KR20170014713A - Method for preparing of bitter gourd extract enhanced biological activity and bitter gourd ethanol extract prepared by the same - Google Patents
Method for preparing of bitter gourd extract enhanced biological activity and bitter gourd ethanol extract prepared by the same Download PDFInfo
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- KR20170014713A KR20170014713A KR1020150108515A KR20150108515A KR20170014713A KR 20170014713 A KR20170014713 A KR 20170014713A KR 1020150108515 A KR1020150108515 A KR 1020150108515A KR 20150108515 A KR20150108515 A KR 20150108515A KR 20170014713 A KR20170014713 A KR 20170014713A
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- ethanol
- bitter gourd
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Classifications
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/02—Acid
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
- A23V2250/2116—Flavonoids, isoflavones
- A23V2250/2117—Rutin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2300/00—Processes
- A23V2300/14—Extraction
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
The present invention relates to a method for preparing a luciferne extract having enhanced physiological activity and a luciferase ethanol extract prepared thereby. More particularly, the present invention relates to a method for producing a luciferase inhibitor having alpha-glucosidase inhibitory activity and antioxidant activity The present invention relates to a method for producing a lily extract having enhanced physiological activity and a lily extract prepared thereby.
Bitter gourd (Bitter gourd; bitter melon; Momordica charantia L.) is a perennial vine plant that is widely cultivated in Asia. It is a bitter melon in China. It is a bitter melon in China, Goya in Japan, Yeoju in Korea, , And so on. It is rich in carotene, carotene, calcium, iron, folic acid, pantothenic acid, plant fiber and mamordecin, including vitamin C, which has antioxidant effect. It has anticancer effect, malaria treatment, weight loss and digestion (Miura T. et al., J. Nut. Sci., Vitaminol., 47: 340-344, 2001), which has been reported in the medical field, to be.
As such, Yeoju has various physiological activities such as antioxidant activity, diabetic effect, and anti-cancer effect, so that the demand for dry powder of Yeoju is increasing, and when the dry powder is subjected to further extraction process, the physiological activity may be significantly increased. Korean Patent No. 10-1074555, for example, discloses that the effect of gout treatment of Yeoju dried powder is enhanced by extracting the Yeoju dried powder with ethanol at 4 ° C. However, since the above-mentioned invention only raises the effect of treating gout, it is necessary to study a method of preparing the yeast extract which can improve other physiological activities.
The inventors of the present invention have found that when extracting with ethanol, the alpha-glucosidase inhibitory activity and the antioxidative activity are enhanced by controlling the extraction temperature while extracting the extract with ethanol to improve the physiological activity of the yeast, And that the physiological activities are further enhanced by controlling the concentration, thus completing the present invention.
Accordingly, it is an object of the present invention to provide a method for preparing a lily extract having enhanced physiological activity, which comprises extracting the dried lily powder with ethanol at an extraction temperature of 55 to 80 ° C.
Another object of the present invention is to provide a method for preparing a fungus containing 6 to 10 mg / g of tannic acid, 6.6 to 10 mg / g of gallic acid and 3 to 13 mg / g of rutin, Ethanol extract of the present invention.
In order to accomplish the object of the present invention, the present invention provides a method for preparing a lily extract having enhanced physiological activity, which comprises extracting the dried lily powder with ethanol at an extraction temperature of 55 to 80 ° C.
The physiological activity may be one or more selected from the group consisting of alpha-glucosidase inhibitory activity and antioxidative activity.
The concentration of ethanol may be 20 to 90% (v / v).
The extracting step may be repeated twice.
The manufacturing method may further include a concentration step or a drying step.
In order to accomplish another object of the present invention, the present invention provides a pharmaceutical composition comprising 6 to 10 mg / g of tannic acid, 6.6 to 10 mg / g of gallic acid, and 3 to 13 mg of rutin / g. < / RTI >
When preparing the ethanol extract of Yeoju according to the production method of the present invention, the antioxidative activity and alpha-glucosidase inhibitory activity of Yeoju can be improved by a simple method of controlling the extraction temperature, and further, the concentration of the aqueous ethanol solution The physiological activity can be further enhanced.
The present invention provides a method for preparing a lady's extract having enhanced physiological activity, comprising extracting the lady's quince with ethanol at an extraction temperature of 55 to 80 ° C.
