Compositions and Methods

Abstract

A composition is disclosed that includes extracts of kale and several other plants in a synergistic mixture.A method of forming metal nanoparticles, and nanoparticles formed using this method, are disclosed.

Description

[Technical Field]

[0001] The present invention relates to a composition. The present invention also relates to a method for preparing the composition, metal nanoparticles, a method for producing the same using the composition, and uses thereof. [Background technology]

[0002] Metal nanoparticles are typically defined as particles less than 100 nm in diameter. They have a wide range of commercial applications, including pharmaceuticals and electronics.

[0003] A variety of processes, including both chemical and physical manufacturing routes, have been developed over the past two decades to produce metal nanoparticles. However, some of these methods raise environmental and safety concerns. It would be desirable to produce these products in a more environmentally friendly manner.

[0004] David L. Johnson et al., Micro & Nano Letters, 2020, 15(2), 110-113, describes the biosynthesis of silver nanoparticles using upland cress, their purification, and characterization. This process uses only upland cress as an antioxidant source. The antioxidant ratio and bioreactor used in this process differ from those of the present invention. Notably, this process is less efficient in that the reaction takes 6 hours at 37°C. Furthermore, a different washing solution is used for centrifugation, and the separation technique is also different. Notably, this process uses AgNO3 as the silver salt, and leaching of nitrate into water supplies is known to be environmentally undesirable.

[0005] Mohammad Pourhassan-Moghaddam et al., Micro & Nano Letters, 2014, 9(5), 345-350, describes watercress-based gold nanoparticles, their biosynthesis, formation mechanism, and their in vitro biocompatibility. This process uses HAuCl4.3H2O to generate gold nanoparticles, with watercress as the only antioxidant source. The bioreactor used is less efficient than the one used in the present invention.

[0006] US Patent Application Publication No. 2012 / 0055873 describes the green synthesis of nanometals using plant extracts. This process uses commercially available plant extracts, such as green tea and coffee, to produce nanoparticles. The primary antioxidants used are polyphenols and caffeine, and the bioreactor is different from that of the present invention. Furthermore, this process also uses AgNO3 as a source of silver ions.

[0007] US Patent Application Publication No. 2011 / 0110723 describes the green synthesis of nanometals using fruit extracts. In this process, fruits (rich in sugar) are used as a source of antioxidants, and the bioreactor is different from that of the present invention. Furthermore, this process also uses AgNO3 as a source of silver ions. Summary of the Invention

[0008] According to one aspect of the present invention, (a) Kale (Brassica oleracea Acephala group), and (b) (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), (v) Sage (Salvia officinalis), (vi) Watercress (Nasturtium officinale), (vii) dog rose (Rosa canina), (viii) mint (Mentha spp.); (ix) Thyme (Thymus spp.), (x) basil (Osimum basilicum), and (xi) Spinach (Spinacia oleracea) at least one other plant selected from the group consisting of A composition is provided comprising an extract from

[0009] According to a further aspect of the present invention, there is provided a method of producing a composition of the present invention, the method comprising the steps of: (a) combining plants to provide a plant preparation as defined herein; and (b) extracting the biologically active component from the combined preparation using a solvent A method is provided that includes one or more of:

[0010] According to a further aspect of the present invention, there is provided a method of producing metal nanoparticles, comprising the steps of: (a) providing dissolved metal ions; and (b) contacting dissolved metal ions with the composition of the present invention such that the dissolved metal ions are reduced to form metal nanoparticles. A method is provided, comprising:

[0011] According to a further aspect of the present invention there is provided metal nanoparticles obtained or obtainable by the method of the present invention.

[0012] According to a further aspect of the present invention, there is provided a metal nanoparticle provided with a coating biopolymer layer.

[0013] According to a further aspect of the present invention, there is provided an article of manufacture comprising the metal nanoparticles of the present invention. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 is a process diagram illustrating a preferred method for producing the compositions of the present invention and metal nanoparticles of the present invention. [Figure 1A] FIG. 1 is a process diagram illustrating an alternative preferred method for producing the compositions of the present invention and metal nanoparticles of the present invention. [Figure 2] 1 is a Pareto frontier analysis showing sensitivity analysis at different constraint configurations for several compositions of the present invention. [Figure 3] 1 shows the results of a Pareto-optimal frontier analysis (antioxidant composition vs. blend cost per 100 g) for nine blend compositions of the present invention. [Figure 4] 1 shows the results of a Pareto-optimal frontier analysis of the relative antioxidant activity against watercress for nine blend compositions of the present invention. [Figure 5] 1 shows the components of nine blend compositions of the present invention. [Figure 6] Example 2 shows the components of five preferred blend compositions of the present invention that were DPPH tested. [Figure 7] 1 shows the inhibition of DPPH by preferred compositions of the present invention as a function of antioxidant concentration / watercress and curly kale concentration. [Figure 8] 1 shows the inhibition of DPPH as a function of antioxidant concentration blend for five preferred blend compositions of the present invention as tested in Example 2. [Figure 9]1 shows the inhibition of DPPH by ascorbic acid as a function of concentration. [Figure 10] 1 shows the linear regression of ascorbic acid inhibition of DPPH as a function of concentration. [Figure 11] 1 shows the linear regression of the inhibition of DPPH in watercress as a function of concentration. [Figure 12] 1 shows the linear regression of the inhibition of DPPH by curly kale as a function of concentration. [Figure 13] 7 shows the linear regression of inhibition of DPPH for the composition of the present invention shown as "Blend 1" in FIG. 6 as a function of concentration. [Figure 14] 6 shows a linear regression of the inhibition of DPPH for the composition of the present invention shown as "Blend 2" in FIG. 6 as a function of concentration. [Figure 15] 6 shows a linear regression of the inhibition of DPPH for the composition of the present invention shown as "Blend 3" in FIG. 6 as a function of concentration. [Figure 16] 6 shows a linear regression of the inhibition of DPPH for the composition of the present invention shown as "Blend 4" in FIG. 6 as a function of concentration. [Figure 17] 6 shows a linear regression of the inhibition of DPPH for the composition of the present invention shown as "Blend 5" in FIG. 6 as a function of concentration. [Figure 18] 1 shows UV-Vis spectrophotometric absorbance as a function of wavelength for the reaction of Example 4 carried out at t=55 minutes and 35° C. [Figure 19] 1 shows UV-Vis spectrophotometric absorbance as a function of wavelength from 0 to 55 minutes to illustrate the kinetics of the reaction of Example 4 carried out at 50° C. [Figure 20] 1 shows UV-Vis spectrophotometric absorbance as a function of wavelength from 0 to 55 minutes to illustrate the kinetics of the reaction of Example 4 carried out at 60° C. [Figure 21] 1 shows UV-Vis spectrophotometric absorbance as a function of wavelength from 0 to 55 minutes to illustrate the kinetics of the reaction of Example 4 carried out at 70° C. [Figure 22]1 shows UV-Vis spectrophotometric absorbance as a function of wavelength over time periods from 4 to 6.5 minutes to illustrate the kinetics of the reaction of Example 4 carried out at 85° C. [Figure 23] 1 shows UV-Vis spectrophotometric absorbance as a function of wavelength at t=55 min to compare reaction kinetics from 50° C., 60° C., and 70° C. for the reaction of Example 4. [Figure 24] 1 shows UV-Vis spectrophotometric peak absorbance as a function of time at t=55 min to compare reaction kinetics from 50° C., 60° C., and 70° C. for the reaction of Example 4. [Figure 25] 1 shows UV-Vis spectrophotometric absorbance as a function of wavelength to illustrate the kinetics of the reaction of Example 4 under different blend concentrations. [Figure 26] 1 is a scanning electron microscope (SEM) image of organic polymer and metal nanoparticles produced according to Example 4. [Figure 27] 1 is a scanning electron microscope (SEM) image of organic polymer and metal nanoparticles produced according to Example 4. [Figure 28] 1 is a scanning electron microscope (SEM) image of organic polymer and metal nanoparticles produced according to Example 4. [Figure 29] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 30] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 31] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 32] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 33] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 34] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 35]1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 36] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 37] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 38] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 39] 1 is a scanning electron microscope (SEM) image of organic polymer and metal nanoparticles produced according to Example 4. [Figure 40] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 41] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 42] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 43] 1 is a scanning electron microscope (SEM) image of metal nanoparticles produced by Example 4. [Figure 44] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 45] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 46] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 47] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 48] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 49] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 50]10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 51] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 52] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 53] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 54] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 55] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4. [Figure 56] 1 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4 showing evidence for a biopolymer protective layer (carbon + oxygen) on the silver nanoparticles. [Figure 57] 1 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4 showing evidence of a biopolymer protective layer (carbon) on the silver nanoparticles. [Figure 58] 1 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Example 4 showing evidence for a biopolymer protective layer (oxygen) on the silver nanoparticles. [Figure 59] 1 is a scanning electron microscope (SEM) image showing the outline of silver nanoparticles on a sampled area produced by Example 4. [Figure 60] 1 is a scanning electron microscope (SEM) image showing silver nanoparticles on a sampled area produced by Example 4. [Figure 61] 59A is a plot of nanoparticle frequency versus diameter showing the size distribution of silver nanoparticles over the sampled area of ​​FIG. 59. [Figure 62] 1 is a plot of nanoparticle frequency versus diameter showing the overall size distribution of silver nanoparticles over multiple sampled areas. [Figure 63]The color of the samples with increasing blend concentration is shown. [Figure 64] The color of the sample along with sampling at different times (every 5 minutes) for a 55 minute reaction at 70° C. is shown. [Figure 65] 1 shows a comparison of curly kale (Set 1) and watercress (Set 6) based blends according to Comparative Example 1. [Figure 66] 1 shows the composition of a watercress-based blend composition according to Comparative Example 1. [Figure 67] 1 is a UV / VIS spectrum of silver nanoparticles produced by batch S1 of Example 5. [Figure 68] 1 is a UV / VIS spectrum of silver nanoparticles produced by batch S2 of Example 5. [Figure 69] 1 is a UV / VIS spectrum of silver nanoparticles produced by Batch S3 of Example 5. [Figure 70] 1 is a UV / VIS spectrum of silver nanoparticles produced by batch S4 of Example 5. [Figure 71] 1 is a UV / VIS spectrum of gold nanoparticles produced by batch S1 of Examples 6 and 7. [Figure 72] 1 is a UV / VIS spectrum of gold nanoparticles produced by batch S2 of Examples 6 and 7. [Figure 73] 1 is a UV / VIS spectrum of gold nanoparticles produced by batch S3 of Examples 6 and 7. [Figure 74] 1 is a plot of nanoparticle frequency versus diameter showing the overall size distribution of gold nanoparticles produced by Examples 6 and 7. [Figure 75] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 76] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 77] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 78] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 79] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 80] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 81] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 82] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 83] 1 is an energy dispersive X-ray spectroscopy image of gold nanoparticles produced according to Examples 6 and 7. [Figure 84] 1 is an energy dispersive X-ray spectroscopy image of gold nanoparticles produced according to Examples 6 and 7. [Figure 85] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Examples 6 and 7, mapping gold atoms on the nanoparticles. [Figure 86] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Examples 6 and 7, mapping the carbon atoms on the biopolymer layer capping the nanoparticles. [Figure 87] 10 is an energy dispersive X-ray spectroscopy image of metal nanoparticles produced according to Examples 6 and 7, mapping oxygen atoms on the biopolymer layer capping the nanoparticles. [Figure 88] 1 is a scanning electron microscope (SEM) image of gold nanoparticles produced according to Examples 6 and 7. [Figure 89] 1 is an energy dispersive X-ray spectroscopy spectrum of gold nanoparticles produced by Examples 6 and 7. DETAILED DESCRIPTION OF THE INVENTION

[0015] definition As used herein, the term "extract" refers to a substance obtained by extraction from a raw material. Typically, the raw material is a naturally occurring substance. In one embodiment, the raw material is a plant, examples of which are described herein.

