KR20210026408A - Eriobotrya japonica leaf extract with antioxidant activity and method for manufacturing the same - Google Patents

Abstract

The present invention relates to an Eriobotrya japonica leaf extract, and a manufacturing method thereof. The manufacturing method comprises: a raw material preparation step of preparing raw materials by processing Eriobotrya japonica leaves; and an extract manufacturing step of manufacturing an extract by extracting the raw materials with ethanol. According to the Eriobotrya japonica leaf extract having antioxidant activity and the manufacturing method thereof of the present invention, the extract are manufactured by using Eriobotrya japonica leaves which are natural materials, thereby being able to obtain a natural antioxidant which can safely and effectively remove active oxygen species.

Description

Loquat leaf extract with antioxidant activity and method for manufacturing the same {Eriobotrya japonica leaf extract with antioxidant activity and method for manufacturing the same}

The present invention relates to a loquat leaf extract having antioxidant activity and a method for producing the same, and relates to a loquat leaf extract having antioxidant activity and a method for producing the same by extracting the leaves of the loquat tree using ethanol.

Phytochemicals extracted from medicinal plants are known to be beneficial to health by acting on the physiological activities of our body such as inflammation, cancer, diabetes, hypertension and anti-aging (Dillard and German, 2000). Among them, phenols and flavonoids are important ingredients that exhibit various physiological activities such as antioxidant, antibacterial, anticancer, and anti-inflammatory, and have antioxidant functions that inhibit not only diabetes and hypertension, but also circulatory disorders and skin aging, so they play an important role in preventing and treating various diseases. (Cicerale et al., 2010). Reactive oxygen species (ROS) refers to substances that have a strong oxidizing power generated in the respiratory process and energy metabolism in our body, and various chronic diseases such as diabetes and cancer by damaging the cells and DNA of our body. It is a substance that causes skin aging and skin aging. Antioxidants such as terpenoid, polyphenol, and flavonoid, which are present in medicinal plants, play an important role in maintaining human health by preventing damage caused by reactive oxygen species. Skin aging can be divided into intrinsic aging and extrinsic aging, which are caused by the decrease in the function of physiological factors. Exogenous aging, which has the greatest effect, is mainly caused by reactive oxygen species generated through ultraviolet rays and respiration. Antioxidants that remove this have been proven to be effective in preventing and delaying skin aging, and research has been actively conducted in various fields such as food, medicine, and beauty.

Antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), polyphenol, flavonoid, and vitamin C are known to be effective in inhibiting the formation and accumulation of harmful active oxygen in the human body (Bowden, 2004; Das et al. , 1999).

However, synthetic antioxidants such as BHA and BHT, which have been widely used for their excellent antioxidant effects, have been reported to have side effects such as increasing the activity of the hepatocyte microsomal enzyme in the body and causing cancer. In particular, in the food industry, concerns about the dangers of using synthetic antioxidants are growing, and interest in natural substances is increasing day by day. Research on natural antioxidants that can remove active oxygen species more safely and effectively than synthetic antioxidants is actively progressing. There is a lot of attention on how to extract effectively from natural products.

Korean Registered Patent Publication No. 10-1135822

The present invention has been invented to solve the above problems, and an object of the present invention is to provide a loquat leaf extract having antioxidant activity obtained by extracting loquat leaves using ethanol, and a method for preparing the same.

The loquat leaf extract having antioxidant activity according to the present invention for achieving the above object is extracted with ethanol from loquat leaves.

It is preferable that the loquat leaf extract having antioxidant activity of the present invention is to extract the loquat leaf using ethanol of 80% concentration.

The loquat leaves are young leaves of loquat trees collected in spring.

On the other hand, the method for producing a loquat leaf extract having an antistatic activity according to the present invention includes a raw material preparation step of preparing a raw material by processing a loquat leaf, and an extract production step of extracting the raw material with ethanol to prepare an extract.

The extract preparation step includes an extraction step of extracting the first extract using the ethanol having a predetermined set ratio of the weight of the raw material, and the ethanol of the weight used in the extraction step. It includes a repeating step of extracting the second extract by using.

In the extraction step, the first extract is extracted using a vacuum low-temperature concentrator for a preset first working time at a preset extraction temperature using the ethanol for the raw material.

