Description:Field of the invention
The invention leads to green synthesis of zinc oxide nanoparticles using Stellaria aquatica leaves extract and their antifungal activity.
Background of the Invention
References which are cited in the present disclosure are not necessarily prior art and therefore their citation does not constitute an admission that such references are prior art in any jurisdiction. All publications, patents and patent applications herein are incorporated by reference to the same extent as if each individual or patent application was specifically and individually indicated to be incorporated by reference.
Several patents have been issued for zinc oxide nanoparticles but none of these are related to the present invention. For example, US9428399B1 relates to a synthesis of nanoparticles of metals and metal oxides using plant seeds extract involves providing a solution comprising a metal ion; providing a plant seeds extract solution that comprises a reducing agent; and combining the metal ion solution and the plant extract solution while stirring at room temperature to produce metal nanoparticles. The plant extract is obtained from Trigonella foenum-graecum seeds, and the metal ion is selected from the group consisting of silver (Ag), gold (Au) and zinc (Zn).
Another patent, US9999600B2 provides a nanoparticle compositions including a plurality of nanoparticles, each of which is comprised of a biodegradable or biocompatible polymer arranged in a nanoparticle structure defining an internal lumen and an external surface and one or more of a preparation of hydrophilic cellular components and a preparation of hydrophobic cellular components. In some embodiments, the preparation of hydrophilic cellular components is encapsulated within the internal lumen and the preparation of hydrophobic cellular components is associated with the external surface. Various methods of making and using disclosed nanoparticle compositions are also provided.
Another patent, US8361324B1 provides a simple, room-temperature process of using zinc oxide nanoparticles was established by reacting zinc nitrate hexahydrate and cyclohexylamine (CHA) in either aqueous or ethanol medium. Particles of polyhedra morphology were obtained for zinc oxide, prepared in EtOH (ZnOE) and zinc oxide prepared in water (ZnOW). The results indicate that there are significant morphological differences between ZnOE and ZnOW. ZnOE showed a regular polyhedral shape, while spherical and chunky particles were observed for ZnOW. The morphology was crucial in enhancing the cyanide ion photocatalytic degradation efficiency of ZnOE by a factor of 1.5 in comparison to the efficiency of ZnOW at equivalent loading of 0.0166 ZnO nanoparticles wt %.
Another patent, US20080248289A1 provides zinc oxide nanoparticles having an average particle size in the range from 3 to 50 nm, dispersed in an organic solvent, according to one or more of claims 1 to 6, characterised in that in a step a) one or more precursors of the nanoparticles are converted into the nanoparticles in an alcohol, in a step b) the growth of the nanoparticles is terminated by addition of at least one modifier, which is a precursor of silica, when the absorption edge in the UV/VIS spectrum of the reaction solution has reached the desired value, in a step c) the silica coating is modified by addition of at least one further surface modifier selected from the group consisting of organofunctional silanes, quaternary ammonium compounds, phosphonates, phosphonium and sulfonium compounds or mixtures thereof, and optionally, in step d), the alcohol from step a) is removed and replaced by another organic solvent, to isolated nanoparticles, and to the use thereof for UV protection in polymers.
Another patent, US10364508B2 provides a method for producing single crystalline zinc oxide nanoparticles that is capable of mass production includes mixing, between processing surfaces which are disposed in a position facing each other so as to be able to approach and separate from each other and rotate relative to each other, a zinc oxide separating solvent prepared by homogeneously mixing an acidic substance with a solvent containing at least alcohol and a raw material solution obtained by mixing a zinc oxide nanoparticle raw material with a basic solvent or a raw material solution that is basic as a result of mixing and dissolving a zinc oxide nanoparticle raw material with and into a solvent, and discharging a mixed fluid in which zinc oxide nanoparticles have separated out from between the processing surfaces. The zinc oxide separating solvent and the raw material solution are mixed between the processing surfaces so that the mixed fluid becomes basic, and zinc oxide nanoparticles are generated by an acid-base reaction due to mixing of the acidic substance and the basic solvent.