[0016] As used herein, the term "biologically active ingredient" refers to a substance obtained from a naturally occurring source (typically an extract as defined above, preferably a plant-based extract) that is capable of acting on metal ions to produce metal nanoparticles as described herein.

[0017] In one embodiment, the biologically active component is a biological reducing agent (i.e., a material obtained from a naturally occurring source, typically a plant-based source) capable of reducing metal ions to produce metal nanoparticles, as described herein.

[0018] In one embodiment, the biologically active component is a biological coating or capping agent, hi one embodiment, the biological coating or capping agent is a carbohydrate.

[0019] As used herein, the term "antioxidant" refers to a substance that inhibits oxidation. In one embodiment, an antioxidant is a substance that can terminate a chain reaction based on free radicals. In one embodiment, the antioxidant is a naturally occurring antioxidant, examples of which are described herein. In some aspects, the term "antioxidant" is synonymous with "biological reducing agent," as defined above.

[0020] As used herein, "nanoparticle" refers to a particle of material having a diameter of 1 nm to 1 μm. In one embodiment, the nanoparticle is 1 nm to 500 nm in diameter. In one embodiment, the nanoparticle is 1 nm to 200 nm in diameter. In one embodiment, the nanoparticle is 1 nm to 100 nm in diameter. In one embodiment, the nanoparticle is 1 nm to 50 nm in diameter. In one embodiment, the nanoparticle is 1 nm to 20 nm in diameter. In one embodiment, the nanoparticle diameter is measured using ultraviolet-visible (UV-Vis) spectroscopy. In one embodiment, the nanoparticle diameter is measured using scanning electron microscopy (SEM). In one embodiment, the nanoparticle diameter is measured using transmission electron microscopy (TEM).

[0021] Benefits and Surprising Findings Surprisingly, it has been found by the inventors that the compositions of the present invention, when used as reducing agents in the preparation of metal nanoparticles from corresponding metal ions, exhibit greater activity than would be expected from additive reducing activity (e.g., when tested using the DPPH assay), thus demonstrating a synergistic relationship between the different botanicals used in the compositions.

[0022] This surprising discovery makes it possible to produce metal nanoparticles in a manner that has much less environmental impact than known processes (including the current processes described in the prior art cited above). The compositions of the present invention have much higher antioxidant activity (and therefore effectiveness as biological reducing agents) and lower cost than extracts from single plants, and because of this synergistic relationship, much less material and energy is required to reduce metal ions to metal nanoparticles. This is particularly important when the process is scaled up.

[0023] The method for producing metal nanoparticles according to the present invention uses a biological reducing agent, allowing for the use of less toxic and more environmentally friendly reagents compared to current manufacturing routes. It also includes recirculation loops before and after crystallization to maximize process conversion at the point where metal ions are introduced, thereby minimizing waste generation throughout the process by avoiding the discharge of significant amounts of metal ions into the environment. Similarly, adding recirculation loops before and after the antioxidant extraction unit reduces waste and minimizes environmental impact.

[0024] It has also been unexpectedly discovered that by forming nanoparticles by the methods of the present invention, the formed nanoparticles have a protective biolayer (described herein) thereon, which protects the nanoparticles without the need to add an additional capping agent.

[0025] composition The composition of the present invention is made from plant extract.Usually, plant is naturally occurring variety.But alternatively, plant can be genetically modified plant.In another alternative, plant can be gene-edited by using technology such as CRISPR-Cas9.

[0026] In one aspect of the present invention, (a) Kale (Brassica oleracea Acephala group), and (b) at least one other plant capable of producing a biological reductant; A composition is provided comprising an extract from

[0027] Many plants are capable of producing antioxidants. Examples of such plants are kale (Brassica oleracea Acephala group), artichoke (Cyanara cardunculus), red cabbage (Brassica oleracea Capitata group var. rubra), oregano (Origanum vulgare), rosemary (Salvia cormanus), sage (Salvia officinalis), watercress (Nasturtium officinale), dog rose (Rosa canina), mint (Mentha spp.), especially spearmint (Mentha spicata). Examples of herbs include, but are not limited to, thyme (Thymus spp., especially Thymus cirtodorus, Thymus hera-barona, Thymus praecox, Thymus pseudolanugionsus, Tymus seryllum, and Thymus vulgaris), basil (Osimum basilicum), and spinach (Spinacia oleracea).

[0028] Thus, in one embodiment, (a) Kale (Brassica oleracea Acephala group), and (b) (i) artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea Capitata group var. rubra), (iii) oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), (v) sage (Salvia officinalis), (vi) watercress (Nasturtium officinale), (vii) dog rose (Rosa canina), (viii) mint (Mentha spp.), (ix) thyme (Thymus spp.). spp.), (x) basil (Osimum basilicum), and (xi) at least one other plant selected from the group consisting of spinach (Spinacia oleracea). A composition is provided comprising an extract from

[0029] In one embodiment, (a) Kale (Brassica oleracea Acephala group), and (b) at least one other plant selected from the group consisting of (i) artichoke (Cyanara cardunculus), (ii) cabbage (Brassica oleracea Capitata group var. rubra), (iii) oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) watercress (Nasturtium officinale). A composition is provided comprising an extract from

[0030] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least two other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), (v) Watercress (Nasturtium officinale), and (vi) Sage (Salvia officinalis); The present invention provides a composition comprising an extract from

[0031] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least two other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) Watercress (Nasturtium officinale); The present invention provides a composition comprising an extract from

[0032] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least two other plants selected from the group consisting of: (i) Oregano (Origanum vulgare), (ii) rosemary (Salvia cormanus), (iii) Watercress (Nasturtium officinale), and (iv) sage (Salvia officinalis); The present invention provides a composition comprising an extract from

[0033] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group), (b) artichoke (Cyanara cardunculus), and (c) Red cabbage (Brassica oleracea var. Capitata group var. rubra) A composition is provided comprising an extract from

[0034] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group), (b) artichoke (Cyanara cardunculus), and (c) Oregano (Origanum vulgare) A composition is provided comprising an extract from

[0035] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group), (b) red cabbage (Brassica oleracea var. Capitata group var. rubra), and (c) Oregano (Origanum vulgare) A composition is provided comprising an extract from

[0036] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least two other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) Watercress (Nasturtium officinale); A composition is provided comprising an extract from

[0037] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least three other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) Watercress (Nasturtium officinale); A composition is provided comprising an extract from

[0038] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group), (b) Red cabbage (Brassica oleracea var. Capitata group var. rubra), (c) oregano (Origanum vulgare), and (d) Rosemary (Salvia cormanus) A composition is provided comprising an extract from

[0039] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least three other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra), and (iii) rosemary (Salvia cormanus); A composition is provided comprising an extract from

[0040] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group); and (b) at least four other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) Watercress (Nasturtium officinale); A composition is provided comprising an extract from

[0041] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group), (b) Artichoke (Cyanara cardunculus), (c) Oregano (Origanum vulgare) (d) rosemary (Salvia cormanus), and (e) Watercress (Nasturtium officinale) A composition is provided comprising an extract from

[0042] In one embodiment, (a) Kale (Brassica oleracea var. Acephala group), and (b) Red cabbage (Brassica oleracea var. Capitata group var. rubra), (c) Oregano (Origanum vulgare) (d) rosemary (Salvia cormanus), and (e) Watercress (Nasturtium officinale) A composition is provided comprising an extract from

[0043] The compositions of the present invention typically contain an extract of kale (Brassica oleracea var. Acephala group). In one embodiment, the composition contains at least 5% kale extract by weight of the total composition. In one embodiment, the composition contains at least 10% kale extract by weight of the total composition. In one embodiment, the composition contains at least 15% kale extract by weight of the total composition. In one embodiment, the composition contains at least 20% kale extract by weight of the total composition. In one embodiment, the composition contains at least 25% kale extract by weight of the total composition. In one embodiment, the composition contains at least 30% kale extract by weight of the total composition. In one embodiment, the composition contains at least 35% kale extract by weight of the total composition. In one embodiment, the composition contains at least 40% kale extract by weight of the total composition. In one embodiment, the composition contains at least 45% kale extract by weight of the total composition. In one embodiment, the composition contains at least 50% kale extract by weight of the total composition. In one embodiment, the composition contains at least 55% kale extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 60% kale extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 65% kale extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 70% kale extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 75% kale extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 80% kale extract by weight, based on the total weight of the composition.

[0044] In one embodiment, the composition contains up to 95% kale extract by weight of the total composition. In one embodiment, the composition contains up to 90% kale extract by weight of the total composition. In one embodiment, the composition contains up to 85% kale extract by weight of the total composition. In one embodiment, the composition contains up to 80% kale extract by weight of the total composition. In one embodiment, the composition contains up to 75% kale extract by weight of the total composition. In one embodiment, the composition contains up to 70% kale extract by weight of the total composition. In one embodiment, the composition contains up to 65% kale extract by weight of the total composition. In one embodiment, the composition contains up to 60% kale extract by weight of the total composition. In one embodiment, the composition contains up to 55% kale extract by weight of the total composition. In one embodiment, the composition contains up to 50% kale extract by weight of the total composition. In one embodiment, the composition contains up to 45% kale extract by weight of the total composition. In one embodiment, the composition contains up to 40% kale extract by weight of the total composition. In one embodiment, the composition contains up to 35% kale extract by weight of the total composition. In one embodiment, the composition contains up to 30% kale extract by weight of the total composition. In one embodiment, the composition contains up to 25% kale extract by weight of the total composition. In one embodiment, the composition contains up to 20% kale extract by weight of the total composition. In one embodiment, the composition contains up to 15% kale extract by weight of the total composition. In one embodiment, the composition contains up to 10% kale extract by weight of the total composition.

[0045] In one embodiment, the composition contains kale extract in an amount of 5% to 95% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 10% to 90% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 20% to 95% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 30% to 90% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 35% to 85% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 40% to 80% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 45% to 75% by weight, based on the total weight of the composition. In one embodiment, the composition contains kale extract in an amount of 50% to 70% by weight, based on the total weight of the composition.

