Vitamin cold-pressed juice saturated with Anti-aging components using liposomes and method for producing same

Abstract

The present invention relates to a production method for obtaining a Nelumbo nucifera fraction by concentrating and fractionating a Nelumbo nucifera extract obtained by immersing Nelumbo nucifera in an organic solvent and then filtering same, and can provide a Nelumbo nucifera fraction which has a high phenolic compound content, thus having antioxidant activity and oxidative DNA damage inhibitory activity, and which, as a plant-derived natural antioxidant, has a very high possibility of being used as a food or cosmetic material.

Description

Vitamin juice with maximized anti-aging properties using liposomes and its manufacturing method

[0001] The present invention relates to a juice extracted from fruits and vegetables containing vitamins, and more particularly, to a vitamin juice that maximizes anti-aging ingredients by utilizing liposomes and a method for producing the same.

[0002] Herein, background information regarding the present disclosure is provided, which does not necessarily imply prior art.

[0003] South Korea is rapidly entering an aging society, driven by economic development, increased life expectancy, and a declining birth rate. The fundamental concept of anti-aging is to improve quality of life and extend lifespan. Consequently, anti-aging is not limited to the elderly; interest in anti-aging, including early anti-aging, is also growing among younger generations.

[0004] Apple juice, made by pressing raw apples, is rich in polyphenols and vitamin C, making it excellent for anti-aging. However, its high sugar content discourages consumption by those with diabetes, metabolic syndrome, and young children. Continued excessive sugar intake increases insulin resistance and can lead to serious health problems, including impaired brain function, dementia, vision loss, and high blood pressure. To address these issues, the development of new products containing ingredients that can both alleviate sugar levels and promote sugar consumption is urgently needed.

[0005] In medicine, the most common factors that contribute to aging include reactive oxygen species, changes in hormone secretion, and protein glycation. Among these, reactive oxygen species are generated by the accumulation of oxygen during the oxidation process. While oxygen plays a vital role in providing energy to the body, this process also generates harmful reactive oxygen species. These reactive oxygen species attack normal cells in the body, contributing to aging and various diseases.

[0006] Free radicals collide with and bind to surrounding molecules, damaging proteins and genes within cells. This damage forces the body to create new cells to replace them. As this happens, damage to DNA (the original cell) gradually accumulates, causing the newly created cells to perform less well than their original counterparts, reducing their regeneration rate, and accelerating aging.

[0007] In order to prevent aging, it is best to maintain an appropriate level of active oxygen. Many antioxidants are being spotlighted and studied as a means of extending lifespan, and among them, it is necessary to manage them so that active oxygen is not generated in large amounts in the first place.

[0008] The present invention aims to provide a vitamin juice with maximized anti-aging properties by utilizing liposomes.

[0009] However, the purposes of the present invention are not limited to the purposes mentioned above, and other purposes not mentioned will be clearly understood by those skilled in the art from the description below.

[0010] The present invention provides a blended vitamin juice comprising an apple juice containing vitamin C, a carrot juice containing vitamin A, and a liposome preparation containing vitamin B group.

[0011] In addition, the apple juice is included in an amount of 40 to 50 wt%, the carrot juice is included in an amount of 30 to 40 wt%, and the liposome formulation is included in an amount of 15 to 25 wt%.

[0012] In addition, the present invention provides a method for producing vitamin juice by combining apple juice containing vitamin C, carrot juice containing vitamin A, and a liposome preparation containing vitamin B group.

[0013] In addition, the liposome preparation is characterized by providing a method for producing vitamin juice, which comprises the steps of: mixing 10 to 30 wt% of an emulsifier, 35 to 45 wt% of a vitamin B group, and 15 to 25 wt% of an organic acid in an aqueous solution; homogenizing the mixture at 1,500 to 2,000 rpm for 100 to 120 minutes at a temperature of 50 to 55°C; stirring the homogenate at 3,000 to 4,000 rpm for 40 to 50 minutes at a temperature of 30 to 40°C; and cooling the stirred mixture.

[0014] In addition, the emulsifier is characterized in that it includes at least one selected from the group consisting of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and hydrogenated compounds thereof, and the organic acid is used at a pH of 3.0 to 3.5.

