This invention relates to a method of water hyacinth flower extracts as a source of anti-oxidants and antimicrobial agents.
Background of the Invention
AU2016325872B2 Food packaging material comprising a polymeric material and a natural antioxidant. The natural antioxidant may be extracted, isolated and/or derived from plant material. A method for forming food packaging material is also provided, comprising forming a mixture comprising a natural antioxidant and a polymeric resin and processing the mixture to form a food packaging material.
Research Gap: No use of flower of water hyacinth reported
CN102458129B Provided are antimicrobial compositions comprising one or more compound components generally recognized as safe for human consumption, and related methods of use, such compositions and methods as can be employed in a wide range of agricultural, industrial, building, pharmaceutical and/or personal care products and applications.
Research Gap: No use of antimicrobial activity of flower of water hyacinth reported
None of the prior art indicate above either alone or in combination with one another disclose what the present invention has disclosed. Present invention is a method of water hyacinth flower extracts as a source of anti-oxidants and antimicrobial agents.
SUMMARY OF THE INVENTION
This summary is provided to introduce a selection of concepts, in a simplified format, that are further described in the detailed description of the invention.
This summary is neither intended to identify key or essential inventive concepts of the invention and nor is it intended for determining the scope of the invention.
To further clarify advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which is illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings.
Eicchornia Crassipes is a monocotyledonous aquatic plant endemic to the Amazon basin. It is a member of the plantae kingdom and the Pontederiaceae family, which is related to the lily family.This plant is found all over the world including India,Africa and many European countries.
It may reach a maximum height of 1 metre above the surface of the water. Leaves are 10-20 cm wide on a stalk and float above the water surface due to buoyant bulb-like nodules at their base. Purple black roots are puffy and hangs freely. Six-petaled flowers vary in colour from lavender to pink. The lavender flowers and sparkling brilliant leaves identify the "beautiful blue devil" water hyacinth, which spreads rapidly.
Although the water hyacinth is usually considered a weed, it takes harmful minerals and organic wastes from the water and so cleanses it. Simultaneously, it is seen as a nuisance, impeding human activity and hampering the efficient use of water resources for different services.
Several studies have found that water hyacinth is a source of various chemical agents with medicinal properties. Phytochemicals found in the leaf extract of E. Crassipes plant include flavonoids (plant pigments), alkaloids (plant chemicals), tannins (plant tannins), and phenols (plant phenols), all of which have biological activities such as antiviral, antifungal, antitumor, and antibacterial agents. In addition, water hyacinth contains a high concentration of oxidative enzymes as well as nonenzymatic antioxidant systems. Researchers are becoming more interested in the antimicrobial and antifungal activities of medicinal plants rather than in common weeds, which are also a source of several antimicrobial and antifungal compounds, because of alternative approach for the detection and development of new and innovative products
DETAILED DESCRIPTION OF THE INVENTION
The detailed description of various exemplary embodiments of the disclosure is described herein with reference to the accompanying drawings. It should be noted that the embodiments are described herein in such details as to clearly communicate the disclosure. However, the amount of details provided herein is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the scope of the present disclosure as defined by the appended claims.
It is also to be understood that various arrangements may be devised that, although not explicitly described or shown herein, embody the principles of the present disclosure. Moreover, all statements herein reciting principles, aspects, and embodiments of the present disclosure, as well as specific examples, are intended to encompass equivalents thereof.