[0046] In one embodiment, the composition of the present invention comprises artichoke (Cyanara cardunculus) extract. In one embodiment, the composition contains at least 1% artichoke extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 2.5% artichoke extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 5% artichoke extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 7.5% artichoke extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 10% artichoke extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 12.5% ​​artichoke extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 15% artichoke extract by weight, based on the total weight of the composition.

[0047] In one embodiment, the composition contains up to 40% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 35% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 30% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 25% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 20% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 15% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 12.5% ​​artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 10% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 7.5% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 5% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 2.5% artichoke extract by weight of the total composition. In one embodiment, the composition contains up to 1% artichoke extract by weight of the total composition.

[0048] In one embodiment, the composition contains artichoke extract in an amount of 0.5% to 40% by weight of the total weight of the composition. In one embodiment, the composition contains artichoke extract in an amount of 10% to 90% by weight of the total weight of the composition. In one embodiment, the composition contains artichoke extract in an amount of 1% to 35% by weight of the total weight of the composition. In one embodiment, the composition contains artichoke extract in an amount of 2.5% to 30% by weight of the total weight of the composition. In one embodiment, the composition contains artichoke extract in an amount of 5% to 25% by weight of the total weight of the composition. In one embodiment, the composition contains artichoke extract in an amount of 10% to 20% by weight of the total weight of the composition.

[0049] In one embodiment, the composition of the present invention comprises an extract of red cabbage (Brassica oleracea Capitata group var. rubra). In one embodiment, the composition contains at least 1% by weight of red cabbage extract, based on the total weight of the composition. In one embodiment, the composition contains at least 2.5% by weight of red cabbage extract, based on the total weight of the composition. In one embodiment, the composition contains at least 5% by weight of red cabbage extract, based on the total weight of the composition. In one embodiment, the composition contains at least 10% by weight of red cabbage extract, based on the total weight of the composition. In one embodiment, the composition contains at least 15% by weight of red cabbage extract, based on the total weight of the composition. In one embodiment, the composition contains at least 20% by weight of red cabbage extract, based on the total weight of the composition. In one embodiment, the composition contains at least 25% by weight of red cabbage extract, based on the total weight of the composition.

[0050] In one embodiment, the composition contains up to 50% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 45% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 40% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 35% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 30% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 25% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 20% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 15% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 10% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 5% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 2.5% by weight of red cabbage extract by total weight of the composition. In one embodiment, the composition contains up to 1% by weight of red cabbage extract by total weight of the composition.

[0051] In one embodiment, the composition contains red cabbage extract in an amount of 0.5% to 50% by weight, based on the total weight of the composition. In one embodiment, the composition contains red cabbage extract in an amount of 1% to 40% by weight, based on the total weight of the composition. In one embodiment, the composition contains red cabbage extract in an amount of 1% to 35% by weight, based on the total weight of the composition. In one embodiment, the composition contains red cabbage extract in an amount of 2.5% to 30% by weight, based on the total weight of the composition. In one embodiment, the composition contains red cabbage extract in an amount of 5% to 25% by weight, based on the total weight of the composition. In one embodiment, the composition contains red cabbage extract in an amount of 10% to 20% by weight, based on the total weight of the composition.

[0052] In one embodiment, the composition of the present invention comprises an oregano (Origanum vulgare) extract. In one embodiment, the composition contains at least 0.5% oregano extract by weight of the total composition. In one embodiment, the composition contains at least 1% oregano extract by weight of the total composition. In one embodiment, the composition contains at least 2% oregano extract by weight of the total composition. In one embodiment, the composition contains at least 3% oregano extract by weight of the total composition. In one embodiment, the composition contains at least 5% oregano extract by weight of the total composition. In one embodiment, the composition contains at least 7.5% oregano extract by weight of the total composition. In one embodiment, the composition contains at least 10% oregano extract by weight of the total composition.

[0053] In one embodiment, the composition contains up to 30% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 25% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 20% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 15% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 10% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 5% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 2.5% by weight of oregano extract by total weight of the composition. In one embodiment, the composition contains up to 1% by weight of oregano extract by total weight of the composition.

[0054] In one embodiment, the composition contains oregano extract in an amount of 0.5% to 30% by weight of the total weight of the composition. In one embodiment, the composition contains oregano extract in an amount of 1% to 20% by weight of the total weight of the composition. In one embodiment, the composition contains oregano extract in an amount of 2% to 15% by weight of the total weight of the composition. In one embodiment, the composition contains oregano extract in an amount of 5% to 10% by weight of the total weight of the composition.

[0055] In one embodiment, the composition of the present invention comprises rosemary (Salvia cormanus) extract. In one embodiment, the composition contains at least 1% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 2.5% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 5% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 10% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 15% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 20% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains at least 25% rosemary extract by weight, based on the total weight of the composition.

[0056] In one embodiment, the composition contains up to 50% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 45% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 40% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 35% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 30% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 25% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 20% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 15% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 10% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 5% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 2.5% rosemary extract by weight, based on the total weight of the composition. In one embodiment, the composition contains up to 1% rosemary extract by weight, based on the total weight of the composition.

[0057] In one embodiment, the composition contains rosemary extract in an amount of 0.5% to 50% by weight, based on the total weight of the composition. In one embodiment, the composition contains rosemary extract in an amount of 1% to 40% by weight, based on the total weight of the composition. In one embodiment, the composition contains rosemary extract in an amount of 1% to 35% by weight, based on the total weight of the composition. In one embodiment, the composition contains rosemary extract in an amount of 2.5% to 30% by weight, based on the total weight of the composition. In one embodiment, the composition contains rosemary extract in an amount of 5% to 25% by weight, based on the total weight of the composition. In one embodiment, the composition contains rosemary extract in an amount of 10% to 20% by weight, based on the total weight of the composition.

[0058] In one embodiment, the composition of the present invention comprises an extract of watercress (Nasturtium officinale). In one embodiment, the composition contains at least 0.5% by weight of the watercress extract, based on the total weight of the composition. In one embodiment, the composition contains at least 1% by weight of the watercress extract, based on the total weight of the composition. In one embodiment, the composition contains at least 2% by weight of the watercress extract, based on the total weight of the composition. In one embodiment, the composition contains at least 3% by weight of the watercress extract, based on the total weight of the composition. In one embodiment, the composition contains at least 5% by weight of the watercress extract, based on the total weight of the composition. In one embodiment, the composition contains at least 7.5% by weight of the watercress extract, based on the total weight of the composition. In one embodiment, the composition contains at least 10% by weight of the watercress extract, based on the total weight of the composition.

[0059] In one embodiment, the composition contains up to 30% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 25% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 20% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 15% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 10% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 5% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 2.5% by weight of watercress extract by total weight of the composition. In one embodiment, the composition contains up to 1% by weight of watercress extract by total weight of the composition.

[0060] In one embodiment, the composition contains watercress extract in an amount of 0.5% to 30% by weight, based on the total weight of the composition. In one embodiment, the composition contains watercress extract in an amount of 1% to 20% by weight, based on the total weight of the composition. In one embodiment, the composition contains watercress extract in an amount of 2% to 15% by weight, based on the total weight of the composition. In one embodiment, the composition contains watercress extract in an amount of 5% to 10% by weight, based on the total weight of the composition.

[0061] In one embodiment, the composition comprises: (i) kale (Brassica oleracea var. Acephala group) in an amount of 60% to 90% by weight as a percentage of the total weight of the composition; (ii) artichoke (Cyanara cardunculus) in an amount of 10% to 20% by weight as a percentage of the total weight of the composition, and (iii) oregano (Origanum vulgare) in an amount of 5% to 15% by weight as a percentage of the total weight of the composition Contains an extract from

[0062] In one embodiment, the composition comprises: (i) kale (Brassica oleracea var. Acephala group) in an amount of 60% to 90% by weight as a percentage of the total weight of the composition; (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra) in an amount of 5% to 15% by weight as a percentage of the total weight of the composition, and (iii) oregano (Origanum vulgare) in an amount of 5% to 15% by weight as a percentage of the total weight of the composition Contains an extract from

[0063] In one embodiment, the composition comprises: (i) kale (Brassica oleracea var. Acephala group) in an amount of 50% to 80% by weight as a percentage of the total weight of the composition; (ii) artichoke (Cyanara cardunculus) in an amount of 15% to 25% by weight as a percentage of the total weight of the composition; (iii) oregano (Origanum vulgare) in an amount of 5% to 20% by weight as a percentage of the total weight of the composition; (iv) rosemary (Salvia cormanus) in an amount of 0.5% to 5% by weight as a percentage of the total weight of the composition, and (v) watercress (Nasturtium officinale) in an amount of 1% to 10% by weight as a percentage of the total weight of the composition Contains an extract from

[0064] In one embodiment, the composition comprises: (i) kale (Brassica oleracea var. Acephala group) in an amount of 50% to 70% by weight as a percentage of the total weight of the composition; (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra) in an amount of 5% to 15% by weight as a percentage of the total weight of the composition; (iii) oregano (Origanum vulgare) in an amount of 1% to 15% by weight as a percentage of the total weight of the composition, and (iv) rosemary (Salvia cormanus) in an amount of 15% to 35% by weight as a percentage of the total weight of the composition Contains an extract from

[0065] In one embodiment, the composition comprises: (i) kale (Brassica oleracea var. Acephala group) in an amount of 10% to 50% by weight as a percentage of the total weight of the composition; (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra) in an amount of 5% to 30% by weight as a percentage of the total weight of the composition; (iii) oregano (Origanum vulgare) in an amount of 0.5% to 10% by weight as a percentage of the total weight of the composition; (iv) rosemary (Salvia cormanus) in an amount of 2.5% to 25% by weight as a percentage of the total weight of the composition, and (v) watercress (Nasturtium officinale) in an amount of 10% to 30% by weight as a percentage of the total weight of the composition Contains an extract from

[0066] The compositions of the present invention contain antioxidants that act as biological reducing agents. In one embodiment, the antioxidants include ascorbic acid, beta-carotene, retinol (vitamin A), phenolic compounds (especially polyphenols such as flavonoids or epicatechin), lutein, all-trans-β-carotene, 9-cis-β-carotene, neoxanthin, violaxanthin, narirutin; apigenin 7-rutinoside, cynarin, 1-caffeoylquinic acid, luteolin, 7-apiosyl-(1->2)-glucoside, cyanaroside, caffeic acid, tannins, chlorogenic acid, luteolin-7-rutinoside, cyanidipine, and the like. The anthocyanins are selected from the group consisting of anthocyanins such as cinnamoyl-3-diglucoside-5-glucoside, kaempferol, p-coumaric acid, ferulic acid, myricetin, resveratrol, rosmarinic acid, carnosol, hesperidin, rosmanol, epirosmanol, rosmadial, carnosic acid, methyl carnosate, gallic acid, quercetin-3-O-rutinoside, dicaffeoyltartaric acid, isorhamnetin, cyanidin-3-glucoside, luteolin-7-O-glucoside, quercetin-3-O-rhamnoside, and gluconasturtiin.