The invention generally relates to the field of chemistry. More specifically, it relates to green synthesis of zinc oxide Nano-particles using Stellaria aquatica plant extract and its biological activity. This invention generally relates to the field of biotechnology and nanotechnology. In this invention various parameters have been worked out for the green synthesis with satisfactory results. This invention provides the novel report on green synthesis of zinc oxide (ZnO) nanoparticles using Stellaria aquatica leaves extract and their antifungal activity. Nanotechnology is highly interdisciplinary, involving physics, chemistry, biology, materials science, and the full range of the engineering disciplines. The word nanotechnology is widely used as shorthand to refer to both the science and the technology of this emerging field. Development of nanotechnology is very active globally, and nanotechnologies are already used in hundreds of products, including sunscreens, cosmetics, textiles, and sports equipment. Nanotechnology is also being developed for use in drug delivery, biosensors, and other biomedical applications. Further, nanotechnologies are also being developed for use in environmental applications, e.g., clean-up of environmental pollutants.
Green syntheses or green methods simply meant synthesis of metal NPs using plants or plant parts or using their extracts, an alternative to chemical and physical methods. Green synthesis is an emerging area in the combined fields of biotechnology and nanotechnology and delivers loads of economic and environmental benefits. Green synthesis though has certain disadvantages such as time-consuming process and reproducibility, yet this approach is very much required to avoid the formation of undesirable or harmful byproducts through the build-up of reliable, sustainable, and eco-friendly synthesis procedures. Green synthesis of NPs aims at minimizing waste generation and implementing sustainable processes.
Stellaria aquatica is a species of flowering plant in the carnation family Caryophyllaceae, known as water chickweed or giant chickweed. It is a perennial herbaceous dicot plant with stems between 10 and 100 cm. high, growing in humid, wet areas, for example, on the sides of gutters or under or between bushes. It occurs naturally in the temperate regions of central and western Europe. The flowers are white. The plants bloom between June and August. Different parts of the plant have been used to treat various gastrointestinal disorders, asthma, diarrhoea, measles, jaundice, renal, digestive, reproductive and respiratory tracts inflammations. They also lessen swelling and used as plasters for broken bones. Stellaria aquatica contains vitamins A, D, B complex, C, Rutin (a bioflavonoid), calcium, potassium, phosphorus, zinc, manganese, sodium, copper, iron and silica.
Zinc oxide nanoparticles (ZnO NPs), as one of the most important metal oxide nanoparticles, are popularly employed in various fields due to their peculiar physical and chemical properties. Zinc oxide (ZnO) nanoparticles (NPs) are a promising platform for use in biomedical research, especially given their anticancer, antifungal and antimicrobial activities. These activities are associated with the ability of ZnO NPs to generate reactive oxygen species (ROS) and induce apoptosis. In addition, ZnO NPs have been successfully exploited as drug carriers for loading and transporting drugs to target sites, thereby reducing unwanted toxicity and off-target effects, and resulting in amplified synergistic effects. The powder ZnO is widely used as an additive in numerous materials and products including ceramics, glass, cement, rubber (e.g., car tyres), lubricants, paints, ointments, adhesives, plastics, sealants, pigments, foods (source of Zn nutrient), batteries, ferrites, and fire retardants. In the Earth crust ZnO is present as zincite mineral but mostly ZnO used for commercial purposes is produced synthetically.
The primary object of the present invention is.
OBJECTIVE OF THE INVENTION: -
• Green synthesis of zinc oxide nanoparticles using Stellaria aquatica leaves extract and their antifungal activity.
• Novel process for the synthesis of zinc oxide nanoparticles using Stellaria aquatica leaves extract.
• In vitro anticancer activity of green synthesized zinc oxide (ZnO) nanoparticles of Stellaria aquatica Applications of ZnO Nanoparticles.
ZnO NPs, as a new type of the low-cost and low-toxicity nanomaterial, have attracted tremendous interest in various biomedical fields, including anticancer, antifungal, antibacterial, antioxidant, antidiabetic, and anti-inflammatory activities, as well as for drug delivery and bioimaging applications.
These and other objects and advantages of the present invention will become readily apparent from the following detailed description.
Summary of Invention
This summary is not a comprehensive overview of the disclosure and does not reflect the main/essential features of the establishment or specify the scope of the establishment. Its sole purpose is to present some of the concepts presented here in a simpler way as a precursor to more detailed explanations presented later.
The primary object of the present invention is green synthesis of zinc oxide nanoparticles using Stellaria aquatica leaves extract and their antifungal activity.
In some embodiments of the present invention, ZnO NPs are exciting inorganic materials. ZnO NPs can be used in various sectors, such as energy conservation, textiles, electronics, healthcare, catalysis, cosmetics, semiconductors, and chemical sensing.
In some embodiments of the present invention, the NPs are nontoxic and biocompatible and display excellent biomedical applications, such as anticancer, anti-inflammatory, and antimicrobial properties, in targeted drug delivery, wound healing, and bioimaging.