[0015] The present invention provides a vitamin B component that helps promote sugar consumption by combining it with liposomes and mixing it into juice, thereby reducing the production of active oxygen and providing an antioxidant effect.

[0016] All technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art, unless otherwise stated. Furthermore, throughout this specification and the claims, unless otherwise stated, the terms "comprise," "comprises," and "comprising" are intended to encompass a stated object, step, or group of objects, and steps, but not to exclude any other object, step, or group of objects, or groups of steps.

[0017] Before describing the present invention in detail below, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention which is limited only by the scope of the appended claims.

[0018] Meanwhile, the various embodiments of the present invention may be combined with any other embodiments unless explicitly stated otherwise. Any feature indicated as particularly desirable or advantageous may be combined with any other feature or features indicated as desirable or advantageous. Hereinafter, embodiments of the present invention and their resulting effects will be described with reference to the accompanying drawings.

[0019] According to one embodiment of the present invention, a vitamin juice is prepared by combining apple juice containing vitamin C, carrot juice containing vitamin A, and a liposome preparation containing vitamin B group to provide an anti-aging effect with maximized anti-aging ingredients.

[0020] The above apple juice is extracted from the liquid portion of the apple by pressing it. It is rich in polyphenols and vitamin C, making it excellent for anti-aging. The apple juice is included in an amount of 40 to 50% by weight. If it is included in an amount less than 40% by weight, the sugar content decreases, making it less sweet and difficult to consume. If it is included in an amount exceeding 50% by weight, continuous consumption can increase insulin resistance and lead to problems such as impaired vision and high blood pressure.

[0021] The carrot juice described above is extracted from the liquid portion of raw carrots by pressing them. Rich in vitamin A, it provides antioxidant benefits and helps prevent aging and cancer. It is also rich in lutein and lycopene, which are beneficial for eye health and help prevent high blood pressure and arteriosclerosis. The carrot juice is included at 30-40% by weight, counteracting the potential problems associated with excessive sugar intake in apple juice.

[0022] The above liposome formulation contains vitamin B complex inside a spherical liposome surrounded by a phospholipid bilayer. Since the interior of the cell is hydrophilic, it is the most useful functional substance transporter in microencapsulation. The capacity of liposomes varies depending on the lipid composition, surface charge, size, and method of forming the liposome. The liposome formulation is included in an amount of 15 to 25 wt%. It is preferable that the liposome formulation be included in an amount of 0.2 to 0.3 times the weight of the raw material (apple, carrot) juice.

[0023] The above liposome preparation is manufactured by including the steps of mixing 10 to 30 wt% of an emulsifier, 35 to 45 wt% of vitamin B group, and 15 to 25 wt% of an organic acid in an aqueous solution; homogenizing the mixture at a temperature of 50 to 55°C for 100 to 120 minutes at 1,500 to 2,000 rpm; stirring the homogenate at a temperature of 30 to 40°C for 40 to 50 minutes at 3,000 to 4,000 rpm; and cooling the stirred mixture.

[0024] The B vitamins mentioned above include eight types: vitamins B1, B2, B6, B12, niacin, pantothenic acid, folic acid, and biotin. Vitamin B is involved in energy metabolism, neurotransmission, and fat metabolism. Because these B vitamins are water-soluble, they are non-toxic and can be excreted without accumulating in the body. Foods containing vitamin B include spinach and buckwheat, but most of the vitamin deficiencies found in our intake are in the B group.

[0025] The above emulsifier is at least one selected from the group consisting of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and hydrogenated compounds thereof. Preferably, phosphatidylglycerol is used.

[0026] The above organic acid is a weak organic acid with a pH of 3.0 to 3.5, and it is preferable to use malic acid.

[0027] The antioxidant ingredients such as polyphenol, lutein, lycopene, vitamins A, B, and C contained in the above vitamin juice have different parts of action depending on their location, and each ingredient has a synergistic effect on the antioxidant effect like gears, so when consumed in the above ratio, the generated active oxygen is converted into H2O and excreted from the body, thereby maximizing the anti-aging effect.

