Description:The present invention is described hereinafter by various embodiments with reference to the accompanying drawing, wherein reference numerals used in the accompanying drawing correspond to the like elements throughout the description.
While the present invention is described herein by way of example using embodiments and illustrative drawings, those skilled in the art will recognize that the
This invention is not limited to the embodiments of drawing or drawings described and are not intended to represent the scale of the various components. Further, some components that may form a part of the invention may not be illustrated in certain figures, for ease of illustration, and such omissions do not limit the embodiments outlined in any way. It should be understood that the drawings and detailed description thereto are not intended to limit the invention to the particular form disclosed, but on the contrary, the invention is to cover all modifications, equivalents, and alternatives falling within the scope of the present invention as defined by the appended claims. As used throughout this description, the word "may" is used in a permissive sense (i.e., meaning having the potential to), rather than the mandatory sense, (i.e., meaning must). Further, the words "a" or "an" mean "at least one” and the word “plurality” means “one or more” unless otherwise mentioned. Furthermore, the terminology and phraseology used herein is solely used for descriptive purposes and should not be construed as limiting in scope. Language such as "including," "comprising," "having," "containing," or "involving," and variations thereof, is intended to be broad and encompass the subject matter listed thereafter, equivalents, and additional subject matter not recited, and is not intended to exclude other additives, components, integers or steps. Likewise, the term "comprising" is considered synonymous with the terms "including" or "containing" for applicable legal purposes. Any discussion of documents, acts, materials, devices, articles and the like is included in the specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters form part of the prior art base or are common general knowledge in the field relevant to the present invention.
This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiment set forth herein. Rather, the embodiment is provided so that this disclosure will be thorough and complete and will fully convey the scope of the invention to those skilled in the art. In the following detailed description, numeric values and ranges are provided for various aspects of the implementations described. These values and ranges are to be treated as examples only and are not intended to limit the scope of the claims. In addition, a number of materials are identified as suitable for various facets of the implementations. These materials are to be treated as exemplary and are not intended to limit the scope of the invention.
In accordance with an embodiment of the present invention, in science, "green" synthesis has drawn a lot of interest as a dependable, sustainable, and environmentally friendly method for creating a variety of materials and nanomaterials, such as hybrid materials, bioinspired materials, metal/metal oxide NPs, and materials. As a result, green synthesis is seen as a crucial tool to lessen the negative impacts connected to the conventional methods of synthesis for NPs that are frequently used in labs and industry. The development of dependable, sustainable, and environmentally friendly synthesis processes is necessary to prevent the formation of undesirable or dangerous by-products. Because green synthesis is less expensive, produces less pollution, and enhances environmental and human health safety, it is more advantageous than standard chemical synthesis.
In accordance with an embodiment of the present invention, figure 1 illustrates a method (100) of preparation of a nano complex using citrus juice. As shown in Figure 1, the method (100) starts at step (110). Firstly, zinc salts are used as a precursor solution. A substance that can be used in a chemical process to create another substance is called a precursor solution. A molecule that takes part in a chemical reaction that results in the production of another component is known as a precursor. Zinc salts used may be, but not limited to, zinc nitrate (Zn (NO3)2, zinc chloride (ZnCl2), zinc sulphate (ZnSO4) to prepare zinc oxide (ZnO) nano particles from a solution of one or more zinc salts. One or more zinc salts are used in the range of, let’s say, for example, 0.05 to 0.01 moles per 100 ml of an aqueous solution. That means, about 0.05 to 0.01 moles of zinc salt is added to 100 ml of an aqueous solution.
Next, at step (120), an aqueous solution is obtained by adding predetermined quantity of a fruit juice at a predetermined temperature. For example, in a preferred embodiment, the fruit juice used is of a citrus fruit such as, but not limited to, lemon, orange, grapefruit, citron and/or clementine. Thus, the aqueous solution is prepared from one or more zinc salts by adding fruit juice and gradually heated in a beaker at a temperature range of about 60°C to 80°C. The gradual heating is done to allow proper mixing of contents in the beaker.