In some embodiments of the present invention, zinc is needed by plants for a variety of metabolic and physicochemical processes. It's one of the most crucial trace elements for plant growth. Through the present work carried out, a method for green synthesis of ZnO NPs from plant of Stellaria aquatica was developed. In this study, the protocol selected for synthesis of ZnO NPs helped to develop targeted nanoparticles and the efficient use of various tools facilitated their characterization.
In some embodiments of the present invention, the synthesized nanoparticles have great properties. Zinc oxide nanoparticles (ZnO NPs) are unique in that they can be produced with high surface areas and with unusual crystal structures. Compared to organic materials, inorganic materials such as ZnO possess superior durability, greater selectivity, and heat resistance.
In some embodiments of the present invention, moreover, zinc is a mineral element essential to human health and ZnO is a form in the daily supplement for zinc. ZnO NPs also have good biocompatibility to human cells. In our study, we reported the synthesis of ZnO NPs using Stellaria aquatica plant extract and their antifungal activities.
In some embodiments of the present invention, the green synthesis technique is bene?cial for plant-based fabrication. Due to less toxicity and chemical-free nature, and signi?cant biological activities, the ZnO NPs synthesized from plant extract of Stellaria aquatica can be considered as bio-nanomaterial. Hence, these ZnO NPs are safe and can be used for medicinal purposes by pharmacological industries.
These and other aspects of the embodiments herein will be better appreciated and understood when considered in concurrence with the following explanation and the accompanying drawings. It should be understood, however, that the following descriptions, while indicating preferred embodiments and numerous specific details thereof, are given by way of illustration and not of limitation. Many changes and modifications may be made within the scope of the embodiments herein without departing from the spirit thereof, and the embodiments herein include all such modifications.
Brief summary of the figures
Figure 1: Collection and preparation of extract from Stellaria aquatica leaves.
Figure 2: Visual observation of ZnO NPs synthesis (a) Extract (b) zinc acetate dihydrate (c) Final color change.
Figure 3: DLS pattern of ZnO NPs.
Figure 4: Antifungal effect of ZnO NPs against pathogenic fungus
Brief summary of the tables
Table 1. Antifungal activity
Detailed Description
These embodiments are described in sufficient detail to enable those skilled in the art to practice the embodiments and all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
In some embodiments of the present invention, Precursor solution was prepared (0.1M) zinc acetate dihydrate. 70 ml of this solution was added to 30 ml of the extract under constant stirring in magnetic stirrer for 3 hours at 50-80? temperature.
In some embodiments of the present invention, the pH of the solution was adjusted between 10-11 using 0.1M NaOH. An intense white coloured precipitate immediately formed. Bio reduction mechanism of these extracts for the synthesis of ZnO NPs was investigated through visual examination (color change), and DLS techniques.
In some embodiments of the present invention, Plant extract 50mg/ml (prepared in 10% DMSO), were added in sterilized potato dextrose agar. A 6mm diameter of the actively growing mycelium disc of the pathogen of 6-7 days old culture (Fusarium spp) was placed in the center of the Petri dish. Plates without extract served as negative control and 5mg/ml of Hygromycin as the positive control.
In some embodiments of the present invention, plates without extract served as negative control. Fusarium spp were incubated at 25°C for 6 days and circular growth of mycelium was measured after 6 days (Fusarium spp) of incubation. After incubation, the circular growth of mycelium was measured. The growth results were compared with the negative control.
In some embodiments of the present invention, dynamic Light Scattering (DLS) is an established and precise measurement technique for characterizing particle sizes in suspensions and emulsions. It is based on the Brownian motion of particles - this states that smaller particles move faster, while larger ones move slower in a liquid. It is a technique suited to the analysis and characterization of nanoparticles. DLS analysis showed the size distribution of particles with maximum intensity at 20-40nm.
In some embodiments of the present invention, the effect of Stellaria aquatica ZnO NPs on different fungus has been investigated, but there is no fungal study related to ZnO NPs synthesized from Stellaria aquatica. The diameter of zone of inhibition was observed with ZnO NPs for Fusarium spp was 55.2%.
A zinc oxide nanoparticles using Stellaria aquatica leaves extract consists of:
Stellaria aquatica plant extract, zinc acetate dihydrate, NaOH.
