along with reference to the NMR spectra obtained by Youchun et al. , 1999, for each of these alkanoyl-L- ascorbic acid esters.

An exemplary HPLC technique suitable for determining the purity of each of the purified alkanoyl-L-ascorbic acid esters is based on using an HPLC system, for example, a Hewlett-Packard HP-1090 instrument with an auto sampler. Such an HPLC system can be used with a 20 ml sample loop, UV detector set at 245 nm, and an Lichrosorb NH2 Column (25 cm x 4.6 mm) of Merck, Darmstadt, Germany. Separation of each alkanoyl-L-ascorbic acid ester is achievable by isocratic elution with an eluent solvent mixture of acetonitrile and a buffer of 0.05 M KH2PO4 (65: 35 v/v).

EXAMPLES Above described novel and inventive aspects and characteristics, and advantages thereof, of the present invention further become apparent to one ordinarily skilled in the art upon examination of the following examples, which are not intended to be limiting.

Additionally, each of the various embodiments and aspects of the present invention as delineated herein above and as claimed in the claims section below finds experimental support in the following examples. In the following description of the Examples, it is noted that room temperature refers to a temperature of 20 +/-2 °C, herein, also referred to as room temperature of about 20 °C. Where relevant, any other temperature is clearly indicated.

SYNTHESIZING THE ALKANOYL-L-ASCORBIC ACID ESTERS' Examples of synthesizing each of the alkanoyl-L-ascorbic acid esters: 6-octanoyl- L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6- decanoyl-L-ascorbate (6-decyl-ascorbate), according to the above described method of the present invention, are provided hereinbelow. Reaction schemes of the alkanoyl-L-ascorbic acid ester synthesis reactions are illustrated in FIG. 1. yntheses Of Tlze 41kanovl-L-Ascorbic Acid Esters L-ascorbic acid was obtained from Roche Products Ltd. , UK. Octanoic acid, decanoic acid, sulfuric acid (98 %), ethyl acetate, acetonitrile, n-hexane, magnesium sulfate, potassium phosphate (KH2PO4), were each obtained from Aldrich Chemical, Israel.

Nonanoic acid was obtained from Merck-Schuchardt, Henbrtmn, Germany. Triple- distilled, de-ionized water was used for the syntheses and for the analytical procedures.

Analvtical Procedures Used For Identifving And Confirming Chemical Structure, And For Determinislv Coznpozcnd Pllritv. Of The Svnthesized Alkanovl-L-Ascorbic Acid Esters An NMR spectroscopy system, Hl-NMR (200 MHz, DMSO-d6), with pure standards, along with reference to the NMR spectra obtained by Youchun et al. , 1999, for each of these alkanoyl-L-ascorbic acid esters, was used for identifying and confirming chemical structure of each of the synthesized alkanoyl-L-ascorbic acid esters. An HPLC system, Hewlett-Packard HP-1090 instrument with an auto sampler was used for determining compound purity of each of the alkanoyl-L-ascorbic acid esters. The HPLC

system was used with a 20 ml sample loop, W detector set at 245 nm, and an Lichrosorb NH2 Column (25 cm x 4.6 mm) of Merck, Darmstadt, Germany. Separation of each alkanoyl-L-ascorbic acid ester was achieved by isocratic elution with an eluent solvent mixture of acetonitrile and a buffer of 0.05 M KH2PO4 (65: 35 v/v).

EXAMPLE 1: SYNTHESIS OF 6-octanoyl-L-ascorbate (6-octyl-ascorbate) To a 1 liter flask were added 31.73 grams (0.22 mole) of octanoic acid and 38.75 grams (0.22 mole) of L-ascorbic acid, thereby forming an equimolar (1/1) mixture of the octanoic acid and the L-ascorbic. An excess quantity, 500 mL, of sulfuric acid having a concentration of 98 % was added to the equimolar mixture, such that the molar ratio of the sulfuric acid to the sum of the saturated fatty acid and the L-ascorbic acid was about 25/1, for forming the reaction mixture. The sulfuric acid functioned as a main reactant and as the solvent of the reaction mixture.

The reaction mixture was stirred for 24 hours at room temperature (about 20 +/-2 °C) only, thereby avoiding the potential occurrence of undesired reactions, especially, of polymerization and/or oxidation of the L-ascorbic acid in the presence of the concentrated sulfuric acid solvent, which more readily take place at reaction temperatures higher than room temperature. During the time period of stirring, the 6-octanoyl-L-ascorbate was formed.

The formed 6-octanoyl-L-ascorbate was separated from the reaction mixture, by the following steps, for obtaining the 6-octanoyl-L-ascorbate. The reaction mixture was carefully poured into about 3 kg of ice water which was sufficient for diluting the concentrated sulfuric acid, for obtaining a diluted reaction mixture including the formed 6- octanoyl-L-ascorbate. The diluted reaction mixture was stirred at room temperature for about 5 min. The formed 6-octanoyl-L-ascorbate was twice extracted from the diluted reaction mixture using 1.5 L of ethyl acetate, for obtaining an organic phase including the formed 6-octanoyl-L-ascorbate. The organic phase was washed two times with about 1 L of water, for obtaining a washed organic phase including the formed 6-octanoyl-L- ascorbate. The washed organic phase including the formed 6-octanoyl-L-ascorbate was dried with anhydrous magnesium sulfate, for removing the water used for washing, for obtaining a dried organic phase including the formed 6-octanoyl-L-ascorbate. The ethyl acetate organic extraction solvent was evaporated from the dried organic phase which

included the formed 6-octanoyl-L-ascorbate, using a rotary evaporator, for obtaining a dry residue including the formed 6-octanoyl-L-ascorbate.

The dry residue including the 6-octanoyl-L-ascorbate was subjected to the following re-crystallization purification procedure. The dry residue was dissolved in about 140 mL of ether. The pure 6-octanoyl-L-ascorbate product was precipitated using about 440 mL of n-hexane, followed by vacuum drying for about 10 hours, for obtaining 48 grams, at a yield of 72 %, of the dry and pure 6-octanoyl-L-ascorbate in the form of white crystals.

The above described NMR spectroscopy analytical procedure was used for identifying and confirming the chemical structure, and the above described HPLC analytical procedure was used for determining compound purity of greater than 99 %, of the synthesized 6-octanoyl-L-ascorbate crystals.