Further, at step (130), the pH of the aqueous solution is maintained by adding alkalizing agents such as, but not limited to, sodium bicarbonate and sodium bicarbonate in the beaker. For example, in case of Sodium Bicarbonate, Bicarbonate ion is produced by the dissociation of an alkalinizing agent. It elevates the pH of the blood and urine while balancing the hydrogen ion concentration. Sodium bicarbonate splits into sodium and bicarbonate ions. In an embodiment of the present invention, the alkalizing agents added are in the range of, for example, 0.05 to 0.02 moles per 100 ml of the aqueous solution. The alkalizing agent may be added in a stirring condition in the beaker.
Next, in step (140), a white precipitate of ZnO-NPs is obtained. A precipitate is an insoluble solid that forms from a liquid solution in chemistry. A solid that is distinct from any of the reactants is referred to as a precipitate. The precipitate frequently appears as a suspension. Precipitation is the process of changing a dissolved substance from a supersaturated solution to an insoluble solid in an aqueous solution. Precipitate refers to the produced solid. The chemical agent that initiates the precipitation in an inorganic chemical process is referred to as the precipitant. The precipitate obtained is suspended in an organic compound called glycine. In order to remove any residues or by-products formed in the aqueous solution, the precipitate is washed using an organic solvent and distilled water several times before being suspended in a glycine solution. On the surface of the precipitate particles, certain contaminants may have been adsorbed. Precipitate is washed with water in order to remove these contaminants since water washes away the residual soluble contaminants. The cleaned liquid may be a substance that won't obstruct the analysis or bring on new precipitation. In order to get the desirable results, the washed liquid is often discarded after washing the precipitate. This process is intended to remove any contaminants that may be adhering to the precipitated substance's surface. To remove the contaminants, ether devoid of ethanol is also employed in addition to water.
Further, in step (150), a solution of the obtained precipitate is mixed with an organic compound to obtain a nano complex. The organic compound used here may be, but not limited to, glycine. Glycine is added so to obtain a nano complex of zinc glycinate. Later in step (160), the nano complex obtained is isolated using for example, but not limited to, methods such as centrifugation method or size exclusion method. In centrifugation method, a centrifugal force is used to separate two liquids from a mixture. During this process, the mixture's denser component moves farther from the axis while its lighter component moves closer to the axis. A centrifuge is used to carry out the centrifugation process. In size exclusion method molecules are filtered over a gel and separated according to their size. The gel is usually made up of, but not limited to, sphere-shaped beads with pores that vary in size. Separation occurs when molecules of different sizes are included or excluded from the pores within the matrix.
Further, in this experiment, various techniques are employed to characterize the zinc-glycine combination produced in order to comprehend its structure, stability, and physical properties. The optical and physical characteristics of the zinc-glycine nano combination have been evaluated using analytical techniques like UV-visible spectroscopy, dynamic light scattering (DLS), and zeta-potential measurement. The zinc-glycine nano complex may be examined using a variety of methods, including X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM).
Particle size distribution
Particle size distribution of a specific material is a crucial analysis parameter in quality control procedures and research applications because many other product qualities are closely correlated with it, the. Particle size distribution affects a variety of material characteristics, including taste, compressibility, reactivity, abrasiveness, solubility, extraction and reaction behaviour, and flow and conveying behaviour (for bulk materials). Various techniques are employed for this depending on the sample material and the breadth of the analysis. These include Sieve Analysis, Dynamic Image Analysis, Dynamic Light Scattering, and Laser Diffraction. Typically, analyses of suspensions, emulsions, and bulk materials are conducted; occasionally, aerosols (sprays) are as well. In nanotechnology, the particle size plays a crucial role in learning the nanoparticle properties and therefore an essential task in property characterization of NPs is particle sizing.
According to the present embodiment, particle size distribution analysis of zinc- glycine nano complex (obtained in step 160) is determined by dynamic light scattering (DLS). The sample containing the isolated nano complex is diluted with a diluent or solvent (also known as liquid) before subjecting to an incident light. The particle size (PS) and zeta potential (ZP) of the complex may be measured using the incident light, incident on the sample. Dynamic light scattering (DLS) also known as photon correlation spectroscopy (PCS), is a particularly effective instrument for examining the diffusion behavior of macromolecules in solution. The incoming laser light is scattered widely if there are particles in the sample. The Stokes-Einstein equation uses the signal from the scattered light, which is monitored over time at a specific angle, to calculate the diffusion coefficient and particle size. The PS (particle size), PDI (polydispersity index), and ZP (zeta potential) of ZnO NPs are determined using DLS (Malvern Zeta Sizer Nano-series) at 25 °C.