It is preferable that the extraction temperature is 90° C., and the first operation time is 4 hours.

The setting ratio is 10 times applied.

The extract manufacturing step may further include a drying step of freeze-drying the second extract after the repeating step.

The drying step includes a freezing step of freezing the second extract at a preset freezing temperature for a preset second working time and then freeze-drying, and after the freezing step, the second extract is stored at a storage temperature higher than the freezing temperature. Including the storage stage of storage.

The freezing temperature is -70°C, and the second working time is applied for 24 hours.

The raw material preparation step includes a leaf washing step in which the loquat leaves are washed and then heated and dried, and after the leaf washing step, the loquat leaves are pulverized to a predetermined size to prepare the raw material.

In the leaf washing step, the washed loquat leaves are dried by heating at 80° C. for 5 hours.

The loquat leaf extract having antioxidant activity and its manufacturing method according to the present invention have the advantage of obtaining a natural antioxidant that can safely and effectively remove active oxygen species because the extract is prepared using the leaves of the loquat tree, which is a natural material. There is this.

1 is a flow chart for a method of manufacturing a loquat leaf extract having antioxidant activity according to the present invention,
2 is a result of measuring DPPH radical scavenging activity of loquat leaves and young leaves extracted with various solvents according to concentration,
3 is a result of comparing the ABTSradical scavenging ability at a concentration of 5-80 μg/mL of loquat leaves and young leaves extracted under various solvent conditions.

Hereinafter, a loquat leaf extract and a manufacturing method thereof according to an embodiment of the present invention will be described in detail with reference to the accompanying drawings. In the present invention, various modifications may be made and various forms may be applied, and specific embodiments will be illustrated in the drawings and described in detail in the text. However, this is not intended to limit the present invention to a specific form disclosed, it should be understood to include all changes, equivalents, or substitutes included in the spirit and scope of the present invention. In describing each drawing, similar reference numerals have been used for similar elements. In the accompanying drawings, the dimensions of the structures are shown to be enlarged than the actual size for clarity of the present invention.

Terms such as first and second may be used to describe various elements, but the elements should not be limited by the terms. The above terms are used only for the purpose of distinguishing one component from another component. For example, without departing from the scope of the present invention, a first element may be referred to as a second element, and similarly, a second element may be referred to as a first element.

The terms used in the present application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, but one or more other features. It is to be understood that the presence or addition of elements or numbers, steps, actions, components, parts, or combinations thereof, does not preclude in advance the possibility of the presence or addition.

Unless otherwise defined, all terms used herein including technical or scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms as defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and should not be interpreted as an ideal or excessive formal meaning unless explicitly defined in this application. Does not.

The loquat leaf extract having antioxidant according to the present invention is an extract obtained by extracting loquat leaves with ethanol. Here, the loquat (Eriobotrya japonica Lindl.) is a leaf of the Rosaceae family. It is a long circular obovate, covered with yellowish brown cilia and is shiny. It grows widely in China, Spain, and Japan. In Korea, it mainly grows in mild climate conditions such as Jeju-do, Gyeongnam and Jeollanam-do. Not only cultivation but also loquat processing industry is active. In addition, in folk remedies, loquat fruits or leaves are effective in Jinhae, expectorant, vomiting, hematopoiesis, diuresis, and soothing breathing and quenching thirst. Loquat contains a large amount of physiologically active ingredients such as terpenoid-based compounds such as various ursolic acid and oleanolic acid, and flavonoid-based compounds such as quercetin, and thus antioxidant, anti-inflammatory, and anti-cancer activities have been studied. In particular, in loquat leaves, in addition to ellagic acid and chlorogenic acid, there are substances with antioxidant activity such as ursolic acid, neochlorogenic acid, and kaempferol.

The loquat leaf extract having antioxidant activity of the present invention extracts loquat leaves using ethanol of 80% concentration, and the loquat leaves are leaves of a loquat tree, and young leaves or June collected in spring, that is, in March or April. Alternatively, leaves harvested in July are used.

On the other hand, Figure 1 is a flow chart for a method for producing a loquat leaf extract having antioxidant activity of the present invention is shown.