[0028]

[0029] Manufacturing Example - Manufacturing of Liposome Formulation

[0030] Phosphatidylglycerol (20 wt%) was added to purified water containing vitamin B complex (40 wt%) and malic acid (20 wt%), and then homogenized and stirred using an inline disperser (μTron-ISP 25). First, the mixture was homogenized at 1,800 rpm at 53°C for 100 minutes using a homogenizer (Homo Emulsifier Disperser; HED) to induce liposome dispersion. Then, the mixture was stirred at 3,500 rpm at 45°C for 45 minutes using a scraper paddle to perform the liposome rolling process. After homogenization, the stirring vessel was immersed in ice water for 30 minutes to prepare a liposome formulation (A).

[0031] Lecithin (20 wt%) was added to purified water containing vitamin B complex (20 wt%) and anhydrous citric acid (5 wt%), and then homogenized and stirred using an inline disperser (μTron-ISP 25). First, the mixture was homogenized at 65°C and 6,000 rpm for 20 minutes using a homogenizer (Homo Emulsifier Disperser; HED) to induce liposome dispersion. Then, the mixture was stirred at 65°C and 60 rpm for 90 minutes using a scraper paddle to perform the liposome rolling process. After homogenization, the stirring vessel was immersed in ice water for 30 minutes to prepare a liposome formulation (B).

[0032]

[0033] Example

[0034] The juice was prepared by mixing the ingredients in the weight ratio shown in Table 1 below and using a mixer for 10 minutes.

[0035] Apple juiceCarrot juiceLiposome preparation ALiposome preparation BExample 1403525-Example 2453520-Example 3503515-Comparative example 14035-25Comparative example 24535-20Comparative example 35035-15

[0036] Experimental example

[0037] (1) Antioxidant activity - DPPH / ABTS radical scavenging activity

[0038] The mechanism of antioxidant activity is to react with free radicals, and free radical scavenging action is used as a measure of antioxidant effect in plants or aging inhibition in the human body by donating electrons to free radicals. DPPH and ABTS are substances with relatively stable radicals, and DPPH radical scavenging activity is a method for measuring the activity of hydrogen donors. It is a method that uses the principle that purple color is discolored when reduced by hydrogen or electrons received by phenolic compounds, aromatic amines, and ascorbic acid, and is widely used to search for antioxidant substances. In order to evaluate the antioxidant activity of the prepared juice, DPPH radical scavenging activity, ABTS radical scavenging activity, and reducing power were evaluated.

[0039] Radical scavenging activity using DPPH was measured with reference to the Bondet method. The DPPH solution was prepared by diluting 300 μM 1,1-diphenyl-2-picryl hydrazyl (DPPH) with ethanol so that the absorbance value at 515 nm was 1.00. 760 μl of the DPPH solution was added to 40 μl of each of the prepared juices, and the mixture was incubated at 37°C for 20 minutes. The absorbance was measured at 515 nm using a UV / Visible spectrophotometer (Human Cop, Xma-3000PC, Korea).

[0040] Scavenging activity (%) = {1-(absorbance of the group with added juice / absorbance of the group without added juice)} × 100

[0041] ABTS radical scavenging activity exhibits scavenging activity by a mechanism that reduces myoglobin radicals through a rapid antioxidant reaction between hydrogen peroxide and metmyoglobin in the presence of ABTS, which generates radicals. The ABTS radical scavenging activity was measured with reference to the Van den Berg method. The ABTS solution was prepared by mixing 7.4 mM 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt and 2.6 mM potassium persulfate to form ABTS radicals for 24 hours, and then diluted with distilled water so that the absorbance value at 734 nm was 0.70. 760 μl of the ABTS solution was added to each 40 μl of the prepared juice, and the mixture was incubated at 37°C for 20 minutes. The absorbance was measured at 734 nm using a UV / Visible spectrophotometer.

[0042] Scavenging activity (%) = {1-(absorbance of the group with added juice / absorbance of the group without added juice)} × 100

[0043] The results of evaluating the DPPH / ABTS radical scavenging activity of the prepared juices are shown in Table 2 below. As shown in Table 2 below, it can be confirmed that the DPPH / ABTS radical scavenging activity of the squeezed juices (Examples 1 to 3) containing the liposome formulation (A) is significantly increased compared to the squeezed juices (Comparative Examples 1 to 3) containing the liposome formulation (B). In particular, the DPPH / ABTS radical scavenging activity of Example 2 containing 0.2 to 0.3 times the liposome formulation (A) compared to the original squeezed juice is the highest, confirming that the antioxidant efficacy is maximized.