[0067] If the composition contains kale, any or all of the following antioxidants may be present in the composition in varying proportions: ascorbic acid (vitamin C), beta-carotene, retinol (vitamin A), phenolic compounds (polyphenols), lutein, all-trans-β-carotene, 9-cis-β-carotene, neoxanthin, violaxanthin.

[0068] If the composition contains artichoke, any or all of the following antioxidants may be present in the composition in varying proportions: narirutin, apigenin 7-rutinoside, cynarin, 1-caffeoylquinic acid, luteolin 7-apiosyl-(1->2)-glucoside, cynaroside, flavonoids, caffeic acid, tannins, chlorogenic acid, luteolin-7-rutinoside.

[0069] If the composition contains red cabbage, any or all of the following antioxidants may be present in the composition in varying proportions: anthocyanins such as cyanidin-3-diglucoside-5-glucoside, kaempferol, vitamin C, vitamin A, lutein.

[0070] If the composition contains oregano, any or all of the following antioxidants may be present in the composition in varying proportions: chlorogenic acid, epicatechin, p-coumaric acid, caffeic acid, ferulic acid, rosemarinic acid, myricetin, resveratrol.

[0071] If the composition contains rosemary, any or all of the following antioxidants may be present in the composition in various proportions: rosmarinic acid, carnosic acid, carnosol, hesperidin.

[0072] If the composition contains sage, any or all of the following antioxidants may be present in the composition in varying proportions: rosmanol, epirosmanol, carnosol, rosmadial, carnosic acid, methyl carnosate.

[0073] If the composition contains watercress, any or all of the following antioxidants may be present in the composition in varying proportions: gallic acid, chlorogenic acid, caffeic acid, quercetin-3-O-rutinoside, dicaffeoyl tartaric acid, isorhamnetin, cyanidin-3-glucoside, luteolin-7-O-glucoside, quercetin-3-O-rhamnoside, gluconasturtiin.

[0074] In one embodiment, the antioxidant is ascorbic acid.

[0075] In one embodiment, the composition comprises at least 5 mmol of antioxidant per 100 g of composition. In one embodiment, the composition comprises at least 15 mmol of antioxidant per 100 g of composition.

[0076] Methods for preparing compositions The compositions of the present invention may be prepared by treating plants such that the active biological reducing agent (antioxidant) components of the composition are extracted from the plant into solution.

[0077] In one embodiment, the method comprises the following steps: (a) providing a plant preparation as defined above, and (b) extracting the biologically active component from the combined preparation using a solvent Contains one or more of the following:

[0078] In one embodiment, the plant preparation step comprises drying the plant. In one embodiment, the plant is blended prior to the drying step. In one embodiment, the individual plant is dried prior to the particle size reduction step.

[0079] The temperature at which the drying step is carried out is typically from room temperature to 80°C, preferably from 30 to 50°C, and more preferably 40°C.

[0080] The pressure at which the drying step is carried out is typically between 0.2 and 10 bar, preferably between 0.5 and 2 bar, more preferably between 0.9 and 1.1 bar.

[0081] The time for carrying out the drying step is typically 10 minutes to 72 hours, preferably 1 to 48 hours, and more preferably 12 to 36 hours.

[0082] In one embodiment, the plant preparation step includes reducing the particle size of the dried plant material. This step can be performed using any method or device suitable for this task. Typically, this is performed in a blender. Typically, the particle size to which the plant material is reduced is in the range of 10 μm to 1 mm, preferably in the range of 20 to 200 μm, and more preferably 100 μm.

[0083] In one embodiment, the extraction step is carried out by leaching. The leaching process allows for the extraction of biologically active components. Typically, this process involves dissolving biologically active components from dried plant particles in a solvent. The active components typically undergo mass transfer diffusion from the internal pores of the particles toward the solid-liquid interface, followed by mass transfer diffusion from the solid-liquid interface into the bulk solution.

[0084] The solvent used in the extraction step can include any solvent that can dissolve biologically active ingredients.Suitable examples of solvents include water, ethers such as diethyl ether and 1,2-dimethoxyethane, hydrocarbons such as hexane, halogenated hydrocarbons such as dichloromethane and chloroform, and mixtures thereof.However, the solvent used in the extraction step is preferably water, because it can dissolve oxygenated active ingredients such as ascorbic acid present in many blended products while avoiding the toxicity and environmental impact of some organic solvents.

[0085] The temperature at which the extraction step is carried out will depend on the nature of the plant, the solvent, and the biological agent requiring extraction, but temperatures are typically between room temperature and the boiling point of the solvent, preferably 30-70°C, more preferably 40-60°C.

[0086] The pressure at which the extraction step is carried out depends on the nature of the plant, the solvent, and the biological agent that requires extraction. However, the pressure is typically between 0.2 and 10 bar, preferably between 0.5 and 2 bar, and more preferably between 0.9 and 1.1 bar.

[0087] The time for carrying out the extraction step is typically 1 minute to 72 hours, preferably 2 minutes to 4 hours, and more preferably 5 to 60 minutes.

[0088] Typically, the extraction process is carried out with stirring.

[0089] In one embodiment, after the extraction step, the solution may be decanted to separate the solution from the solids. The decanter may form three phases (scum, clarified liquid, and sludge). The sludge may be recycled back to the mixer for further processing to extract the maximum amount of biologically active components until the concentration in the solid particles is minimized. At this point, the spent plant particles may be discarded from the process.

[0090] In one embodiment, after decantation but before filtration, the filtrate can be passed through a hydrocyclone to recover biologically active components and / or disrupt cells, also using centrifugal force to remove solids, but continuously.

[0091] In one embodiment, after the extraction step or after the decanting step, the composition may be filtered to produce a filtrate and a residue. Depending on the solids content, the filtration may be performed under pressure, under vacuum, or a combination thereof. The pressure at which the filtration step is performed depends on the nature of the plant, the solvent, and the biological agent requiring extraction. The pressure at which the filtration step is performed is typically 0.1 to 100 bar, preferably 0.3 to 10 bar, and more preferably 0.3 to 5 bar. In one embodiment, the filtration step is performed at a pressure of 0.001 to 0.1 bar.

[0092] In one embodiment, after the extraction step or after the decanting step, the composition may be filtered through a screen to remove larger particles.

[0093] In one embodiment, the filtrate may be centrifuged after the filtration step. Centrifugation allows for further cell disintegration, removal of relatively small particles, and dissolution of a relatively large amount of biologically active components. In one embodiment, centrifugation is performed at 500 to 20,000 rpm. In one embodiment, centrifugation is performed at 500 to 15,000 rpm. In one embodiment, centrifugation is performed at 300 to 5000 rpm. In one embodiment, centrifugation is performed at 100 to 3000 rpm.

[0094] In one embodiment, centrifugation is performed at 2500 rpm. This centrifugation speed is particularly preferred because it allows the removal of many broken cells in the precipitate, which may interfere with the nucleation of nanoparticles, and allows a large amount of impurities in the crystals. By removing cells from the filtrate, more spherical nanoparticles with a particle size distribution can be obtained.

[0095] In one embodiment, centrifugation is carried out for 10 seconds to 1 hour. In one embodiment, centrifugation is carried out for 30 seconds to 30 minutes. In one embodiment, centrifugation is carried out for 1 to 20 minutes. In one embodiment, centrifugation is carried out for 10 minutes.

[0096] In an alternative embodiment, the biologically active component may be recovered from the solution by chromatography, which may be thin layer chromatography, column chromatography, liquid chromatography, especially high pressure liquid chromatography, or gas chromatography.

[0097] In an alternative embodiment, the biologically active component may be recovered from the solution by passing the solution through a membrane. Examples of such techniques include micro- and nanofiltration techniques.

[0098] In alternative embodiments, the biologically active component may be recovered from solution by precipitation or crystallization. In alternative embodiments, the biologically active component may be recovered from solution by liquid-liquid extraction. In alternative embodiments, the biologically active component may be recovered from solution by drying. In alternative embodiments, the biologically active component may be recovered from solution by osmotic shock. In alternative embodiments, the biologically active component may be recovered from solution by electrophoresis. All of these techniques are well known to those skilled in the art.

[0099] Metal nanoparticles The present invention also includes metal nanoparticles obtained or obtainable by the process of the present invention.

[0100] It has been unexpectedly discovered that forming nanoparticles by the methods of the present invention results in the formation of nanoparticles having a protective biolayer thereon, thereby protecting the nanoparticles without the need to add an additional capping agent.

[0101] Thus, in another aspect, there is provided a metal nanoparticle provided with a coating layer. In one embodiment, the coating layer comprises carbon. In one embodiment, the coating layer comprises oxygen. In one embodiment, the coating layer comprises nitrogen. In one embodiment, the coating layer comprises sulfur. In one embodiment, the coating layer comprises a biopolymer. In one embodiment, the coating layer comprises a carbohydrate. In one embodiment, the coating layer comprises a compound identified above as an antioxidant in the biological reducing agent of the composition of the present invention described and exemplified above, or a compound derived therefrom.

[0102] The nanoparticles can comprise a single metal or a mixture of metals. In one embodiment, the nanoparticles comprise a single metal. In one embodiment, the nanoparticles comprise a mixture of metals.

[0103] The metal forming the nanoparticles can be any metal that is capable of forming nanoparticles and is stable to the above process. Examples of suitable metals include, but are not limited to, gold, silver, copper, nickel, platinum, iron, zinc, gadolinium, titanium, cerium, thallium, manganese, aluminum, zirconium, chromium, palladium, and cobalt, or any combination thereof.

[0104] In one embodiment, the metal is selected from the group consisting of silver, gold, platinum, aluminum, zirconium, and iron, or any combination thereof.

[0105] In one embodiment, the metal is selected from the group consisting of silver, gold, and copper, or any combination thereof. In one embodiment, the metal is silver. In one embodiment, the metal is gold.

[0106] In one embodiment, the nanoparticles are multilayer nanoparticles. Typically, such multilayer nanoparticles comprise an inner core and an outer shell. The metals comprising the core and shell may be the same or different and may be any of the metals listed above. Illustratively, the nanoparticles may comprise an iron core and a silver outer shell.

[0107] The metal nanoparticles of the present invention can be formed into a variety of shapes, depending on the exact nature of the process used to form them and their intended use. Typical examples of suitable shapes include spheres, ovals, cylinders, rods, cubes and rectangular prisms, prisms (e.g., triangular, pentagonal, or hexagonal), cones, and pyramids.

[0108] Method for producing metal nanoparticles In another aspect, the present invention provides a method for producing metal nanoparticles, the method comprising: (a) providing dissolved metal ions; and (b) contacting dissolved metal ions with the composition of the present invention such that the dissolved metal ions are reduced to form metal nanoparticles. The present invention provides a method comprising:

[0109] Typically, the dissolved metal ions are present in the form of a solution of metal salts. The salts preferably contain counterions that have low toxicity to humans and low environmental impact. Examples of suitable salts include chlorides, sulfates, carbonates, gluconates, and acetates. In one embodiment, the metal salt is an organometallic salt. In one embodiment, the metal salt is an acetate salt.

