1. A nano-food composition comprises; a) bioactive compounds extracted from beet greens, wherein, the said bioactive compounds are encapsulated by preparing a nanoemulsion and b) surfactants.

2. The composition as claimed in claim 1, wherein the oil phase (dispersed phase) of the nanoemulsion is cold-pressed sunflower oil.

3. The composition as claimed in claim 1, wherein the aqueous phase (dispersion/ continuous phase) of the nanoemulsion is bioactives extract.

4. The composition as claimed in claim 1, wherein tween 80 is used as a surfactant.

5. The composition as claimed in claim 1, wherein soy lecithin is used as a co-surfactant.

6. The composition as claimed in claim 1, wherein the surfactant (tween 80) and co-surfactant (soy lecithin) are used in a ratio of 3:1.

7. The composition as claimed in claim 1, wherein the nanoemulsion are used for the preparation of the nano food “focaccia”.

8. A method of preparation of a nano-food comprises the steps of: a) extracting bioactive compounds from beet greens, in which 50 g of dried sample of beet greens is dissolved in 100 ml of diluted methanol (1:1 v/v); the solution is let sit for 48 hours in room temperature (25-30?C); b) exposing the solution of a) in ultrasonic water-bath for 2 hours and evaporating the solvent using rotary evaporator at 69 rpm with bath temperature 45?C till it gets concentrated; c) drying the extract in an air circulated oven at 45-50?C to remove the excess moisture; d) encapsulating the bioactives extracted from beet greens; e) weighing surfactant, co- surfactant and aqueous phase (bioactive extract) in a 200 ml beaker and mixing by using a magnetic stirrer at 50? C; f) homogenising the mixture by adding the oil phase drop by drop and further homogenising for 30 minutes at 4000 rpm to produce a coarse emulsion; g) sonicating the coarse emulsion using probe sonicator at 20 kHz in ice bath for 15 minutes; and h) preparing a focaccia enriched with the beet green bioactives nanoemulsion.

9. The method as claimed in claim 8, wherein the extraction of bioactive from beet greens is carried out by using an ultrasonicated assisted extraction method.

