KR20010107182A - Development of anti-stress drinks using pine needie extract - Google Patents

Abstract

The present invention relates to the production of anti-stress beverages using pine needle extract, pine needle extract (pine needle extract: PNE) is added to the water or yogurt (yogurt) to 1 to 3, respectively, and as a functional enhancer, arginine (arginine) 0.1 , Taurine 0.2, tyrosine and tryptophan, respectively, were added 0.05, and ethanol from bamboo shoots, soybean germ and shiitake (P'yogo: Lentinus edodes S _ING ) Anti-stress beverages were prepared by adding 0.55 extracts each of 0.05 dry extract powders. Taking antistress beverages significantly reduces plasma corticosteroid (CS) secretion, increases the secretion of noradrenaline (NA) in brain tissues, and decreases the secretion of MHPG-SO ₄ . The number was increased, and as a result of EEG measurement, the brain wave (α + θ) to relieve stress 30 minutes after taking the effect was significantly more than 20.

Description

Preparation of antistress drink using pine needle extract {DEVELOPMENT OF ANTI-STRESS DRINKS USING PINE NEEDIE EXTRACT}

The present invention relates to a method for producing an anti-stress drink using pine needle extract.

Until now, stress had been known for physical, chemical, and mental stress, but it was true that the development of stress-relieving beverages was not conceived because stress was not so seriously regarded as a disease. However, as stress has recently been found to be the cause of all illnesses, the need for stress relief is emphasized.

But the question is: 1) How do you measure stress? It is very difficult to study the problem of human beings and those who have complicated social life. 2) How to load stress through animal experiments and 3) How to evaluate the stress relief. Until now, there has been a problem of lack of validity and complexity of research because it has been focused on the fact that under stress, gastrointestinal disorders occur.

An object of the present invention is the pine needle (Pin needle densiflora Sieb et Zucc.) Of the leaf extract (pine needle extract: PNE) as a predominant agent soybean germ (soybean germ), bamboo shoots (bamboo shoots), shiitake (P ') It is intended to provide anti-stress functional beverages by adding and preparing food ingredients such as yogo) and drinking them as beverages.

According to the Chinese literature, pine needles have been used to treat diseases used to treat diseases since ancient times and researched on the development of beverages using the same, as a result of the invention of a new anti-stress functional beverage manufacturing method.

That is, pine needle extract (pine needle extract (PNE)) is added to water or yogurt (yogurt) to 1 to 3, respectively, and as functional enhancers, arginine, arginine, taurine, tyrosine and typtophan (tryptophan). A predetermined amount of anti-stress beverage is added to the ethanol-derived dry powder of bamboo shoot, soybean germ, and shiitake (P'yogo: Lentinus edodes S _ING ). Prepared.

The anti-stress beverage of the present invention was tested using an animal laboratory apparatus and an experimental method for sociopsyhological stress developed by Masahiro Ishikawa, a team of professors of medicine at Ehime University, Japan in 1992, and sympathetic as an autonomic nervous system. Neuronal activity and physiological changes in the endocrine system were evaluated.

As a result, compared with the experimental group taking the functional antistress beverage of the present invention, the amount of corticosterone (CS) in plasma was significantly decreased, and the amount of noradrenaline (NA) in brain tissue was increased, and MHPG- The amount of SO ₄ secretion decreased, the number of cells increased, and in human experiments, EEG results showed that EEG (α + θ), which releases stress after 30 minutes, was markedly secreted by 20 or more.

Therefore, the anti-stress drink based on the pine needle extract of the present invention is a functional anti-stress drink that can be effectively taken by those who are stressed in the complex modern social life.

Pine needle extract (PNE) is predominantly added to the water or yogurt (0.2 to 10 parts by weight of pine needle extract so that 1 to 3, respectively) Arginine 0.1, taurine (taurine) as a functional enhancer ) 0.2, tyrosine and tryptophan are added 0.05, respectively, and ethanol-derived dry powder of bamboo shoot, soybean germ and shiitake (P'yogo: Lentinus edodes S _ING ) A total of 0.55 was added each 0.05, and centrifuged at 1,000 x g for 10 minutes to prepare a superstressed beverage.

