KR101414741B1 - The manufacturing method of green tea nut extract and compositions for anti-obesity containing green tea nut extract - Google Patents

Abstract

The present invention relates to a method for preparing green tea fruit extract and an anti-obesity composition containing the green tea fruit extract as an active ingredient. More particularly, the present invention relates to a method for preparing a green tea fruit extract, And extracting the residue of the defatted green tea supercritical extract with water or alcohol, and a method for producing an antiobesity composition containing green tea fruit extract prepared by the above method as an active ingredient . Accordingly, the antiobesity composition containing the green tea extract as an active ingredient has an effect of inhibiting body fat accumulation.

Description

Description: TECHNICAL FIELD The present invention relates to a method for producing an extract of green tea and an antiobesity composition containing green tea extract as an active ingredient,

The present invention relates to a method for preparing green tea fruit extract and an anti-obesity composition containing the green tea fruit extract as an active ingredient. More particularly, the present invention relates to a method for preparing a green tea fruit extract, And extracting the residue of the defatted green tea supercritical extract with water or alcohol, and a method for producing an antiobesity composition containing green tea fruit extract prepared by the above method as an active ingredient .

Tea is a drink that has been popular among humans for many years and consumes much in modern times. The tea is made by fermenting the leaves of a tea tree with the scientific name Camellia sinenisi. The types of tea are classified into green tea, oolong tea and black tea due to difference in fermentation process, and the taste, aroma, color and composition of tea are different according to the manufacturing process. Green tea is one of the oldest and most consumed foods in the world and has been used as a treatment for various diseases since ancient times. The beneficial effects of green tea on the human body have been variously characterized by classical and modern science.

In other words, the efficacy of green tea can be seen in the fixation of the new nerve, the main bone gangmok, the multi gyeong, and the bronze gyeongbokgam. Especially, the green tea shows the effect of green tea, It is said to brighten the eyes, brighten the hair, wash away the worries, and prevent obesity.

In addition, there are the following known effects on the human body of green tea. First, by stimulating the cerebral central nervous system with arousal and excitement, it cleanses the mind and restores fatigue, as well as enhances memory, judgment, and endurance, and gives mental vitality to refresh the mood. Second, anti-inflammation and bacterial growth inhibition inhibits the growth of bacteria such as Diphtheria, Typhoid and Staphylococcus, prevents food poisoning and cold, and especially after eating, suppresses the growth of mouth bacteria. Third, catechins of green tea significantly reduce serum cholesterol content and efficiently excrete to prevent hypertension and arteriosclerosis, and vitamin C and pigment components in green tea leaves have also been reported to inhibit the development of hypertension and arteriosclerosis . Fourth, the polyphenol component in the tannins of green tea is combined with unstable terminals having carcinogenic properties to prevent cancer and inhibit the proliferation of cancer cells. In fact, the city of Stich in the UK reported that the polyphenol component of green tea, black tea, and oolong tea has an inhibitory effect on carcinogens, and Hashayihiko of Japan reported that green tea extract was directly administered to cancer cells It also proved the anti-proliferation and antimutagenic effects of solid tumors. In the 2nd International Green Tea Symposium, which was held on September 1st, 1993, green tea inhibited the proliferation of cancer cells and inhibited AIDS virus as well as heavy metal detoxification, It has been reported that it is effective to remove bad breath and to have sterilization effect.

Although tea leaves are widely used in various forms, oils extracted from fruits and berries are not available in a variety of applications and are of limited use. Recently, it has become known that green tea fruit extract has efficacy and efficacy such as antibacterial and antitumor, and thus the interest in green tea is increasing. According to the results of the study of "Nah et al. (Na Hyo-hwan, Baek Soon-ok, Han Sang Bin, Bum Jin Young, General composition of green tea seeds, Journal of Korean Agricultural Science, Vol 35, p272-275, 1992) The presence of unsaturated fatty acids such as linolenic acid and tocopherol has not been found to be significantly different from the seeds of other plants, but it has been reported that the saponin content is very high. Also, Han LK. The saponin content of green tea extracts was significantly higher than that of the lipolytic enzymes secreted from the pancreas of the human body (Fig. lipase activity and inhibits digestion and absorption of fat, thereby preventing obesity.

