KR100536083B1 - Mixture of pine tree leaves supernatant juice fermentation solution and Lespedeza cuneate G. Don extract and health food containing the same - Google Patents

Abstract

The present invention relates to a fermentation broth of pine needle juice supernatant and a non-aqueous alcohol extract having excellent anticancer, antibacterial and vascular relaxation effects, and a health supplement containing the same as an active ingredient, and more specifically, a fermentation broth of pine needle juice supernatant: Non-repair alcohol extract = 10 to 20% by weight: 80 to 90% by weight of the anti-cancer, antibacterial and vascular relaxation effect prepared by mixing after aging and a healthy supplement containing the same as an active ingredient. Mixed fermentation broth of fermentation broth and non-aqueous alcohol extract of pine needle juice supernatant of the present invention can be useful as a health supplement because the main components are well preserved and excellent anti-cancer, antibacterial and vasodilation effect.

Description

Mixture of pine tree leaves supernatant juice fermentation solution and Lespedeza cuneate G. Don extract and health food containing the same}

The present invention relates to a mixed fermentation broth of fermentation broth and non-aqueous alcohol extract of pine needle juice supernatant and a dietary supplement containing the same as an active ingredient.

Pine tree (Pinus densiflora Siebold et Zuccarini) is a coniferous tree belonging to the Pinaceae family and is wild in the mountains of Korea (Kim Il-hyuk, Ph.D., Jinmyeong Publishing Co., 1981, 71). Pine needles have long been known to be effective in preventing blood stroke, arteriosclerosis, hypertension, diabetes, etc. (Kim Wan-hee, Hyeon Dong Bogam, Pacific Publishing Co., 1983, 960).

Terpene, an active ingredient of pine needles and volatile antibiotics, contains about 7-12% of pine needles and is composed of a large amount of isoprene, called monoterpene, sequiterpene, and diterpene. It also has a unique pharmacological effect, such as lowering blood pressure, stimulating hormone secretion, and antioxidants. Pine needles contain 16 essential amino acids, including 8 essential amino acids, which are very effective for body growth and vitality, improve blood flow and stimulate hormone secretion. Excellent effect on hyperlipidemia and cholesterol lowering.

Among the active ingredients of pine needles, vitamin C has a blood purification and anti-neuropathy, and resistance to stress, vitamin A functions to strengthen the mucous membranes, and is used to treat night blindness, and resin and tannin help the digestive function. In addition, alcohols and esters promote body metabolism by releasing waste products in the body, and are effective in treating atherosclerosis due to the components of calcification. Iron is effective in treating anemia, and chlorophyll (chlorophyll) reduces accumulated cholesterol. In addition, glycogenin has a potentiating effect on diabetes and is effective in diabetes. In addition, apienic acid is good for detoxifying opioids and nicotine, pulmonary tuberculosis and pleurisy, and gastrointestinal diseases.It is rich in vitamin C and iron, which is effective for anemia. Vitamin P (rutin) helps to prevent aging by strengthening capillaries.

In addition, non-suri (night view) means a herb that opens the bar at night, and when it is eaten, it is said to shine even outside of Cheonli (千里光). It is also called the Great King (大力 王) because of its great power, and it is also called Satemcho (하여 草) for chasing snakes. The rhinoceros has an excellent effect in treating various male diseases, lack of nourishment, premature ejaculation, oil well, and vulgaris. In particular, it has excellent effects on the lack of yang of elderly people with weak kidney function. The leaves, roots, and stems of the rhinoceros contain flavonoids, finitol, phenol, tannin, and cytosterol, which remove inflammation, sputum, and kill or inhibit staphylococcus aureus, pneumococci, streptococci, and catarrhals. (Kim Tae-Jung, Resources Plant of Korea, Seoul National University Press, 1996; Jin-Kyu Choi, Our Herb, Flower, Tree, Korean Culture, 2001).