[0110] In one embodiment, the dissolved metal ions are present in the form of a solvate of a metal salt. The solvent that forms the solvate preferably has low toxicity to humans and low environmental impact. In one embodiment, the dissolved metal ions are present in the form of a hydrate in which the solvent is water.

[0111] When the metal is silver, preferred salts include silver chloride, silver carbonate, and silver acetate, or a solvate of either of these. When the metal is copper, preferred salts include copper(II) sulfate and copper(II) gluconate, or a solvate of either of these. When the metal is gold, preferred salts include gold(I) chloride and gold(III) chloride, or a solvate of either of these.

[0112] The solvent used in the contacting step can include any solvent that can dissolve biologically active ingredients.Examples of suitable solvents include water and alcohols such as methanol, ethanol and isopropanol, and mixtures thereof.However, the solvent used in the contacting step is preferably water, because it can dissolve both metal ions and oxygenated biological reducing agents such as ascorbic acid present in many blended products, while avoiding the toxicity and environmental impact of some organic solvents.

[0113] The contacting step can be carried out by any means known to those skilled in the art to produce metal nanoparticles from dissolved metal ions. Preferably, the composition of the present invention is added to a solution of metal ions. In an alternative embodiment, a solution of metal ions is added to the composition of the present invention.

[0114] In one embodiment, a smaller volume of a more concentrated composition of the present invention is added to a larger volume of a less concentrated solution of metal ions. In one embodiment, the volume of the metal ion solution used is 2 to 50 times the volume of the composition of the present invention. In one embodiment, the volume of the metal ion solution used is 5 to 20 times the volume of the composition of the present invention.

[0115] It is preferred to preheat the solution of metal ions prior to addition of the composition of the present invention. When this process step is used, nucleation to produce nanoparticles has been found to be essentially instantaneous. In one embodiment, the solution of metal ions is preheated to a temperature of 40-100°C. In one embodiment, the solution of metal ions is preheated to a temperature of 60-80°C.

[0116] Typically, the concentration of the metal ions used in the contacting step is 0.1 to 5 g / L, preferably 0.5 to 1.5 g / L.

[0117] Typically, the concentration of the composition of the present invention used in the contacting step is 0.5 to 7 g / L, preferably 5 to 7 g / L.

[0118] In one embodiment, the contacting step is carried out at a temperature between room temperature and the boiling point of the solvent. In one embodiment, the contacting step is carried out at a temperature between 25 and 90°C. In one embodiment, the contacting step is carried out at a temperature between 35 and 70°C.

[0119] In one embodiment, the contacting step is carried out at a pressure of 0.3 to 1.2 bar. In one embodiment, the contacting step is carried out at a pressure of 0.9 to 1.15 bar.

[0120] In one embodiment, the contacting step is carried out for a time period of 30 seconds to 2 hours. In one embodiment, the contacting step is carried out for a time period of 1 minute to 1 hour. In one embodiment, the contacting step is carried out for a time period of 5 minutes to 40 minutes.

[0121] In one embodiment, the contacting step is carried out at a pH of 2 to 12. Without wishing to be bound by theory, it is believed that the pH of the reaction can affect the size of the resulting metal nanoparticles. Preferably, the contacting step is carried out at a pH of 5 to 8.

[0122] In one embodiment, the contacting step is carried out under stirring, hi one embodiment, the stirring is carried out at 200 to 1000 rpm.

[0123] In one embodiment, a seed solution of metal nanoparticles (typically produced using the methods of the present invention) is added to the solution of metal ions before the composition of the present invention is added in the contacting step. Without wishing to be bound by theory, it is believed that adding the seed solution of metal nanoparticles allows for better control of nanoparticle growth. Typically, a volume of 500 nL to 1 mL of seed solution is added before the contacting step.

[0124] In one embodiment, the contacting step is followed by quenching of the reaction mixture. This can be done by any means known in the art. Typically, this is done by cooling, such as using a jacketed reactor, which allows heat transfer around the reactor without direct contact with the product solution.

[0125] After the contacting step, the reaction mixture is worked up to purify the metal nanoparticles, which can be done by methods known to those skilled in the art.

[0126] In one embodiment, the product is purified by ion exchange chromatography. As known to those skilled in the art, ion exchange chromatography separates molecules based on their respective charged groups by passing them through a matrix (typically a column) capable of ion exchange (typically an ion exchange resin). Ion exchange chromatography retains analyte molecules on the column based on Coulomb (ionic) interactions. The ion exchange chromatography matrix consists of positively and negatively charged ions. Essentially, the molecules undergo electrostatic interactions with the opposite charge on the stationary phase matrix. The stationary phase consists of an immobile matrix containing charged, ionizable functional groups or ligands.

[0127] In one embodiment, the product is purified by crystallization, which, as known to those skilled in the art, typically involves forming a crystalline solid phase from a solution, typically from a supersaturated solvent.

[0128] In one embodiment, a portion of the output from the crystallization step is recycled to the input to the contacting step. It has been found that including a recycle loop before and after the contacting step minimizes waste generation throughout the process and maximizes process conversion at the point where the metal ions are introduced, thereby avoiding the discharge of significant amounts of metal ions into the environment.

[0129] In one embodiment, the product is purified by centrifugation. Centrifugation can be performed in a single centrifugation step or multiple centrifugation steps (such as two, three, four, or five steps). When centrifugation is performed in multiple centrifugation steps, the centrifugation speed and the centrifugation time for each step can be the same or different.

[0130] In one embodiment, the centrifugation is carried out at 500 to 100,000 rpm. In one embodiment, the centrifugation is carried out at 500 to 16,000 rpm. In one embodiment, the centrifugation is carried out at 100 to 5000 rpm. In one embodiment, the centrifugation is carried out at 2000 to 3000 rpm. In one embodiment, the centrifugation is carried out at 1000 rpm. In one embodiment, the centrifugation is carried out at 2500 rpm. In one embodiment, the centrifugation is carried out at 5000 rpm. In one embodiment, the centrifugation is carried out at 7000 rpm.

[0131] Differential centrifugation can be used to tailor the nanoparticle size as a function of centrifugation speed. Multiple centrifugations can be used to increase the purity of the metal nanoparticles.

[0132] In one embodiment, the centrifugation is performed in two steps. In one embodiment, the first centrifugation step is performed at 500-100,000 rpm, and the second centrifugation step is performed at 500-100,000 rpm. In one embodiment, the first centrifugation step is performed at 500-2000 rpm, and the second centrifugation step is performed at 5000-20,000 rpm. In one embodiment, the first centrifugation step is performed at 5000-20,000 rpm, and the second centrifugation step is performed at 500-2000 rpm. In one embodiment, the first centrifugation step is performed at 1000 rpm, and the second centrifugation step is performed at 10,000 rpm. In one embodiment, the first centrifugation step is performed at 10,000 rpm, and the second centrifugation step is performed at 1000 rpm.

[0133] In one embodiment, centrifugation is carried out for 20 seconds to 2 hours. In one embodiment, centrifugation is carried out for 1 minute to 1 hour. In one embodiment, centrifugation is carried out for 10 to 30 minutes. In one embodiment, centrifugation is carried out for 20 minutes. In one embodiment, centrifugation is carried out for 2 minutes. In one embodiment, centrifugation is carried out for 5 minutes. In one embodiment, centrifugation is carried out for 10 minutes.

[0134] In one embodiment, the product is purified by ultrasonication. In one embodiment, ultrasonication is performed at 30 to 50 kHz. In one embodiment, ultrasonication is performed at 35 to 45 kHz. In one embodiment, ultrasonication is performed at 0 to 50°C. In one embodiment, ultrasonication is performed at 15 to 40°C. In one embodiment, ultrasonication is performed for 1 to 30 minutes. In one embodiment, ultrasonication is performed for 10 to 20 minutes. In one embodiment, ultrasonication is performed for 15 minutes.

[0135] In one embodiment, the sonication is performed in two steps. The frequency, temperature, and time of each step follow the general sonication procedure described above. In one embodiment, the first sonication step is performed for 2 to 15 minutes, and the second sonication step is performed for 1 to 10 minutes. In one embodiment, the first sonication step is performed for 10 minutes, and the second sonication step is performed for 5 minutes.

[0136] In one embodiment, the method includes a shaking step following the or each sonication step. In one embodiment, the shaking is for between 10 seconds and 30 minutes. In one embodiment, the shaking is for between 30 seconds and 15 minutes.

[0137] In one embodiment, the method comprises two sonication steps, each followed by a shaking step, with the conditions for each sonication step and each shaking step following the general sonication and shaking procedure described above.

[0138] In one embodiment, the method includes a step of increasing the pH between the centrifugation and sonication steps. Typically, the pH is increased to 10-14. This can be done using any suitable base, provided it does not affect the nanoparticles. Examples include alkali metal hydroxides, such as sodium hydroxide.

[0139] In one embodiment, the product is purified by microfiltration, ultrafiltration, and / or nanofiltration. As known to those skilled in the art, microfiltration is a membrane filtration-based method that uses membranes with micrometer-sized pores to filter particulate matter. Furthermore, as known to those skilled in the art, ultrafiltration is a type of membrane filtration in which separation through a semipermeable membrane is achieved by forces such as pressure or a concentration gradient. Furthermore, as known to those skilled in the art, nanofiltration is a membrane filtration-based method that uses nanometer-sized through-pores that pass through the membrane. Microfiltration membranes typically have pore sizes of 1 to 100 μm, and nanofiltration membranes typically have pore sizes of 1 to 100 nm, e.g., 1 to 10 nm. In one embodiment, the product is purified by microfiltration. In one embodiment, the product is purified by microfiltration followed by ultrafiltration. In one embodiment, the membrane has a pore size of 1 nm to 1 μm. In one embodiment, the membrane has a pore size of 100 nm to 500 nm.

[0140] In one embodiment, the method includes a step of adjusting the pH after the sonication step. Typically, the pH is adjusted to 7-9. This can be done using any suitable acid, provided that it does not affect the nanoparticles. Weak acids, especially organic acids, are preferred, with acetic acid being particularly preferred.

[0141] In one embodiment, the method includes drying the nanoparticles. In one embodiment, the drying is by vacuum drying. In a preferred embodiment, the drying is by vacuum distillation.

[0142] The preferred process is shown in more detail in Figure 1. The plant solution, mixed in the appropriate ratio, enters drying unit 10 via line 12. Air is pumped via line 14 to compressor 16 and then via line 18 to heater 20. The heated air is then fed to drying unit 10 via line 22 and exits the drying unit via line 24.

[0143] Dried plants exit drying unit 10 and are fed via line 26 to blender 28. The blended plant solution exits the blender and is fed via line 30 to leaching unit 32. Water is heated in heater 34, exits the heater via line 36, and is pumped via pump 38 and line 40 to a heating jacket 41 around leaching unit 32. Deionized water is introduced into the top of leaching unit 32 via line 42.