H1-NMR (200MHz, DMSO-d6), 5, ppm: 0.82 ( t, 3H, J=6.7Hz), 1.24 ( m, 8H), 1.53 ( m, 2H), 2.31 ( t-2H, J= 7.3Hz), 3.99-4. 06 (m, 6H), 5.31 (m, 1H), 5.74 ( s, 1H, OH), 8.40 (br. s, 1H, OH), 11. 11 (br. s, 1H, OH, acidic). The NMR spectra matched those obtained by Youchun et al. , 1999.

EXAMPLE 2 : SYNTHESIS OF 6-nonaizoyl-L-ascorbate (6-nonyl-ascorbate) Using the same synthesis procedure as described for 6-octanoyl-L-ascorbate in Example 1,13. 9 grams (0.044 mole) of nonanoic acid, 7.7 grams (0.044 mole) of L- ascorbic acid, and 100 mL of concentrated sulfuric acid (98 %), were used for forming the reaction mixture.

There was obtained 9.48 grams, at a yield of 60 %, of the dry and pure 6-nonanoyl- L-ascorbate in the form of white crystals, with a purity of greater than 98 %, as determined by using the same analytical procedure as used in Example 1.

H1-NMR (200MHz, DMSO-d6), 5, ppm: 0.83 (3, d, J=5.64Hz), 1.22-1. 5 (m, 12 aliphatic), 2.30 ( t, 2, J=7.2Hz), 3.45 ( 1, d, J=6.3Hz), 4.05 (br s, 2), 4.65 ( 1, br s), 8.40 (br s, OH), 11.11 (S, OH). The NMR spectra matched those obtained by Youchun et al., 1999.

EXAMPLE 3: SYNTHESISOF6-deca7loyl-L-asco7bate (6-decyl-ascorbate) Using the same synthesis procedure as described for 6-octanoyl-L-ascorbate in Example 1,37. 9 grams (0.22 mole) of decanoic acid, 38.75 grams (0. 22 mole) of L-

ascorbic acid, and 500 mL of concentrated sulfuric acid (98 %), were used for forming the reaction mixture.

There was obtained 44.2 grams, at a yield of 60.9 %, of the dry and pure 6- decanoyl-L-ascorbate in the form of white crystals, with a purity of greater than 99 %, as determined by using the same analytical procedure as used in Example 1.

Hl-NMR (200MHz, DMSO-d6), 8, ppm: 8. 37 (3, d, J=5.64Hz), 1. 22-1. 5 (m, 14 aliphatic), 2.30 ( t, 2, J=7.2Hz), 3.45 ( 1, d, J=6. 3Hz), 4.05 (br s, 2), 4.65 ( 1, br s), 8.40 (br s, OH), 11.11 (S, OH). The NMR spectra matched those obtained by Youchun et al. , 1999. <BR> <BR> <BR> <BR> <BR> <BR> <P>TESTING FUNGICIDAL ACTIVITY OF THE ALKANOYL-L-ASCORBIC ACID ESTERS Procedure and results of testing the fungicidal activity for each of the alkanoyl-L- ascorbic acid esters: 6-octanoyl-L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6-decyl-ascorbate), are provided hereinbelow.

6-octanoyl-L-ascorbate, 6-nonanoyl-L-ascorbate, and 6-decanoyl-L-ascorbate, separately, were each dissolved in water, and the fungicidal activity as relating to growth inhibition (%) of Penicillium Digitatum, Penicillium Exponsum, and Botrytis, fungi, in an in-vitro system, including potato dextrose broth (PDB) as the growth medium, was measured for each ester. The results are presented in Tables 1,2, and 3, respectively. The results show that each alkanoyl-L-ascorbic acid ester exhibits a significant fungicidal activity, even at a relatively low concentration. The fungicidal results were also used, in part, for determining optimum concentrations of the alkanoyl-L-ascorbic acid esters as the antioxidation active ingredient in the alkanoyl-L-ascorbic acid ester compositions used for testing the harvested plant matter.

Table 1 : Fungicidal activity of different concentrations, 0-50 mM, of 6-octanoyl-L- ascorbate in water, as relating to growth inhibition of Penicillium Digitatum, Penicillium Exponsum, and Botrytis, fungi. P. dig. P. exp. Botrytis Concentration of 6- Growth Growth Growth octanoyl-L-ascorbate inhibition (%) inhibition (%) inhibition (%) 0 0 0 0 10 mM 20.3 10.1 63.4 20 mM 41. 1 73 72. 5 30 mM 53. 1 78. 6 100 40 mM 55. 2 86. 7 100 50 mM 100 92. 8 100

Table 2: Fungicidal activity of different concentrations, 0-50 mM, of 6-nonanoyl-L- ascorbate in water, as relating to growth inhibition of Penicillium Digitatum, Penicillium Exponsum, and Botrytis, fungi. P. dig. P. exp. Botrytis Concentration of 6- Growth Growth Growth nonanoyl-L-ascorbate inhibition (%) inhibition (%) inhibition (%) 0 0 0 0 10 mM 100 100 100 20 mM 100 100 100 30 mM 100 100 100 40 mM 100 100 100 50 mM 100 100 100

Table 3: Fungicidal activity of different concentrations, 0-50 mM, of 6-decanoyl-L- ascorbate in water, as relating to growth inhibition of Penicillium Digitatum, Penicillium Exponsum, and Botrytis, fungi. P. dig. P. exp. Botrytis Concentration of 6- Growth Growth Growth decanoyl-L-ascorbate inhibition (%) inhibition (%) inhibition (%) 0 0 0 0 10 mM 47. 8 51. 8 18 20 mM 62. 4 66 46. 3 30 mM 92. 9 100 64. 7 40 mM 95. 6 100 100 50 mM 98. 4 100 100

TESTING HARVENTED PLANTMi1TTER WITH COMPOSITIONS OF THE SLKANOYL- L-ASCORBIC ACID ESTERS Chemicals used for the testing The alkanoyl-L-ascorbic acid ester compositions used in the experiments of the following Examples of testing harvested plant matter were prepared as described hereinabove, by using the alkanoyl-L-ascorbic acid esters synthesized as described hereinabove and exemplified in Examples 1,2, and 3, for 6-octanoyl-L-ascorbate (6-octyl- ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl-ascorbate), and 6-decanoyl-L-ascorbate (6- decyl-ascorbate), respectively, in accordance with the present invention.

Potassium sorbate (K-Sorbate), gum arabic, and calcium chloride (CaCl,), were included individually or in combination, as additive ingredients in selected alkanoyl-L- ascorbic acid ester compositions used for testing the harvested plant matter. Potassium sorbate and calcium chloride were obtained from Sigma or Aldrich Chemicals, Israel or USA. Gum arabic was obtained from Merck, Germany. Local tap or faucet water was used for preparing the alkanoyl-L-ascorbic acid ester compositions, and for preparing the 'comparative'compositions of reference solutions or of water alone.