Accordingly, the present inventors separated pine needle juice supernatant without loss and change of major components from pine needles, fermented and matured it, and then mixed with the non-aqueous distilled liquor extract and aged, and completed the present invention by preparing a dietary supplement comprising the same. .

It is an object of the present invention to provide a fermentation broth of pine needle juice supernatant and a mixed fermentation broth and a dietary supplement comprising the same.

In order to achieve the above object, the present invention provides a mixed aging solution of fermentation broth and non-repair alcohol extract of pine needle juice supernatant.

The present invention also provides a dietary supplement containing the mixed aging liquid as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a mixed ripening liquid of fermentation broth and non-repair alcohol extract of pine needle juice supernatant.

The fermentation broth of the pine needle juice supernatant and the non-aqueous alcohol extract fermentation broth of the pine needle juice supernatant: non-alcoholic alcohol extract = 10 to 20% by weight: preferably 80 to 90% by weight, fermentation of pine needle juice supernatant: non-aqueous alcohol extract = 15 wt%: more preferably 85 wt%. In addition, it is preferable to mix the fermentation broth and non-aqueous extract of pine needle juice supernatant and then aged at 25 ℃ for more than 3 months.

The pine needle extract fermentation broth is

1) washing the pine needles;

2) pulverizing and washing the pine needles washed;

3) separating the supernatant from the juice; And

4) It is preferable that the pine needle supernatant of step 3 is prepared, including the step of fermentation at 37 ℃.

In the step 1, it is preferable that pine needles are selected from healthy pine trees in a clean area grown at more than 350 m above sea level to minimize contamination of sulfurous acid gas. .

After dipping in the sterilized clear water to dissolve the impurities of the pine needles selected above, it is preferable to wash 3-4 times until clear water comes out, and then washed once with water soaked with charcoal.

In the step 2, the step of crushing the pine needles washed, characterized in that the dehydration until the moisture of the surface of the pine needles washed in step 1 is completely removed and then pulverized with a juicer to juice. The most important thing in the process of grinding pine needles with green juice is to prevent heat from being generated during green juice and to make the pressing point precise.

In the step 3, the step of separating the supernatant from the juice solution is characterized in that to obtain a separated supernatant after separating the fibrous layer and the supernatant by crushed in step 2 and left the juice juice juice at 4 ℃ for 3 hours. . In the present invention, the separated supernatant was named "sol gold".

In step 4, fermenting the pine needle supernatant at 37 ℃ is characterized in that the fermentation for 6 months by incubating at 270 rpm in a shaking incubator at 37 ℃.

In addition, the non-repair alcohol extract is

1) harvesting the ground and roots of the non-repair;

2) selecting and washing the nutrients having good impurities and impurities from the collected non-repair; And

And 3) extracting the washed non-repair with alcohol solution.

In the above, the alcohol solution of step 3 is preferably 40 to 100% alcohol solution, more preferably 40 to 100% ethanol solution.

In the present invention, a mixed aging solution of fermentation broth and non-aqueous alcohol extract of pine needle juice supernatant prepared by the above method was prepared, which was named "Solbi Suri". As a result of analyzing the effect using the solbi repair, it was found that the anti-cancer (see Table 1 and Table 2), antibacterial (see Figure 1) and vascular relaxation (see Figure 2) is excellent.

The present invention also provides a dietary supplement containing a fermentation broth of the pine needle juice supernatant and a non-repair alcohol extract as an active ingredient.

The fermentation broth of the pine needle juice supernatant prepared by the method of the present invention and the mixed aging solution of the non-aqueous alcohol extract are excellent anticancer, antibacterial and vasodilation effect, it can be used to prepare a health supplement for improving health.

