Description:DESCRIPTION:
Field of the invention:
[0001] The present disclosure generally relates to the technical field of pharmaceutical and nutraceutical formulations and, in specific, relates to a method for extracting sulforaphane, from broccoli to determine whether the sulforaphane has antibiotic properties against bacterial cultures at different concentrations.
Background of the invention:
[0002] Sulforaphane is a naturally occurring compound found in cruciferous vegetables such as broccoli, cauliflower, and kale. It has garnered significant attention due to its potent health benefits, including anticancer and antidiabetic properties. The primary source of sulforaphane is the precursor glucoraphanin, which, when acted upon by the enzyme myrosinase during plant damage or chewing, is converted into sulforaphane. This bioactive compound is of particular interest in the field of nutraceuticals and functional foods.

[0003] The sulforaphane acts as a powerful antioxidant, enhances detoxification processes in the body, and has been shown to induce apoptosis in cancer cells. Additionally, the sulforaphane exhibits anti-inflammatory properties and improves insulin sensitivity, making it a valuable compound in managing conditions like diabetes and metabolic syndrome. These properties make sulforaphane a promising candidate for developing natural health products.

[0004] The rise of antibiotic-resistant bacteria poses a significant challenge to public health worldwide. Overuse and misuse of synthetic antibiotics have led to the emergence of multidrug-resistant strains of bacteria, rendering many conventional treatments ineffective. This growing threat underscores the urgent need for alternative antimicrobial agents that can effectively combat bacterial infections without contributing to the development of resistance. The limitations of synthetic antibiotics, there is a renewed interest in exploring natural products for their antimicrobial properties. Natural compounds often have complex structures and mechanisms of action that can be less prone to resistance development. Additionally, they tend to have fewer side effects compared to synthetic drugs. The Sulforaphane, with its well-documented health benefits, presents a promising candidate for examination as a natural antibiotic.

[0005] Currently, the super bacterial antibiotic Zyvox is on the market, but super bacterial antibiotics such as Zyvox have side effects and MRSA (Methicillin-resistant Staphylococcus aureus) bacteria are beginning to develop resistance and the drug efficacy is rapidly falling, the development of super bacterial antibiotics that have a wide effect on super bacteria such as vancomycin, penicillin, methicillin resistant strains are urgently developed.

[0006] In existing methodology, a composition containing sulforaphane for preventing from antibiotics-resistant strain is provided to suppress gram-positive bacteria and gram-negative bacteria having resistance to antibiotics. The composition for preventing from antibiotics-resistant strain contains sulforaphane as an active ingredient. The sulforaphane is artificially synthesized or extracted from plants. The antibiotics is beta lactam or non-beta lactam. The strain is gram-positive or gram negative. However, the composition with sulforaphane does not adequately compare the effectiveness of sulforaphane with established synthetic antibiotics.

[0007] In existing methodology, a simple solid-phase extraction (SPE) method for the determination of the sulforaphane in broccoli has been developed. The optimal conditions are found to be use of a silica SPE cartridge, and ethyl acetate and dichloromethane as washing and eluting solvents, respectively, which could eliminate interferences originating from the broccoli matrix. The extracts are sufficiently clean to be directly injected into high-performance liquid chromatography (HPLC) for further chromatographic analysis. Good linearity is obtained from 0.05 to 200 µg/mL (r = 0.998) for the sulforaphane with the relative standard deviations less than 3.6%. The mean recoveries of the sulforaphane from broccoli are more than 90.8% and the detection limit (S/N = 3:1) is 0.02 µg/mL. The SPE method provides a higher yield of sulforaphane from crude extracts compared to conventional liquid-liquid extraction. However, the SPE method requires high dosage of the sulforaphane to achieve high inhibition of bacterial growth.

[0008] By addressing all the above-mentioned problems, there is a need for a method extracts sulforaphane from broccoli to determine whether the sulforaphane has antibiotic properties against bacterial cultures at different concentrations. There is also a need for a method that compares the effectiveness of sulforaphane with established synthetic antibiotics adequately. There is also a need for a method that develops a natural alternative to synthetic antibiotics to reduce dependency on chemically synthesized drugs.