Referring to the drawings, the method of manufacturing a loquat leaf extract having antioxidant activity includes a raw material preparation step (S110) and an extract manufacturing step (S120).

The raw material preparation step (S110) is a step of preparing raw materials by processing loquat leaves, and includes a leaf washing step (S111) and a crushing step (S112).

The leaf washing step (S111) is a step of washing the loquat leaves and then heating and drying them. Here, the loquat leaves are the leaves of the loquat trees, and young leaves harvested in spring, that is, in March or April, or leaves harvested in June or July, are used. After washing fresh loquat leaves, it is dried by heating at 80°C for 5 hours in a dry oven.

The grinding step (S112) is a step of manufacturing the raw material by grinding the loquat leaves to a predetermined size after the leaf washing step (S111). The heat-dried loquat leaves are pulverized to a size of 1 cm or less in diameter to prepare raw materials.

The extract manufacturing step (S120) is a step of preparing an extract by extracting the raw material with ethanol, and includes an extraction step (S121), a repeating step (S122), and a drying step (S123).

The extraction step (S121) is a step of extracting the first extract by using the ethanol having a weight of the raw material at a predetermined ratio of the weight of the raw material. Here, 10 times the setting ratio is applied. In addition, it is preferable to extract the first extract using a vacuum low-temperature concentrator (COSMOS660-50L, Kyungseo Machines Co, Incheon, Korea) for a preset first working time at a preset extraction temperature using the ethanol for the raw material. . Here, the extraction temperature is 90°C, and the first working time is applied for 4 hours. That is, the first extract is extracted from 1.5 kg of loquat leaf raw materials in ethanol at a concentration of 10 times 80%, using an ultra-high-speed vacuum low-temperature concentrator at 90° C. for 4 hours.

The repeating step (S122) is a step of extracting the second extract by using the ethanol of the weight used in the extraction step (S121). That is, the first extract is extracted in ethanol at a concentration of 10 times 80% per 1.5 kg using an ultra-high vacuum low-temperature concentrator at 90° C. for 4 hours. At this time, it is preferable that the operator repeats the repeating step (S122) twice. Here, it is preferable to concentrate the second extract to about 30 Bx with an ultra-high vacuum low-temperature concentrator.

The drying step (S123) is a step of freeze-drying the second extract after the repeating step (S122), and includes a freezing step (S124) and a storage step (S125).

The freezing step (S124) is a step of freezing the second extract at a predetermined freezing temperature for a predetermined second working time and then freeze drying. Here, the freezing temperature is -70°C, and the second working time is applied for 24 hours.

Meanwhile, the second extract may be frozen at a freezing temperature and then dried through a drying device. Although not shown in the drawing, the drying device includes a main body in which the second extract frozen therein is accommodated, a gripping member installed inside the main body and holding the second extract away from the bottom surface of the main body, and the main body A hygroscopic material installed therein to absorb moisture, and cold air supply means for providing cold air at a low temperature corresponding to a freezing temperature into the body.

The cold air supply means includes a cold air supply pipe communicating with the inside of the main body, a blowing fan installed in the cold air supply pipe to forcibly blow outside air into the body, and a blowing fan installed in the cold air supply pipe and blown into the body by a blowing fan. And a cooling unit that cools the outside air to a low temperature. Since the cooling unit is a cooling means generally used in the related art for cooling air, a detailed description thereof will be omitted.

The gripping member is rotatably installed inside the body at a position spaced a predetermined distance upward from the bottom surface of the body, and a gripping arm provided with tongs for gripping the frozen second extract at an end thereof, and rotating the gripping arm In order to be provided with a rotary motor installed on the gripping arm. Since the holding member rotates the frozen second extract, drying efficiency for the second extract is improved.

On the other hand, the holding member is installed inside the body at a position spaced a predetermined distance upward from the bottom surface of the body, the second extract frozen thereon is seated, a support net having a plurality of through holes penetrating in the vertical direction and , It may be provided with a vibrator installed on the support net for applying vibration to the support net at predetermined unit time intervals. Meanwhile, the drying apparatus is not limited thereto, and a drying means conventionally used for freeze drying may be applied.

As described above, since it is freeze-dried using a drying device, it is possible to obtain a natural antioxidant having a higher effect by preventing evaporation or disappearance of substances related to the antioxidant activity contained in the second extract.