The health supplement may be tea, raw food, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products, soups, drinks or drinks, etc. It is not. In a preferred embodiment of the present invention, tea, beverages, raw foods, candy, chewing gum was prepared using a mixed fermentation broth of fermentation broth and non-aqueous alcohol extract of pine needle juice.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

<Example 1> Preparation of fermentation broth (sol gold) of pine needle juice supernatant

To minimize contamination of sulfur dioxide, healthy pine trees were grown in a clean area over 350 m above sea level. After soaking the impurities of the selected pine needles soaked in sterile water sterilized by a sterile sterilizer and washed 3-4 times until clear water comes out. The washed pine needles were washed once with water soaked with charcoal again. Only dehydrated pine needles were pulverized by crushing with a green juice machine (Fhwahwa Precision, Samkwang Armed Forces Corporation, Myeongshin Corp.). The juicer used in the present invention has an improved performance compared to conventional juicers, which uses a separate grinder and a compressor, consumes less power when juiced, and the juice part is a polycarbonate which is an FDA-approved product, and noise and vibration are reduced by using a reduction gear. It is less, the process is less forging and the loss of nutrients, the parts are broken down, the repair cost is low, the ratio of juice is about three times higher than conventional. During the crushing process of the pine needles so that heat does not occur so that the compression point is precise. The crushed juice was left for 3 hours at 4 ℃ to separate the fibrin layer and the supernatant. The isolated supernatant was obtained and named as "sol gold" and stored in a freezer at −70 ° C. or lower so that no crystals formed. In addition, the sol gold was incubated for 6 months at 37 ° C. shaking incubator at 270 rpm for fermentation and was named “sol gold fermentation broth”.

Example 2 Preparation of Non-Repairing Alcohol Extract

The ground and roots of non-eagles were harvested from uncontaminated habitats. From the collected non-repair, the nutritional state was good and no impurities were selected. The selected non-repair was soaked in clear water to dissolve impurities and washed 3-4 times until clear water came out. The washed non-repair was dried in a shaded and airy place. The dried non-repair was cut into 2-3 cm in size with a cutter. The cleaved non-repair was put in 40% to 100% ethanol in an amount immersed in the ethanol solution and extracted for at least 3 months.

Example 3 Preparation of Mixed Aging Solution of Solgold Fermentation Solution and Non-Repairing Alcohol Extract

After mixing 15% of the sol gold fermentation broth prepared in Example 1 and 85% of the non-aqueous alcohol extract prepared in Example 2 and aged at room temperature for 3 months or more, this was named "Solbi Suri".

Example 5 Anticancer Effect Analysis of Solbi Repair

<5-1> anticancer effect 1

Human ovarian cancer cell line SK-OV-3 (ATCC, USA) was suspended in 100 μl of cell culture (GIBCOBRL, USA) in 1 × 10 ⁴ cell numbers and then inoculated into each well of a 96 well plate. 40 μl of the sol gold prepared in Example 1 or the sorbbi prepared in Example 3 was dried on a 45 ° C. hot plate and dissolved in DMSO. Solgold or sorbabius dissolved in the prepared DMSO was diluted 250, 500, 1000 or 2500 times with DMSO and added to each well inoculated with the cells and incubated for 2 days. After incubation, cytotoxicity experiments were performed by MTT method (Pieters et al ., 1988, Cancer Letters ., 41, 323-332).

As a result, compared with ovarian cancer cells cultured without adding sol gold or sorbbi, cultivation by adding sol gold or sorbbi repair resulted in increased cytotoxicity depending on the added concentration and decreased survival rate. It was found that solgold had higher cytotoxicity (Table 1).

Dilution factor 1/250 1/500 1/1000 1/2500 Sol gold 9% 39% 37% 67% Solbi Repair 55% 58% 73% 99%

<5-2> Anticancer Effect 2

The sol gold prepared in Example 1 or the solbi repair prepared in Example 3 was dissolved in DMSO. The dissolved samples were diluted 250, 500, 1000 or 2500 times in DMSO and then treated with MEL-2 skin cancer cell lines (ATCC, USA) to measure cell viability in the same manner as in Example <5-1>.