Imazalil, a systemic imidazole fungicide used to control a wide range of fungi on fruit, vegetables and ornamentals, including powdery mildew on cucumber and black spot

on roses, and also used as a seed dressing and for postharvest treatment of citrus, banana, and other fruit to control storage decay, was used in an aqueous solution standard 'comparative'composition in the experiments of Example 9-Bananas. Trade names of products containing imazalil include Bromazil, Deccozil, Fungaflor, Freshgard, and Fungazil. Imazalil has a CAS # of 35554-44-0, and a formal chemical name of 1- [2- (2, 4- dichlorophenyl)-2- (2-propenyloxy) ethyl]-IH-imidazole. A known manufacturer of Imazalil is Janssen Pharmaceutica Plant Protection Division, Piscataway, NJ, USA.

Harvested plant matter tested A variety of different types of exemplary harvested plant matter was used for demonstrating effectiveness of the alkanoyl-L-ascorbic acid ester compositions of the present invention, for use in extending the shelf life of the harvested plant matter. This included fruits (melons, apples, grapes, and bananas), and a vegetable (lettuce).

Treatment and application compositions, procedures, and conditions Different concentrations of the alkanoyl-L-ascorbic acid ester compositions of the present invention, were applied, at room temperature (about 20 °C), by dipping, by spraying onto, or by brushing onto, for different periods of time, the above indicated different types of harvested plant matter.

As a basis of comparison for determining the effectiveness of the compositions of the present invention, additionally, correspondingly same concentrations of'comparative' compositions of reference solutions, or of water alone, were similarly applied, at room temperature (about 20 °C), by dipping, by spraying onto, or by brushing onto, for the correspondingly same periods of time, the above indicated different types of harvested plant matter.

In each of the following Examples, harvested plant matter controls were used for comparative purposes. In most of the Examples, the controls were simply the indicated harvested plant matter, untreated, without being subjected to a dipping, spraying, or brushing, treatment of an alkanoyl-L-ascorbic acid ester composition, along with shelf life storage at room temperature, or, the controls were subjected to shelf life storage at 6 °C or 2 °C.

Specific treatment and application compositions, procedures, and conditions, depended upon the particular harvested plant matter being tested, and are clearly indicated in each specific Example below.

Storage procedures and conditions In some of the following Examples of harvested plant matter, following the dipping, spraying, or brushing, treatment, the treated harvested plant matter was first allowed to dry at room temperature (about 20 °C) for a period of time of up to about two to three hours, then stored at room temperature (about 20 °C) for different periods of time, on the order of days or weeks, along with determining the shelf lives of the treated and untreated (control) harvested plant matter, by measuring, evaluating, and analyzing several parameters directly relating to properties, characteristics, and behavior, of the tested harvested plant matter, as described immediately below.

Alternatively, in other Examples of harvested plant matter, following the dipping, spraying, or brushing, treatment, the treated harvested plant matter was first allowed to dry at room temperature (about 20 °C) for a period of time of up to about two to three hours, directly followed by'cold'storage at either 2 °C or 6 °C in a cold room for a period of time, followed by'shelf life'storage at room temperature (about 20 °C) for different periods of time, on the order of days or weeks, along with determining the shelf lives of the treated and untreated (control) harvested plant matter, by measuring, evaluating, and analyzing, several parameters directly relating to properties, characteristics, and behavior, of the tested harvested plant matter, as described immediately below.

Specific periods of time of drying, cold storing, and shelf life storing, the treated and untreated (control) harvested plant matter, depended upon the particular harvested plant matter being tested, and are clearly indicated in each specific Example below.

Detere ationofShelffife-parameters procedllres andmeasllreme7lts Several well known and reliably used parameters directly relating to properties, characteristics, and behavior, of plant matter, in general, and of harvested plant matter, in particular, were measured and evaluated, using various different chemical, visual, organoleptic, and/or microbiological, measurements, tests, and methods of evaluation, according to the following described procedures, and analyzed for accurately and reproducibly determining the shelf life of the tested harvested plant matter. These results were then used for describing, characterizing, and confirming, the high effectiveness of the alkanoyl-L-ascorbic acid ester compositions of the present invention for use in extending the shelf life of harvested plant matter. These parameters, each of which is described immediately below, are (a) total sugars, (b) weight loss, (c) decay, and (d) color change (s).

Total sugars Total sugars (%) of the tested harvested plant matter was measured and evaluated using 100 ul of filtered juice in a refractometer-REF 103 instrument, and a Brix 0-32 % scale. For this measurement, fresh juice was prepared from 5 grams of each of 8 to 10 fresh fruits, and then filtered.

Weight loss Weight loss (%) of the tested harvested plant matter was measured and evaluated by recording the differences between the weight of the tested harvested plant matter prior to a given treatment, and the weights of the tested harvested plant matter at selected time intervals during the period of shelf life storage at room temperature.

Decay In some of the following Examples, decay (%) of the tested harvested plant matter was measured and evaluated by visually observing effects, in particular, fungal growth, cracks, change in surface color to brown and/or black, of decay, decomposition, or rotting, of the tested plant matter. The results were recorded using a scale of 0 % (no decay) to 100 % (severe or maximum decay). Alternatively, in several Examples, photographs were taken, using a camera, for measuring and evaluating decay of the harvested plant matter.

Alternatively, in the Example of (cut) Apples, decay was measured and evaluated in terms of total microbial count, total molds count, and total yeast count, expressed in units of colony forming units per gram formula weight, or CFU/gr-FW.

Color change (s) Color change (s) (%) of the tested harvested plant matter was measured and evaluated by visually observing any change (s) in surface color of the tested plant matter.

For example, visually observing development of brown and/or black spots or browning in the surface, skin, or peel, color of the tested plant matter. The results were recorded using a scale of 0 % (no surface color change) to 100 % (significant surface color change (s)).

Additionally, in Example 10-iceberg lettuce heads, a browning level index or scale of 1- 5 was used for comparing the extent of browning. A value of 1 indicates an undesirable high extent of browning, while a value of 5 indicates a desirable low extent of browning, of the iceberg lettuce heads. In several Examples, photographs were taken, using a camera, for measuring and evaluating color change (s) of the harvested plant matter.

Specific measurement and evaluation parameters, procedures, and measurements, used for determining the shelf life depended upon the particular harvested plant matter being tested, and are clearly indicated in each specific Example below.