[0009] There is also a need for a method that extracts sulforaphane from widely available cruciferous vegetables like broccoli to provide a more affordable treatment option for consumers and healthcare providers. There is also a need for a method that reduces side effects, lowers the risk of resistance, provides cost-effective production, provides multifunctional health benefits, and aligns with consumer preferences for natural and sustainable products. There is also a need for a method that prepares the sulforaphane antibiotic medication to enhance the body’s natural immune defenses, thereby providing an additional layer of protection against infections. There is also a need for a method that incorporates the sulforaphane antibiotic medication into various formulations, including capsules, tablets, powders, and topical applications, thereby improving patient convenience and adherence.
Objectives of the invention:
[0010] The primary objective of the present invention is to provide a method a method that extracts sulforaphane from broccoli to determine whether the sulforaphane has antibiotic properties against bacterial cultures at different concentrations.

[0011] Another objective of the present invention is to provide a method that utilizes a dosage of 25 ul of sulforaphane at a concentration of 10 mg/ul of Escherichia coli (E. coli) bacteria to exhibit more than 96% inhibition of bacterial growth.

[0012] The other objective of the present invention is to provide a method that compares the effectiveness of sulforaphane with established synthetic antibiotics adequately.

[0013] The other objective of the present invention is to provide a method that develops a natural alternative to synthetic antibiotics to reduce dependency on chemically synthesized drugs.

[0014] The other objective of the present invention is to provide a method that extracts sulforaphane from widely available cruciferous vegetables like broccoli to provide a more affordable treatment option for consumers and healthcare providers.

[0015] The other objective of the present invention is to provide a method that reduces side effects, lowers the risk of resistance, provides cost-effective production, provides multifunctional health benefits, and aligns with consumer preferences for natural and sustainable products.

[0016] Yet another objective of the present invention is to provide a method that prepares the sulforaphane antibiotic medication to enhance the body’s natural immune deficiency, thereby providing an additional layer of protection against infections.

[0017] Further objective of the present invention is to provide a method that incorporates the sulforaphane antibiotic medication into various formulations, including capsules, tablets, powders, and topical applications, thereby improving patient convenience and adherence.
Summary of the invention:
[0018] The present disclosure proposes a method for extracting antibacterial properties of sulforaphane from broccoli. The following presents a simplified summary in order to provide a basic understanding of some aspects of the claimed subject matter. This summary is not an extensive overview. It is not intended to identify key/critical elements or to delineate the scope of the claimed subject matter. Its sole purpose is to present some concepts in a simplified form as a prelude to the more detailed description that is presented later.

[0019] In order to overcome the above deficiencies of the prior art, the present disclosure is to solve the technical problem to provide a method that extracts sulforaphane from broccoli to determine whether the sulforaphane has antibiotic properties against bacterial cultures at different concentrations.

[0020] According to one aspect, the invention provides a method for extracting the antibacterial properties of sulforaphane. At one step, broccoli is pulverized and the pulverized broccoli is dried to obtain a fine powder. The broccoli is pulverized by using grinding tools. At one step, the fine powder is mixed with 40 ml of an acidic solution to obtain a mixture. The acidic solution is a hydrochloric acid (HCl) with a pH value of 3.

[0021] At one step, the mixture with an organic solvent is extracted for at least three times, thereby separating an organic mixture from an aqueous mixture. The mixture is extracted with the organic solvent for at least three times. The organic solvent is dichloromethane.

[0022] At one step, the organic mixture with a drying agent is dried at a temperature at least 27 °C for evaporating the organic solvent from dried organic mixture to extract sulforaphane. The drying agent is anhydrous sodium sulfate. The organic mixture is dried with the drying agent at a room temperature upon detecting moisture.