As a result, the survival rate of the cells was reduced in a concentration-dependent manner treated with sol gold and sorbbi (Table 2).

Dilution factor 1/250 1/500 1/1000 1/2500 Sol gold 3% 15% 58% 65% Solbi Repair 68% 75% 77% 91%

Example 6 Antibacterial Effect Analysis of Sol Gold and Solbi Repair

The supernatant was obtained after centrifuging the sol gold prepared in Example 1 and the sorbbi prepared in Example 3 at 4000 rpm for 20 minutes. The obtained sample was soaked in 40 μl or 120 μl of paper disk, respectively, and then placed on nutrient agar coated with staphyllococcus aureus or E. coli. Place and incubate overnight at 37 ° C. After incubation, the diameter of the transparent ring (developing ring) formed around the paper disk was measured.

As a result, it was found that the growth of Staphylococcus aureus and Escherichia coli was inhibited in a paper disk soaked with 40 μl and 120 μl of sol gold, thus having a high antibacterial effect (FIG. 1).

Example 7 Analysis of Vascular Relaxation Effect of Solbi Repair

Solvari repair prepared in Example 3 was to determine whether there is a vascular relaxation effect. Specifically, the rats of 150-200 g body weight were intraperitoneally injected with 40 mg of pentobarbital-Na per kg of body weight, anesthetized, and then opened, and the thoracic aorta was extracted. The isolated vessels were Krebs-ringer solution (NaCl 112 mM, KCl 5 mM, NaHCO ₃ 25 mM, KH ₂ PO ₄ 1.2 mM, MgSO saturated with 95% O ₂ and 5% CO ₂ at room temperature). ₄ 1.2 mM, EDTA 0.026 mM, CaCl ₂ 2.5 mM, glucose 11.5 mM, pH 7.4) Remove blood from the blood vessel and clean the surrounding tissues with fine scissors to remove the 5 mm long ring ( vascular sections were made. These sections were transferred to a 15 ml vertical laboratory container containing Krebs-Ringer solution saturated with 95% O ₂ and 5% CO ₂ at 37 ° C, with one end of the muscle anchor and the other end of the force transducer. , Grass FT03), to record isotonic contraction with a physiological recorder. The bath solution was replaced with fresh nutrient solution every 20 minutes and the section length was extended to 3 g of driven tension. After a 1-hour recovery of the aortic section, shrinkage was introduced by incorporating phenylephrine (PE) ^10-6 M. After inducing the contraction, solvy repair was added to record the isotonic contraction to measure the degree of vascular relaxation.

As a result, the aortic section was treated with phenylephrine, a contractile substance, to induce a vasoconstriction reaction, and then the treatment of sorbabiis at a concentration of 0.15 mg / ml resulted in a vasorelaxation reaction and a concentration of 0.3 mg / ml. More powerful vasorelaxation reactions were shown (FIG. 2). Thus, it can be seen that the sorbabiri of the present invention can have a vascular relaxation action.

Preparation Example 1 Preparation of Health Supplements Containing Pine Cone (Saturated Fermentation Solution of Fermented Liquid of Pine Needle Juice Symbol Solution)

<1-1> Preparation of Tea Containing Solbi Repair

The present inventors manufactured the health promoting tea by the conventional manufacturing method of the tea using the sorbbi repair prepared in Example 3. Tea was prepared as a liquid tea using solbi shu or granule tea in a granule form solbi suri.

First, liquid form tea using solbi suri was generally prepared as a can drink that is easy to drink, solbi suri was properly diluted, and additives were added to increase taste and taste. Additives include 0.02 to 0.04% (w / v%) of vitamin C and 0.02 to 0.06% (v / v%) of liquid flavoring, heated to 90 ± 5 ℃ in a mixed planar heat exchanger, and then the can is hot. Charged. Nitrogen gas was injected into the headspace of the filled can to reduce the dissolved oxygen content of the product, and then the lid was sealed, sterilized and cooled to prepare a liquid sorbabi tea.