In recent years, the pharmacological effects of various ingredients have been proved, and the value as a health drink has been newly recognized and widely consumed, and research has been conducted in various fields such as functional foods using natural aroma, color, and taste of green tea, and cosmetics. However, the fruit of the tea is mostly disposed of without being used for special purposes. Tea seeds are effective for asthma, brain, etc. They are used as a sensitive therapy. They are high in yield and rich in oil, so they are used for some food in China. Considering such various physiological activities, the use value is considered to be sufficient.

Accordingly, the inventors of the present invention completed the present invention by studying a composition for anti-obesity containing green tea extract, confirming that the defatted green tea extract obtained by supercritical extraction of green tea has an anti-obesity effect.

It is an object of the present invention to provide a method for producing green tea fruit extract in which oil is separated from green tea fruit through a supercritical fluid extraction method.

Another object of the present invention is to provide a composition for anti-obesity containing green tea fruit extract prepared by the above method as an active ingredient.

In order to solve the above-mentioned problems, the present invention provides a method for manufacturing a green tea product, comprising the steps of: (1) crushing green tea fruit to a predetermined size; Extracting the green tea powder with supercritical carbon dioxide together with an auxiliary solvent (step 2); And extracting and purifying the supercritical extract with water or alcohol (step 3).

The step 1 is a step of grinding the green tea fruit to a certain size after hot-air drying the green tea to increase the efficiency of degreasing extraction of the green tea fruit.

The term 'hot air drying' used in the present invention means a drying method in which drying is performed at a constant temperature using a hot air drier, and generally, a constant temperature means a temperature of 60 ° C. The hot air drying can be carried out for several days, preferably for 1 to 5 days. More preferably for 3 days.

The pulverization is preferably carried out by using a pin mill to pulverize the green tea fruit into a size of 0.5 mm to 2 mm. If the particle size during the above milling is outside the above range, there is a drawback that extraction efficiency of green tea fruit is deteriorated. Particularly, when the particle size is less than 0.5 mm, the extraction efficiency is lowered due to the pressure load during the supercritical extraction. When the particle size is larger than 2 mm, the effective substance contained in the particles is difficult to be extracted to the outside by the supercritical fluid. There is a disadvantage in that extraction efficiency of an effective substance is inferior.

The step 2 is a step for extracting and degreasing the oil of the green tea, and extracting the green tea fruit by the supercritical extraction under the predetermined extraction condition through the step 1. The supercritical extraction is preferably carried out at a temperature of 40 ° C to 60 ° C and a pressure of 200 bar to 400 bar.

The supercritical extraction in step 2 is preferably carried out at a flow rate of 2 ml / min to 70 ml / min using supercritical carbon dioxide for 60 minutes to 240 minutes.

Step 3 is a step of extracting the anti-obesity active ingredient from the residue of the defatted green tea extract obtained after supercritical extraction with water or alcohol. The alcohol is preferably methanol or ethanol. In addition, the extraction can be carried out by a conventional method known in the art.

The present invention also provides a composition for anti-obesity containing the green tea extract prepared by the above-described method as an active ingredient.

According to one embodiment of the present invention, an anti-obesity composition containing the green tea extract as an active ingredient is in vivo And in and it was confirmed that there was an effect of inhibiting obesity in vitro .

The production method according to the present invention can extract a composition having anti-obesity effect from green tea fruit, and can be applied to the development of an anti-obesity product using the composition.

