COMPOSITIONS AND METHODS FOR TREATING PRODUCE

Abstract

The present invention discloses the use of winter melon wax as a preservative and an antimicrobial agent. In addition, the present invention also discloses a kind of powder of artificial winter melon wax, which is obtained by spray drying the solution of following components with ratios by weight: 30-55% triterpenenol, 10%-20% triterpenenol acetate and 35-60% wax. Experiments proved that the natural winter melon wax and the artificial Winter melon wax powder prepared by the invention had antiseptic and anti-microbial effects, could significantly improve the shelf life of fruits, and had a promoting effect on wound healing.

Claims

1. A method of treating produce to reduce the amount of microbial contamination on the surface of the produce and to reduce spoilage of the produce, comprising contacting the surface of the produce with a compound comprising: triterpenenol in concentration from 30-55%; triterpenenol acetate in concentration from 10%-20%, and wax in concentration from 35-60%.

2. The method of claim 1, wherein the triterpenenol is Betulin or a structural analog thereof.

3. The method of claim 1, wherein the triterpenenol acetate is -Amyrin acetate, or a structural analog thereof.

4. The method of claim 1, wherein the wax is beeswax.

5. The method of claim 1, wherein the amount of compound that is applied to the produce is between 0.0001% to 5% of weight of the produce.

6. The method of claim 1, wherein the amount of compound that is applied to the produce is between 0.01% to 2.5% of the weight of the produce.

7. The method of claim 1, wherein the compound is applied to the surface of the produce.

8. The method or creating a powder for preserving produce, the method comprising: extracting triterpenenol from a winter melon or a winter melon extract; extracting triterpenenol acetate from a winter melon or a winter melon extract; obtaining wax; and combining triterpenenol in concentration from 30-55% with triterpenenol acetate in concentration from 10%-20% and wax in concentration from 35-60% to form the powder.

9. The method of claim 8, wherein the triterpenenol is Betulin or a structural analog thereof.

10. The method of claim 8, wherein the triterpenenol acetate is -Amyrin acetate, or a structural analog thereof.

11. The method of claim 8, wherein the wax is beeswax.

Description

BRIEF DESCRIPTION OF THE DRAWING FIGURES

[0043] The accompanying drawings illustrate several embodiments and, together with the description, serve to explain the principles of the invention according to the embodiments. It will be appreciated by one skilled in the art that the particular arrangements illustrated in the drawings are merely exemplary and are not to be considered as limiting of the scope of the invention or the claims herein in any way.

[0044] FIG. 1 illustrates a chemical structure for triterpenenol and triterpenenol acetate in accordance with one embodiment of the invention.

[0045] FIG. 2a illustrate an exemplary Winter melon wax (WMW) in accordance with one embodiment of the invention as observed using an electronic microscope. The magnification illustrated in this figure is at 5000.

[0046] FIG. 2b illustrate an exemplary Winter melon wax (WMW) in accordance with one embodiment of the invention as observed using an electronic microscope. The magnification illustrated in this figure is at 7000.

[0047] FIG. 2c illustrate an exemplary Winter melon wax (WMW) in accordance with one embodiment of the invention as observed using an electronic microscope. The magnification illustrated in this figure is at 20000.

[0048] FIG. 3a illustrates the antiseptic effect of WMW on lemon fruit in accordance with one embodiment of the invention. Item 1 in FIG. 3a illustrates a fresh lemon, item 2 in FIG. 3a illustrates a lemon after 15 to 20 without any application of WMW, and item 3 in FIG. 3a illustrates a lemon after 15 to 20 days with an application of WMW.

[0049] FIG. 3b illustrates the antiseptic effect of WMW on lemon fruit in accordance with one embodiment of the invention. Item 1 in FIG. 3b illustrates a partially decayed lemon, item 2 in FIG. 3b illustrates a partially decaled lemon after 15 to 20 without any application of WMW, and item 3 in FIG. 3b illustrates a partially decayed lemon after 15 to 20 days with an application of WMW.