Description:FIELD OF THE INVENTION
The present invention relates to the field of nanotechnology. More specifically, the invention relates to nano-encapsulation of bioactive compounds extracted from beet greens. The present invention also focuses on the preparation of focaccia with the nanoencapsulated bioactives extracted from beet greens
BACKGROUND
Use of nanotechnology in development of nano-food is an emerging concept as encapsulation of bioactive compounds such as vitamins, antioxidants, and essential fatty acids in nanostructures, enable their controlled release and better absorption in the body. This not only enhances the nutritional value of foods but also allows for the creation of functional foods with targeted health benefits, such as fortified beverages and snacks. But there is still a large gap in fully utilizing this technique to develop new value-added innovative nano-foods.
There were lot of food products being made with the beetroot, beetroot extracts, beet root powder etc., however, there is little to no attention given towards the beet greens, which are generally discarded as waste but, are full of bioactive compounds. This is a sustainable approach towards utilizing such underutilized byproducts. The present invention focuses on utilizing the bioactive compounds extracted from beet greens, and formulating a nanoemulsion to develop value added focaccia (bread).
Although, there has been a lot of variants of bread in the market that fortifies with minerals but none of it was ever been ever developed in ordered to enhance its bioactive profile. The present invention enhances the bioactive profile of the focaccia bread by nanoencapsulating the bioactives extracted from the beet greens.
OBJECTIVE OF THE INVENTION
The objective of the present invention is to extract the bioactives from Beet greens.
Another objective of the present invention is to nanoencapsulate the bioactives extracted from Beet greens.
Yet another objective of the present invention is to prepare focaccia enriched with nanoencapsulated bioactive compounds extracted from beet greens.
DISCUSSION OF THE PRIOR ART
CN113368255A titled ‘Betacyanin nano-liposome as well as preparation method and application thereof’ discloses a betacyanin nano-liposome and a preparation method and application thereof, wherein the preparation method comprises the following steps: dissolving soybean lecithin and cholesterol in an organic solvent to obtain a solution 1; dissolving betacyanin in PBS solution to obtain betacyanin solution(ii) a Mixing the solution 1 and the betacyanin solution, and carrying out short-time ultrasonic treatment; removing the organic solvent by rotary evaporation, adding a Tween 80 PBS solution after reaching the colloidal state, and carrying out reduced pressure hydration; and carrying out short-time ultrasonic treatment again to obtain the betacyanin nano liposome solution. The liposome prepared by the invention has higher stability, and lipid molecules are not easy to hydrolyze or oxidize; the shape is regular, the grain size is small, and the product is safe and nontoxic; EC for human liver cancer cell HepG250EC of value compared to betacyanin50The value is improved by at least 18 times, and the anti-tumor effect is good.
BR102016006496A2 titled ‘Process of preparation of nanoparticles of pigments encapsulated in gelatin and nanoparticles of pigments encapsulated in gelatin’ discloses a process of preparation of gelatin encapsulated pigment nanoparticles and gelatin encapsulated pigment nanoparticles. The present invention relates to a process for preparing gelatin encapsulated pigment nanoparticles and the nanoparticles thus obtained, providing a means for stabilizing dyes and pigments. allowing the application of extracts of natural, hydrophobic or water soluble origin, preserving the vibrant color that they present at the time of extraction from their natural sources. This will solve the problems related to the toxicology of artificial colors and the poor stability of colorings and extracts of natural pigments, thus increasing the potential for application in food and other products. The product can be applied to fast food. In addition, it can also be used for pharmaceutical, cosmetic and biomedical purposes.
It is yet another object of the present invention the process to comprise natural dyes selected from a group comprising phycobiliproteins, pyrrolic heterocyclic structure, flavonoids, betalains, quinoidal pigments and ribloflavin. And comprising artificial dyes selected from a group containing identical natural dyes, inorganic dyes and caramel.
Noor Mohammed, Padma Ishwarya S, and Prakasan Nisha titled ‘Nanoemulsion Versus Microemulsion Systems for the Encapsulation of Beetroot Extract: Comparison of Physicochemical Characteristics and Betalain Stability’ beetroot extract is a widely used food colorant. Betalain is the major red pigment present in it, which is unstable at high temperature and pH. In this study, the edge of nanoemulsion over microemulsion in improving the thermal and pH stability of betalain has been demonstrated. The micro- and nanoemulsions of beet extract were prepared by rotor-stator homogenization and rotor-stator homogenization followed by ultrasonication, respectively. The resultant emulsions were compared for their physicochemical and rheological properties, storage stability (for 15 days, at refrigeration temperature: 4 ± 1 °C; room temperature: 30 ± 5 °C; 40 °C and 100 °C), and release kinetics. Compared to microemulsion, nanoemulsion resulted in a sustained release and higher thermal and pH stabilities of betalain. During storage, nanoemulsion showed a lower rate of droplet coarsening and improved color stability than microemulsion, due to its nanoscale droplet diameter (701.53 ± 27.2 nm), high viscosity (0.089 Pa s, at 100 s-1), and non-Newtonian rheological behavior.
In the present invention, beet greens are used that are valuable source of bioactive components such as pigments (betacyanin, and betaxanthies) and phytochemicals (phenolic compounds, flavonoids, carotenoids) that can be extracted to boost plant utilization and they can be utilized as an economical source of vital compounds which could be incorporated in fortifying food products.
SUMMARY OF THE INVENTION
The present invention relates to focaccia enriched with nanoencapsulated bioactive compounds extracted from beet greens.
According to an embodiment of the present invention, bioactive compounds are extracted from beet greens. Beet greens are a valuable source of bioactive components such as pigments (betacyanin, and betaxanthies) and phytochemicals (phenolic compounds, flavonoids, carotenoids). These bioactives can be extracted and used as food additives because of their antioxidant properties. Bioactives are susceptible to degrading circumstances such as temperature alterations, oxidation, photosensitivity, variations in pH level, undesired interactions with other components, etc. to overcome it the process of nanoencapsulation helps in conserving bioactive components by providing a safe medium for its delivery of the active compound. Nanoemulsion is one such technique for nanoencapsulation bioactive compound to develop a nano-food “Focaccia”.
According to another embodiment of the present invention, to prepare bioactive extract, 50 g of dried sample was dissolved in 100 ml of diluted methanol (1:1 v/v); the solution was let sit for 48 hours in room temperature (25-30?C).
Later it was exposed in ultrasonic water-bath for 2 hours and then solvent was evaporated using rotary evaporator at 69 rpm with bath temperature 45?C till it was concentrated. The extracts were dried in an air circulated oven at 45-50?C to remove the excess moisture.
According to another embodiment of the present invention, to formulate the nanoemulsion, surfactant, co- surfactant and aqueous phase (bioactive extract) were initially weighed in a same 200 ml beaker and the mixture was mixed using magnetic stirrer at 50? C. To produce coarse emulsion, mixture was then homogenised while adding the oil phase drop by drop and it was homogenised further for 30 minutes at 4000 rpm.
Later this coarse emulsion was sonicated using a probe sonicator at 20 kHz in ice bath for 15 minutes. After ultrasonication the samples were collected for further analysis.
According to another embodiment of the present invention, the bioactives extracted from beet greens are encapsulate by preparing a nanoemulsion. It includes, oil phase (dispersed phase) of cold-pressed sunflower oil, aqueous phase (dispersion/ continuous phase) of bioactives extract, tween 80 as a surfactant and soy lecithin as a co-surfactant. The surfactant (tween 80) and co-surfactant (soy lecithin) are used in a ratio of 3:1.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 shows process of making the focaccia enriched with nanoencapsulated bioactive compounds extracted from beet greens.
DETAILED DESCRIPTION OF THE INVENTION
The following description includes the preferred best mode of one embodiment of the present invention. Reference now will be made in detail to embodiments of the invention, one or more examples of which are illustrated in the drawings. Each example is provided by way of explanation of the invention, not limitation of the invention.
In the present invention, beet greens are used that are valuable source of bioactive components such as pigments (betacyanin, and betaxanthies) and phytochemicals (phenolic compounds, flavonoids, carotenoids) that can be extracted to boost plant utilization and they can be utilized as an economical source of vital compounds which could be incorporated in fortifying food products. These bioactives can be extracted and used as food additives because of their antioxidant properties. Bioactives are susceptible to degrading circumstances such as temperature alterations, oxidation, photosensitivity, variations in pH level, undesired interactions with other components, etc. to overcome it the process of nanoencapsulation helps in conserving bioactive components by providing a safe medium for its delivery of the active compound. Nanoemulsion is one such technique for nanoencapsulation bioactive compound to develop a nano-food “Focaccia”. Focaccia is a type of a bread baked in a flat sheet pan and used to accompany soups, stew, and pasta.
Extraction of bioactive compounds from beet greens:
Prior to encapsulation, bioactive were extracted from beet greens using an ultrasonicated assisted extraction method. The extract was then analyzed for the determination of several bioactives such as betacyanin, betaxanthin, total phenolic compounds, total flavonoid content, carotenoids, alkaloids, tannins, and saponins.
To prepare bioactive extract, 50 g of dried sample of beet greens was dissolved in 100 ml of diluted methanol (1:1 v/v); the solution was let sit for 48 hours in room temperature (25-30?C).
Later it was exposed in ultrasonic water-bath for 2 hours and then solvent was evaporated using rotary evaporator at 69 rpm with bath temperature 45?C till it was concentrated. The extracts were dried in an air circulated oven at 45-50?C to remove the excess moisture.
The extracts were stored in an airtight container for further analysis of bioactive compounds present in it.
Table 1: Analysis of bioactives extracted from beet greens.
Betaxanthin (mg/100g) Betacyanin (mg/100g) Carotenoids (mg/100g) Flavonoids (mg/g) Alkaloids (mg/g) Saponins % Tannins (mg/l) Terpenoids (mg/g)
Beet stalk and greens 9.18 7.27 6.27 9.29 7.98 0.52 275 3.48