[0044] DPPH scavenging activity (%) ABTS scavenging activity (%) Example 184.47±4.1483.85±8.06 Example 287.02±0.6886.42±0.25 Example 383.62±3.7181.42±4.61 Comparative example 148.13±1.9456.44±0.15 Comparative example 255.61±7.4659.15±7.14 Comparative example 349.73±3.1155.67±3.61

[0045] (2) Inhibitory activity against DNA damage caused by oxidative stress

[0046] Cells maintain multiple antioxidant systems to defend against reactive oxygen species. However, when the total amount of reactive oxygen species exceeds the antioxidant defense system's ability to maintain equilibrium, disrupting the redox balance, cells are inevitably exposed to oxidative stress. Given that oxidation-induced mutations are responsible for the breakdown and resynthesis of proteins and lipids, the development of cancer due to DNA damage is significantly influenced by oxidative stress.

[0047] In order to evaluate the inhibition activity of the above-mentioned juice on DNA damage caused by oxidative stress, DNA damage was comparatively analyzed using oxidative stress through the Fenton reaction of FeSO4 (ferrous sulfate) and H2O2, and evaluated using a non-cellular system using the φX-174 RF I plasmid DNA cleavage assay.

[0048] φ X-174 RF I plasmid DNA oxidative stress damage inhibitory activity was measured using the method of Jung and Surh.

[0049] 40 μl of each of the prepared juices and 760 μl of a 1:1 mixture of 0.5 mM FeSO4 and 0.5 mM hydrogen peroxide (H2O2) were added, and the mixture was reacted at 37°C for 15 minutes. 20 μl of the reactant and 5 μl of φX-174 RF I plasmid DNA were added, and the mixture was reacted at 37°C for 3 minutes, mixed with 10X loading buffer, and electrophoresis was performed on a 1% agarose gel to measure the DNA damage inhibition activity (%).

[0050] The results of evaluating the activity of inhibiting DNA damage caused by oxidative stress of the prepared juices are shown in Table 3 below. As shown in Table 3 below, in the case of the juices containing liposome preparation (A) (Examples 1 to 3), the OH derived from FeSO4 was higher than in the juices containing liposome preparation (B) (Comparative Examples 1 to 3). - It can be confirmed that the oxidative damage inhibition effect against radicals is significantly increased. In particular, it can be confirmed that the DNA damage inhibition activity caused by oxidative stress of Example 2, which contains 0.2-0.3 times the amount of liposome preparation (A) compared to the raw juice, is the highest, thereby maximizing the antioxidant efficacy.

[0051] DNA damage inhibition activity (%) Example 170.16 Example 277.85 Example 372.43 Comparative Example 142.60 Comparative Example 250.26 Comparative Example 325.18

[0052] The features, structures, effects, etc. exemplified in each of the aforementioned embodiments can be combined or modified to implement other embodiments by those skilled in the art. Therefore, the contents related to such combinations and modifications should be construed as being included within the scope of the present invention.

Claims

1. A blended vitamin juice comprising apple juice containing vitamin C, carrot juice containing vitamin A, and a liposome preparation containing vitamin B group.

2. In paragraph 1, The above apple juice is contained in an amount of 40 to 50 wt%, The above carrot juice is contained in an amount of 30 to 40 wt%, Vitamin juice characterized in that the liposome formulation is contained in an amount of 15 to 25 wt%.

3. A method for producing vitamin juice comprising mixing apple juice containing vitamin C, carrot juice containing vitamin A, and a liposome preparation containing vitamin B group.

4. In paragraph 3, A method for producing vitamin juice, characterized in that the liposome preparation is produced by including the steps of: mixing 10 to 30 wt% of an emulsifier, 35 to 45 wt% of a vitamin B group, and 15 to 25 wt% of an organic acid in an aqueous solution; homogenizing the mixture at a temperature of 50 to 55°C for 100 to 120 minutes at 1,500 to 2,000 rpm; stirring the homogenate at a temperature of 30 to 40°C for 40 to 50 minutes at 3,000 to 4,000 rpm; and cooling the stirred mixture.

5. In paragraph 3, The above emulsifier comprises at least one selected from the group consisting of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and hydrogenated compounds thereof. A method for producing vitamin juice, characterized in that the organic acid has a pH of 3.0 to 3.5.