Experimental method and test results for the functional anti-stress beverages based on the pine needle extract of the present invention are as follows.

1. Experimental Materials

(1) Pine needle extract antistress drink

Drinks prepared by adding 70 ° C.-ethanol extract of pine needles to purified water or yogurt to 1, 2 and 3 respectively were WPNE-1, WPNE-2, WPNE-3, and YPNE-1, YPNE-2 and YPNE-3, respectively. Add 0.1 arginine, taurine 0.2, tyrosine and tryptophan as 0.05 functional enhancers, and add 0.055 each of ethanol-extracted dry powder of bamboo shoots, soybean embryos and shiitake, and add 0.55 citric acid and sorbitol as taste and sweetener. , Anti-stressed beverage was prepared by adding white sugar.

(2) Animals and devices for the test of effectiveness

Experimental animals: 4 weeks old ICR mice (mouse: 25 ± 3g, ♂), 7 per group

Experimental device: Socio-psychological stress box (communication box) → 2.0mA

Allow 10 seconds of current, followed by 120 seconds of rest

(3) Reagent

As a reagent used in this experiment, enzymes such as noradrenaline (NA), MHPG-SO ₄ (3-methoxy-4-hydroxyphenylethyleneg lycol sulfate), cortosterone, and SOD for measuring and evaluating stress relief effects The reagent used was a special reagent made by Sigma, and the reagent used was a reagent of grade 1 or higher.

2. Experiment Method

(1) Collection and Separation of Blood

Seven animals per experimental animal group were anesthetized with ether, blood was collected from the heart, left at low temperature for 2 hours, centrifuged at 700 x g for 10 minutes, and the supernatant was added with sodium heparin 0.05 per 1.0 ml of blood. The cells were treated in ml-treated CBC bottles (complete blood cell count, green cross), and frozen and stored at −70 ° C. to be used for the analysis of corticosterone.

(2) fraction of brain tissue

Seven animals per group of animals were collected and then emptied. Brain tissues were aliquoted and placed in a buffer solution (1.15 KCl / 10 mM phosphate buffer + 5 mM EDTA, pH 7.4) and frozen and stored at -70 ° C. Before analyzing the stress relief effect, the contents of noradrenaline (NA) and MHPG-SO ₄ were analyzed according to the following pretreatment, respectively.

(3) Culture and Survival of Splenocytes

After evaluating 7 animals per group of animals, spleen was isolated according to the method of Coligan et al. (1991), and then dulbecco's modified eagle's medium (DMEM, Gibco) medium was prepared using slide glass in the plate. Was released. The cells were separated by centrifugation (1,000 rpm, 5 minutes), and the cell density was adjusted to a concentration of 1 × 10 ⁶ / ml by cell counting in DMEM medium containing 10 fetal bovine serum (FBS, Gibco). . Cell suspensions were dispensed into 24well plates, and then cultured for 24h, 48h, 72h in a CO ₂ incubator maintained at 37 ° C and 5CO ₂ . Cell of viability was measured during the incubation period. The tryphan blue exclusion test was performed for each cell culture period and the number of living and dead cells was measured to compare the proliferation and death of splenocytes. The measurement was performed by counting the stained and unstained cells using a hemocytometer to calculate the survival rate according to the following equation.

Survival rate () = 100 × number of viable cells / total cells

(4) measurement of corticosterone

Corticosterone content of plasma isolated from blood was measured according to the method of Sliber et al. (1958). After shaking 1.0 ml of plasma vigorously for 30 seconds in three times the amount of petroleum ether, the solvent layer is discarded after centrifugation. After diluting the purified plasma with 5.0 ml of distilled water, the mixture was extracted by shaking with 15.0 ml of methylene chloride for 30 seconds in a centrifuge tube. The extract is added to a tube containing 1.0 ml of 0.1N NaOH solution, shaken quickly and centrifuged.