[0050] FIG. 3c illustrates the antiseptic effect of WMW on orange fruit in accordance with one embodiment of the invention. Item 1 in FIG. 3c illustrates a partially decayed orange, item 2 in FIG. 3c illustrates a partially decaled orange after 15 to 20 without any application of WMW, and item 3 in FIG. 3c illustrates a partially decayed orange after 15 to 20 days with an application of WMW.

[0051] FIG. 3d illustrates the antiseptic effect of WMW on orange fruit in accordance with one embodiment of the invention. Item 1 in FIG. 3d illustrates a fresh orange, item 2 in FIG. 3d illustrates an orange after 15 to 20 without any application of WMW, and item 3 in FIG. 3d illustrates an orange after 15 to 20 days with an application of WMW.

DETAILED DESCRIPTION

[0052] One or more different embodiments may be described in the present application. Further, for one or more of the embodiments described herein, numerous alternative arrangements may be described; it should be appreciated that these are presented for illustrative purposes only and are not limiting of the embodiments contained herein or the claims presented herein in any way. One or more of the arrangements may be widely applicable to numerous embodiments, as may be readily apparent from the disclosure. In general, arrangements are described in sufficient detail to enable those skilled in the art to practice one or more of the embodiments, and it should be appreciated that other arrangements may be utilized and changes may be made without departing from the scope of the embodiments. Particular features of one or more of the embodiments described herein may be described with reference to one or more particular embodiments or figures that form a part of the present disclosure, and in which are shown, by way of illustration, specific arrangements of one or more of the aspects. It should be appreciated, however, that such features are not limited to usage in the one or more particular embodiments or figures with reference to which they are described. The present disclosure is neither a literal description of all arrangements of one or more of the embodiments nor a listing of features of one or more of the embodiments that must be present in all arrangements.

[0053] Headings of sections provided in this patent application and the title of this patent application are for convenience only and are not to be taken as limiting the disclosure in any way.

[0054] A description of an aspect with several elements does not imply that all such components are required. To the contrary, a variety of optional elements may be described to illustrate a wide variety of possible embodiments and in order to more fully illustrate one or more embodiments. Similarly, although process steps, and method steps or the like may be described in a sequential order, such process steps, and method steps may generally be configured to work in alternate orders, unless specifically stated to the contrary. In other words, any sequence or order of steps that may be described in this patent application does not, in and of itself, indicate a requirement that the steps be performed in that order. The steps of described processes may be performed in any order practical. Further, some steps may be performed simultaneously despite being described or implied as occurring non-simultaneously (e.g., because one step is described after the other step). Moreover, the illustration of a process by its depiction in a drawing does not imply that the illustrated process is exclusive of other variations and modifications thereto, does not imply that the illustrated process or any of its steps are necessary to one or more of the embodiments, and does not imply that the illustrated process is preferred. Also, steps are generally described once per aspect, but this does not mean they must occur once, or that they may only occur once each time a process, or method is carried out or executed. Some steps may be omitted in some embodiments or some occurrences, or some steps may be executed more than once in a given aspect or occurrence.

[0055] When a single article is described herein, it will be readily apparent that more than one article may be used in place of a single article. Similarly, where more than one article is described herein, it will be readily apparent that a single article may be used in place of the more than one article.

[0056] Techniques and mechanisms described or referenced herein will sometimes be described in singular form for clarity. However, it should be appreciated that particular embodiments may include multiple iterations of a technique or multiple instantiations of a mechanism unless noted otherwise. Alternate implementations are included within the scope of various embodiments in which, for example, functions may be executed out of order from that shown or discussed, including substantially concurrently or in reverse order, depending on the functionality involved, as would be understood by those having ordinary skill in the art.