[0023] At one step, the sulforaphane is dissolved in a dimethyl sulfoxide (DMSO) and drying the dissolved sulforaphane at the room temperature, thereby collecting dried sulforaphane at one or more concentrations. The sulforaphane is formulated for oral administration. At one step, the antibiotic test is conducted on a gram negative bacteria using the one or more concentrations of collected sulforaphane. The gram negative bacteria include Escherichia coli (E. coli) bacteria. The one or more concentrations of the collected sulforaphane includes 10 ul and 25 ul. The 25 ul of collected sulforaphane at 10mg/ul concentration of E. coli bacteria exhibits at least 96 percent of inhibition of bacterial growth.

[0024] At one step, the antibiotic test is conducted on a gram positive bacteria using the one or more concentrations of collected sulforaphane. The gram positive bacteria include Lactobacillus bacteria. The one or more concentrations of the collected sulforaphane includes 20 ul and 40 ul. The 40 ul of collected sulforaphane at 0.033 mg/ul concentration of Lactobacillus bacteria exhibits at least 74.8 percent of inhibition of bacterial growth.

[0025] Further, objects and advantages of the present invention will be apparent from a study of the following portion of the specification, the claims, and the attached drawings.
Detailed description of drawings:
[0026] The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate an embodiment of the invention, and, together with the description, explain the principles of the invention.

[0027] FIG.1 illustrates a flow chart of a method for extracting the antibacterial properties of sulforaphane, in accordance to an exemplary embodiment of the invention.

[0028] FIG. 2 illustrates a schematic representation of the method for extracting the antibacterial properties of sulforaphane, in accordance to an exemplary embodiment of the invention.

[0029] FIGs. 3A-3B illustrate pictorial representations of an antibiotic test on Escherichia coli (E. coli) bacteria using at least one concentration of sulforaphane, in accordance to an exemplary embodiment of the invention.

[0030] FIGs. 4A-4B illustrate pictorial representations of the antibiotic test on Lactobacillus bacteria using at least one concentration of sulforaphane, in accordance to an exemplary embodiment of the invention.
Detailed invention disclosure:
[0031] Various embodiments of the present invention will be described in reference to the accompanying drawings. Wherever possible, same or similar reference numerals are used in the drawings and the description to refer to the same or like parts or steps.

[0032] The present disclosure has been made with a view towards solving the problem with the prior art described above, and it is an object of the present invention to provide a method that extracts sulforaphane from broccoli to determine whether the sulforaphane has antibiotic properties against bacterial cultures at different concentrations.

[0033] According to one exemplary embodiment of the invention, FIG. 1 refers to a flow chart 100 of a method for extracting antibacterial properties of sulforaphane. The method prepares the sulforaphane antibiotic medication to enhance the body’s natural immune deficiency, thereby providing an additional layer of protection against infections. The method extracts sulforaphane from widely available cruciferous vegetables like broccoli to provide a more affordable treatment option for consumers and healthcare providers. The method for extracting the antibacterial properties of sulforaphane. At step 102, broccoli is pulverized and the pulverized broccoli is dried to obtain a fine powder. The broccoli is pulverized by using grinding tools. At step 104, the fine powder is mixed with 40 ml of an acidic solution to obtain a mixture. The acidic solution is a hydrochloric acid (HCl) with a pH value of 3.

[0034] At step 106, the mixture with an organic solvent is extracted for at least three times, thereby separating an organic mixture from an aqueous mixture. The mixture is extracted with the organic solvent for at least three times. The organic solvent is dichloromethane.

[0035] At step 108, the organic mixture with a drying agent is dried at a temperature at least 27 °C for evaporating the organic solvent from dried organic mixture to extract sulforaphane. The drying agent is anhydrous sodium sulfate. The organic mixture is dried with the drying agent at a room temperature upon detecting moisture.

[0036] At step 110, the sulforaphane is dissolved in a dimethyl sulfoxide (DMSO) and drying the dissolved sulforaphane at the room temperature, thereby collecting dried sulforaphane at one or more concentrations. The sulforaphane is formulated for oral administration. At step 112, the antibiotic test is conducted on a gram negative bacteria using the one or more concentrations of collected sulforaphane. The gram negative bacteria include Escherichia coli (E. coli) bacteria. The one or more concentrations of the collected sulforaphane includes 10 ul and 25 ul. The 25 ul of collected sulforaphane at 10mg/ul concentration of E. coli bacteria exhibits at least 96 percent of inhibition of bacterial growth.