After centrifugation, the alkali layer is discarded and 10.0 ml of methylene chloride is placed in a tube containing 2.0 ml of 30N sulfuric acid. After shaking for about 30 seconds, after centrifugation for 1 to 2 minutes, the upper solvent layer was discarded and the lower layer was 2.0 ml and left at room temperature for 30 to 90 minutes, and measured at excitation 470 nm and emission 530 nm. Corticosterone standard solution was dissolved in 5.0 ml of corticosterone in 5.0 ml ethanol, diluted with 1,000 ml of distilled water, and then corticosterol content (ug / dl plasma) by the above standard calibration procedure between 0.1 and 0.5 ug. Was quantified.

(5) measurement of noradrenaline

The content of noradrenaline (NA) in the brain was measured according to the method of Kohno (1979). Brain tissue is triturated in a low temperature room with 4.0 ml of 0.2 N sulfuric acid containing 0.1 methabis sulfate (Na ₂ S ₂ O ₅ ), and centrifuged at 8000 × g for 5 minutes. The supernatant is taken 2.0 ml each to simultaneously measure NA and MHPG-SO ₄ . Next, 3.0 ml of 0.4N PCA containing 0.1Na ₂ S ₂ O ₅ and 0.05EDTA was added.

After centrifugation at 8000 × g for 10 minutes, the supernatant was transferred to a cap tube and stored at −45 ° C. until the experiment. The pyrex column (diameter 6 mm, height 25 cm) for NA measurement is filled with glass wool and filled with alumina activated at pH 7.5-8.0 at a height of exactly 6.0 cm. NA is extracted from the column by 0.05 ml PCN 2.0 ml. After extraction, the solution is converted to a fluorescent compound by 2.0 ml of iodine reagent. Fluorescence of NA was measured at excitation 380 nm and emission 495 nm. The NA standard was prepared at a concentration of 100 ug / ml, and the content of noradrenaline (ng / g brain) was quantified according to the above procedure by a standard calibration curve.

(6) Measurement of MHPG-SO ₄

The content of MHPG-SO ₄ in brain tissue was measured according to the method of Kohno (1979). Brain tissue is triturated in a low temperature room with 4.0 ml of 0.2N sulfuric acid containing 0.1 metabisbissulphate (Na ₂ S ₂ O ₅ ) and centrifuged at 8000 × g for 5 minutes. The supernatant is taken 2.0 ml each to simultaneously measure NA and MHPG-SO ₄ . Each sample is adjusted to pH 6.0-6.5 with 0.3 N barium hydroxide solution, and then all samples are made to the same amount with distilled water. After centrifugation at 8000 x g for 10 minutes, the supernatant was transferred to a cap tube and stored at -45 ° C until the experiment. The pyrex column (diameter 6 mm, height 25 cm) for the measurement of MHPG-SO ₄ is covered with glass wool and filled with DEAE sephadex A-25 at exactly 6.0 cm. MHPG-SO ₄ is extracted from the column with 2.0 ml of 0.4N PCA containing 0.1 metabisbissulphate.

Immediately after the separation of MHPG-SO ₄ , 0.1 ml of 0.2 cysteine and 0.1 ml of 70PCA are added, and these reaction mixtures are boiled in a water bath at 90-95 ° C. for 25 minutes. In this case, blank is added without heating. After adding 0.3 ml of ethylenediamine, all samples were placed in a hot bath for 5 minutes and cooled. Fluorescence of MHPG-SO ₄ was measured at excitation 325 nm and emission 465 nm. MHPG-SO ₄ standard solution (100 ug / ml) was prepared at a concentration of 0.5 metabisbissulphate (Na ₂ S ₂ O ₅ ) and the content of MHPG-SO ₄ (ng / g brain) was obtained by the above procedure according to the standard calibration curve. Quantification

(7) Measurement of EEG through Clinical Trials

According to the experimental design as shown in Fig. 3, EEG was measured by clinical experiments. The EEG was measured by Q-Jump (Changsei, Korea Institute of Psychiatry). Seventy-five minutes were conducted for each university student, and the EEG region was measured at the frontal lobe. In addition, the measurement was made after one hour after eating to minimize the effect after eating. The measurement method was cleaned for 5 minutes to stabilize the mind and body during the first 5 minutes of the total 75 minutes, and then the development prototype was taken.