Fig. 1 shows the appearance of (a) a green tea powder before grinding and (b) a 0.5 mm particle size green tea fruit powder according to an embodiment of the present invention. (C) sample after supercritical extraction at 40 ° C at 200 bar of 0.5 mm particle size green tea powder, (d) sample after supercritical extraction at 50 ° C, and (e) sample after supercritical extraction at 60 ° C. will be. (F) sample after supercritical extraction at 40 ° C, (g) sample after supercritical extraction at 50 ° C, and (h) sample after supercritical extraction at 60 ° C. will be. (I) the sample after supercritical extraction at 40 ° C, (j) the sample after supercritical extraction at 50 ° C, and (k) the appearance of the green tea extract after supercritical extraction at 60 ° C at 400 bar of 0.5 mm particle size green tea fruit pulverized product. Lt; / RTI >
FIG. 2 is a graph showing oil yields obtained after supercritical fluid extraction in a green tea fruit having a particle size of 0.5 mm according to an embodiment of the present invention. FIG.
FIG. 3 is a photograph showing (a) defatted green tea fruit after supercritical extraction and (b) hot water extraction according to an embodiment of the present invention.
FIG. 4 is a graph showing the effect of green tea fruit extract on the production of TG in adipocytes by using hot-water extract and non-degummed hot-water extract and hot-water extract after extracting supercritical fluid according to an embodiment of the present invention.
FIG. 5 is a graph showing the effect of green tea extract on glycerol release in primary cultured adipocytes using non-defatted hot-water extract, alcohol extract, and hot-water extract after supercritical fluid extraction according to an embodiment of the present invention.
FIG. 6 is a graph showing the effect of the HSL (hormone sentitive lipase), ATGL (Adipose triglyceride lipase), LPL (Lipoprotein lipase) of green tea fruit extract.
FIG. 7 shows the effect of green tea extract on the efflux of FAS (Fatty acid synthase) using unhatched hot-water extract, alcoholic extract, and hot-water extract after supercritical fluid extraction according to an embodiment of the present invention.
FIG. 8 is a graph showing the results of the H & E (Hematoxylin and Eosin) treatment of fat pad after 13 weeks of ingestion of the non-defatted hot water extract, the alcoholic extract and the hot water extract prepared after the supercritical fluid extraction, according to an embodiment of the present invention, staining and microscopic observation of the size and number.

Hereinafter, the present invention will be described in more detail with reference to the following examples. These embodiments are only for describing the present invention more specifically, and the scope of the present invention is not limited by these examples.

Example  1 to Example  9: Crushed green tea Supercritical  Extract preparation

In order to obtain antioxidant component efficiently from green tea fruit, it was thoroughly degreased. Degreasing was performed by using a supercritical fluid extraction method which can efficiently extract oil components. In addition, pure carbon dioxide and co-solvent were mixed to maximize oil extraction efficiency. The green tea was dried at 60 ° C for 5 days at room temperature and then pulverized to a size of 0.5 mm to 2 mm using a pin mill. Supercritical extraction (ISA-SCFE-0050-B) was performed under the conditions of 40 ℃, 50 ℃ and 60 ℃ and the pressure was adjusted to 200 bar, 300 bar and 400 bar at each temperature condition. 0100-080-re, Ilshin Autoclave, Korea), and the supernatant was extracted for 90 minutes per each green tea sample. Supercritical extraction (SFE extraction) was carried out with supercritical carbon dioxide at a flow rate of 60 ml / min for 90 minutes. Subsequently, the supercritical carbon dioxide was transferred to the separation tank together with the green tea oil, and the lower valve was opened to lower the extraction tank pressure of the supercritical extraction apparatus, thereby releasing the supercritical fluid state and recovering the extract. The supercritical extraction conditions are shown in Table 1 below.

division Extraction condition Extraction temperature (캜) Extraction pressure (bar) Extraction time (min) Particle Size (mm) Example 1 SFE 40 200 90 0.5 mm Example 2 SFE 50 200 90 0.5 mm Example 3 SFE 60 200 90 0.5 mm Example 4 SFE 40 300 90 0.5 mm Example 5 SFE 50 300 90 0.5 mm Example 6 SFE 60 300 90 0.5 mm Example 7 SFE 40 400 90 0.5 mm Example 8 SFE 50 400 90 0.5 mm Example 9 SFE 60 400 90 0.5 mm

Example  10: Green tea fruit Supercritical  After extraction of defatted green tea Heat number  Extract preparation

The samples of Example 8 were subjected to hot water extraction through the degreased green tea extracts of Examples 1 to 9. Using the completely degreased green tea fruit of Example 8, hot water extraction was performed using an ultrahigh vacuum low temperature extractor. 400 g of the defatted green tea was added to the concentrated extraction tank with 8 L of distilled water and then heated to maintain the temperature of the extraction tank at 55 to 60 DEG C while maintaining the temperature at 50 to 60 DEG C, And extracted for 3 hours. After the extraction was completed, the green tea extract and the extract were separated and vacuum concentrated. The concentrate was lyophilized to a powder form.