The storage step (S125) is a step of storing the second extract at a storage temperature higher than the freezing temperature after the freezing step (S124). Here, the storage temperature is 4℃. Since the dried second extract is stored at a low temperature, it can be stored in a fresh state for a long time without destroying the active substance.

On the other hand, in order to confirm the efficacy of the loquat leaf extract prepared by the manufacturing method according to the present invention, the loquat leaf extract of the present invention and the extracts extracted using hot water and 20% ethanol concentration of the loquat leaf under the same conditions were total polarized. An experiment was conducted to verify the bonoid content, DPPH radical scavenging, ABTS radical scavenging, and HPLC method. The experiment was also conducted on the loquat leaf extract of the present invention using young loquat leaves collected in spring, and extracts extracted using hot water and ethanol at a concentration of 20%.

First, the total flavonoid content analysis was measured by applying the experimental method of Moreno et al. (2000). For analysis, 100 μL of each extract was diluted with 990 μL of 80% ethanol, and 100 μL was mixed with 4.3 mL of 80% ethanol containing 10% aluminumnitrate and 1M potassium acetate, and allowed to stand at room temperature for 40 minutes. Absorbance was measured at 415 nm using a photometer (Neogen, Optizen 2120 UV, Sejong, Korea). At this time, the total flavonoid content was obtained from the standard curve prepared using the standard quercetin (mg QE/sample of 100 g) (Absorbance=0.0468 μg quercetin+0.0195 (R2=0.99)).

DPPH radical scavenging measurement is one of the methods of measuring the antioxidant activity of extracts by measuring DPPH radical scavenging activity, and was measured by applying the experimental method of Blois (1958). Using 99.9% ethanol, 100 μL of DPPH (2.0*?*10*?*4 M) solution and 100 μL of each extract for each concentration were added to a 96 well plate, mixed well, and allowed to react at room temperature for 30 minutes. It was measured at nm using an ELISA reader (MECASYS, Optizen 2120 UV, Daejeon, Korea). L-ascorbic acid was used as a positive control. Antioxidant activity was expressed as a percentage by the following method.

ABTS radical scavenging measurement is one of the methods of measuring the antioxidant activity of extracts through the scavenging activity of ABTS radicals, and was measured by applying the experimental method by Pellegrini et al. (1999). By mixing 7 mM ABTS and 2.45 mM potassium persulfate in a ratio of 1:1 (v/v), radicals were generated by reacting for 24 hours in a dark place at 37oC to minimize the consumption of radicals by light. Radical stock solution was diluted with Phosphate Buffer Saline (pH 7.4) so that the absorbance value at 734 nm was 0.68±0.020. For each concentration, 990 μL of the ABTS radical solution diluted in 10 μL of the extract was added and left in the dark for 1 minute, and then measured with a spectrophotometer (Neogen) at 734 nm. L-ascorbic acid was used as a positive control. Antioxidant activity was expressed as a percentage by the following method.

To this end, a standard solution was prepared, and the preparation of the standard solution is as follows. As the ellagic acid standard, distilled water containing 1 N NaOH was used, and this solution was diluted to prepare a standard solution to a concentration of 10-100 μg/mL. Chlorogenic acid standards were dissolved in distilled water and stored in a refrigerator at 4°C, and a standard solution was prepared to a concentration of 10-80 μg/mL.

In addition, high performance liquid chromatography (HPLC) equipped with an autosampler and DAD detector was used to measure the content of ellagic acid and chlorogenic acid in phenol components to be analyzed, using the Agilent 1100 series (Agilent Technologies, Santa clara, CA, USA). I did. SHISEIDO CAPCELL PAK (4.6x250 mm, Tokyo, Japan) was used as an analytical column. The mobile phase was analyzed using tertiary distilled water (solvent A) and 100% Acetonitrile (solventB) containing 0.1% formic acid, and the analysis conditions are shown in Table 1 below.

In addition, the validation of the analysis method for indicator components is linearity, limit of detection (LOD), and limit of quantitation (LOQ) in accordance with the validation guidelines (KFDA, 2004) for analysis methods such as pharmaceuticals. , Accuracy and precision were judged to verify the analysis method.