The physiological activity may be one or more selected from the group consisting of alpha-glucosidase inhibitory activity and antioxidative activity. Materials exhibiting alpha-glucosidase inhibitory activity exhibit antidiabetic, anticancer and antiviral activity (Dennis et al., Science , 236, 5825 (1987); Goss et al ., Clin Cancer Res , 1,93544 , FEBS Lett, 430,1722 (1998) ; van de Laar , etc., Diabetes Care, 28,15463 (2005) ; Lee DS , etc., Int J Mol Med, 20,37983 ( 2007)). In particular, harmful viruses including dengue virus, HIV, hepatitis virus and the like are known to be very sensitive to alpha-glucosidase activity inhibitory substances (Courageot et al ., J Virol , 74, 56472 (2000)). Thus, improving alpha-glucosidase inhibitory activity is a very important effect because it improves the effects of improving or treating various diseases.
The liposome-inhibiting activity, that is, the anti-obesity activity, is significantly weaker than that of the dried powder of Yeoju produced by the method according to the present invention, which means that the physiological activity which is improved by the method of producing the extract is very selective .
The dried powder may be at least one selected from the group consisting of flower, leaf, fruit, stem, root, ground part and outpost, and may be a dry powder of fruit.
In the production process of the present invention, the extraction temperature is 55 to 80 ° C, preferably 60 to 80 ° C, and most preferably 65 to 75 ° C. When the extraction temperature is lower than 55 ° C, the alpha-glucosidase inhibitory activity and antioxidant activity of the Y. rumen extract are more or less increased than that of the dried powder. In addition, when the extraction temperature exceeds 80 캜, the physiological activity is not significantly increased even if the extraction temperature is further increased, which is inefficient.
The concentration of the ethanol, more specifically, the concentration of the aqueous ethanol solution may be 20 to 90% (v / v), preferably 40 to 90% (v / v) And more preferably 60 to 90% (v / v). When the concentration of the aqueous ethanol solution is more than 20% (v / v), the alpha-glucosidase inhibitory activity and antioxidative activity of the Y. japonica extract are remarkably increased, especially when the concentration is more than 40% (v / v) The content of phenol and total flavonoid components is significantly increased. However, when the concentration of the aqueous ethanol solution is more than 90% (v / v), the extraction yield of the extract is much lowered. Therefore, the concentration of the aqueous ethanol solution is preferably 90% (v / v) or less.
The ethanol is not limited thereto, but may be added 10 to 20 times (v / w) of the dried powder. In addition, the extraction may be repeated twice, and in the second iteration, ethanol may be added at 5 to 15 times (v / w) of the dried powder. Repeating twice can extract more water-soluble and lipid-soluble components in Yeoju as well as extract more bioactive substances in Yeoju. However, the extract prepared by repeating 3 times is inefficient because it has no significant synergistic effect on the bioactivity and extraction yield than the extract prepared by repeating the extract twice.
The production method of the present invention may further include a concentration step or a drying step after extraction with ethanol, and may preferably include both a concentration step and a drying step. The concentration step may be, but is not limited to, heat condensation, reduced pressure condensation or reduced pressure heat condensation, and preferably reduced pressure condensation. In addition, the drying step may be hot air drying, freeze drying or spray drying, and preferably freeze drying.
The present invention relates to a method of preparing a yeast ethanol extract comprising 6 to 10 mg / g of tannic acid, 6.6 to 10 mg / g of gallic acid, and 3 to 13 mg / g of rutin, to provide.
The ethanol extract of Yeoju may preferably contain tannic acid in a concentration of 6.5 to 10 mg / g, gallic acid in a concentration of 7 to 10 mg / g, and rutin in a concentration of 4 to 13 mg / g, ) Is the content of extracts extracted with ethanol at 40 ~ 90% (v / v) concentration in the preparation of Yeoju extract.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, and it is to be understood by those skilled in the art that the present invention is not limited thereto It will be obvious.
≪ Examples and Comparative Examples > Preparation of Yeast Extracts >
1. Preparation of raw materials for yaju
Yeomju ( Momordica) grown in April, 2014 in Hamyang-gun, Gyeongsangnam- charantia L.) was harvested in the middle of June, and the immature and fruit were washed with water and then hot-air dried at 40 to 50 ° C to have a moisture content of 5 to 7% (w / w) Were obtained from a food branch.