Comparative Example  1: Green tea berries Supercritical  Preparation of alcohol extract of defatted green tea after extraction

Using the completely degreased green tea fruit of Example 8, alcohol extraction was carried out using a rotary vacuum concentrator. 400 g of degreased green tea was added to 8 liters of concentrated ethanol after the extraction of supercritical fluid, followed by heating to maintain the temperature of the extraction tank at 55 ° C to 55 ° C. rpm and extracted for 3 hours. After the extraction was completed, the green tea extract and the extract were separated and concentrated in vacuo. The concentrate was lyophilized to give a powder.

Comparative Example  2: Supercritical  Manufacture of defatted green tea powder after fluid extraction

The supernatant fluid of Example 8 was extracted, and the remaining fully defatted green tea was firstly differentiated to prepare a direct sample. This was prepared to compare whether the defatted green tea powder had an anti-obesity effect.

Comparative Example  3: Green tea using ancillary solvent Supercritical  Extract preparation

Extraction was carried out using an auxiliary solvent in order to determine whether the extract obtained by mixing pure supercritical fluid and ethanol co - solvent from green tea fruit had an anti - obesity effect. The green tea fruits were dried in hot air at 60 ° C for 5 days and then pulverized to a size of 0.5 mm to 2 mm using a pin mill. The supercritical extraction conditions were adjusted to 50 ° C and a pressure of 400 bar. Using the supercritical extraction device (ISA-SCFE-0050-0100-080-re, Ilshin Autoclave, Korea), the pre- And supercritical fluid extraction was performed. The supercritical extraction was performed in five steps. Supercritical extraction (SFE extraction) using pure supercritical carbon dioxide at a flow rate of 60 ml / min for 90 minutes; Extracting the supercritical carbon dioxide at a flow rate of 60 ml / min and a co-solvent ethanol at a flow rate of 2 ml / min for 60 minutes; Transferring the green tea fruit extract and the supercritical fluid to the separation tank while flowing the mixture; When the supercritical fluid extract is passed to the separation tank, opening the lower valve to lower the extraction tank pressure of the supercritical extraction system to release the supercritical fluid state to recover the extract; And a mixture of green tea extract and ethanol was concentrated with a rotary vacuum concentrator to recover the extract material to prepare a supercritical green tea extract. The final extraction amount was 46.2 g, and the extraction yield was 30.8%.

Comparative Example  4: Green tea berries Heat number  Extract preparation

In order to confirm the antiobesity effect of the hot - water - extracted material without the degreasing in the green tea, the hot - water extract of green tea was prepared. The inside of the green tea material was washed with tertiary distilled water, and 400 g of the raw material for the finely ground green tea was added to 8 liters of tertiary distilled water in the concentrate extracting tank. Heat was applied to the extracting tank while the extraction tank was rotated at 55 rpm And extracted for 3 hours. After the extraction was completed, the green tea extract and the extract were separated and vacuum concentrated. The concentrate was lyophilized to prepare powder form.

Experimental Example  1: Analysis of extraction yield by temperature and pressure extraction conditions

The extraction yields of the oils obtained from the green tea supercritical fluid extraction obtained in Examples 1 to 9 were examined. Supercritical fluid extraction from green tea leaves is intended to completely remove the oil components from the final hot water extract. The extraction yield of the extract is shown in Table 2 and FIG. 2 below.

division Extraction condition Extraction temperature (캜) Extraction pressure (bar) Extraction time (min) Particle Size (mm) Extraction amount (g) Extraction yield (%) Example 1 SFE 40 200 90 0.5 mm 21.0 14.0 Example 2 SFE 50 200 90 0.5 mm 17.0 11.3 Example 3 SFE 60 200 90 0.5 mm 7.4 4.9 Example 4 SFE 40 300 90 0.5 mm 34.9 23.3 Example 5 SFE 50 300 90 0.5 mm 38.9 25.9 Example 6 SFE 60 300 90 0.5 mm 35.6 23.7 Example 7 SFE 40 400 90 0.5 mm 42.2 28.1 Example 8 SFE 50 400 90 0.5 mm 44.1 29.4 Example 9 SFE 60 400 90 0.5 mm 43.8 29.2

As shown in Table 2, it was confirmed that the yield of supercritical green tea extract obtained at 50 ° C and 400 bar extraction condition of Example 8 was the highest.