It may be observed that by increasing the concentration of glycine in the glycine-zinc nano complex the size of the complex increased (refer Table 1) (Figure 2). As shown in Table 1, the particle size of Zn-glycine complex increased from 596±23.8 to 799±22.34 when the concentration of zinc salt 1 increased from 0.5M to 1M and the complex size increased from 713±54.2 to 835±29.36 when the concentration of zinc salt 2 increased from 0.5M to 1M. Similarly, the particle size of Zn-glycine complex increased from 788±17.2 to 906±23.45 when the concentration of zinc salt 3 increased from 0.5M to 1M. Thus, the larger size of the complex will allow for a higher concentration of zinc to be held in the complex, leading to increased zinc uptake by living organisms. Thus, as the concentration of glycine increases, the size of the nano complex will also increase, allowing for an increased capacity for zinc uptake.

Table 1. The PS, PDI and ZP assessment of Zn-glycine nano complex
X-ray photoelectron spectroscopy (XPS) analysis
The uniformity of nanoparticle surfaces may be profiled using XPS, which can also provide details about the surface chemistry there and provide elemental data on the chemical makeup of the nanoparticle. Nanomaterials can be analyzed and characterized using XPS analysis. By examining the X-rays that are released from a sample when it is exposed to an X-ray beam, it is possible to determine the elemental composition of various materials. The composition of thin films, surfaces, and interfaces between various materials have all been analyzed using XPS. Additionally, it can be used to assess the degree of oxidation and detect the chemical bonding of the sample's constituent elements.
XPS analysis of the Zn-glycine nano complex shows peak intensities of zinc Zn, oxygen O, nitrogen N, and carbon C peaks in each spectrum which are indicative of the composition of the ZnO-NPs. Figure 3 exhibited the typical XPS-wide survey spectra of ZnO-NPs biosynthesized from different zinc salts namely zinc nitrate (Zn (NO3)2), zinc chloride (ZnCl2), and zinc sulphate (ZnSO4). Zn, C, O and N peaks may be detected as shown in the wide survey XPS spectra in Figure 3 (A), (B), (C), and (D). The peaks corresponding to Zn and O may be identified to have different peak intensities in each spectrum. The peak corresponding to C is also detected in each spectrum but at varying intensities, which suggests that the ZnO-NPs synthesized from different zinc salts had different levels of carbon impurities. The peak corresponding to N is at a low intensity in all spectra with the lowest peak intensity in (A) and (D) which suggests that zinc salts used contain low nitrogen levels.
In the spectrum of Figure 3(A), as shown in the figure the intensity of the Zn peak is found to be the highest, followed by the O peak and then the C peak whereas N peak is lowest or negligible. This may suggest, but not limited to, that the NPs synthesized from this zinc salt i.e., zinc nitrate, had a higher concentration of zinc, with a low level of O and C impurities. In the spectrum of Figure 3 (B), the peak of O is found to be the highest, followed by the C peak and the Zn peak, and at last the N peak. This may suggest, but not limited to, the NPs synthesized from this zinc salt i.e., zinc sulphate had a higher concentration of oxygen, with a low level of Zn, C and N impurities. In the spectrum of Figure 3 (C), the O peak is found to be the highest, followed by the C peak and Zn peak and N peak is observed to be the lowest. This may suggest, but not limited to, that the NPs synthesized from this zinc salt i.e., zinc chloride had a higher concentration of oxygen, with a low level of Zn, C and N impurities. In the spectrum of Figure 3 (D), the Zn peak is found to be the highest and is followed by O and C peaks with negligible peak of N. This may suggest, but not limited to, that the NPs synthesized from this zinc salt i.e., pure zinc oxide had a higher concentration of O, with a low level of Zn, C and N impurities.
Fourier Transform Infrared Spectroscopy (FTIR)
The term "Fourier transform infrared" (FTIR) refers to the most popular kind of infrared spectroscopy. All infrared spectroscopies operate under the premise that some IR energy is absorbed when it passes through a material. It is noted which radiation enters the sample. For the identification and characterization of diverse organic and inorganic NPs, FTIR is a commonly utilized analytical technique. The results of the technique FTIR gives information about the binding structure between zinc ions and glycine molecules. The chemical and physical characteristics of NPs can be investigated using FTIR. The interactions between NPs and their surroundings, particularly their interactions with other molecules, can also be revealed by FTIR.