The average value of cumulative EEG data was determined based on the mean value of cumulative EEG data 10 minutes before taking 10 minutes, and the change of EEG expression was analyzed by comparing the data before and after taking the average value of cumulative EEG data for 10 minutes after taking 10 minutes. EEG rate calculation is based on [(α + θ) / (β-high + β-) of α waves and θ waves related to physical and mental stability and stress among the emergent brain waves (β-high, β-low, α, θ) for 2 minutes. low + α + θ)] × 100.

3. Evaluation of Experimental Results

(1) Increased effect of noradrenaline

Noradrenaline (NA) is a hormone secreted with adrenaline and is secreted as a neurotransmitter at the sympathetic nerve endings of brain tissue. Among these neurotransmitters, noradrenaline is used as an indicator of stress. In other words, noradrenaline is significantly reduced in physical stress such as electric shock (FS), non-foot stress (NFS) and sociopsychological stress (NFS). Increasing the amount of sugar can be an effective way to relieve stress.

Prototype WPNE-1, WPNE-2, WPNE-3 and YPNE-1, YPNE-2, and YPNE-3 developed by dissolving pine needle extracts 1, 2, and 3 in water and yogurt as functional antistress beverages. Daily administration was performed to measure the amount of noradrenaline (NA) secretion in the brain cells, the results are shown in Table 1. As shown in Table 1, WPNE-I, WPNE-II, and WPNE-III increased to 10.6, 18.1, and 21.7, respectively, compared to the control group, and YPNE-I, YPNE-II, and YPNE-III, respectively, were 12.2 and 15.8, respectively. As a result, no significant difference was found between water and yogurt. Therefore, when seen in the increase of NA, a very good stress relief effect is expected in the administration of two or more pine needle extract.

Effects of Anti-Stress Beverages WPNE and YPNE on the Secretion of Noradrenaline and MHPG-SO ₄ by Socio-psychological Stress division Noradrenaline content (ng / g brain) MHPG-SO ⁴ content (ng / g brain) NA / MHPG-SO ₄ ratio Control non-foot shock 179.40 ± 9.18 ^* 100.0 197.60 ± 13.41 100.0 0.91 100.0 Pine needle extract / water-beverage ^** WPNE-Ⅰ 198.41 ± 17.51 ^a 110.6 182.01 ± 9.40 92.1 1.09 119.8 WPNE-Ⅱ 211.90 ± 14.90 ^b 118.1 178.09 ± 9.21 ^a 90.1 1.19 130.8 WPNE-Ⅲ 218.39 ± 17.01 ^c 121.7 173.31 ± 12.60 ^b 87.7 1.26 138.5 Control non-foot shock 180.27 ± 10.95 ^* 100.0 177.78 ± 13.62 100.0 1.01 100.0 Pine needle extract / yogurt-beverage ^** YPNE-Ⅰ 201.84 ± 14.09 ^a 112.2 171.80 ± 12.02 96.7 1.17 115.8 YPNE-Ⅱ 208.68 ± 15.31 ^b 115.8 162.85 ± 11.95 ^a 91.6 1.28 126.7 YPNE-Ⅲ 213.19 ± 10.73 ^c 118.3 158.83 ± 11.93 ^a 89.3 1.34 132.7