Experimental Example  2: Supercritical  Defatted green tea after extraction Heat number  Extract Crude saponin  Content survey

In order to examine the crude saponin content of the above Example 10, Comparative Example 1, Comparative Example 2, Comparative Example 3 and ground green tea (raw material), methanol was added to each of the above samples repeatedly extracted three times and then concentrated under reduced pressure. The concentrate was degreased by heating with diethyl ether for 30 minutes, and then treated with water-saturated n-butanol again to obtain an n-butanol layer, followed by concentration under reduced pressure. Each of the residues was measured for the amount of crude saponin according to the following formula after the constant amount was measured.

Crude saponin content;

In the above equation, A represents the weight (g) of the flask after the water-saturated butanol layer is concentrated and dried, B represents the weight (g) of the empty flask with constant weight, and S represents the weight (g) of the sample.

The content of crude saponin in Example 10, Comparative Example 1, Comparative Example 2, Comparative Example 3 and pulverized green tea (raw material) is shown in Table 3 below.

Green tea (raw material) Comparative Example 1 Comparative Example 2 Comparative Example 3 Example 10 Crude saponin (mg / g) 42.52 204.6 43.20 178.1 208.5

As shown in Table 3, green tea fruit contains crude saponin equivalent to 42.52 mg / g. The crude saponin contents of the extracts obtained by freeze-drying the respective extracts obtained by defatting through fluid extraction, hot water extraction (Example 10) or degreasing and extracting the alcohol (Comparative Example 1) were 208.5 and 204.6, respectively, It was confirmed that crude saponin was contained about 4.5 times or more than the fruit (raw material) itself. In addition, the amount of crude saponin similar to that of the defatted green tea extract after the supercritical extraction of Comparative Example 1 was also confirmed in the extract obtained by using the supernatant fluid and the auxiliary solvent obtained in Comparative Example 3 together.

Experimental Example  3: Supercritical  Defatted green tea after extraction Heat number  Extract in vitro Anti-obesity  Effect analysis

In order to confirm the anti-obesity effect of Comparative Example 4 (green tea fruit extract), Example 10 (defatted green tea fruit extract after supercritical extraction) and Comparative Example 1 (defatted green tea extract) Experiments were performed by culturing adipocytes from adult rats. 40% high-fat diet for 16 weeks induced mast intended for C57BL / 6 mouse by extracting the fat tissue in DMEM containing 10% FBS after removing the fat in primary culture by In vitro anti - obesity experiments were performed. MTT assay was performed to determine the concentration of the adipocytes in rats. The amount of the cell extract-safe extract was determined by adding 200 μg / ml (hereinafter referred to as "hot water extract 200") of green tea fruit extract of Comparative Example 4 (hereinafter, referred to as "hot water extract 200") per 1 ml of nutrient medium for growing cells, Ug / ml (hereinafter, referred to as "alcohol extract") and 140 μg / ml of defatted green tea fruit extract after supercritical extraction of Example 10 (hereinafter, supercritical extract 140). The levels of triglyceride, glycerol, hormone sensitive lipase (HSL), adipose triglyceride lipase (ATGL) and lipoprotein lipase (LPL) in adipocytes were measured using an adipocyte- Respectively.

The results of measuring the TG content in adipocytes are shown in Fig. 4 to confirm the anti-obesity effect. As shown in FIG. 4, when the fat was decomposed in general, the TG value decreased. Experimental results showed that the TG values decreased in all the green tea extract extracts. In particular, the defatted green tea extract and the extract After 10 supercritical extraction, the TG value was significantly decreased in the defatted green tea fruit juice extract. Based on these results, it can be confirmed that the defatted green tea juice extract after the supercritical extraction of Comparative Example 1 and the defatted green tea juice extract after the supercritical extraction of Example 10 have an anti-obesity effect.