[0037] At step 114, the antibiotic test is conducted on a gram positive bacteria using the one or more concentrations of collected sulforaphane. The gram positive bacteria include Lactobacillus bacteria. The one or more concentrations of the collected sulforaphane includes 20 ul and 40 ul. The 40 ul of collected sulforaphane at 0.033 mg/ul concentration of Lactobacillus bacteria exhibits at least 74.8 percent of inhibition of bacterial growth.

[0038] According to another exemplary embodiment of the invention, FIG. 2 refers to a schematic representation 200 of the method for extracting the antibacterial properties of sulforaphane containing natural extracts from the broccoli. At step 202, the broccoli is taken. At step 204, the broccoli is pulverized, thereby drying the pulverized Broccoli and make it into the fine powder using grinding tools. At step 206, 10 g of resultant powder is taken with 40 ml of pH 3 of HCl.

[0039] At step 208, the resultant mixture is extracted 3 times with 40 ml of Dichloromethane i.e., separation of the organic mixture from the aqueous mixture. At step 210, the organic mixture is dried with Anhydrous Sodium Sulphate at temperature between 40°C and 60°C. At step 212, the organic mixture should be dried again at the room temperature if moisture is still detected to ensure complete removal of water. At step 214, the organ solvent from the dried organic mixture is evaporated to extract the sulforaphane. At step 216, the dried extract of the sulforaphane is dissolved in the dimethyl sulfoxide (DMSO) and drying the dissolved sulforaphane at the room temperature. At step 218, the one or more concentrations of sulforaphane are taken for further antibiotic testing process in an agar plate.

[0040] According to another exemplary embodiment of the invention, FIGs. 3A-3B refer to pictorial representations (300, 302) of an antibiotic test on Escherichia coli (E. coli) bacteria using at least one concentration of sulforaphane. The antibiotic test is conducted on the E.coli bacteria and here are the results, the sulforaphane is used at a concentration of 25 mg/ul as shown in FIG. 3A. The sulforaphane is used at a concentration of 10 mg/ul as shown in FIG. 3B. The one or more concentrations of the collected sulforaphane includes 10 ul and 25 ul. The 25 ul of collected sulforaphane at 10 mg/ul concentration of E. coli bacteria exhibits at least 96 percent of inhibition of bacterial growth. The Antibiotic test is performed using the laboratory E. coli bacterial cells. The 25 ul of collected sulforaphane prevents effective bacterial growth when compared with the 10 ul of collected sulforaphane.

[0041] In one embodiment herein, the Escherichia coli (E. coli) bacteria is genus of the gram-negative bacteria. The E. coli bacteria that are commonly found in the lower intestine of warm-blooded organisms. The E. coli bacteria causes diarrhea and food poisoning, or more serious illnesses like pneumonia. The E. coli infection spreads through contact with contaminated food or water, or by contact with people who are sick. The E. coli infection is reduced by avoiding risky foods and practicing good hygiene.

[0042] According to another exemplary embodiment of the invention, FIGs. 4A-4B refer to pictorial representations (400, 402) of the antibiotic test on Lactobacillus bacteria using at least one concentration of sulforaphane. The antibiotic test is conducted on the Lactobacillus bacteria and here are the results, the sulforaphane is used at a concentration of 20 mg/ul as shown in FIG. 4A. The sulforaphane is used at a concentration of 40 mg/ul as shown in FIG. 4B. The one or more concentrations of the collected sulforaphane includes 20 ul and 40 ul. The 40 ul of collected sulforaphane at 0.033 mg/ul concentration of Lactobacillus bacteria exhibits at least 74.8 percent of inhibition of bacterial growth. The Antibiotic test is performed using the laboratory Lactobacillus bacterial cells. The 40 ul of collected sulforaphane prevents effective bacterial growth when compared with the 20 ul of collected sulforaphane.