[0144] The solution exits the bottom of leaching unit 32 and is pumped via pump 48 and line 46 to decanter unit 44, where it forms a separated blended solution and blended sludge. Some sludge exits the decanter unit and is recycled back to leaching unit 32 via line 49. The blended sludge is removed via line 51.

[0145] The decanted solution of blended botanicals is fed from decantation unit 44 via line 50 and pump 52 to filtration unit 54. Additional sludge is removed via line 53 to line 51. The solution exits filtration unit 54 via line 56 and pump 58 and enters centrifuge 60, which separates the solution and removes additional solids. Pellet waste exits centrifuge unit 60 via line 62.

[0146] The blended plant solution exits centrifuge 60 via line 64 and enters reactor 66. A solution of metal ions is introduced into the top of reactor 66 via line 68, where the biological reducing agent in the blended plant solution reacts with the metal ions to form metal nanoparticles.

[0147] Water is introduced into heater 63 via line 61. The heated water is supplied via pump 65 and line 67 to heating jacket 59 around reactor 66. Water exits reactor 66 via line 69, enters cooler 71, and is recycled back to reactor 66 via line 73. Water also exits jacket 59 via line 75, enters heater 63, and is recycled back to jacket 59 via pump 65 and line 67.

[0148] The metal nanoparticles exit reactor 66 and are fed via line 70 and pump 74 to centrifuge 72. In centrifuge 72, the reaction mixture is separated into nanoparticles and a solution of metal ions.

[0149] The metal ion solution exits centrifuge 72 and is fed via line 76 to an ion exchange chromatography unit 78. An acid solution is fed via line 80 to unit 78 to regenerate the ion exchange resin. The purified metal ions exit ion exchange chromatography unit 78 and are recycled back to reactor 66 via line 82. A waste liquid exits ion exchange chromatography unit 78 and is removed via line 84.

[0150] The metal nanoparticles exit centrifuge 72 and are fed to mixer 83 via line 86 and pump 88. A base, preferably a sodium hydroxide solution, is introduced to mixer 83 via line 90. The pH is adjusted in mixer 83, and the metal nanoparticles exit mixer 83 and are fed to sonicator 92 via line 94.

[0151] Following sonication, the metal nanoparticles exit sonicator 92 and are fed to membrane 96 via line 98 and pump 100. Oversized nanoparticles (typically greater than 200 nm) are removed by membrane 96 and exit via line 102.

[0152] Metal nanoparticles that pass through membrane 96 are fed to mixer 104 via line 106. An acid solution is introduced into mixer 104 via line 108 to adjust the pH. Colloidal nanoparticles are removed from mixer 104 via line 110.

[0153] The metal nanoparticles exit mixer 104 and are fed to dryer 112 via line 114. Water is removed in dryer 112 and exits dryer 112 via line 116 and pump 118. The finished nanoparticle powder exits dryer 112 via line 120.

[0154] An alternative process is shown in Figure 1 A. This process embodiment differs from that of Figure 1 in that unit 92 is a hybrid unit in which both sonication and mixing occur.

[0155] Purpose The metal nanoparticles of the present invention can be incorporated into a wide variety of products and processes.

[0156] Thus, in another aspect of the present invention, there is provided an article of manufacture comprising the metal nanoparticles of the present invention.

[0157] In one embodiment, a cosmetic product is provided comprising the metal nanoparticles of the present invention.

[0158] In one embodiment, a battery is provided that includes the metal nanoparticles of the present invention.

[0159] In one embodiment, a pharmaceutical product (which may be an active pharmaceutical ingredient, a pharmaceutical excipient, or a drug delivery agent) is provided that comprises the metal nanoparticles of the present invention.

[0160] In one embodiment, a plant protection product, such as an agricultural chemical product (which may be an active pesticide ingredient, a pesticide excipient, or a pesticide delivery agent) is provided that comprises the metal nanoparticles of the present invention.

[0161] In one embodiment, a building material is provided comprising the metal nanoparticles of the present invention.

[0162] In one embodiment, a detergent or cleaning product is provided comprising the metal nanoparticles of the present invention.

[0163] In one embodiment, there is provided a textile or clothing item, footwear, or accessory made therefrom (such as jewelry, bags, or other fashion items) comprising the metal nanoparticles of the present invention.

[0164] In one embodiment, a biosensor is provided that includes the metal nanoparticles of the present invention.

[0165] In one embodiment, a lubricant is provided comprising the metal nanoparticles of the present invention.

[0166] In one embodiment, an optical or optoelectronic device is provided comprising the metal nanoparticles of the present invention.

[0167] In one embodiment, a conductive ink is provided comprising the metal nanoparticles of the present invention.

[0168] In one embodiment, an antimicrobial agent is provided comprising the metal nanoparticles of the present invention.

[0169] In one embodiment, there is provided a biocide (including an insecticide) comprising the metal nanoparticles of the present invention.

[0170] In one embodiment, a catalyst is provided comprising the metal nanoparticles of the present invention.

[0171] In one embodiment, a reinforced material (such as a polymer, which may be a nanoparticle-embedded polymer, or may be a functionalized material or derivative) is provided, comprising the metal nanoparticles of the present invention.

[0172] In one embodiment, a preservative (such as a food preservative) is provided comprising the metal nanoparticles of the present invention.

[0173] In one embodiment, a filter (eg, a filter for air and / or water purification) is provided that comprises the metal nanoparticles of the present invention.

[0174] In one embodiment, a machine (such as a robot, which may be a nanorobot) is provided that comprises the metal nanoparticles of the present invention.

[0175] In one embodiment, there is provided the use of the metal nanoparticles of the present invention in biomedical imaging.

[0176] In one embodiment, there is provided the use of the metal nanoparticles of the present invention in gene sequencing.

[0177] In one embodiment, there is provided the use of the metal nanoparticles of the present invention in information storage and processing, including nanocomputing and quantum computing applications.

[0178] In one embodiment, there is provided the use of the metal nanoparticles of the present invention in nanoelectronics. [Example]

[0179] overview UV / VIS spectra were recorded on a visible spectrophotometer (721 LDC Digital Lab Spectrophotometer). Scanning electron microscopy (SEM) measurements were performed using Hitachi field emission SEMs SU8230 and SU8200. Energy dispersive X-ray spectroscopy (EDX) spectra were recorded on Hitachi SEMs SU8230 and SU8200 using a Bruker XFlash detector.

[0180] [Example 1] -GAMS modeling to develop optimal blends The optimal composition of the blend was obtained by a multi-objective linear optimization program using the ε-constraint method in GAMS, which resulted in the construction of a Pareto frontier. The two variables optimized were antioxidant capacity per 100g and cost per 100g.

[0181] Antioxidant capacity was used as a pro-environmental variable. Higher antioxidant capacity for the same mass of blend means less plant material is needed for the biosynthesis of metal nanoparticles, resulting in reduced material and energy usage. This is due to the reduced CO2 and SO2 emissions in energy production, as outlined in the article "Design Through the 12 principles of Green Engineering" (Paul T. Anastas et al., Environ. Sci. Technol. 2003, 37(5), 94A-101A). x This would result in environmental benefits through reduced direct and indirect resource consumption from reduced emissions and reduced water for growing plants. This would therefore reduce upstream environmental impacts. The process was evaluated as a whole and its impact on the environment under a life cycle analysis framework. The water, sugar, zinc, iron, and carbohydrate contents per 100 g were used as lower optimization constraints, and the vitamin C (ascorbic acid), lutein, zeaxanthin, and carotene contents per 100 g were used as upper constraints. Because drying is the first step in the production process, the amount of water in the blend was minimized to reduce downstream energy costs. Given the high enthalpy of evaporation of water (40.65 kJ / mol), this resulted in a significant reduction in energy requirements during process scale-up.

[0182] The amount of sugars and carbohydrates could also be reduced to inhibit potential microbial growth in the blend, making it shelf-stable for a longer period. The amount of zinc and iron was minimized to avoid the formation of other metal nanoparticles (MNPS) and the disruption of the zeta potential after nanoparticle generation, which could lead to aggregation of MNPS and render them unsuitable for use. Finally, the amount of water-soluble antioxidants (ascorbic acid, xanthophylls) and fat-soluble antioxidants (carotenes) was maximized to the extent reasonable, taking into account economic cost constraints.

[0183] The base case constraints developed are set as outlined in Table 1.

[0184] [Table 1]

[0185] Plants and spices readily available in the UK were included in the blending model: artichoke, curly kale, red cabbage, oregano, rosemary, sage, and watercress. Their nutritional composition was obtained from various online databases, primarily myfooddata.com. Because a linear program was used, all optima (minima in this case) were considered global optima if they belonged to the feasible set. The cost per 100g was based on local supermarket prices. These results are shown in Table 2.

[0186] [Table 2]

[0187] Finally, nine blends were obtained using programming software, which allowed the construction of a Pareto frontier, as shown in Figure 3. As can be seen in Figure 5, the main component of each blend is curly kale, with either artichoke or red cabbage used as a "filler." The spices oregano, sage, and rosemary are used as rich sources of antioxidants. The antioxidants were used to reduce metal ions to nanoparticles. It is noteworthy that the higher the antioxidant capacity of a blend, the higher its cost per 100g. Finally, from Figure 4 and Table 2, it can be seen that almost all blends, except for Blend 9, are superior (in economic terms) to a watercress-only solution, which has been widely used in previous technologies. This is because Blend 9 contains an even distribution of each plant, rather than a preferred [rich in the main filler] composition. Therefore, it was recommended to test blends that are both economically and chemically favorable for reactive synthesis. However, it should be noted that an even distribution of plant varieties allows for maximum antioxidant power, thus requiring less material and saving overall economic resources in processing.

[0188] [Example 2] -DPPH assay - experimental validation and synergistic interactions To validate the results obtained from the optimization algorithm, the antioxidant content of the resulting blends could be experimentally verified. To do so, individual plant extracts and blend extracts were tested to evaluate their relative potency in reducing the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). Pure ascorbic acid was used as the benchmark antioxidant. A colorimetric assay protocol using DPPH was developed and used to evaluate antioxidant activity. The corresponding 50% inhibitory concentration of free radicals in solution was determined for each blend and individual plant. The 50% inhibition point (IC 50 ) to IC 50The concentrations were determined by linear regression, which allowed prediction of the concentrations (Figures 7-17). All experiments were replicated and averaged. The following formula was used for the calculation: DPPH scavenging activity (%)=[(Ab-At) / Ab]×100 Ab: blank absorbance At: measured absorbance of the sample Relative intensity = IC 50 (Sample) / IC 50 (Ascorbic Acid) (May be performed in comparison with other antioxidants such as Watercress WC)

[0189] [Table 3]

[0190] [Table 4]

[0191] From Table 4, all the blends tested in Figure 5 had better DPPH IC than Watercress. 50 It can be observed that curly kale performed the least well when used alone, but was more potent when used as the primary antioxidant source in the blends. All blends ended up with higher relative intensities than most single ingredients, and although oregano and rosemary were minor ingredients, Blend 4 (without watercress) had the second highest single IC after ascorbic acid. 50 It had a higher relative intensity than oregano, which is a concentration of

[0192] This was unexpected. Therefore, the theoretical IC 50We decided to calculate the concentrations and compare these to the experimental concentrations. In Table 3, we see that nearly all blends have a higher reducing potential than predicted using linear programming. This can be the case for Blend 4, which can be over 150% stronger than theoretically predicted using mathematical modeling.