2. Preparation of Examples and Comparative Examples
(30%, 50%, 70%, 95%) of ethanol was used as the extraction solvent and the extraction temperature was 50 ° C or 70 ° C in order to establish the manufacturing process for producing the yeast extract having excellent physiological activity. Lt; 0 > C. The extraction was carried out twice. The first extraction was carried out at a rate of 15 times (v / w) of the sample for 4 hours, the second extraction was carried out for 2 hours at a rate of 10 times (v / w) And extracted. After the extraction was completed, the extract was concentrated at 50 to 55 ° C with a vacuum concentrator (Heidolph Laborata 20 control, Germany), and then lyophilized (-75 ° C, 5 mTorr) was used as a sample.
The conditions of Examples and Comparative Examples are summarized in Table 1 below.
<Experimental Example>
Experimental Example 1. Measurement of extraction yield
The weight of the lyophilized concentrated extract powder was measured by a moisture meter (MA50C, Satorius Co., Germany), and the extraction yields of the respective extracts were calculated by the following equations, and the results are shown in Table 2 below.
(° C)
(Comparative Example 2)
(Comparative Example 3)
(Comparative Example 4)
(Comparative Example 5)
(Comparative Example 6)
(Comparative Example 1)
(Example 1)
(Example 2)
(Example 3)
(Example 4)
In Example 4, which was extracted with a solvent having an ethanol concentration of 95%, the extraction yield was significantly smaller than those of the other Examples. Comparing Examples 1 to 3 and Comparative Examples 3 to 5 having the same extraction solvent but different extraction temperature, The yield was higher than the comparative example. From the above results, it is preferable to use purified water or ethanol having a concentration of less than 95% as the extraction solvent for extracting the Y. japonica extract, and it is preferable that the extraction temperature is about 70 ° C rather than 50 ° C.
Experimental Example 2. Measurement of α-glucosidase inhibitory activity
α-glucosidase inhibitory activity was performed using the method of measuring the glass (cleavage) of p NPG (p-nitrophenyl-α -D-glucoside) by the α-glucosidase with a spectrophotometer (spectrophotometer).
10 μl of 1 U / ml α-glucosidase (Sigma-Aldrich Corp., USA) and 20 μl of each of the above Example and Comparative Example at a concentration of 1,000 μg / ml were added to 50 μl of a buffer (0.1 M phophate buffer, pH 6.9) After the addition, the reaction was carried out at 37 DEG C for 30 minutes. Thereafter, 20 μl of p NPG at a concentration of 1 mM was added thereto, followed by reaction at 37 ° C for 15 minutes. To stop the reaction, 50 μl of stop solution (0.1 M sodium carbonate, Na 2 CO 3 ) was added and the α-glucosidase inhibitory activity was calculated using the average of the values obtained after 3 repetitions and the equation described below And the results are shown in Table 3 below.
* A c1 : Absorbance of (buffer + α-glucosidase + p NPG + stop solution)
* A c2 : Absorbance of (buffer + p NPG + stop solution)
* A s : absorbance of (buffer + α-glucosidase + p NPG + sample + stop solution)
* A b : absorbance of (buffer + α-glucosidase + sample + stop solution)
The positive control group was acarbose (Sigma Co., USA), a diabetic agent, and the control group was a freeze-dried powder.
In Table 3, it can be seen that the alpha-glucosidase inhibitory activity of the example extracted at 70 ° C is significantly superior to the comparative example extracted at 50 ° C. The comparative example extracted at 50 占 폚 shows lower inhibitory activity than the dry powder of the control yeast.