In addition, for the measurement of linearity and the limit of detection and quantification, an experiment was conducted by preparing a solution of standard solutions of ellagic acid and chlorogenic acid in 5 concentrations, and these standard solutions were used in HPLC to obtain retention time and regression equation (y=ax+by). : peak area, x: concentration (μg/mL)) was used to determine the Determination coefficient (R2). The detection and quantitation limits for each component were calculated based on the standard deviation and slope of the calibration curve using the chromatogram of the standard solution.

And, both accuracy and precision were evaluated for accuracy and precision in order to find out the inter-day and intra-day fluctuations of the index components of the same concentration.

In addition, for content analysis, the peak area value was substituted based on the calibration curve prepared through standard analysis and calculated using a proportional equation. As the extract, a solution filtered with a syringe filter (0.45 μM, Hyundaimicro Co., Ltd, Seongnam, Korea) was used as a test solution.

And, all the experimental results presented in this experiment are expressed as the mean value ± standard deviation (Mean ± D) measured repeatedly three times. Statistical processing was performed using the Statistical Package for the Social Sciences (SPSS), and a one-way analysis of variance (ANOVA) was performed for the statistical significance test between each experimental group. A range test was performed. When p<0.05, it was judged to be significant.

The results of this experiment are as follows. The yield of the extract acts as an important factor in the measurement of antioxidant activity, and even if the physiological activity of the extract is excellent, if the yield of the extract is low, it is practically inferior in the economical use of the sample, so the extraction yield is suitable for various commercialization of functional extracts. It is an important factor to be considered (Ham et al., 2015). The yield of each extract is shown in Table 2 below. Here, the 80% EtOH item of E.japonica Lindl.leaf and E.japonica Lindl.young leaf is a loquat leaf extract obtained by the manufacturing method of the present invention for loquat leaves and young loquat leaves, and subsequent experimental results also correspond The item applies the extract obtained by the manufacturing method of the present invention.

Referring to Table 2, the extraction yields of 80% ethanol extracts of loquat leaves and young leaves were relatively high at 22.0% and 23.9%, and 20% ethanol extracts of loquat leaves showed the lowest yield at 9.4%. This is considered to show the difference in extraction yield as a variable of the mixing ratio of water and ethanol to the sample, and it is known that it is economical if the extraction yield is more than 10% (Park et al., 2003), so loquat leaves and young leaves are economically utilized. It seems to be a possible plant material.

Flavonoids, known to inhibit oxidation in the body, are important compounds in determining the antioxidant activity of natural products. They are the most helpful substances for improving immunity. They have the effect of scavenging free radicals. Can be used. In this experiment, the content of flavonoids according to the extraction solvent conditions of loquat leaves and young leaves was measured, and the content of the total flavonoid compound was converted to the amount of quercetin, and is shown in Table 3 below.

Referring to the above table, in the loquat leaves, 80% ethanol extract was the highest at 2,149.7 mg QE/g, followed by 20% ethanol extract and hot water extract. In the case of young leaves, like loquat leaves, 80% ethanol extract showed the highest value at 502.3 mg QE/g, followed by hot water extract and 20% ethanol extract, which contained total flavoids. Compared to the previously studied rose family bokbunja 95% ethanol extract, which showed a total flavonoid content of 249.8 mg QE/g, loquat leaves and loquat young leaves are thought to exhibit high antioxidant efficacy. . It also suggests that 80% ethanol extract of loquat leaves and young leaves is a more effective extraction method than 20% ethanol and hot water extract.

Meanwhile, FIG. 2 shows the results of measuring DPPH radical scavenging activity of loquat leaves and young leaves extracted with various solvents according to concentration. Here, FIG. 2(a) is an extract using loquat leaves harvested in June or July, and FIG. 2(b) is an experiment result for an extract using young loquat leaves harvested in March or April. Referring to FIG. 2, DPPH radical scavenging activity of loquat leaves and young leaves extracted with various solvents was measured according to concentration, and as a result, at a concentration of 0.8-100 μg/mL, higher scavenging activity was observed. Lascorbic acid, known as an existing antioxidant, showed the highest scavenging activity of 43.2-91.0% at 1.5625-6.25 μg/mL (data not shown), and more than 50% scavenging activity (IC ₅₀ ) at the concentration of 2.45 μg/mL. (Table 4). All extract samples showed significantly higher scavenging activity of 70% or more at 25 μg/mL.