EMBODIMENTS

[0057] The invention will be further described in conjunction with the specific embodiment, through the description advantages and features of the invention will become more apparent. However, these examples are merely exemplary and do not constitute any limitation on the scope of the invention. It should be understood by those skilled in the art that the details and forms of the present invention may be modified or substituted without departing from the spirit and scope of the invention, and such modifications and substitutions fall within the scope of the present invention.

Embodiment 1: Preparation and Analysis of Winter Melon Wax (WMW) Powder

[0058] Collect 10 to 25 kilograms powder of natural WMW from mature winter melon using a special removable automatic cleaner. The wax powder is sieved to remove unwanted contaminants and chips. Then, the powder was extracted with distilled water and 95% ethanol. The extract was analyzed by high performance liquid chromatography and confirmed by the method of the literature (High Performance Liquid Chromatography of Triterpenols, Gerard J. Niemann and Wim J. Baas Journal of chromatographic science 16 (6): 260-262., June 1978). The triterpenoids were also analyzed by thin layer chromatography (TLC) and color reaction with carbazole. The results were confirmed by retention time and fragments with gas chromatography and mass spectrometry (GC-MS). The results showed that WMW contained a large amount of Triterpenoids (30-55%) and triterpeneacetal (about 10%-20%), also contain a small amount of aliphatic hydrocarbons. The nanostructure of the WMW was observed using microscopy, and the main superfines could be found. Most of the structure consists of tiny bundles (FIG. 1).

[0059] By experiments we tested whether coatings, including that with WMW, on some common fruits have the effect of protecting the fruit from fungal decay. One experiment was to wrap the lemon with WMW, the experiments were carried out in a well ventilated room at the temperature of 15 to 27 C. with a humidity range of 30% to 90%. The results showed that the coating would prevent the decay effect of lemons by the common rotten fungus green moldisewis type 3. Even partially rotten lemons can be protected by WMW coatings as compared to uncoated samples, as shown in FIGS. 2A-B.

[0060] Similar results have been obtained from the same experiments on oranges, WMW also exerted an antifungal protection on oranges, as shown in FIG. 2C-D.

[0061] Common solvents: 8 common solvents were studied. They are: acetone (A), ethanol (B), cyclohexane (C), dimethyl sulfoxide (D), dichloromethane (E), chloroform (F), mineral oil (G) and distilled water (H). The results showed that distilled water (H) does not dissolve WMW at all. Dimethyl sulfoxide (D), chloroform (F) and mineral oil (G) readily dissolve WMW, the solubilities of WMW in these three solvents were very high. On the other hand, acetone (A), cyclohexane (C) and dichloromethane (E) only partially dissolved WMW, solubilities of WMW were low in these solvents. These results are very useful in the purification of WMW, as well as in researches and WMW applications.

Embodiment 2: Preparation of Artificial WMW Powder

[0062] On the basis of the above research, we prepared artificial WMW powder, named Guaguoxian, which is composed of 40% Betulin (which is illustrated as Formula I in FIG. 1), 15% -Amyrin acetate (which is illustrated as Formula II in FIG. 1), and 45% of beeswax, all by weight of total composition, by spray drying of their solution. The product had an ultrastructure similar to that of natural WMW.

[0063] The synthesized WMW powder was sprayed on the wax gourds with their wax layer removed, and the antiseptic ability of the artificial WMW powder was compared with the natural WMW. The experimental results showed that the artificial WMW powder and the natural WMW had the same preventive effect on the decay of the wax gourd body.

Embodiment 3: Microbiological Analysis of Antibacterial and Antifungal Effects of Artificial WMW Powder

[0064] The antibacterial activities of the artificial WMW powder against Escherichia coli and Staphylococcus aureus were examined according to the method provided by the literature (CRAMP Analogues Having Potent Antibiotic Activity against Bacterial, Fungal, and Tumor Cells without Hemolytic Activity, Shin S Y, Kang S w, Lee D G, Eom S H, Song W K, Kim J I., BiochemBiophys Res Commun, 2000 Sep. 7; 275(3) 904-909). E. coli and S. aureus were cultured at different concentrations of artificial WMW at 37 C. for 20 h. and the minimum inhibitory concentration (MIC) was determined. The results showed that the artificial WMW powder had minimal inhibition concentrations (MIC) against E. coli and S. aureus at 6.4 M and 14 M, respectively.