In the tables of the experimental results accompanying the following description of the Examples of the present invention, the term refers to the'standard error'associated with the average or mean values of the previously described measured parameters.

Average or mean values and standard error (SE) values thereof were calculated as follows.

For each Example of the harvested plant matter tested, a set of at least two, typically, three or more, separate experiments, was performed, with each experiment involving a sizeable plurality, of the same type of individual samples or units (for example, a plurality of melons, a plurality of apples, or a plurality of lettuce heads) of the indicated tested harvested plant matter. The previously described parameters were measured and evaluated from at least three of the same type of individual samples or units (for example, at least three melons, at least three apples, or at least three lettuce heads, respectively) of the tested harvested plant matter of each experiment. An average or mean value, and the standard deviation thereof, were calculated for each parameter of each experiment.

A single representative experiment, from the original set of at least two, typically, three, separate experiments, was selected for determining reproducibility of the average or mean values of the parameters of the representative experiment. For an additional at least two times, the same parameters were again measured and evaluated from another at least three of the same type of individual samples or units of the tested harvested plant matter of the representative experiment, along with calculating the average or mean value, and the standard deviation thereof, for each parameter. The average of the standard deviations, that is, the average standard deviation, of each parameter of the representative experiment was then used for calculating the associated standard error (i), by dividing the average standard deviation by the square root of the number, n, of the total number of times, typically, three times, a particular parameter was measured and evaluated. The standard error ( : L) was then associated with the average or mean value of the number, n, of the total number of average or mean values of the particular parameter of the representative experiment.

The average or mean value of the total number of average or mean values of each parameter of the representative experiment, along with the associated standard error ( : L),

was used as the representative value of each parameter of the representative experiment.

The representative value of each parameter and the associated standard error () thereof, were selected for inclusion in the tables of the experimental results accompanying the following description of the Examples, and were used as the basis of detailed presentation and analysis of the Examples of the present invention, in support of the various embodiments and aspects of the present invention as described herein above and as claimed in the claims section below.

EXAMPLE 4: MELONS (FRUIT)/SPRAYING Treatment and application compositions, procedures, and conditions - Frutavit D: 6-octanoyl-L-ascorbate, 40 mM; in water.

- Frutavit D-4: 6-octanoyl-L-ascorbate, 20 mM + gum arabic, 5 %, wt/vol + K-sorbate, 2 %, wt/vol; in water.

- Standard : rinsing in cold water, then in hot water at 56-57 °C.

Harvested melons, in separate groups, were sprayed, without first rinsing in cold or hot water, with one of the above indicated compositions at room temperature. Following the spraying treatments, all the treated melons were first allowed to dry at room temperature, then treated with wax, followed by cold storage at 5 °C in a cold room for a period of 14 days. Following the cold storage, the treated melons were stored at room temperature for periods of 3 days and 7 days, along with determining the shelf lives of the treated and untreated (standard) melons, by measuring, evaluating, and analyzing, the previously described parameters directly relating to properties, characteristics, and behavior, of the tested melons.

Reslilts aizd Atzalvsis The results are presented in Tables 4-7, and in FIGS. 2-4.

Melons treated with the 6-octanoyl-L-ascorbate compositions exhibited higher degrees of firmness (Table 4, FIGS. 2 and 3) and lower percentages of decay and color change in the form of brown spots (Table 5, FIG. 4). However, the treated melons had no effect on total sugars or on weight loss (Tables 6 and 7, respectively). Table 4: Effect of different treatments on firmness (%) of melons after spraying with different compositions, then cold storage at 5 °C for 14 days, followed by shelf life at 20 °C for 3 days (FIG. 2) and for 7 days (FIG. 3).

14 days/5 °C + 14 days/5 °C + Treatments Firmness 14 days/5 °C 3 days shelf 7 days shelf life/20 °C life/20 °C Firm 48 ~ 5. 9 29 5. 6 13 ~ 4. 1 Medium 40 ~ 4.1 43 ~ 4.3 38 ~ 3.4 Standard 49 # 4.7 Soft 12 ~ 3.6 28 ~ 3.9 Firm 73 ~ 3.8 59 ~ 2.9 22 ~ 4. 3 Frutavit D Medium 25 ~ 3. 4 33 ~ 3. 1 49 5. 8 Soft 2 ~ 1. 1 8 ~ 2. 2 29 ~ 4. Firm 66 4. 3 51 3. 6 28 ~ 4. 4 Medium 27 ~ 4. 1 30 ~ 3. 2 384 Frutavit D-4 Soft 7 ~ 3. 2 19 ~ 3. 8 345. 5

Table 5 : Effect of different treatments on brown spots (%) of melons after spraying with different compositions, then cold storage at 5 °C for 14 days, followed by shelf life at 20 °C for 3 days and for 7 days (FIG. 4). 14 days/ 5 °C + 3 14 days/ 5 °C + 7 Treatments 14 days/5 °C days shelf life/days shelf life/ 20 °C 20 °C Standard 10. 5 47. 4 71. 4 Frutavit D 16. 6 20 50 Frutavit D-4 7. 1 14. 3 39

FIGS. 5-10 are photographs illustrating the effect of different treatments on initiation and development of brown spots and discoloration of melons after spraying with different compositions, then cold storage at 5 °C for 14 days, followed by shelf life at 20 °C for 3 days (FIGS. 5-7) and for 7 days (FIGS. 8-10).

Table 6: Effect of different treatments on total sugars (%) of melons after spraying with different compositions, then cold storage at 5 °C for 14,3, and 7 days, followed by shelf life at 20 °C. 14 days/5 °C + 3 14 days/5 °C + 7 Treatments 14 days/5 °C days shelf life/days shelf life/ 20 °C 20 °C Standard 11. 50. 24 12.8 ~ 0. 09 ~ 0. 2912.87 Frutavit D 12. 6 ~ 0.28 12.33 ~ 0. 3 12. 80. 19 Frutavit D-4 10 ~ 0. 74 13. 33 ~ 0. 27 13. 53 ~ 0. 20

Table 7: Effect of different treatments on weight loss (%) of melons after spraying with different compositions, then cold storage at 5 °C for 14,3, and 7 days, followed by shelf life at 20 °C.