Referring to Table 4 below, the 80% ethanol extract IC ₅₀ of loquat leaves and young leaves showed the lowest values of 13.9 and 20.7 μg/mL, respectively, and young leaves were 20.7-27.4 μg/mL in all extracts than loquat leaves. It showed a low value.

The radical scavenging activity of 80% ethanol extracts of loquat leaves and young leaves showed the highest scavenging activity of 72.1-90.1% and 58.9-89.9% at concentrations of 12.5-50 μg/mL, respectively, which was found to contain a large amount of flavonoids. It is presumed that a relatively high scavenging activity was seen.

As an experimental method highly correlated with actual antioxidant activity, ABTS is _{an active cation radical reacted with peroxidase, H 2} O _2, and antioxidant activity is measured by utilizing the principle that the blue-green color peculiar to the radical is decolorized when removed by an antioxidant.

3 shows the results of comparing the ABTSradical scavenging ability at a concentration of 5-80 μg/mL of loquat leaves and young leaves extracted under various solvent conditions. Here, FIG. 3(a) is an extract using loquat leaves harvested in June or July, and FIG. 3(b) is an experiment result for an extract using young loquat leaves harvested in March or April.

Referring to FIG. 3, the results of comparing the ABTSradical scavenging activity at a concentration of 5-80 μg/mL of loquat leaves and young leaves extracted under various solvent conditions showed a result consistent with the DPPH radical scavenging activity experiment. In each extract sample, as the concentration increased, the scavenging ability increased significantly, and the scavenging activity was higher than 50% at the concentration of 20 μg/mL.

L-ascorbic acid showed a high ABTS radical scavenging activity of 53.8-96.5% at 2.5-4.5 μg/mL, and referring to Table 5 below, the IC ₅₀ value was confirmed to be 2.36 μg/mL.

In particular, the hot water, 20% ethanol, and 80% ethanol extracts of young loquat leaves (extracts of the present invention) had IC ₅₀ values of 21.0, 19.0, and 17.3 μg/mL, respectively, and the antioxidant activity of the hot water extracts of loquat leaves was similar or low. Showed the results. In this study, the results of ABTS radical scavenging activity showed relatively higher activity than DPPH radical scavenging activity, because ABTS can measure both the radical scavenging activity of hydrophilic and hydrophobic samples and thus exhibits high antioxidant activity. In addition, in terms of natural products, in hot water extracts of loquat leaves and young leaves, IC ₅₀ values were 18.0 and 21.0 μg/mL, respectively, showing high antioxidant activity (see Table 4). In addition, in the study of Lee and Kim (2009), 80% ethanol extract of loquat leaves was consistent with the results that showed the most effective antioxidant scavenging ability SD50 of 1.71 mg/mL. Such antioxidant activity can inhibit the oxidation of food ingredients and can be used as a natural antioxidant that can effectively remove active oxygen, and it can be used as a potential value.

Linearity was evaluated to confirm whether a linear measurement value could be obtained for the amount of ellagic acid and chlorogenic acid in a certain concentration range, and the standard solution of ellagic acid and chlorogenic acid was repeatedly measured by HPLC to obtain a calibration curve. The coefficient of determination (R2) value is 0.9994-0.9999, which is almost the same as 1, showing high linearity. Referring to Table 6 below, the detection limit of ellagic acid was verified by the test method showing the detection limit and the quantitative limit lower than the analyzed concentration of each component, and thus the detection limit and the quantitative limit for the analysis of the index component were verified.

In order to obtain the accuracy and precision of this experiment, standard solutions of ellagic acid and chlorogenic acid were added at low and high concentrations, respectively, and then collected by analysis, intra-day and inter-day. It was confirmed with reference to Table 7 of. The accuracy of ellagic acid was found to be within 100.0-101.0%, and chlorogenic acid was found to be within 99.7-108.0%.

This satisfies within 90-110% of the recovery rate error, which is the standard value of the Ministry of Food and Drug Safety. Referring to Table 8 below, precision shows good values of 0.20-5.31% of ellagic acid and 0.29-0.73% of chlorogenic acid as a cv, coefficient variation. % Or less was satisfied.