The FTIR spectrum of ZnO-NPs, as shown in Figure 4 depicts the presence of several peaks at wavenumbers between 500-4000 cm-1. The stretching of Zn-O bonds is thought to be responsible for the peak at about 500 cm-1. The stretching of the Zn-O and O-H bonds is what causes the peaks at about 890 cm-1 and 1020 cm-1, respectively. O-H bond stretching is thought to be responsible for the peak at about 3178 cm-1. The pure ZnO FTIR spectrum reveals many peaks with wavenumbers between 500 and 4000 cm-1. The stretching of Zn-O bonds is thought to be responsible for the peak at about 500 cm-1. The stretching of the Zn-O and O-H bonds is what causes the peaks at about 890 cm-1 and 1020 cm-1, respectively. The stretching of O-H bonds is thought to be responsible for the peak at about 3100 cm-1.
UV-Vis spectroscopy
Ultraviolet-visible (UV-Vis) spectroscopy is a widely used technique. The foundation of UV-Visible Spectroscopy is the idea that chemical compounds can absorb ultraviolet or visible light, creating unique spectra in the process. The basis of spectroscopy is the interaction of light and matter. The number of discrete wavelengths of UV or visible light that are absorbed by or transmitted through a sample in contrast to a reference or blank sample is measured by the analytical technique known as UV-Vis spectroscopy. ZnO-NPs optical characteristics are studied using UV-visible spectroscopy. ZnO-NPs ' UV-vis spectra may be obtained using a spectrophotometer (Simadzu-1800), which may be used to scan the NPs between 300 and 700 nm. In the ultraviolet and visible portions of the electromagnetic spectrum, a sample's absorption of light is measured via UV-visible spectroscopy.
This method can be used to measure the optical absorption, emission, and excitation spectra of ZnO-NPs. The purified ZnO-NPs (Fig. 5A) and the Zn-glycine nanocomplex (Fig. 5B) may be found to have UV-Vis absorption spectra of 367 nm and 335 nm, respectively. Information regarding the electronic structure of the NPs and their interactions with the environment can be found in the emission and excitation spectra (Figure 5).
Morphological characterization of Zn-glycine nano complex study by TEM
Using HRTEM (Tecnai G2 20 TWIN, FEI, Czech Republic), zinc-glycine nano particles or ZnO NP (Pure) is morphologically characterized (Figure 6A). The average particle size of the spherical particles is between 35 and 6.93 nm (Figure 6A). (Figure 6B) depicts the structure of the Zn-glycine nanocomplex formed. The SAED pattern of Zn-glycine nanocomplex depicts the selected area electron diffraction (SAED) patterns (Figure 6C).
X-ray diffraction pattern of Zn- glycine nano complex
Constructive interference between monochromatic X-rays and a crystalline sample is the foundation of X-ray diffraction. The XRD technique is used to understand the pattern of the complex formed that can reveal information about the crystalline nature of the material, and the bond formations. A cathode ray tube produces the X-rays, which are then filtered to produce monochromatic radiation, focused by collimation, and pointed at the sample. A versatile non-destructive analytical technique called X-ray diffraction (XRD) is used to examine the physical characteristics of powder, solid, and liquid materials, including their phase composition, crystal structure, and orientation. A potent method for examining the structure of materials, including nanomaterials, is X-ray diffraction (XRD). An XRD analysis can reveal details about the crystalline structure, size, and shape of NPs. ZnO-NPs crystalline structure, size, shape, and crystallite size have all been determined by XRD analysis. The presence of contaminants and flaws in the material can also be detected using XRD.
Zinc-glycine nanocomplex may be produced using citrus fruit juice, and their purity and composition may be evaluated by XRD. Figure 7 displays a typical XRD pattern of ZnO NPs shown by several Bragg reflections with 2? values (2 theta values) of 31.827, 34.497, 47.611, and 64.026 which correspond to 100, 101, 102, 103 planes (JCPDS card No. 89-7102) against intensity of the X-ray light. ZnO is the only substance to exhibit characteristic peaks that showed excellent crystallinity (Figure 7).