14 days/ 5 °C + 3 14 days/ 5 °C + 7 Treatments 14 days/5 °C days shelf life/days shelf life/ 20 °C 20 °C Standard 2. 66 ~ 0. 14 4. 56 0. 17 7.41 : L 0. 31 Frutavit D 2. 52 i 0. 06 4.18 ~ 0.11 6.83 ~ 0. 18 Frutavit D-4 2. 32 ~ 0. 05 3.91 ~ 0. 08 6. 68 i 0. 11

EXAMPLE 5: APPLES (FRUIT)/DIPPING Treatment and application compositions, procedures, and conditions - Frutavit D-2: 6-octanoyl-L-ascorbate, 20 mM + K-sorbate, 2 %, wt/vol; in water.

- Control : no treatment at room temperature.

Harvested apples, in separate groups of three different species (golden apples, green apple, and red apple), were cut into pieces and then dipped into the Frutavit D-2 composition at room temperature, for a period of about 3-10 seconds. Following the

dipping treatment, the treated cut apples were stored at either room temperature for up to 7 days, or in cold storage at 4 °C in a cold room for up to 7 days, followed by storage at room temperature for up to an additional 4 days, along with determining the shelf lives of the treated and untreated (control) cut apples, by measuring, evaluating, and analyzing the previously described parameters directly relating to properties, characteristics, and behavior, of the tested cut apples.

Results and Snalysis The results are presented in Tables 8-10, and in FIGS. 11-25.

The results indicate that Frutavit D-2 solution, compared to the Control, significantly prevented decay and microbial growth and browning of all three species of cut apples.

Table 8: Effect of momentary dipping of cut apples in 6-octanoyl-L-ascorbate solution on total microbial count (CFU/g-FW). Treatments 0 days/20 4 days/20 7 days/20 7 days/4 °C 7 days/4 °C °C °C °C + 4 days/20 °C Control 665 3000000 3000000 290000 3000000 Frutavit D-2 265 < 10 280 < 10 110 Table 9: Effect of momentary dipping of cut apples in 6-octanoyl-L-ascorbate solution on total molds count (CFU/g-FW). Treatments 0 days/20 4 days/20 7 days/20 7 days/4 °C 7 days/4 °C °C °C °C + 4 days/20 °C Control 100 20000000 30000000 29000 6000000 55 < 10 < 10 < 10 425 Frutavit D-2 Table 10: Effect of momentary dipping of cut apples in 6-octanoyl-L-ascorbate solution on total yeast count (CFU/g-FW).

Treatments 0 days/20 4 days/20 7 days/20 7 days/4 °C 7 days/4 °C °C °C °C +4days/20 oc Control 105 2100000 1500000 10 1400000 Frutavit D-2 < 10 <10 <10 < 10 75 EXAMPLE 6: GRAPES (FRUIT)/SPRAYING Treatment and application compos1rtions, procedllres and conditions - Frutavit D: 6-octanoyl-L-ascorbate, 40 mM; in water.

- Frutavit D-3: 6-octanoyl-L-ascorbate, 40 mM + gum arabic, 5 %, wt/vol; in water.

- Gum arabic : gum arabic, 5 %, wt/vol, in water.

- Control : no treatment at room temperature.

Harvested grapes (petiole), in separate groups, were sprayed with one of the above indicated compositions at room temperature. Following the spraying treatment, the treated grapes were stored at room temperature for periods of 2 days, 4 days, and 6 days, along with determining the shelf lives of the treated and untreated (control) grapes, by measuring, evaluating, and analyzing the previously described parameters directly relating to properties, characteristics, and behavior, of the tested grapes.

Restllts and Analvsis The results are presented in FIGS. 26-31.

The results indicate that the Frutavit D and Frutavit D-3 6-octanoyl-L-ascorbate compositions, were highly effective for preventing decay and browning of the petiole grapes, as shown in FIGS. 26 and 27,28 and 29, and, 30 and 31, for shelf life at 20 °C for 2 days, 4 days, and 6 days, respectively.

EXAMPLE 7: LETTUCE (VEGETABLE)/DIPPING Treatment and applicatio7t compositions, procedures and conditions - Preparation 7 : Frutavit D-2 : 6-octanoyl-L-ascorbate, 20 mM + K-Sorbate, 2 %, wt/vol ; in water.

- Preparation 8: Frutavit D-1 : 6-octanoyl-L-ascorbate, 20 mM, in water.

Work Procedures Plant Material The heads of'Iceberg'lettuce were taken directly from a grower's field in the Western Negev region of Israel, brought to the laboratory, and pre-cooled overnight at 1 °C. On the next day, the lettuce was subjected to the minimal processing procedure.

P70cessie The processing was conducted at the pilot laboratory of the Dept. of Postharvest Science, in accordance with the technological scheme and rules applied in the food industry. The process included removal of external leaves, preliminary manual dissection of lettuce heads, and shredding the leaves using a vegetable cutting machine (GS-10, Kronen, Germany), which is the type of machine commonly used in the fresh-cut industry.

The width of leaf strips was set at 10 mm.

The shredded lettuce was dipped (washed) in different ways. Part of the shredded lettuce was decontaminated for one minute in the sterilizing solution of sodium hypochlorite at the concentration commonly used in the industry (150 ppm of free chlorine), and afterwards was subjected for one minute to an additional wash. One of the following materials was used in this additional wash: water, ascorbic acid solution (0.5 %), solution of one of the two Frutavit preparations (compositions) tested, marked by numbers 7 and 8. Solutions of the Frutavit preparations were prepared in accordance with the company's instructions. Another part of the shredded lettuce was treated only in the chlorine solution, without an additional wash, and served as a control. The third part of the shredded lettuce was washed directly in the solutions of the Frutavit preparations, without the preliminary chlorine decontamination. Altogether, the trial study included 19 treatments, which were arranged in two experimental series tested in two stages. The

series of the Frutavit preparations 7 and 8 was tested on Oct. 29, '02. This series included control treatments (sodium hypochlorite with or without subsequent water wash).

After washing, the excessive liquid was removed from the shredded lettuce in a centrifuge (Kronen, Germany). The shredded lettuce was packaged in polypropylene bags (30 micron thick), 200 grams fresh mass per bag. Bags of this material are used commercially to pack fresh-cut vegetable salads.

Storage The shredded lettuce was stored for up to 14 days at 6 °C, similar to the conditions during the commercial marketing chain of fresh-cut vegetables, and subsequent storage in a home refrigerator. Samples were taken for the analysis immediately after the treatments and after 5,8, and 12, days of storage. In some cases, an additional test was conducted after 14 days of storage.

Qzcalit evaluation The assessment of the shredded lettuce quality included (a) visual and organoleptic evaluation, and (b) microbiological analysis.