Phenol is one of the antioxidant substances that protects DNA, proteins and enzymes damaged by oxygen free radicals, and has recently attracted attention due to its biological and various pharmacological effects. Representative polyphenol compounds, ellagic acid and chlorogenic acid, are natural compounds and are known as major active ingredients of loquat leaves, and their anti-inflammatory, antioxidant and antiviral effects have been reported. In particular, it has been reported that ellagic acid exhibits anticancer activity against carcinogenesis induced by benzo[alpha]pyrene, and that chlorogenic acid can regulate fat metabolism and glucose. To confirm the correlation between flavonoid content and antioxidant activity and polyphenols, ellagic acid and chlorogenic acid content were analyzed in 80% ethanol extract of loquat leaves with the highest antioxidant activity (see Table 5). The 80% ethanol extract of loquat leaves contained 4.21±0.04 mg/g of ellagic acid and 5.13±0.06 mg/g of chlorogenic acid.

Therefore, as a result of this study, when examining the extraction yield of loquat leaves, antioxidant properties, and active ingredients, the yield, flavonoid content, and antioxidant activity (DPPH, ABTS) are the most excellent in the 80% ethanol extract as in the manufacturing method of the present invention. Can be seen.

The loquat leaf extract having antioxidant activity according to the present invention and its manufacturing method according to the present invention constituted as described above, is prepared by using the leaf of the natural material loquat tree to prepare a natural antioxidant that can safely and effectively remove active oxygen species. There is an advantage that can be obtained.

The description of the presented embodiments is provided to enable any person skilled in the art to use or practice the present invention. Various modifications to these embodiments will be apparent to those of ordinary skill in the art, and general principles defined herein can be applied to other embodiments without departing from the scope of the present invention. Thus, the present invention is not to be limited to the embodiments presented herein, but is to be construed in the widest scope consistent with the principles and novel features presented herein.

S110: raw material preparation step
S111: Leaf washing step
S112: grinding step
S120: extract preparation step
S121: extraction step
S122: Repeat step
S123: drying step
S124: freezing phase
S125: storage phase

Claims (13)

The loquat leaves were extracted with ethanol,
Loquat leaf extract with antioxidant activity. The method of claim 1,
Extracting the loquat leaves using 80% ethanol,
Loquat leaf extract with anti-oxidant activity. The method of claim 1,
The loquat leaves are young leaves of loquat trees collected in spring,
Loquat leaf extract with anti-oxidant activity. Raw material preparation step of preparing raw materials by processing loquat leaves; And
Containing; an extract manufacturing step of extracting the raw material with ethanol to prepare an extract
Method for producing a loquat leaf extract having an antioxidant activity. The method of claim 4,
The extract manufacturing step
An extraction step of extracting a first extract by using the ethanol having a weight of the raw material in a predetermined ratio of the weight of the raw material; And
Including; a repeating step of extracting the second extract using the ethanol of the weight used in the extraction step of the first extract;
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 5,
In the extraction step, extracting the first extract using a vacuum low-temperature concentrator for a preset first working time at a preset extraction temperature using the ethanol for the raw material,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 6,
The extraction temperature is 90° C., and the first operation time is 4 hours,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 5,
The setting ratio is 10 times,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 5,
The extract manufacturing step
After the repeating step, a drying step of freeze-drying the second extract; further comprising,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 9,
The drying step
A freezing step of freezing the second extract at a predetermined freezing temperature for a predetermined second working time and then freeze drying; And
After the freezing step, a storage step of storing the second extract at a storage temperature higher than the freezing temperature; including,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 10,
The freezing temperature is -70°C, and the second working time is 24 hours,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 4,
The raw material preparation step
A leaf washing step of washing the loquat leaves and then drying them by heating; And
After the leaf washing step, a grinding step of manufacturing the raw material by grinding the loquat leaves to a predetermined size; including,
Method for producing a loquat leaf extract having antioxidant activity. The method of claim 12,
In the leaf washing step, drying the washed loquat leaves by heating at 80° C. for 5 hours,
Method for producing a loquat leaf extract having antioxidant activity.

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