A method of green synthesis of zinc oxide nanoparticles using Stellaria aquatica leaves extract and their antifungal activity comprising the steps of:
adding 10gm of Stellaria aquatica to 150 ml distilled water;
keeping it on hot plate about 30 min at 50? temperature;
filtering the sample using whatman filter paper, and Stored at 4?;
adding 70 ml of zinc oxide solution to 30 ml of the extract under constant stirring in magnetic stirrer for 3 hours at 50-80? temperature;
adjusting the pH of the solution between 10-11 using 0.1M NaOH; and
forming intense white colored precipitate of ZnO NPs immediately.
The method as claimed in claim 2, wherein precursor solution was prepared using (0.1M) zinc acetate dihydrate.
EXAMPLE 1
EXPERIMENTAL SECTION
Sample collection and identification of specimen: -
Samples were collected from Bharmour region and were identified from the Botanical Survey of India (BSI) Dehradun and submitted in the Herbarium of Shoolini University, Solan Bajhol under the voucher number given (Stellaria aquatica - SUBMS/BOT-2071).
Preparation of plant extract from Stellaria aquatica: -
• 10gm of sample (Stellaria aquatica) added to 150 ml distilled water
• Keep it on hot plate about 30 min at 50? temperature
• Filter the sample using whatman filter paper
• Store at 4?
Synthesis of zinc oxide nanoparticles:
Precursor solution was prepared (0.1M) zinc acetate dihydrate. 70 ml of this solution was added to 30 ml of the extract under constant stirring in magnetic stirrer for 3 hours at 50-80? temperature. The pH of the solution was adjusted between 10-11 using 0.1M NaOH. An intense white coloured precipitate immediately formed. Bio reduction mechanism of these extracts for the synthesis of ZnO NPs was investigated through visual examination (color change), and DLS techniques (Fig 1-2).
Characterisation of ZnO NPs:
The primary detections of ZnO NPs were carried out by visual observation of the change in the color of the precursor (Fig 3).
Assessment Antifungal activity: -
Plant extract 50mg/ml (prepared in 10% DMSO), were added in sterilized potato dextrose agar. A 6mm diameter of the actively growing mycelium disc of the pathogen of 6-7 days old culture (Fusarium spp) was placed in the center of the Petri dish. Plates without extract served as negative control and 5mg/ml of Hygromycin as the positive control. Plates without extract served as negative control. Fusarium spp were incubated at 25°C for 6 days and circular growth of mycelium was measured after 6 days (Fusarium spp) of incubation. After incubation, the circular growth of mycelium was measured. The growth results were compared with the negative control (Fig 4). The percent inhibition of the fungus in the experiment was calculated using the following formula:
L=C-T/C×100
Where L is the percent inhibition; C is the colony radius in the control plate, and T is the radial growth of the pathogen in the plant extracts.
EXAMPLE 2
Results and discussion
Dynamic Light Scattering (DLS):
Dynamic Light Scattering (DLS) is an established and precise measurement technique for characterizing particle sizes in suspensions and emulsions. It is based on the Brownian motion of particles - this states that smaller particles move faster, while larger ones move slower in a liquid. It is a technique suited to the analysis and characterization of nanoparticles. DLS analysis showed the size distribution of particles with maximum intensity at 20-40nm.
Antifungal activity:
The effect of Stellaria aquatica ZnO NPs on different fungus has been investigated, but there is no fungal study related to ZnO NPs synthesized from Stellaria aquatica. The 55.2% diameter of zone of inhibition was observed with ZnO NPs for Fusarium spp (Table 1).
Table 1. Antifungal activity
S.no Fungus control Test Positive control Zone of inhibition
1. Fusarium spp 3.8cm 1.7cm ND 55.2%

, Claims:
1. A zinc oxide nanoparticles using Stellaria aquatica leaves extract consists of:
Stellaria aquatica plant extract, zinc acetate dihydrate, NaOH.
2. A method of green synthesis of zinc oxide nanoparticles using Stellaria aquatica leaves extract and their antifungal activity comprising the steps of:
a. adding 10gm of Stellaria aquatica to 150 ml distilled water;
b. keeping it on hot plate about 30 min at 50? temperature;
c. filtering the sample using whatman filter paper, and Stored at 4?;
d. adding 70 ml of zinc oxide solution to 30 ml of the extract under constant stirring in magnetic stirrer for 3 hours at 50-80? temperature;
e. adjusting the pH of the solution between 10-11 using 0.1M NaOH; and
f. forming intense white colored precipitate of ZnO NPs immediately.
3. The method as claimed in claim 2, wherein precursor solution was prepared using (0.1M) zinc acetate dihydrate.