Nanoencapsulation of bioactive compounds extracted from beet greens:
Later, to encapsulate the bioactives extracted from beet greens, a nanoemulsion was formulated. Components of nanoemulsion:
Oil phase (dispersed phase): cold-pressed sunflower oil
Aqueous phase (dispersion/ continuous phase): bioactives extract
(To prepare bioactive extract, beet green extract was dissolved (1g/ 2ml distilled water) for 24 hours. The solution is then filtered using whatman filter paper, to remove the residue and collect the filtrate. The filtrate was used as bioactive extract.)
Stabilizer: Surfactant (tween 80) and a co-surfactant (soy lecithin) was used in a ratio of 3:1. The optimization for the amount of surfactant and co surfactant to be used in the preparation of nanoemulsion was carried out using 10 formulations to find out the best suited composition.
Method of formulation of beet green bioactive nanoemulsions
As per the composition given in table-2, surfactant, co- surfactant and aqueous phase (bioactive extract) were initially weighed in a same 200 ml beaker and the mixture was mixed using magnetic stirrer at 50? C.
To produce coarse emulsion, mixture was then homogenised while adding the oil phase drop by drop and it was homogenised further for 30 minutes at 4000 rpm.
Later this coarse emulsion was sonicated using probe sonicator at 20 kHz in ice bath for 15 minutes. After ultrasonication the samples were collected for further analysis.
Table 2: Composition of oil, surfactants, and Aqueous (Bioactive Extract) for the formulation of beet green bioactive nanoemulsions.