In vitro experiments were conducted to determine the changes in the glycerol levels in the adipocytes of the green tea extract. The results are shown in FIG.

As shown in FIG. 5, when the fat is decomposed, the glycerol level increases. As a result, the glycerol level of the alcohol extract of supernatant 80 and supercritical extract 140 was greatly increased. However, the increase in hot water extract 200 was small. These results confirmed that the alcohol extract and defatted supercritical extract had an anti-obesity effect.

The results of confirming the levels of HSL (hormone sensitive lipase), ATGL (Adipose Triglyceride lipase) and LPL (Lipoprotein lipase) in adipocytes are shown in FIG. As shown in FIG. 6, when fat was decomposed, HSL and ATGL levels increased and LPL levels decreased. Experimental results showed that the HSL and ATGL levels were significantly increased and the LPL levels were greatly decreased in the alcohol extract of supernatant 80 and the supercritical extract 140. However, the increase / decrease in hot water extract 200 was small. These results suggest that the alcohol extract and defatted supercritical extract have an anti - obesity effect.

In addition, an evaluation test was conducted to examine the effect of FAS (Fatty acid synthase) of green tea extract on the extract, and the results are shown in FIG. The biosynthesis of fatty acids is caused by the acetyl CoA carboxylase and FAS (Fatty acid synthase). FAS is an enzyme that synthesizes fatty acids by using NAD (P) H as the coenzyme in acetyl CoA and malonyl CoA. Therefore, the degree of degradation of intracellular fat can be indirectly confirmed by checking the amount of FAS released.

As shown in FIG. 7, the hydrothermal extract 200, the alcohol extract 80 and the supercritical extract 140 were treated to show the FAS efflux reduction effect in the order of supercritical extract 140? Alcohol extract 80? Hot water extract 200. In the group treated with Supercritical Extract 140, 55% of the FAS was decreased compared to the control group. In the group treated with the extract of the alcoholic extract 80, 45% of the treated group and 21% of the treated group with FAS 200 Effect. There was no significant difference between the group treated with the alcohol extract of Supernatant 80 and Supercritical Extract 140, but all of them including the hydrothermal extract 200 showed the lipolytic effect.

In the above four experimental results, it was confirmed that the defatted green tea extract and the defatted green tea fruit juice extract after supercritical extraction from green tea extract had an anti - obesity effect. On the other hand, the effect was not significant in non - degreased green tea extracts without supercritical extraction.

Experimental Example  4: Defatted green tea fruit Heat number  Extract in vivo Anti-obesity  Effect analysis

Green tea extracts were found to have anti - obesity effect in vitro and further confirmed by animal experiments. The samples were prepared by dissolving the defatted green tea fruit hot water extract (Example 10) after supercritical green tea extract extraction of Example 10 and Comparative Example 1, Comparative Example 2 and Comparative Example 3 (Example 10), and defatted green tea extract (Comparative Example 1), a defatted green tea powder after supercritical extraction (Comparative Example 2), and a concentrate of supercritical extract extracted with carbon dioxide and a co-solvent (Comparative Example 3).

Experimental groups were divided into 12 groups: wild type (wild type) and normal group (normal group). The other 11 groups were fed with obesity diet to induce obesity. For the anti-obesity test, the samples were prepared at a concentration of 200 ㎎ and 500 ㎎ per kg body weight of rats, respectively, and the experiments were carried out using 200, 500, and so on (hereinafter referred to as hot water 200, hot water 500, 500, supercritical degreasing 200, supercritical degreasing 500, supercritical enrichment 200 and supercritical enrichment 500). In the obesity induction group, the control group was fed a mixture of a normal diet and an obesity diet. In addition, the other 10 groups were fed a diet containing a normal diet and an obesity diet, , Defatted green tea extract (hereinafter referred to as "500", "200") after supercritical extraction, defatted green tea after supercritical extraction (Supercritical water extract (hereinafter, referred to as supercritical degreasing 500, supercritical degreasing 200), supercritical green tea oil (supercritical water concentration 500, supercritical water concentration 200), and catechin .