[0193] Therefore, in light of these results, it was concluded that there is a synergistic underlying mechanism during DPPH reduction when using blends compared to individual plants. This was surprising, as deviations from the theoretical model were expected to be less than 5%. 50 In addition to the variation in the amount of curly kale in these particular combinations, the fact that curly kale can exhibit stronger reducing activity than when used alone, despite remaining the major component in the blend, may still point to the enhancement and optimization according to the present invention.

[0194] [Example 3] -Method of producing the composition The selected plant material was then dried at 40°C and 1 bar for 24 hours. The dried plant material was then collected and blended in various ratios using a blender until particles smaller than 100 μm were obtained. After blending, the powder was collected and infused in a tank by adding deionized water at 50°C and 1 bar for 20 minutes with continuous stirring. The stirred tank was connected to a decanter, which itself formed three phases (scum, clarified liquid, and sludge). The sludge was recycled back to the blender for further processing to extract the maximum amount of antioxidants until the concentration in solid particles was minimized. At this point, the spent plant particles were discarded from the process. The clarified liquid was then collected and filtered under pressure (microfiltration) through various grades of filter membranes to remove particles in suspension down to 1 μm. The filtrate was then subjected to differential centrifugation (also at 2500 RPM for 10 minutes) to pellet and discard smaller particles.

[0195] [Example 4] -Method for producing silver metal nanoparticles An ionic solution is prepared at the desired concentration of 0.5 g / L to 1.5 g / L and preheated to 70°C at 1 bar for 1 hour in a reactor under continuous stirring. Once a stable temperature value is reached, a blend concentration of 0.5 g / L to 7 g / L is added to the reactor and nucleation begins instantly.

[0196] Silver acetate (AgCH3COO) was used as a substitute for silver nitrate (AgNO3), which allows for the removal of NO3 in toxic and environmentally unfriendly solutions. - The presence of ions is avoided. Crystallization lasts from 5 minutes to 2 hours, depending on the desired conversion and nanoparticle size. The nanoparticles can be seen to grow over time, thus demonstrating the tuning of nanoparticle size with reaction time. This is when bioreduction occurs and the metal nanoparticles grow by primary heterogeneous nucleation. During this period, a color change from clear to brown / black can be observed.

[0197] The reaction is then quenched using 4°C chilled water until the internal temperature of the mixture reaches 10°C. The mixture of nanoparticles and plant extract is then collected and subjected to differential centrifugation (2500 RPM for 20 minutes). To increase conversion, the supernatant is recycled back to the reactor. This passes through a cation exchange resin to maximize the recovery of pure metal ions, which are then recycled back. This purges the inactivated antioxidants from the process. The pellet (with traces of water) is collected, and NaOH is added to increase the pH to 10-14. The pellet is then sonicated for 15 minutes at 25°C and 40 kHz.

[0198] The colloid is then filtered through a micromembrane to remove large aggregates or deformed particles, ensuring high-quality functional nanoparticles with a high surface area to volume ratio, a property particularly useful for antibacterial applications, conductive inks, and catalysis. Prior to shipping, the pH of the solution is adjusted to neutral or slightly alkaline (pH=7–pH=9) using a weak organic acid such as acetic acid to prevent the solution from becoming corrosive to the user.

[0199] The colloids can also be sent to a vacuum dryer where the water is removed under vacuum (less than 3000 Pa) at room temperature. This allows for the formation of nanopowders which can then be redissolved to produce other nano-enabled products such as conductive inks which require relatively high concentrations to become conductive.

[0200] Silver nanoparticles are finally obtained in colloidal solution. As highlighted by the energy dispersive X-ray spectroscopy results, the naturally occurring biological agents in the extract (possibly including carbohydrates) acted as capping agents for the nanoparticles (see Figures 56-58).

[0201] We were able to verify that the chemical reaction was possible from room temperature (25°C). Tests were conducted at 35°C, 40°C, 50°C, 60°C, 70°C, and 85°C (Figures 18-22). Metal nanoparticles can be produced at atmospheric pressure or higher. In each case, sufficient nanoparticles were produced and identified using spectral analysis (UV / VIS). We were also able to test the effect of blend concentration on nanoparticle formation (Figure 25). A 500 mg / L blend produced Ag. + It could be observed that 0.1% of the HCl solution is a threshold concentration for sufficient reduction of 0.1% to form nanoparticles.

[0202] In Figure 24, the cyclical nature of crystallization following the LaMer nucleation mechanism can be observed. After nucleation occurs, the nanoparticles increase in size, which in turn increases the concentration of nanoparticles in solution until a critical concentration is reached. This concentration is reached 30 minutes after the start of the reaction. This concentration decreases until it reaches a stable value (reaching Gibbs free energy equilibrium), which corresponds to the thermodynamic stability achieved by colloidal solutions, which is described by a complex balance of attractive and repulsive forces acting on the nanoparticles.

[0203] From the UV-Vis analysis it was possible to determine (by the method of Paramelle et al. Analyst, 2014, 139, 4855-4861) that the nanoparticles produced at the outlet of the reactor had a diameter of 38-50 nm.

[0204] After the reactor, the nanoparticles were filtered using microfiltration. A 200 nm membrane was used to discard any large aggregates that may have remained after sonication. SEM of the filtered sample revealed nanoparticles ranging in size from 5 nm to 30 nm, as shown in Figures 26-43 and 59-62. Figure 62 shows that a large sampling of nanoparticles yielded a normal distribution with a mean diameter of 12.59 nm and a standard deviation of 5.64 nm. The nanoparticles were well distributed, non-agglomerated, and had good sphericity (>70%).

[0205] Energy dispersive X-ray spectroscopy (Figures 44-58) further confirmed the formation of silver nanoparticles. Even more interesting is the discovery of biopolymers in solution, as highlighted in Figures 56-58. Carbon and oxygen atoms can be seen to be present within and between the particles. This suggests that the presence of carbohydrates capping the nanoparticles in solution prevents nanoparticle aggregation through steric hindrance and electrostatic repulsion. No other polymers or organic stabilizers were added to the solution; these molecules originate from the original blend. This has the advantage that no additional stabilizers are needed when using our blend, thus saving material and production costs from the process.

[0206] Finally, several key advantages can be seen from Tables 5 and 6 from the process of the present invention compared to other chemical reduction routes. First, use of the cost-optimized blends disclosed herein can result in significant cost savings on reducing agents, as outlined in Table 6. Furthermore, the blends are not toxic to humans or present flammable or reactivity hazards, making the product inherently safer for the end user compared to current market alternatives. Another key advantage is that up to five times the commercial concentration can be produced by using the process of the present invention in less than 30 minutes.

[0207] [Table 5]

[0208] [Table 6]

[0209] It can also be seen in the UV-Vis spectrum that absorbance is detectable from t = 0 min. Therefore, the reaction can be inferred to be quasi-instantaneous upon contact of the reagents, thus opening up the possibility of continuous flow crystallization, as opposed to the current industry standard of batch flow. This would be a major advantage for scale-up, turning previously known batch processes into continuous processes. Furthermore, we do not use any VOCs (volatile organic compounds) or organics in our process, either as solvents or for liquid-liquid extraction.

[0210] The process of the present invention is a greener and safer biochemical alternative to current market products, from inorganic reduction pathways to current plant-based reductant pathways in the prior art.

[0211] Comparative Example 1 The possibility of substituting curly kale with other plants was evaluated. Watercress was chosen as a possible replacement because it has similar nutritional properties to curly kale. As can be seen in Figures 65 and 66, the watercress-based blend uses sage, oregano, and rosemary as high-quality antioxidant carriers and red cabbage as a filler. Nevertheless, it can be observed that to achieve the same results shown (in terms of antioxidant activity), costs could increase by 30% to 70% compared to the curly kale model (see Table 7). This further validates the choice of curly kale over watercress, compared to currently available literature findings.

[0212] [Table 7]

[0213] [Example 5] Further methods for producing silver metal nanoparticles The method generally described in Example 4 was repeated to produce silver nanoparticles in four batches. The exact conditions are designated S1-S4 in Table 8 below, with concentrations varying from 0.25 g / L to 0.9 g / L. Stirring was at 600 RPM. Any of Blends 1-5, with compositions shown in Figure 6, are suitable for carrying out this method.

[0214] Following reaction at 70°C and 600 RPM stirring, the S1, S2, and S3 reaction mixtures were quenched with ice-cold water after 5 minutes, and the S4 reaction mixture after 30 minutes. It was noted that the reactions were very fast, i.e., the conversion of silver ions to silver nanoparticles was observed in less than 5 minutes for reaction mixtures S1, S2, and S3.

[0215] After recovery of the pellet, it was then sonicated first for 10 minutes at 25°C and 40 kHz, then shaken for 5 minutes, then sonicated for another 5 minutes at 25°C and 40 kHz, and then shaken again for 5 minutes.

[0216] [Table 8]

[0217] The diameter of the nanoparticles was estimated by UV-VIS spectroscopy using the same method detailed in Example 4. The UV-VIS spectra are shown in Figures 67-70. The molar concentration of the nanoparticles was also measured and compared to that of commercially available nanoparticles. The estimated diameter and concentration are shown in Table 9.

[0218] [Table 9]

[0219] This example demonstrates that silver nanoparticles having concentrations that are more than 2.9 times, and in some cases more than 4.5 times, that of commercially available concentrations can be produced using the process of the present invention.

[0220] [Example 6] - A general method for producing gold metal nanoparticles An ionic solution is prepared at the desired concentration of 0.1 g / L to 1.5 g / L and preheated to 70 °C for 1 hour at 1 bar in a reactor with continuous stirring at 600 RPM. Once a stable temperature is achieved, a blend concentration of 0.5 g / L to 10 g / L is added to the reactor, and nucleation begins instantly. Any of Blends 1 to 5, with compositions shown in Figure 6, are suitable for this purpose. Gold(III) chloride hydrate (HAuCl4.HO) was used.

[0221] Crystallization lasted from 5 minutes to 2 hours, depending on the desired conversion and nanoparticle size. Nanoparticles can be seen to grow over time, thus demonstrating tuning of nanoparticle size with reaction time. This is when bioreduction occurs and metal nanoparticles grow by primary heterogeneous nucleation. During this period, a color change from clear to wine red / violet can be observed.

[0222] The reaction was then quenched using 4°C chilled water until the internal temperature of the mixture reached 10°C. The nanoparticle and plant extract mixture was then collected and centrifuged (10,000 rpm for 20 minutes). To increase conversion, the supernatant was recycled back to the reactor. This was passed through a cation exchange resin to maximize the recovery of pure metal ions, which were then recycled back. This purged the deactivated antioxidants in the process. The pellet (with traces of water) was collected, and NaOH was added to increase the pH to 10-14. The pellet was then sonicated at 40 kHz at 25°C for 10 minutes, followed by shaking for 5 minutes, and then sonicated at 40 kHz at 25°C for another 5 minutes, followed by shaking for another 5 minutes.