^* Mean ± SD; ^** WPNE-I, II, III and YPNE-I, II, III: Antistress beverage ^a p <0.05 in which pine needle extract (PNE) 1, 2 and 3 were dissolved in water and yogurt, respectively; ^b p <0.01; ^c p <0.001: Significance test versus control group

(2) the effect of reducing the amount of MHPG-SO ₄ produced

MHPG-SO ₄ (3-methoxy-4-hydroxyphenylethyleneglycol sulfate) is a metabolite of noradrenaline (NA) that is secreted as a neurotransmitter at the sympathetic nerve endings of brain tissue, and is used as an indicator of stress like NA. . This is because MHPG increases significantly during physical stress (foot shock (FS), non-foot shock (NFS), and sociopsychological stress (NFS)) such as electric shock. Reducing the content of -SO ₄ may be an effective way to relieve stress. As a functional antistress beverage, prototypes WPNE-1, WPNE-2, WPNE-3 and YPNE-1 YPNE-2 YPNE-3, developed by dissolving pine needle extracts 1, 2, and 3 in water and yogurt, were administered to ICR mice for 17 days. By measuring the secretion amount of MHPG-SO ₄ in the brain tissues are shown in Table 1. As shown in Table 1, WPNE-I, WPNE-II, and WPNE-III decreased to 7.9, 9.9, and 12.3, respectively, compared to the control group, and YPNE-I, YPNE-II, and YPNE-III, respectively, 3.3 and 8.4, respectively. In addition, it was reduced to 10.7, and water DF was slightly more effective than solvent. Therefore, when seen in the reduced amount of MHPG-SO ₄ , a significantly good stress relief effect is expected in the administration of pine needle extract 3 or more.

(3) increase effect of NA / MHPG-SO ₄ ratio

Under stress, NA is reduced and its metabolite MHPG-SO ₄ is increased. Therefore, the effect of functional stress relief drink can be evaluated by the increase effect of NA / MHPG-SO ₄ ratio. Therefore, as shown in Table 1, not only did WPNE-I, WPNE-II, and WPNE-III significantly increase to 19.8, 30.8, and 38.5, respectively, but also YPNE-I, YPNE-II, and YPNE-III, respectively, compared to the control group. Significant increases to 15.8, 26.7, and 32.7 are expected to resolve stress very effectively, regardless of solvent.

(4) Comparison of the amount of corticosteroid secretion

Corticosterone, which is present in plasma, is a hormone involved in the metabolism of glucose in the adrenal cortex and is used as an indicator of stress. In other words, corti- tic stress increases significantly during physical stress (foot shock (FS), non-foot shock (NFS), and sociopsychological stress (NFS)) loads. Decreasing the amount of cholesterol can be a way to relieve stress.

Prototype WPNE-1, WPNE-2, WPNE-3 and YPNE-1, YPNE-2, and YPNE-3 developed by dissolving pine needle extracts 1, 2, and 3 in water and yogurt as functional antistress beverages. Daily administration was performed to measure the amount of corticosteroid secretion in the blood. Table 2 shows the results. As shown in Table 2, WPNE-I, WPNE-II, and WPNE-III significantly decreased to 7.2, 11.2, and 12.8, respectively, compared to the control group, and YPNE-I, YPNE-II, and YPNE-III, respectively, compared to the control group. 6.8, 8.3, and 9.5% were significantly reduced, so that the solvent is water, the anti-stress effect is expected to be slightly larger than yogurt.