Next, lactose, corn starch, glucose and vitamin C were added as 60.0%, 20.0%, 1.5%, and 1.0%, respectively, so that lactose, corn starch, glucose, and vitamin C were 82.5% of the total granular tea content. Weighed, mixed, ground and sifted to 80 mesh size. The additive pulverization and sorbabii of the present invention prepared in the above was added and mixed at a volume ratio of 17.5%: 82.5%, and granules were formed by using a reverse rotating granulator. Granules formed in the above granules were granulated with an extruded granulator using a No. 35 sieve, and the granules were placed in a tray with 2 cm thickness and dried in a hot air dryer for 24 hours at 60 to 80 ° C. The dried granules were placed in a vibrating apple and sifted into 250 mesh sizes using sieves No. 35 and 80. The granular sorbabi tea was packaged in an aluminum foil tea bag (Al foil tea bag) heat-bonded at 3.0 g / bag.

<1-2> Preparation of Drinking Water Containing Solbi Repair

The inventors of the present invention prepared the beverage for health promotion using the sorbabi prepared in Example 3 using a conventional beverage preparation method with the composition described below.

522 mg of honey

Chioctosanamide 5 mg

Nicotinamide 10 mg

Riboflavin Sodium Hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

Inositol 30 mg

Orthoic acid 50 mg

Solbi Repair 0.5 ~ 1.5 ㎎

200 ml of water

<1-3> Preparation of raw foods containing sorbabie

The inventors of the present invention prepared the health-promoting reproduction by using the sorbbi repair prepared in Example 3 using a conventional method for producing a raw food with the composition described below.

Solbi Repair 10%

Brown rice 40%

12%

15% sorghum

Black Bean 8%

Kale 5%

Perilla 10%

<1-4> Preparation of Candy Containing Solbi Repair

The present inventors manufactured the candy for health promotion using the sorbabi suri prepared in Example 3 using a conventional candy manufacturing method with the composition described below.

50 to 60% sugar

Starch syrup 39.26-49.66%

Solbi Repair 0.24 ~ 0.64%

Orange 0.5%

<1-5> Preparation of Chewing Gum Containing Solbi Repair

The inventors of the present invention were prepared using the chewing gum production method using the cheolbisu prepared in Example 3 to improve health for the composition described below.

Gum base 20%

Sugar 76.36-76.76%

Solbi Repair 0.24 ~ 0.64%

1% fruit flavor

Water 2%

As described above, the fermentation broth mixed with the fermentation broth of the pine needle juice supernatant prepared in the present invention (non-repair alcohol) is a preservation of the main components of pine needles and non-repair well, anti-cancer, antibacterial and vascular relaxation effect It is excellent and can be used as a health supplement.

1 is a graph showing the growth inhibitory effect on Staphylococcus aureus and Escherichia coli by using a paper disk moistened with 40 μl or 120 μl of solgold and sorbobia prepared by the method of the present invention.

Figure 2 is a graph showing the effect of vascular relaxation when treated with rat thoracic aortic sections prepared by the method of the present invention at 0.15 mg / ml and 0.3 mg / ml concentration.

Claims (7)

(correction)  Pine needle juice supernatant, characterized in that the fermentation broth and non-aqueous alcohol extract of pine needle juice supernatant showing anti-cancer, antibacterial and vasorelaxant effect is mixed by 10 to 20% by weight: 80 to 90% by weight and aged at 37 ° C. for 3 months. Health foods containing fermentation broth and non-alcoholic alcohol extracts as a active ingredient. (delete) (delete) (delete) (delete) (delete) (correction)  The method of claim 1, wherein the health food is selected from the group consisting of tea, raw food, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, noodles, gum, dairy products, soups, drinks, drinks Health food characterized in that