[0065] The antifungal activity of artificial WMW powder was determined by Aspergillus fumigatus. A. fumigatus and different concentrations of artificial WMW powder were incubated at 37 C. for 20 h, and the minimum inhibitory concentration (MIC) was determined. The results showed that the MIC of artificial WMW powder against A. fumigatus was 32 M.

[0066] The above results indicate that the artificial WMW powder proposes by the present invention has strong antibacterial and fungal activity.

Embodiment 4: Antiseptic Effect of Artificial WMW on Fruits and Grains

[0067] The artificial WMW powder of the present invention is used for the preservation of the following fruits and grains: the Winter melon containing the intact natural WMW layer as the positive control 1 (+), and the sample washed with 0.1% hydrogen peroxide as the positive control 2 (+), the surface The bare melon with the wax powder completely removed as a negative control (): 1. Citrus fruits such as oranges, tangerines, lemons, limes and grapefruits; 2. Other edible fruits such as apples, pears, melons, peaches and others; 3. Grains such as wheat, rice, corn and others; 4. Beans and other crops.

[0068] The results showed that the use of the artificial WMW powder of the present invention can provide a longer protection period.

[0069] It should be noted that the corrosion protection properties of MWM depend mainly on environmental factors such as temperature, humidity and ventilation. The data obtained in the experimental observations were obtained in a well ventilated room at 15 C. to 27 C. and a humidity range of 30% to 90%. However, the use of WMW can increase the storage time of the fruits and crops listed above by 4 to 20 times, with large differences between different fruits and crops.

Embodiment 5: Safety Test of Artificial Winter WMW Powder

[0070] The artificial WMW powder of the present invention was analyzed for its safety by a large number of toxicity tests and tests, the results showed that the artificial WMW powder of the present invention is safe in use. The hemolysis assay of human red blood cells was negative, and the Ames test showed no carcinogenic activity of the present invention.

Embodiment 6: Application of Artificial WMW Powder in the Treatment of Diabetic Wounds

[0071] Using the rat model of diabetic wound infection as described by J J Mendes et al. (Joo J Mendes*, Clara I Leandro, Dolores P Bonaparte, and Andreia L Pinto, Comparative Medicine, Vol 62, No. 1, February 2012, Pages 37-48), the antimicrobial activity of WMW was studied, with beeswax powder as a negative control.

[0072] The experimental results showed that the wound area was reduced by 2030% 910 days after applying WMW, while the negative control group using beeswax powder reduced the wound area by 5%0 or less. This result indicates that WMW treats diabetic wounds effectively.

Embodiment 7: Anti-Microbial Treatment for Diabetic Foot Ulcer

[0073] An amount of 50 mg to 200 mg of purified and sterilized winter melon wax (WMW) powder was filled into a 7 cm9.25 cm rectangular teabag, and the powder was spreader into a thin and even layer inside the teabag. The teabag was closed and readied for application to diabetic foot ulcer.

[0074] The WMW powder bag was placed over the diabetic foot ulcer wound and covered with a slightly larger rectangular protective plaster to securely fix it on the skin. The WMW powder bag can be changed every twenty-four (24) hours. The area of the ulcer wound was then measured every three days for a 21-day treatment regiment.

[0075] In one case, the diabetic foot ulcer wound was treated for 41 days by this protocol to a 64 years old male diabetic patient with foot ulcer. No adverse event was observed during and after the treatment. The area of infection of the diabetic foot ulcer was measured and found to reduced by 12 to 16 percent, suggesting the effective anti-microbial and anti-infectious activity of the WMW powder treatment.

[0076] The skilled person will be aware of a range of possible modifications of the various embodiments described above. Accordingly, the present invention is defined by the claims and their equivalents.