Effects of Anti-Stress Beverages WPNE and YPNE on the Secretion of Corticosterone by Socio-psychological Stress division Corticosterone content ^* (ug / dl plasma) prepare() Control non-foot shock 368.51 ± 12.01 100.0 Pine needle extract / water-beverage ^** WPNE-Ⅰ 341.80 ± 18.19 92.8 WPNE-Ⅱ 327.18 ± 22.68 ^a 88.8 WPNE-Ⅲ 321.31 ± 12.30 ^b 87.2 Control non-foot shock 387.20 ± 17.10 100.0 Pine needle extract / yogurt-beverage ^** YPNE-Ⅰ 360.75 ± 14.65 93.2 YPNE-Ⅱ 355.10 ± 12.70 ^a 91.7 YPNE-Ⅲ 350.35 ± 10.05 ^a 90.5

^* Mean ± SD; ^** WPNE-I, II, III and ^*** YPNE-I, II, III: Antistress beverages in which pine needle extract (PNE) 1, 2 and 3 were dissolved in water and yogurt, respectively ^a p <0.05; ^b p <0.01: Significance test compared to the control group

(5) evaluation of immune capacity

Prolonged exposure to stress significantly reduces the immune system's immune function. Therefore, in order to evaluate the effect of the developed functional antistress beverage, fermented pine needle extract was added and fermented for 17 days as a yogurt drink. After collecting blood cells, the splenocytes were isolated and cultured for 3 days to increase the immune function as viable counts of splenocytes. The evaluation results are shown in Table 3. When comparing the control group in Table 3, when the number of cells after 1 day of culture was 100, the number of cells decreased almost 20 after 2 days, and nearly 40 after 3 days.

However, in the group receiving the developed antistress drink, the YPNE-I, YPNE-II, and YPNE-III groups after 1 day increased 4.7, 7.8, 12.1 or more cell numbers, respectively, and the YPNE-Ⅰ, YPNE after 2 days. In the ⅡNE and YPNE-III groups, the number of cells increased by 6.8, 13.0 and 16.4, respectively, compared to the control group, and in the YPNE-I, YPNE-II, and YPNE-III groups after 3 days, the number of cells by 5.5, 13.7, 16.4, respectively, Increased. Therefore, a significant increase of 12 to 16 was observed in the case of the 3 added beverages after the development of antistress beverages.

Effect of Antistress Drink YPNE on Immune Activity of ICR Mice by Socio-psychological Stress Load division Cell number (× 10 ⁴ ) / ml 1 day later 2 days later 3 days later Control non-foot shock 85.33 ± 6.11100.0 100.0 69.00 ± 6.0880.9 100.0 48.67 ± 6.4357.0 100.0 Pine needle extract / yogurt-beverage ^** YPNE-Ⅰ 89.33 ± 5.30 104.7 73.67 ± 3.21 106.8 51.33 ± 4.16 105.5 YPNE-Ⅱ 92.00 ± 2.00 107.8 78.00 ± 6.00 ^a 113.0 55.33 ± 5.77 ^a 113.7 YPNE-Ⅲ 95.67 ± 2.08 ^a 112.1 80.33 ± 4.04 ^b 116.4 56.67 ± 5.03 ^b 116.4

^* Mean ± SD; ^** YPNE-I, II, III: antistress beverage ^a p <0.05 in which pine needle extract (PNE) 1, 2, 3 was dissolved in yogurt; ^b p <0.01: Significance test compared to the control group

(6) Evaluation of clinical trial results

Change-up, which was previously released by measuring the effect of stress relief through clinical trials of YPNE-3, which was developed by dissolving pine needle extract 3, which proved to be the most effective as a functional antistress beverage, in yogurt. Table 4 shows the EEG results compared with the clinical trial results (Yoon et al., 1997). As shown in Table 4, as a result of clinical experiments for university students, EEG measurement results of the anti-stress beverages developed, EEG (α + θ) to relieve stress after 30 minutes of administration is secreted more than 20, already stress relief drink It has also been demonstrated in clinical trials that it is a much better stress relieving beverage than the change-up that has been released.