In accordance with the present invention, the advantage of the present invention is that Zn-glycine nanocomplex is biosynthesized from a citrus natural product. The method of the present invention is eco-friendly. The obtained zinc amino corrosive chelate nano complex is characterized using various techniques such as FTIR spectroscopy, UV-visible optical spectrum, XRD pattern. The zinc glycinate nano complex obtained is used to improve solubility and bioavailability in the agriculture and food industries. Zinc is an essential mineral for human health and plays an important role in various enzyme systems and in maintaining the integrity of cell membranes. It can be used to produce zinc glycinate nano complex in large quantities thus zinc can be available at large quantities for uptake by crops in agriculture and thus also can be used to increase nutrient in the food industry. Zinc glycinate complexes are utilised in dietary supplements as an alternative to inorganic zinc salts to give zinc in a more accessible form. Due to their capacity to accelerate wound healing and due to their antibacterial, antineoplastic, and antigenic capabilities, ZnO-NPs offer a variety of intriguing applications in veterinary research. The agriculture and food industries have been very interested in ZnO-NPs as a means of eliminating or limiting the activity of microorganisms. ZnO-NPs' antimicrobial qualities may enhance food quality, which directly affects human health.
Various modifications to these embodiments are apparent to those skilled in the art from the description and the accompanying drawings. The principles associated with the various embodiments described herein may be applied to other embodiments. Therefore, the description is not intended to be limited to the embodiments shown along with the accompanying drawings but is to be providing broadest scope of consistent with the principles and the novel and inventive features disclosed or suggested herein. Accordingly, the invention is anticipated to hold on to all other such alternatives, modifications, and variations that fall within the scope of the present invention and the appended claims.
, C , Claims:We claim:

1. A method (100) for preparation of a nano complex using citrus juice, the method comprising steps of:
preparing (110) a mixture of nano particles from a precursor solution of one or more zinc salts;
obtaining (120) an aqueous solution by adding predetermined quantity of a fruit juice at a predetermined temperature;
adding (130) aqueous alkalizing agents in a predetermined quantity in the aqueous solution;
obtaining (140) a predetermined precipitate;
mixing (150) a solution of the predetermined precipitate with an organic compound to obtain a nano complex; and
isolating (160) the nano complex obtained.
2. The method (100) for preparation of a nano complex using citrus juice as claimed in claim 1, wherein the zinc salts are in the range of 0.05 to 0.01 moles per 100 ml of the aqueous solution; and
wherein zinc salts are used as a precursor solution to prepare zinc oxide (ZnO) nano particles.
3. The method (100) for preparation of a nano complex using citrus juice as claimed in claim 1, wherein the predetermined temperature is in the range of 60°C to 80°C; and
wherein the aqueous solution is heated in a beaker gradually.
4. The method (100) for preparation of a nano complex using citrus juice as claimed in claim 1, wherein the predetermined quantity of the fruit juice is 25 to 50 ml per 100 ml of the aqueous solution; and
wherein the fruit juice used is of a citrus fruit.
5. The method (100) for preparation of a nano complex using citrus juice as claimed in claim 1, wherein the predetermined quantity the alkalizing agents is in the range of 0.05 to 0.02 moles per 100 ml of the aqueous solution; and
wherein the alkalizing agents in the aqueous solution manages pH of the solution.
6. The method (100) for preparation of a nano complex using citrus juice as claimed in claim 1, wherein the predetermined precipitate obtained is zinc oxide nano particles or ZnO NPs, it is white in colour; and
wherein the precipitate obtained is suspended in an organic compound called glycine.
7. A nano complex composition using citrus juice, the composition comprising:
one or more zinc salts;
atleast one aqueous alkalizing agent;
a glycine solution; and
a solution of a citrus fruit juice.
8. The nano complex composition using citrus juice as claimed in claim 7, wherein the zinc salts used are zinc nitrate (Zn (NO3)2), zinc sulphate (ZnSO4), zinc chloride (ZnCl2) and pure zinc oxide.
9. The nano complex composition using citrus juice as claimed in claim 7, wherein the fruit juice used is of a citrus fruit such as lemon, orange, grapefruit, citron and clementine.
10. The nano complex composition using citrus juice as claimed in claim 7, wherein the alkalizing agent used such as sodium bicarbonate and sodium hydroxide.