(° C)
Experimental Example 3. Fat Absorption Inhibitory Activity
The lipid absorption inhibition activity experiment was performed with pancreatic lipase which is widely used for anti - obesity material screening. Porcine pancreatic lipase (Sigma Co., USA) was dissolved in 0.5 g / 200 ml of enzyme buffer (10 mM MOPS, 1 mM EDTA, pH 6.8) at 4 ° C and centrifuged at 4,000 rpm to remove the supernatant. ㎕ Tris buffer were mixed (100 mM Tris-HCl, 5 mM CaCl 2, pH 7.0) and 6 ㎕ enzyme buffer. Samples were dissolved in 70% alcohol and diluted to a concentration of 1.0 mg / ml. After p- nitrophenyl butyrate ( p- NPB, Sigma Co., USA) was dissolved in dimethylformamide to 10 mM, pancreatic lipase and sample were first reacted at 37 ° C for 15 minutes. Substrate was added and incubated at 37 ° C for 30 minutes Lt; / RTI > The absorbance at 405 nm was measured using an ELISA reader (Thermo Fisher Scientific Inc., USA). The inhibitory activity of pancreatic lipase was shown by the absorbance reduction rate of the sample solution added group and the no-added group, and the results are shown in Table 4 below. The positive control group was orlistat (Sigma Co., USA) at a concentration of 100 ㎍ / ㎖, which is a lipase inhibitor drug of the stomach and pancreas.
In Table 4, it is seen that the lipase inhibitory activity of the negative control group is significantly higher than that of the Comparative Examples and Examples, which means that the production method of the present invention does not enhance the fat absorption inhibitory activity.
(° C)
Experimental Example 4. Analysis of total polyphenol and total flavonoid content
4-1. Total polyphenol content analysis
Folin-denis'reagent was used for total polyphenol content measurement. 100 μl of folin-denis'reagent (Sigma Co., USA) was added to 20 μl of a 1 g-concentration sample, followed by reaction at 40 ° C. for 1 minute. After the reaction, 80 7.5 of a 7.5% Na 2 CO 3 solution was added, reacted at 40 캜 for 15 minutes, and absorbance was measured at 765 nm. The measured absorbance values were assigned to standard curves and their contents were determined. The standard solutions were gallic acid (Sigma Co., USA) and tannic acid (Sigma Co., USA). Total polyphenol content was expressed as mg gallic acid equivalent (GAE, dry basis) and ㎎ tannic acid equivalent (TAE, dry basis) in 1 g of sample. Are shown in Table 3 below.
4-2. Total flavonoid content analysis
Total flavonoid contents were measured by diethylene glycol colorimetric method. 200 di of diethylene glycol (Samchun Co., Korea) and 20 1 of 1N NaOH were added to 20 시 of a 1 g-concentration sample, followed by reaction at 37 캜 for 1 hour. After the reaction, the absorbance was measured at 420 nm, and the absorbance value was substituted into the standard curve to determine its content. The standard solution was rutin (Sigma Co., USA). The total flavonoid content was expressed as mg rutin equivalent (RE, dry basis) in 1 g of the sample, and the result was expressed as an average value of the results of three repeated experiments. The results are shown in Table 3 below.
(° C)
density(%)
When the results of Table 3 are analyzed, it is preferable that the extraction temperature is about 70 ° C in order to increase the total polyphenol and total flavonoid content. When the ethanol concentration is about 50% and 70% The content of polyphenols and total flavonoids can be remarkably increased, and particularly when the ethanol concentration is about 70%, the content of the routine as a flavonoid can be remarkably increased.
Experimental Example 5. Measurement of Antioxidative Activity
ABTS cation radical scavenging assay was performed to measure the antioxidant activity of the examples and comparative examples. This test method measures the extent to which the color of the reagent is reduced by eliminating the ABTS cation radical by the sample. Bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS, Sigma Co., USA) was added to 20 μl of a sample having a concentration of 0.25 mg / ml, 0.5 mg / Mu] l of the mixture was reacted at room temperature for 7 minutes, and the absorbance was measured at 734 nm. The average value of the derived values was substituted into the following equation to calculate the radical scavenging ability, and the results are shown in Table 4 below.
* Blank: absorbance of the control without sample
* Test: absorbance of the test group containing the sample
(° C)
In Table 4, it can be seen that the radical scavenging ability of the example having an extraction temperature of 70 占 폚 is much higher than that of Comparative Example where the extraction temperature is 50 占 폚. When the sample treatment concentration is 1 mg / ml, Which is about 2.5 times better than the comparative example. These results show that the control of extracting temperature alone can significantly enhance the antioxidant activity of Yeoju extract.
Claims (6)
Wherein the physiological activity is at least one selected from the group consisting of alpha-glucosidase inhibitory activity and antioxidative activity.
Wherein the ethanol concentration is 20 to 90% (v / v).
Wherein the step of extracting is repeated twice.
Wherein the production method further comprises a concentration step or a drying step.
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