Effect of pineal leaf extract triadditive antistress beverage (YPNE-3) on EEG secretion in 15 university students division Before taking After taking antistress drink 10 minutes 10 minutes 20 minutes 30 minutes 40 minutes 50 minutes 60 minutes YPNE-Ⅲ *** 54.1 ± 8.6 ^* 100.0 55.9 ± 6.9 ^* 103.3 60.8 ± 5.6 ^a 112.4 65.6 ± 7.7 ^c 121.3 63.4 ± 9.0 ^b 117.2 59.1 ± 8.6 ^a 109.2 56.9 ± 8.4105.2 change-up *** 38.6 ± 5.3100.0 39.8 ± 6.2103.1 39.4 ± 5.4102.1 41.1 ± 6.2106.5 39.0 ± 6.7101.0 40.6 ± 5.7105.2 40.0 ± 5.8103.6

^* Mean ± SD; ^** YPNE-III: antistress beverage prepared by adding pine needle extract (PNE) 3 to yogurt; ^*** Results of Yoon Sang-won and Kim Sung-il (1997); ^a p <0.05; ^b p <0.01; ^c p <0.001: Significance test compared to the control group

Example 1

Anti-stress beverages added to water and yogurt with pine needle extract (PNE) as the main ingredient, respectively, to WPNE-1, WPNE-2, WPNE-3, YPNE-1, After preparing YPNE-2 and YPNE-3, 0.1 parts by weight of arginine, 0.2 parts by weight of taurine, 0.05 parts by weight of tyrosine and tryptophan, respectively, were added as reinforcing agents. 0.05 parts by weight of ethanol-derived dry powder of bamboo shoot, soybean germ, and shiitake (P'ygo: Lentinus edodes S _ING ), and 0.05 parts by weight of ginger juice as a deodorant, citric acid as a flavor enhancer and sweetener 0.07 parts by weight, 0.5 parts by weight of sorbitol, 2.5 parts by weight of white sugar and 4.5 parts by weight of fructose were added and mixed, and then centrifuged at 1,000 × g for 10 minutes to prepare a functional stress-relieving beverage as a supernatant. Prepared by weight part of ingredients And the combined parts by weight were the most preferable.

Prescription Ingredients and Amount of Stress Relief Beverage Ingredients \ parts by weight 0.01 0.05 0.07 0.1 0.2 0.5 1.0 2.0 2.5 3.0 4.0 4.5 5.0 10.0 Pine needle extract Arginine Taurine Tyrosine Tryptophan Bamboo shoot Soybean embryo Shiitake Extract Powder Ginger juice Citric acid Sorbitol White sugar Fructose

: Displays the actual name and quantity of ingredients

The pine needle extract antistress beverage of the present invention has a decrease in the secretion of corticosteroid (CS) in the plasma and noradrenaline (NA) in the brain tissue as a stress indicator component through physical and social and psychological stress animal experiments. It has been proved that the release of MHPG-SO ₄ is very effective in relieving stress.

In addition, the immune capacity is significantly reduced when stress is applied. In the experimental group to which the present anti-stress beverage was administered, the number of cells increased in the culture experiment of splenocytes of ICR mice, and the remarkable enhancement effect of the immune capacity was recognized.

In addition, as a result of clinical experiments with university students, EEG measurement results of the anti-stress drink of this development, EEG (α + θ), which releases stress after 30 minutes of administration, was secreted more than 20, and has been released as a stress relief drink. Clinical trials have shown that stress-relieving beverages are much better than change-up.

Therefore, anti-stress drink based on pine needle extract has an effect that can be effectively taken by those who are physically and mentally stressed in the complex modern city life.

Claims (1)

Pine needle extract (PNE) is predominantly added to water or yogurt 0.2 to 10 parts by weight of pine needle extract to 1 to 3, respectively, and arginine, taurine, tyrosine and tryptophan, respectively, 0.01 to 5.0 parts by weight Next, add 0.01 to 5.0 parts by weight of bamboo shoots, soybean embryos, and shiitake extract powder, add 0.01 to 5.0 parts by weight of ginger juice, citric acid, sorbitol, and 0.1 to 10 parts by weight of white sugar and fructose, respectively, and mix at 1,000 x g. Preparation of anti-stress beverages using pine needle extract, characterized in that to prepare a supernatant by centrifugation for 10 minutes.