Visual and organoleptic checks were conducted by a constant panel including 3-4 trained evaluators. The following quality parameters were checked: browning severity- according to the scale from 0 (no browning) to 4 (severe browning); wilting severity- according to the scale from 0 (no wilting) to 3 (severe wilting); decay severity-according to the scale from 0 (no decay) to 4 (severe decay); off-odor-according to the scale from 0 (no off-odor) to 3 (severe off-odor) ; off-taste severity-according to the scale from 0 (no off-taste) to 3 (severe off-taste). The apparently spoiled (decayed) samples were not tasted. General quality assessment was made on the basis of the complex of all visual and organoleptic quality parameters according to the scale from 1 (poor quality) to 5 (excellent quality), while the sample getting the mark below 2.5 was considered non-marketable.

The microbiological analysis included determination of total microbial count. 10 gram samples of the shredded lettuce were homogenized in a Stomacher instrument within 190 ml of sterile water, and inoculated with various dilutions onto a solid PCA medium (plate count agar), 100 microliter per each of a Petri dish. The microbial count was calculated per gram fresh matter.

The composition of headspace atmosphere inside the plastic packages was measured by gas chromatography.

Results and Analvsis Ahnosphere composition Table 11: Headspace atmosphere composition in the packages of shredded lettuce. Preliminary Atmosphere composition Wash decontamin-Acetald-Ethanol, Ethylene, C02, % 02, % ation hyde, ppm ppm ppm Cl 150 ppm 7. 29 8. 07 0. 00 0. 26 0. 59 Water Cl 150ppm 6. 70 8. 85 0. 00 0. 18 0. 72 Ascorb. acid Cl 150 ppm 8. 47 6. 32 0. 00 0. 00 0. 93 Cl 150 ppm 10.77 3.18 12.08 21.55 4.35 Prep. 7 - 11.02 3.03 12.49 21.12 0.00 Cl 150 ppm 10.093.496.0824.35 Prep. 8 11. 60 2.32 6.17 15.66 2.25 Table 11 represents the composition of atmosphere inside the lettuce packages after 7 days of storage. At that stage, the control packages contained relatively high concentration of oxygen (8-9 %) together with about 7 % of carbon dioxide and low level of ethylene, as well as ethanol and acetaldehyde vapors. The ascorbic acid treatment slightly reduced the oxygen level, and to the same extent, increased the level of carbon dioxide. Washing the shredded lettuce in the Frutavit preparations 7 and 8 was associated with significantly lower oxygen level and significantly higher level of carbon dioxide and ethylene than in the control, possibly due to the enhanced physiological activity of tissues.

The Frutavit preparations 7 and 8 enhanced ethanol and acetaldehyde concentrations inside the packages. The headspace concentrations of acetaldehyde were 6-13 ppm, and of ethanol, 8-25 ppm. The analysis of headspace atmosphere composition was conducted also after 5 and 12 days of storage (data not shown). They have demonstrated the same trends as reported above.

Visual and organoleptic quality of the produce during storage The effect of various treatments on the visual and organoleptic quality is presented in Tables 12-15, and in FIGS. 32 and 33.

Table 12: Effect of washing on the quality of shredded lettuce immediately after treatment. Preliminary Quality parameters Wash decontami-General Off odor Decay Browning Wilting Off-taste nation quality - Cl 150 ppm 0 0 0 0 0 5 Water Cl 150 ppm 0 0 0 0 0 5 Asc. acid Cl 150ppm 0 0 0 0 0 5 Cl 150 ppm Prep. 7 - 0.5 0 0 0 0.3 4 Cl 150 ppm 0. 5 0 0 0. 2 0. 3 3. 7 Prep. 8 - 0. 2 0 0 0 0.3 4.8

Table 13: Effect of washing on the quality of shredded lettuce after 5 days of storage at 6 °C Preliminary Quality parameters Wash decontami-General Off-odor Decay Browning Wilting Off-taste nation quality - Cl 150 ppm 0 0 0.1 ~ 0.1 0 0 4.7 ~ 0. 2 Water Cl 150 ppm 0 0 0.1 ~ 0.1 0 0 4.8 ~ 0. 2 Asc. acid Cl 150 ppm 0 0 0. 2 0. 1 0. 1 0. 1 0 4. 6 0. 2 C1150ppm 0. 6 : L 0. 2 0. 2 0. 1 0 2. 8 0. 3 not 1. 9 0. 2 tasted Prep. 7 - 9 L 0. 3 0.2 ~ 0. 1 0. 5 0. 3 3 not 1. 3 t 0. 2 tasted Cl 150 ppm 0 ~ 0.1 0 0 0 0 ~ 0.1 4.9 ~ 0.2 Prep. 8 - 0.1 0 0 0.1 0 4.8

The effects of each treatment on visual and organoleptic quality of the shredded Iceberg lettuce are summarized below.

Control. Shredded lettuce, which underwent chlorine decontamination without additional wash had good visual quality and no off-flavor in the three first checks (up to 8 days of storage at 6 °C). During this period, only negligible signs of browning were seen on the produce. However, considerable reddish discoloration ('browning') was observed in this treatment in the fourth check after 12 days of storage, which rendered the produce unmarketable (general quality mark below 2.5). Water wash after the chlorine decontamination did not improve the produce quality and at late stages of the storage even slightly enhanced the browning and reduced the general quality mark. Usually, this reduction was not statistically significant.

Table 14: Effect of washing on the quality of shredded lettuce after 8 days of storage at 6 °C. Preliminary Quality parameters Wash decontami-General Off odor Decay Browning Wilting Off-taste nation quality - Cl 150 ppm 0 0 0.2 ~ 0.1 0 0 4.1 ~ 0. 2 Water Cl 150 ppm 0 0 0. 4 0. 1 0 0 3. 6 0. 2 Asc. acid Cl 150 ppm 0 0 0.4 ~ 0.1 0.1 ~ 0.1 0 3.3 ~ 0. 2 Cl 150 ppm 0. 5 0. 3 0. 5 0. 3 0. 5 ~ 0.3 3 not 1.5 tasted Prep. 7 - 0. 5 1 1 3 not 1.25 tasted Cl 150ppm 0. 1 0 0 0 0 4. 6i0. 2 Prep. 8 0. 1 zt O. l O 0. 1 ~ 0.1 0.1 0. 1 0 4. 1 0. 1

Ascorbic acid. Washing lettuce in 0.5% solution of ascorbic acid after the chlorine decontamination reduced to a certain extent the severity of browning after 12 days of storage. As a result, the treated produce at this stage was found still marketable, although it demonstrated already some signs of browning. No significant difference was found between ascorbic acid-treated lettuce and the control at earlier stages of storage. It is noted that applying the ascorbic acid alone brought better results than its combined application with citric acid, which was tested in our previous trials and was found to enhance the browning.