[0223] The colloids were then filtered through a micromembrane to remove large aggregates or deformed particles, ensuring high-quality functional nanoparticles with a high surface area-to-volume ratio—a particularly useful property for medical, pharmaceutical, antibacterial applications, conductive inks, and catalysis. Prior to shipping, the pH of the solution is adjusted to neutral or slightly alkaline (pH=7–pH=9) using a weak organic acid such as acetic acid to prevent the solution from becoming corrosive to users.

[0224] Gold nanoparticles were finally obtained in a colloidal solution. In each case, sufficient nanoparticles were produced and identified using spectral analysis (UV / VIS). From the UV-Vis analysis, it was possible to determine by known methods (described at https: / / www.sigmaaldrich.com / GB / en / technical-documents / technical-article / materials-science-and-engineering / biosensors-and-imaging / gold-nanoparticles) that the nanoparticles produced at the reactor outlet had a diameter of 40–60 nm.

[0225] After the reactor, the nanoparticles were filtered using microfiltration: a 200 nm membrane was used to discard any large aggregates that may remain after sonication.

[0226] [Example 7] - A specific method for producing gold metal nanoparticles The method generally described in Example 6 was carried out to produce gold nanoparticles in three batches. The exact conditions are designated S1-S3 in Table 11 below. Any of Blends 1-5, with the compositions shown in Figure 6, are suitable for carrying out this method.

[0227] The particles were made in three batches S1 to S3 with the concentrations in Table 10 below.

[0228] [Table 10]

[0229] Following reaction at 70°C and 600 RPM stirring, the S1 and S2 reaction mixtures were quenched with ice-cold water after 3 minutes, and the S3 reaction mixture after 30 minutes. It was noted that the reactions were very fast, i.e., the conversion of gold ions to gold nanoparticles was observed in less than 3 minutes for reaction mixtures S1 and S2.

[0230] The diameter of the nanoparticles was estimated by UV-VIS spectroscopy using the same method detailed in Example 6. The UV-VIS spectra are shown in Figures 71-73.

[0231] The size distribution of the gold nanoparticles is shown in Figure 74. From this figure, it can be observed that a large sampling of the nanoparticles yielded a normal distribution with a mean diameter of 21.65 nm and a standard deviation of 10.49 nm.

[0232] The gold nanoparticles were examined by scanning electron microscopy (SEM), as shown in Figures 75-82, and by energy dispersive X-ray spectroscopy (EDX), as shown in Figures 83 and 84. From EDX, the formation of gold nanoparticles could be further verified, and the positions of the gold nanoparticles and carbon and oxygen atoms could be overlaid, as shown in Figures 85-88. This further confirms the presence of at least one carbohydrate biocapping layer on the gold nanoparticles. Figure 89 is an EDX spectroscopy spectrum of gold nanoparticles, further confirming the presence of elemental gold and carbon and oxygen on the nanoparticles. This further confirms the presence of a biocapping agent present on the metal nanoparticles.

[0233] The molar concentration of the nanoparticles was also measured and compared with that of commercially available nanoparticles. The estimated diameter and concentration are shown in Table 11.

[0234] [Table 11]

[0235] This example demonstrates that gold nanoparticles with concentrations more than three times higher than those available commercially can be produced using the process of the present invention.

[0236] All publications mentioned in the foregoing specification are herein incorporated by reference. Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the present invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in chemistry or related fields are intended to be within the scope of the following claims. [Explanation of symbols]

[0237] 10 Drying Unit 12 lines 14 lines 16 Compressor 18 lines 20 Heater 22 lines 24 lines 26 lines 28 Blender 30 lines 32 Leaching Unit 34 Heater 36 lines 38 Pump 40 lines 41 Heating jacket 42 lines 44 Decanter unit 46 lines 48 Pump 49 lines 50 lines 51 Line 52 Pump 54 Filtration unit 56 lines 58 Pump 59 Heating Jacket 60 Centrifuge 61 Line 62 lines 63 Heater 64 lines 65 Pump 66 Reactor 67 Line 70 lines 71 Cooler 72 Centrifuge 73 Line 74 Pump 75 lines 76 lines 78 Ion Exchange Chromatography Unit 80 lines 82 Line 83 Mixer 84 Line 86 Line 88 Pump 90 lines 92 Ultrasonic Processor 94 lines 96 Membrane 98 lines 100 pumps 102 Line 104 Mixer 106 Line 108 Line 110 Line 112 Dryer 114 lines 116 lines 118 Pump 120 lines The inventions described in the original claims of this application are set forth below. [1] 1. A method for producing metal nanoparticles, said method comprising: (a) providing dissolved metal ions; (b) reducing the dissolved metal ions to form the metal nanoparticles; (a) Kale (Brassica oleracea Acephala group); and (b) at least one other plant selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), (v) Sage (Salvia officinalis), (vi) Watercress (Nasturtium officinale), (vii) dog rose (Rosa canina), (viii) mint (Mentha spp.); (ix) Thyme (Thymus spp.), (x) basil (Osimum basilicum), and (xi) Spinach (Spinacia oleracea); and contacting the composition with a composition comprising an extract from A method comprising: [2] 10. The method of claim 1, wherein the metal is selected from the group consisting of gold, silver, copper, nickel, platinum, iron, zinc, gadolinium, titanium, cerium, thallium, manganese, aluminum, zirconium, chromium, palladium, and cobalt, or any combination thereof. [3] The method according to [1], wherein the metal is silver. [4] The method according to [1], wherein the metal is gold. [5] The method according to any one of [1] to [4], wherein the dissolved metal ions are present in the form of a solution of a metal salt. [6] [5] The method according to [5], wherein the metal salt is an acetate salt. [7] The composition comprises: (a) Kale (Brassica oleracea Acephala group); and (b) at least one other plant selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) Watercress (Nasturtium officinale); The method according to any one of [1] to [6], comprising an extract from [8] The composition comprises: (a) Kale (Brassica oleracea var. Acephala group); and (b) at least two other plants selected from the group consisting of: (i) Artichoke (Cyanara cardunculus), (ii) red cabbage (Brassica oleracea var. Capitata group var. rubra); (iii) Oregano (Origanum vulgare), (iv) rosemary (Salvia cormanus), and (v) Watercress (Nasturtium officinale); The method according to any one of [1] to [6], comprising an extract from [9] 10. The composition of claim 1, wherein the composition comprises: (i) kale (Brassica oleracea var. Acephala group) in an amount of 30% to 90% by weight as a percentage of the total weight of the composition; (ii) artichoke (Cyanara cardunculus) in an amount of 0% to 20% by weight as a percentage of the total weight of the composition; (iii) red cabbage (Brassica oleracea var. Capitata group var. rubra) in an amount of 0% to 30% by weight as a percentage of the total weight of the composition; (iv) oregano (Origanum vulgare) in an amount of 0% to 20% by weight as a percentage of the total weight of the composition; (v) rosemary (Salvia cormanus) in an amount of 0% to 30% by weight as a percentage of the total weight of the composition, and (vi) watercress (Nasturtium officinale) in an amount of 0% to 30% by weight as a percentage of the total weight of the composition The method according to any one of [1] to [6], comprising an extract from

[10] The method according to any one of [1] to [9], wherein a solution of metal ions is added to the composition.

[11] 11. The method of claim 10, wherein the solution of metal ions is preheated before addition to the composition.

[12] Metal nanoparticles obtained or obtainable by the method according to any one of [1] to

[11] .

[13] Metal nanoparticles provided with a coating biopolymer layer.

[14] The metal nanoparticles according to

[13] , wherein the biopolymer is a carbohydrate.

[15] A product comprising the metal nanoparticles according to any one of

[12] to

[14] .

Claims

1. 1. A method for producing metal nanoparticles having a diameter between 1 nm and 1 μm, said method comprising: (a) providing dissolved metal ions; (b) reducing the dissolved metal ions to form the metal nanoparticles; (a) Kale (Brassica oleracea acephala group) and oregano (Origanum vulgare); and (b) at least one other plant selected from the group consisting of: (i) artichoke (Cynara cardunculus); (ii) Red cabbage (Brassica oleracea capitata group var. rubra); (iii) rosemary (Salvia rosmanis); (iv) sage (Salvia officinalis); (v) Watercress (Nasturtium officinale); and contacting the composition with a composition comprising an extract from A method comprising:

2. The method of claim 1, wherein the metal of the metal nanoparticles is selected from the group consisting of gold, silver, copper, nickel, platinum, iron, zinc, gadolinium, titanium, cerium, thallium, manganese, aluminum, zirconium, chromium, palladium, and cobalt, or any combination thereof.

3. The method of claim 1 , wherein the metal of the metal nanoparticles is silver.

4. The method of claim 1 , wherein the metal of the metal nanoparticles is gold.

5. 5. The method of claim 1, wherein the dissolved metal ions are present in the form of a solution of metal salts.

6. The method of claim 5 wherein the metal salt is an acetate salt.

7. The composition comprises: (a) Kale (Brassica oleracea acephala group) and oregano (Origanum vulgare); and (b) at least one other plant selected from the group consisting of: (i) artichoke (Cynara cardunculus); (ii) Red cabbage (Brassica oleracea capitata group var. rubra); (iii) rosemary (Salvia rosmanis), and (iv) Watercress (Nasturtium officinale); 7. The method of claim 1 , comprising extracting from

8. The composition comprises: (a) Kale (Brassica oleracea var. acephala group) and oregano (Origanum vulgare); and (b) at least two other plants selected from the group consisting of: (i) artichoke (Cynara cardunculus); (ii) Red cabbage (Brassica oleracea var. capitata group var. rubra), (iii) rosemary (Salvia rosmanis), and (iv) Watercress (Nasturtium officinale):

7. The method of claim 1 , comprising extracting from

9. The composition comprises: (i) kale (Brassica oleracea var. acephala group) in an amount of 30% to 90% by weight, as a percentage of the total weight of the composition; (ii) artichoke (Cynara cardunculus) in an amount of 0% to 20% by weight as a percentage of the total weight of the composition; (iii) red cabbage (Brassica oleracea var. capitata group var. rubra) in an amount of 0% to 30% by weight as a percentage of the total weight of the composition; (iv) oregano (Origanum vulgare) in an amount of 1% to 20% by weight as a percentage of the total weight of the composition; (v) rosemary (Salvia rosmanis) in an amount of 0% to 30% by weight as a percentage of the total weight of the composition, and (vi) watercress (Nasturtium officinale) in an amount of 0% to 30% by weight as a percentage of the total weight of the composition 7. The method of claim 1 , comprising extracting from

10. 10. The method of claim 1, wherein a solution of metal ions is added to the composition.

11. 11. The method of claim 10, wherein the solution of metal ions is preheated prior to addition to the composition.

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