[0043] In one embodiment herein, the Lactobacillus bacteria is a genus of lactic acid bacteria and gram positive bacteria. The Lactobacillus bacteria is commonly found in yogurt and fermented foods, The Lactobacillus bacteria prevents or treats infectious diarrhea in children and adults. The Lactobacillus bacteria is most effective in treating rotavirus in children and campylobacter infections in adults.

[0044] Numerous advantages of the present disclosure may be apparent from the discussion above. In accordance with the present disclosure a method for extracting antibacterial properties of sulforaphane from broccoli is disclosed. The proposed invention provides a method that extracts sulforaphane from broccoli in order to find whether the sulforaphane have antibiotic nature on the bacterial cultures at different concentrations. The method utilizes less dosage of sulforaphane at some concentration of Escherichia coli (E. coli) bacteria to exhibit higher inhibition of bacterial growth. The method compares the effectiveness of sulforaphane with established synthetic antibiotics adequately. The method that develops a natural alternative to synthetic antibiotics to reduce dependency on chemically synthesized drugs.

[0045] The proposed invention provides the method that extracts sulforaphane from widely available cruciferous vegetables like broccoli to provide a more affordable treatment option for consumers and healthcare providers. The method reduces side effects, lowers the risk of resistance, provides cost-effective production, provides multifunctional health benefits, and aligns with consumer preferences for natural and sustainable products. The method prepares the sulforaphane antibiotic medication to enhance the body’s natural immune deficiency, thereby providing an additional layer of protection against infections. The method incorporates the sulforaphane antibiotic medication into various formulations, including capsules, tablets, powders, and topical applications, thereby improving patient convenience and adherence.

[0046] It will readily be apparent that numerous modifications and alterations can be made to the processes described in the foregoing examples without departing from the principles underlying the invention, and all such modifications and alterations are intended to be embraced by this application.
, Claims:CLAIMS:
I/We Claim:
1. A method for extracting the antibacterial properties of sulforaphane, comprising:
pulverizing broccoli and drying pulverized broccoli to obtain a fine powder;
mixing 10 g of the fine powder with 40 ml of an acidic solution to obtain a mixture;
extracting the mixture with an organic solvent for at least three times, thereby separating an organic mixture from an aqueous mixture;
drying the organic mixture with a drying agent at a temperature of at least 27 °C for evaporating the organic solvent from dried the organic mixture to extract sulforaphane;
dissolving the sulforaphane in a dimethyl sulfoxide (DMSO) and drying the dissolved sulforaphane at a room temperature, thereby collecting dried sulforaphane at one or more concentrations;
conducting an antibiotic test on a gram negative bacteria using one or more concentrations of collected sulforaphane, wherein the one or more concentrations of the collected sulforaphane includes 10 ul and 25 ul,
wherein the 25 ul of collected sulforaphane at 10 mg/ul concentration of the gram negative bacteria exhibit at least 96 percent of inhibition of bacterial growth; and
conducting an antibiotic test on a gram positive bacteria using one or more concentrations of collected sulforaphane, wherein the one or more concentrations of the collected sulforaphane includes 20 ul and 40 ul;
wherein the 40 ul of collected sulforaphane at 0.033 mg/ul concentration of the gram positive bacteria exhibit at least 74.8 percent of inhibition of bacterial growth.
2. The method as claimed in claim 1, wherein the broccoli is pulverized by using grinding tools.
3. The method as claimed in claim 1, wherein the acidic solution is a hydrochloric acid (HCl) with a pH value of 3.
4. The method as claimed in claim 1, wherein the mixture is extracted with the organic solvent for at least three times.
5. The method as claimed in claim 1, wherein the organic solvent is dichloromethane.
6. The method as claimed in claim 1, wherein the drying agent is anhydrous sodium sulfate.
7. The method as claimed in claim 1, wherein the organic mixture is dried with the drying agent at the room temperature upon detecting moisture.
8. The method as claimed in claim 1, wherein the gram negative bacteria include Escherichia coli (E. coli) bacteria.
9. The method as claimed in claim 1, wherein the gram positive bacteria includes Lactobacillus bacteria.
10. The method as claimed in claim 1, wherein the sulforaphane is formulated for oral administration.