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments. As used herein, the singular forms “a",” “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprises,” “comprising,” “includes” and/or “including,” when used herein, specify the presence of stated features, integers, steps, operations, elements and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components and/or groups thereof.
It should also be noted that in some alternative implementations, the functions/acts noted may occur out of the order noted in the figures. For example, two figures shown in succession may, in fact, be executed concurrently or may sometimes be executed in the reverse order, depending upon the functionality/acts involved.
In addition, the descriptions of "first", "second", “third”, and the like in the present invention are used for the purpose of description only, and are not to be construed as indicating or implying their relative importance or implicitly indicating the number of technical features indicated. Thus, features defining "first" and "second" may include at least one of the features, either explicitly or implicitly.
Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which example embodiments belong. It will be further understood that terms, e.g., those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.
These and other advantages of the present subject matter would be described in greater detail with reference to the following figures. It should be noted that the description merely illustrates the principles of the present subject matter. It will thus be appreciated that those skilled in the art will be able to devise various arrangements that, although not explicitly described herein, embody the principles of the present subject matter and are included within its scope.
The current research comprises investigating the antioxidant and antibacterial properties of extracts from flower of Eicchornia Crassipes (commonly known as Water Hyacinth).
In this study, the antioxidant and anti-microbial activities of water hyacinth are being investigated in its flowers.
According to the claims, flower extract has significant anti-oxidative and anti-microbial properties, making it suitable for usage as a herbal medication to provide anti-oxidant and anti-ageing properties.
Many work has been done on water hyacinth but no one till now has reported about the use of water hyacinth flower extract as food packaging or herbal medication material to provide anti-oxidant and anti-ageing properties.
Best Method of working:
Material and Methods
Assan Barrage in Uttarakhand, India, was visited by researcher to collect fresh, disease-free leaves of the plant E. crassipes. The leaves were thoroughly washed three times with running water and once with sterile distilled water before being dried in the open air to remove any remaining moisture.
Methanol (Technical Grade) and Acetone was used for solvent extraction. All the microorganisms for activity were taken from Biotech Department, School of Applied and Life Sciences (SALS).
Plant extracts
The flowers of the plant were dried at room temperature before grinding them into a fine powder with a particle mesh size of one millimetre in a mechanical grinder. Process was used to extract two portions (10 g each) of the powdered plant at room temperature using two solvents: acetone (3 x 100 mL) and analytical grade methanol 80 percent (3 x 100 mL). The extracts from each solvent were mixed and allowed to evaporate and dry under reduced pressure at 45°C in order to create aqueous and hydro-methanolic extracts that contained less solvent than the original extracts. The yields of extraction for the acetonic and methanolic extracts were determined to be 1.2 and 2.9 percent, respectively, for the two types of extracts.
Chemical analyses of E. crassipes flower were carried out in triplicate.
Results and Discussion
Antioxidant Activity
The free radical-scavenging activity was measured by 1,1 diphenyl-2-picryl hydrazyl (DPPH). For this purpose, 1ml of DDPH solution (0.1 mM) in methanol was added to the extract. Thirty minutes later, absorbance was measured at 517 nm using a UV-Visible spectrophotometer. A control was prepared without adding extract. Ascorbic acid at various concentrations (50-250µg mL-1) was used as a standard. The percentage of DPPH-scavenging activity was calculated as DPPH Scavenged (%) = A Control – A Test/ A Control X 100, where A control represents absorbance of control reaction and A test represents absorbance in presence of the extract.
DPPH Scavenging Assay
Concentration (µg mL-1) Methanol Extract Acetone Extract Ascorbic Acid
50 41 45 53
100 60 62 69
150 65 68 72
200 70 71 79
250 78 79 83