S/O Oil (g) Surfactant (g)
(S1+ S2) S1 (g) S2 (g) Aqueous (Bioactive Extract) (g)
0.3 5 1.5 1.125 0.375 93.5
0.35 5 1.75 1.3125 0.4375 93.25
0.4 5 2 1.5 0.5 93
0.45 5 2.25 1.6875 0.5625 92.75
0.5 5 2.5 1.875 0.625 92.5
0.55 5 2.75 2.0625 0.6875 92.25
0.6 5 3 2.25 0.75 92
0.65 5 3.25 2.4375 0.8125 91.75
0.7 5 3.5 2.625 0.875 91.5
0.75 5 3.75 2.8125 0.9375 91.25

The characterization of nanoemulsion was carried out to find out the applicable nanoemulsion for the formulation of the nano food “focaccia”.
Development of Focaccia:
It is a flat bread made up with flour and yeast. A good amount of olive oil in used to give itself a distinct flavour. For this study, focaccia will be enriched with the beet green bioactives nanoemulsion, which will therefore enhance its bioactive profile than the standard focaccia made.
, Claims:WE CLAIM
1. A nano-food composition comprises; a) bioactive compounds extracted from beet greens, wherein, the said bioactive compounds are encapsulated by preparing a nanoemulsion and b) surfactants.
2. The composition as claimed in claim 1, wherein the oil phase (dispersed phase) of the nanoemulsion is cold-pressed sunflower oil.
3. The composition as claimed in claim 1, wherein the aqueous phase (dispersion/ continuous phase) of the nanoemulsion is bioactives extract.
4. The composition as claimed in claim 1, wherein tween 80 is used as a surfactant.
5. The composition as claimed in claim 1, wherein soy lecithin is used as a co-surfactant.
6. The composition as claimed in claim 1, wherein the surfactant (tween 80) and co-surfactant (soy lecithin) are used in a ratio of 3:1.
7. The composition as claimed in claim 1, wherein the nanoemulsion are used for the preparation of the nano food “focaccia”.
8. A method of preparation of a nano-food comprises the steps of:
a) extracting bioactive compounds from beet greens, in which 50 g of dried sample of beet greens is dissolved in 100 ml of diluted methanol (1:1 v/v); the solution is let sit for 48 hours in room temperature (25-30?C);
b) exposing the solution of a) in ultrasonic water-bath for 2 hours and evaporating the solvent using rotary evaporator at 69 rpm with bath temperature 45?C till it gets concentrated;
c) drying the extract in an air circulated oven at 45-50?C to remove the excess moisture;
d) encapsulating the bioactives extracted from beet greens;
e) weighing surfactant, co- surfactant and aqueous phase (bioactive extract) in a 200 ml beaker and mixing by using a magnetic stirrer at 50? C;
f) homogenising the mixture by adding the oil phase drop by drop and further homogenising for 30 minutes at 4000 rpm to produce a coarse emulsion;
g) sonicating the coarse emulsion using probe sonicator at 20 kHz in ice bath for 15 minutes; and
h) preparing a focaccia enriched with the beet green bioactives nanoemulsion.
9. The method as claimed in claim 8, wherein the extraction of bioactive from beet greens is carried out by using an ultrasonicated assisted extraction method.