Eight groups of rats fed each diet were divided into 12 groups. TG (triglyceride), TC (total cholesterol) and HDL-C blood glycerol levels were measured before and after ingestion of each diet using experimental rats. In addition, TG and TC levels were measured in the liver, and the changes in the number and size of lipids were observed to confirm the anti - obesity effect. The body weight change before and after the ingestion of 12 groups of the experimental group was measured, and the results are shown in Table 4 below.

Initial weight (g) Final weight (g) Weight gain (g / 13 weeks) Dietary intake (g / day) Dietary intake (kcal / day) Dietary intake (kcal / 13 weeks) FER wild type 16.41 ± 0.196 31.29 + 1.343 14.88 ± 1.299 3.20 ± 0.243 13.14 1178.24 0.01 ± 0.001 control 16.26 ± 0.421 50.34 + - 2.244 34.07 ± 2.347 3.04 + 0.265 15.95 1416.32 0.02 0.002 positive control (CLA 0.5%) 16.08 + - 0.462 46.54 ± 2.141 30.46 + 2.044 3.28 ± 1.455 17.21 1556.97 0.02 0.001 Number of columns 500 15.66 + 0.588 48.71 ± 3.713 33.05 ± 3.692 3.32 ± 0.364 17.41 1518.53 0.02 0.002 Number of columns 200 15.59 + 0.476 51.34 + 1.45 35.75 ± 1.348 3.32 ± 0.430 17.38 1528.72 0.02 0.001 Alcohol 500 15.48 + 0.477 45.53 + - 2.726 30.05 ± 2.593 3.81 ± 0.856 19.97 1768.41 0.02 0.001 Alcohol 200 15.39 + - 0.356 48.55 + 1.634 33.16 ± 1.39 3.13 ± 0.376 16.40 1488.00 0.02 0.001 Supercritical degreasing 500 15.50 + - 0.689 48.90 ± 2.803 33.40 ± 2.326 2.74 ± 0.182 14.38 1276.03 0.03 0.002 Supercritical degreasing 200 15.54 + 0.358 44.06 ± 2.132 28.53 + 2.087 3.15 ± 0.499 16.53 1547.57 0.02 0.001 Supercritical enrichment 500 15.45 + 0.251 48.21. + -. 3.646 32.76 ± 3.768 4.83 ± 1.391 25.33 2181.57 0.02 0.002 Supercritical enrichment 200 15.33 + - 0.456 46.66 ± 2.704 31.34 ± 2.811 3.74 ± 1.404 19.61 1710.14 0.02 0.002 catechin 0.25% 15.55 + 0.16 44.84 ± 3.163 29.29 ± 3.101 5.28 ± 0.554 27.67 1687.67 0.02 0.001

As shown in Table 4, weight gain was suppressed in all of the defatted green tea extract (supernatant 500 and 200) after supercritical extraction, and the defatted green tea extract (supercritical defat 200) after supercritical extraction It was confirmed that weight control effect similar to that of diet was obtained.

TG, TC, HDL-C, and glycerol levels were confirmed to confirm the anti-obesity effect. The results are shown in Table 5 below.

Control group Experimental group (-) Wild type (+) Cont. (+) P.cont (CLA) (+) P.cont (Catechin) Number of columns 200 Number of columns 500 Alcohol 200 Alcohol 500 Degreasing after supercritical extraction 200 Degreasing after supercritical extraction 500 Enrichment after supercritical extraction 200 Enrichment after supercritical extraction 500 serum lipid TG (mg / dl) 104.57 ± 15.85 274.39 + - 31.75 153.24 ± 23.48 144.03 ± 1911 241.51 ± 19.11 219.47 ± 14.1 191.81 + - 34.29 162.18 ± 27.34 140.33 + - 19.41 168.22 + - 16.63 164.73 ± 20.4 190.15 ± 28.94 TC (mg / dl) 76.21 + - 10.33 138.020.847 ± 90.21 + - 13.28 85.17 + - 11.44 140.64 + - 31.42 131.47 ± 22.67 128.61 + - 14.63 93.41 + - 11.36 86.27 ± 14.74 107.35 + - 24.32 101.7 ± 18.55 110.44 ± 37.21 HDL-C (mg / dL) 68.17 ± 14.79 120.81 + - 10.27 85.34 + - 10.27 83.09 ± 14.42 129.37 ± 28.54 114.65 + - 22.47 104.76 ± 16.84 92.89 ± 10.69 78.67 ± 17.63 97.54 ± 20.32 95.31 + - 22.23 119.42 ± 31.21 Glyderol (mg / dl) 785.16 + - 56.66 875.06 + - 40.31 984.61 + - 38.14 975.08 ± 49.14 891.72 + - 72.81 861.29 + - 79.38 904.38 ± 63.48 926.34 + - 41.84 944.36 ± 27.54 894.21 + - 34.69 898.14 ± 50.21 854.32 + - 41.21