Table 15: Effect of washing on the quality of shredded lettuce after 12 days of storage at 6 °C. Preliminary Quality parameters Wash decontami-General Off odor Decay Browning Wilting Off-taste nation quality - Cl 150 ppm 0 0 1.4 ~ 0.4 0.3 ~ 0.2 0 2.4 ~ 0. 2 Water Cl 150 ppm 0 0 1.5 ~ 0. 2 0. 2 0. 1'02. 3 0. 2 Asc. acid Cl 150 ppm 0 0 0.8 ~ 0.3 0.25 0 2. 7 ~ 0. 2 Cl 150 ppm 0. 8 0. 2 3. 3 0. 3 2. 2 ~ 0.3 3 not 1 tasted Prep. 7 - 0.8 ~ 0.2 3.7 ~ 0. 2 3. 0 i 0. 3 3 not 1 tasted Cl 150 ppm 0.1 ~ 0.1 0 ~ 0.1 0 ~ 0.1 0.1 ~ 0.2 0 3.6 ~ 0. 2 Prep. 8 Prep. 8 0. 3 0. 2 0. 2+0. 1 0. 3+0. 2 0. 4 0. 3 0 2. 4 0. 2

Preparation 7: Frutavit D-2: 6-octanovl-L-ascorbate. 20 mM + K-Sorbate, 2 %, wt/vol ; in water. This material showed strong phytotoxic effect, so that the treated lettuce was found unmarketable already after 5 days of storage. The treatment, either with or without chlorine decontamination, caused tissue breakdown, decayed appearance, complete loss of turgor (wilting), brown discoloration, together with uncharacteristic odor and taste.

Preparation 8: Frutavit D-1 : 6-octanovl-L-ascorbate, 20 mM, in water. This preparation inhibited the oxidative browning of the shredded lettuce without any side effect. The best visual results were obtained when the lettuce was treated with preparation 8 after chlorine decontamination. The shredded lettuce maintained excellent quality for at least two weeks.

On the other hand, the shredded lettuce treated with preparation 8 without preliminary chlorine decontamination showed after 12 days of storage the initial symptoms of microbial spoilage: first signs of decay, some off-odor, etc..

Microbiological qzlality of the produce during storage The results of microbiological analysis are presented in Table 16. The initial microbial load on the lettuce leaves was about 104. Decontamination with sodium hypochlorite (150 ppm free chlorine) reduced this level approximately by one order.

Additional water wash did not change the microbial load, but washing in the solution of ascorbic acid (0.5%) enhanced the efficacy of decontamination approximately by one order, in addition to the effect of chlorine.

Treating lettuce with the Frutavit preparations 7 or 8, instead of chlorine, in most of the cases, reduced the amount of microorganisms to a certain extent (usually by one order).

Combination of the Frutavit preparations with preliminary chlorine decontamination improved the microbiological quality of shredded lettuce as compared with the Frutavit preparations alone. Preparation 8, showed promising results in visual and organoleptic evaluations. Treatment with each of these preparations, especially preparation 8, is preferably combined with a decontamination agent, such as chlorine or another sterilizing agent, for increasing effectiveness.

Table 16: Effect of washing with various additives on the total microbial count (CFU/g fresh mass) on shredded Iceberg lettuce during storage. Preliminary Storage duration, days Wash decontami- 0 5 8 12 nation 2. 9* 104 Cl 150 ppm 5 0*10 2 5*10 5. 7*10 2. 3*107 Water Cl 150 ppm 3. 1*103 2.0*106 5.2*106 1.5*107 Asc. acid Cl 150 ppm 6. 0*102 3.7*105 2.2*106 3.4*106 Cl 150 ppm 6. 0*1O2 4. 9*105 7. 0*105 6. 6*106 Prep. 7 - 1. 4*104 1. 2*106 1. 3*107 1.0*108 Cl 150 ppm 9. 2*103 3.8*105 8.0*106 2.7*107 Prep. 8 - 8.0*103 9. 5*106 2. 0*107 1.0*108

EXAMPLE 8: LETTUCE (VEGETABLE)/BRUSHING Treatment and application compositions, procedures, and conditions Frutavit D-1 : 6-octanoyl-L-ascorbate, 20 mM, in water.

- Control : no treatment at room temperature.

Harvested heads of'Iceberg'lettuce were cut and then brushed with the above indicated 6-octanoyl-L-ascorbate composition at room temperature. Following the brushing treatment, the treated lettuce heads were stored at 6 °C in a cold room for periods of 0,18, 48,72, 96,120, and 216 hours, along with determining the shelf lives of the treated and untreated (control) lettuce heads, by measuring, evaluating, and analyzing the previously described parameters directly relating to properties, characteristics, and behavior, of the tested lettuce heads.

Results and Analysis The results are presented in FIG. 34.

The results indicate that the Frutavit D-1, 6-octanoyl-L-ascorbate composition, was highly effective for preventing decay and browning of the lettuce head leaves, as shown in FIG. 34, for shelf life at 6 °C for 0,18, 48,72, 96,120, and 216 hours.

EXAMPLE 9: BANANAS (FRUIT)/DIPPING Treatnleslt and application compositions, procedures, and conditions - Frutavit D: 6-octanoyl-L-ascorbate, 40 mM, in water.

- Frutavit D-a: 6-octanoyl-L-ascorbate, 40 mM + K-Sorbate, 2 %, wt/vol; in water.

- Frutavit F-a: 6-decanoyl-L-ascorbate, 40 mM + CaCl2, 0. 1 %, wt/vol ; in water.

- Standard : imazalil (fungicide), 500 ppm, in water.

- Control : no treatment at room temperature.

Harvested ripened bananas absent of blemishes or spots, in separate groups, were dipped into one of the above indicated compositions at room temperature, for a period of about 3 minutes. Following the dipping treatment, the treated bananas were dried at room temperature, packed in perforated nylon bags, and stored at room temperature for periods of 0 days, 4 days, 5 days, 7 days, and 8 days, along with determining the shelf lives of the treated, standard, and untreated (control), bananas, by measuring, evaluating, and analyzing the previously described parameters directly relating to properties, characteristics, and behavior, of the tested bananas.

Reslllts andAnalvsis The results are presented in Table 17, and in FIGS. 35 and 36.