MIC (minimum inhibitory concentration)
Minimum inhibitory concentration (MIC) is defined as the lowest concentration that resulted in the preservation or decrease of inoculum viability according to current standards. This approach was used to measure the minimum inhibitory concentration (MIC) of the extracts of E. crassipes against bacteria in extracts. The stock solution was prepared by dissolving the extract (0.512 mg) in 2 mL distilled water (2 tween-80 drops were added to aid dissolution). Using test organism suspensions (107 organisms per mL), 1 drop of suspension (0.02 mL) was added to each broth dilution after it had been prepared. After an 18-hour incubation period at 37 degrees Celsius, the tubes were checked for growth. The minimum inhibitory concentration (MIC) of the extract was determined as the lowest concentration at which no growth was observed. Growing cultures were seen in the tubes containing lower concentrations of the extract than the inhibitory threshold, and the broth medium was noted to be muddy in appearance.
MIC (minimum inhibitory concentration) (µg mL-1)
Bacteria Methanol Extract Acetone Extract
S. Aureus 22 18
Pseudomonas Aeruginosa 32 41
Bacillus Subtilis 18 17

Antimicrobial Or Antibacterial Activity
It is very crucial in any food packaging material in order to maintain the sanitation of the food that has been packaged in the container. Moreover, herbal ointment too needs to be effective against various bacterias and micro-organisms. Active packaging prevents the development of germs, bacteria, or fungus on the surface of food as a result of direct contact between the packing material and the food product. Efficacy of extracted substance was tested against S. Aureus, Pseudomonas Aeruginosa and Bacillus Subtilis.
Into each sterile petriplate, a layer of the sterilised nutrient-agar and potato dextrose-agar mixture was poured and allowed to form. Fresh bacterial and fungal cultures were dispersed across the plates using sterile cotton swabs in accordance with the spread plate procedure, which was followed. With the use of a sterile cork borer, one 5 mm well was created in the agar plates for each sample. The wells were filled with 200 mL of flower extracts of E. crassipes. For bacteria, the plates were incubated for 48 hours at 350 Celsius. Each experiment was carried out five times, and the mean diameter of the inhibition zones surrounding the wall in millimetres (mm) was determined for each.
Zone of Inhibition (diameter in mm)
Bacteria Methanol Extract Acetone Extract Control Streptomycin
S. Aureus 16.2 14.1 18.2
Pseudomonas Aeruginosa 14.4 13.2 15
Bacillus Subtilis 13.2 12.1 14.2

ADVANTAGES OF THE INVENTION:
Extracts of flower of plant water hyacinth can be produced in ethanolic/methanolic solvent and its extract can be used for antimicrobial, antibacterial and anti-oxidant property.
• Antioxidant Property by SOD/DPPH or any other method
• Antioxidant property of flower extract of Water Hyacinth is highly significant and can be used as free radical scavenger.
• Anti-microbial property of flower extract of water hyacinth is more as compared to other extracts.

We Claim:

1. A method of water hyacinth flower extracts as a source of anti-oxidants and antimicrobial agents comprising the steps of:
a) drying the flowers of the plant at room temperature before grinding them into a fine powder with a particle mesh size of one millimetre in a mechanical grinder;
b) Processing used to extract two portions (10 g each) of the powdered plant at room temperature using two solvents: acetone (3 x 100 mL) and analytical grade methanol 80 percent (3 x 100 mL);
c) mixing the extracts from each solvent and allowed to evaporate and dry under reduced pressure at 45°C in order to create aqueous and hydro-methanolic extracts that contained less solvent than the original extracts; The yields of extraction for the acetonic and methanolic extracts were determined to be 1.2 and 2.9 percent, respectively, for the two types of extracts;
wherein Chemical analyses of E. crassipes flower are carried out in triplicate.
2. The method as claimed in claim 1, wherein the free radical-scavenging activity is measured by 1,1 diphenyl-2-picryl hydrazyl (DPPH); For this purpose, 1ml of DDPH solution (0.1 mM) in methanol is added to the extract; Thirty minutes later, absorbance is measured at 517 nm using a UV-Visible spectrophotometer;
Wherein ascorbic acid at various concentrations (50-250µg mL-1) is used as a standard.
3. The method as claimed in claim 1, wherein for Antimicrobial Or Antibacterial Activity; each sterile petriplate, a layer of the sterilised nutrient-agar and potato dextrose-agar mixture was poured and allowed to form; and Fresh bacterial and fungal cultures were dispersed across the plates using sterile cotton swabs in accordance with the spread plate procedure, which was followed; wherein with the use of a sterile cork borer, one 5 mm well was created in the agar plates for each sample; and the wells are filled with 200 mL of flower extracts of E. crassipes; For bacteria, the plates are incubated for 48 hours at 350 Celsius; and each experiment is carried out five times, and the mean diameter of the inhibition zones surrounding the wall in millimetres (mm) is determined for each.