As shown in the above Table 5, as the blood TG and TC values decrease and the HDL-C and glycerol levels increase, it means that the anti-obesity effect is present. Experimental results showed that the antiobesity effect was found in all groups consuming the green tea extracts compared to the normal rats. However, it was confirmed that the rats fed with the defatted green tea fruit juice extract after supercritical extraction had excellent antiobesity effect.

In addition, TG and TC values in the liver were confirmed. In general, the lower the TG and TC values in the liver, the greater the anti-obesity effect. The TG and TC values in rats fed with the alcohol extract 500 and supercritical degreasing 200 were found to be decreased compared with the control. These two substances were found to have an anti-obesity effect. The results are shown in Table 6 below.

Control group Experimental group (-) wild type (+) Cont. (+) P.cont (CLA) (+) P.cont (Catechin) Number of columns 200 Number of columns 500 Alcohol 200 Alcohol 500 Degreasing after supercritical extraction 200 Degreasing after supercritical extraction 500 Enrichment after supercritical extraction 200 Enrichment after supercritical extraction 500 Hepatic lipid TG (mg / dl) 189.54 ± 23.74 349.46 + 54.37 242.78 + - 38.61 201.36 ± 29.21 329.17 ± 21.54 294.06 ± 34.69 274.27 + - 56.31 235.47 ± 30.65 247.59 + 24.65 269.54 ± 37.91 267.05 ± 20.82 286.31 + - 31.33 TC (mg / dl) 139.41 + - 34.04 324.87 ± 78.69 164.26 ± 15.33 156.17 ± 27.37 273.52 + - 104.18 254.98 + - 54.68 238.52 + - 34.07 178.54 ± 24.99 184.33 + - 42.31 204.36 ± 52.17 256.04 + - 69.82 156.17 ± 27.67

Cells stained with H & E (Hematoxylin & Eosin) staining method were observed with a microscope. The results are shown in FIG. 8.

As shown in FIG. 7, when the fat is decomposed, the size and number of the fat globes are decreased and the weight of the fat tissue is decreased. Thus, it can be said that the antiobesity effect is obtained. The green tea extract, the defatted supercritical extract, It was confirmed that the size of lipid sphere decreased in all groups. Especially, it was confirmed that the size of lipid sphere was small and the number of adipose tissue was decreased in the hydrothermal 500, ethanol 200, supercritical degreasing 500, supercritical degreasing 200, and supercritical enrichment 200.

As a result of the above in vitro and in vivo tests, it was confirmed that the ethanol extract and the defatted supercritical hot water extract in the green tea extract had a large anti-obesity effect. This result shows that green tea extract has anti - obesity effect.

Claims (6)

1) crushing the green tea to a predetermined size;
2) supercritical extraction of the green tea powder with supercritical carbon dioxide; And
3) extracting the residue of the supercritical extract with water or alcohol, and then separating and purifying the residue. The composition for an anti-obesity containing green tea extract as an active ingredient.
The antiobesity composition according to claim 1, wherein the pulverization is performed by pulverizing the green tea fruit to a size of 0.5 to 2 mm.
The antiobesity composition according to claim 1, wherein the supercritical extraction is carried out at a temperature of 40 ° C to 60 ° C and a pressure of 200 bar to 400 bar.
The method of claim 1, wherein the supercritical extraction of step 2)
Characterized in that supercritical carbon dioxide is used for supercritical extraction at a flow rate of 2 ml / min to 70 ml / min for 60 to 240 min.
The antiobesity composition according to claim 1, wherein the alcohol in step 3) is methanol or ethanol. delete

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