The results indicate that the Frutavit D and Frutavit D-a, 6-octanoyl-L-ascorbate compositions, and the Frutavit F-a, 6-decanoyl-L-ascorbate composition, were each highly effective for preventing decay, discoloration (browning, appearance of dark or black spots), and spoilage, of the banana peels, compared to the Control and to the imazalil Standard composition, as indicated in Table 17 and as shown in FIGS. 35 and 36, for shelf life at 20 °C for 0 days, 4 days, 5 days, 7 days, and 8 days.

Table 17: Percentage of black spots appearing on banana peels following various treatments and storage of the bananas at room temperature. Day 4 Day 5 Day 7 Day 8 Treatment 20 40 55 65 Control Standard 9 30 50 60 Frutavit D 0 0 5 10 Frutavit D-a 0 0 1 6

Frutavit F-a 0 0 6 20 EXAMPLE 10: LETTUCE (VEGETABLE)/BRUSHING Treatmeszt and application compositions, procedures, and conditions -Frutavit D-1-a : 6-octanoyl-L-ascorbate, 20 mM + CaCl2, 0.1 %, wt/vol; in water.

- Frutavit D-1: 6-octanoyl-L-ascorbate, 20 mM, in water.

- Water : water alone at room temperature.

- Control : no treatment at room temperature.

Harvested heads of'Iceberg'lettuce were cut and then brushed with one of the above indicated compositions at room temperature. Following the brushing treatment, the treated lettuce heads were directly covered with nylon pieces and stored at 2 °C in a cold room for periods of 2 days, 3 days, 4 days, 6 days, and 7 days, along with determining the shelf lives of the treated and untreated (control) lettuce heads, by measuring, evaluating, and analyzing the previously described parameters directly relating to properties, characteristics, and behavior, of the tested lettuce heads.

Restllts and Anal The results are presented in Tables 18-22, and in FIG. 37.

The results indicate that the Frutavit D-1-a and Frutavit D-l, 6-octanoyl-L- ascorbate compositions, were highly effective for preventing decay and browning of the lettuce heads, as indicated in Tables 18-22, for storage at 2 °C for periods of 2 days, 4 days, and 6 days, respectively, and shown in FIG. 37, for storage at 2 °C for a period of 7 days. As a matter of fact, the Control exhibited a high level of browning following only 4 hours of storage at 2 °C.

In Tables 18-22, a browning level index or scale of 1-5 is used for comparing the extent of browning of the lettuce heads subjected to the different treatments. A value of 1 indicates an undesirable high extent of browning, while a value of 5 indicates a desirable low extent of browning, of the iceberg lettuce heads. Table 18: Effect of different treatments on the percent of lettuce heads exhibiting an indicated browning level (1-5), of brushing the lettuce heads, followed by cold storage at 2 °C for 2 days. browning level Treatments 1 2 3 4 5 Control 62. 8% 34.9% 2.3% Water 50% 50% Frutavit 16.3% 39.5% 44.2% D-1 Frutavit 33.3% 50% 16.7% D-1-a

Table 19: Effect of different treatments on the percent of lettuce heads exhibiting an indicated browning level (1-5), of brushing the lettuce heads, followed by cold storage at 2 °C for 3 days.

browning leve Treatments 1 2 3 4 5 Control 74. 4% 20.9% 4.7% Water 75% 20. 8% 4.2% Frutavit 62.8% 30.2% 7% D-1 Frutavit 83. 3% 16.7% D-1-a

Table 20: Effect of different treatments on the percent of lettuce heads exhibiting an indicated browning level (1-5), of brushing the lettuce heads, followed by cold storage at 2 °C for 4 days. browning level Treatments 1 2 3 4 5 Control 23. 3% 46. 5% 30. 2% Water83. 3% 12. 5% 4. 2% Frutavit 79% 21 % D-1 Frutavit 90% 10% D-1-a Table 21 : Effect of different treatments on the percent of lettuce heads exhibiting an indicated browning level (1-5), of brushing the lettuce heads, followed by cold storage at 2 °C for 6 days. browning level Treatments 1 2 3 4 5 Control 25.6% 62.8% 11.6% Water 29.2% 70.8% Frutavit 41.9% 55. 8% 2.3% D-1 Frutavit 30% 63.3% 6.7% D-1-a

Table 22: Effect of different treatments on the percent of lettuce heads exhibiting an indicated browning level (1-5), of brushing the lettuce heads, followed by cold storage at 2 °C for 7 days. browning level Treatments 1 2 3 4 5 Control 37.2% 55.8% 7% Water 37.5% 62.5% Frutavit 51.2% 48. 8% D-1 Frutavit 40% 60% D-1-a Thus, it is clearly understood that the present invention, as illustratively described and exemplified hereinabove, has the following beneficial and advantageous aspects, characteristics, and features.

First, the method, and various different embodiments thereof, for extending the shelf life of harvested plant matter using the compositions of the alkanoyl-L-ascorbic acid esters: 6-octanoyl-L-ascorbate (6-octyl-ascorbate), 6-nonanoyl-L-ascorbate (6-nonyl- ascorbate), and 6-decanoyl-L-ascorbate (6-decyl-ascorbate), are relatively simple and inexpensive to implement. Second, the solution or suspension compositions of the alkanoyl-L-ascorbic acid esters are relatively simple and inexpensive to manufacture. In particular, the method of using the disclosed compositions, and the compositions themselves, in which a selected alkanoyl-L-ascorbic acid ester functions as an antioxidation active ingredient, may further include, but do not require, additional or

separate treatment steps, and/or, additional components or ingredients, for being highly effective for use in extending the shelf life of the harvested plant matter.

Third, the present invention is generally applicable prior to, during, or following, commercial activities such as handling, packaging, storing, transporting, physical treatment, and chemical treatment, which are part of a variety of numerous processes and sequences, before the harvested plant matter is eventually sold in commercial wholesale and retail environments.

Fourth, the method for synthesizing the alkanoyl-L-ascorbic acid esters is suitable and cost effective for commercial large-scale manufacturing applications.

Fifth, the present invention is environmentally safe, and non-hazardous to humans or animals during handling and/or consumption of the treated harvested plant matter.

Sixth, the method of using the disclosed compositions, and the compositions themselves, are generally applicable for use in extending the shelf life of harvested plant matter, such as, but not limited to, fruits, vegetables, and flowers.

Based upon the above stated beneficial and advantageous aspects, characteristics, and features, the present invention widens the scope of the related art.

It is appreciated that certain aspects and characteristics of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various aspects and characteristics of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention.

While the invention has been described in conjunction with specific embodiments and examples thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

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