KR20150141028A - Method for production of small colored potato extract with antioxidative materials - Google Patents

Abstract

The present invention relates to a method for producing a small colored potato extract with an anti-oxidative material and, more specifically, to a method for producing a small colored potato extract with an anti-oxidative material, comprising: a step of obtaining a supercritical pressure extract by putting a small colored potato in a extracting device using supercritical pressure and extracting a small colored potato at 200 to 400 MPa pressure; and a step of hot-water extracting the supercritical pressure extract. The small colored potato extract containing a large amount of anti-oxidative material is produced by using the method of the present invention.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing an antioxidant,

The present invention relates to a method for preparing an extract of colorant potato comprising an antioxidant, and more particularly to a method for preparing an extract of a colorant potato having an antioxidant substance further enhanced by further comprising an ultra-high pressure extraction step.

The active oxygen generated in the body was generally able to inhibit and defend by the antioxidant system present in the body. However, environmental factors such as various kinds of soot, environmental hormones, alcohol and smoking due to industrialization have increased the amount of active oxygen, so that the antioxidant system in the body can not adequately defend the damage caused by oxidative stress (Gutteridge JMC, Halliwell B (1994) Antioxidants In nutrition, Health, and Disease. Oxford University Press. London, UK.

Reactive oxygen species (ROS) are unstable and highly reactive, damaging polymer proteins and DNA, damaging biological membranes, damaging tissues and organs, causing diseases such as cancer and causing aging Fridovich I (1989) Superoxide dismutase. An adaptation to a paramagnetic gas. J Biol Chem 264 (14): 7761-7764).

Human cells are exposed to oxidative damage every day, and the DNA of each cell in the human body receives about 10,000 oxidative attacks per day. It is reported that some of these damages are recovered, but unrecovered damage accumulates and causes diseases such as aging, cancer, heart disease (Ames BN (1984) Dietary carcinogen and anti-carcinogens. Clin Toxicol 22 (3): 291-301, Block G, Langseth L (1994) Antioxidant vitamins and disease prevention. Food Technol 48 ((7): 80-84).

To remove ROS, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), which are highly effective and economical synthetic antioxidants, have been widely used in the past. However, toxicity of these compounds to human body (2004). Changes in the total polyphenol content and antioxidant activity of Aster scaber thunb extracts with different (Kim, YE, Nahmang, B, 2004) . microwave assisted extraction conditions Korean J Food Preserv 11 (1):. 88-95; Kim TK, Shin HD, Lee YH (2003) Stabilization of polyphenolic antioxidants using inclusion complexation with cyclodextrin and their utilization as the fresh-food preservative Korean J Food Sci Technol 35 (2): 266-272). Studies on the development of new natural antioxidants using safe natural products have been actively conducted.

On the other hand, the color potato (color potato) is developed by complementing the disadvantages of general potato which is difficult to store for a long time because of high moisture content. It is stronger than the general potatoes and is more resistant to pests, has a unique taste and color, . In addition, it is possible to reduce the loss of nutrients due to cooking because it is possible to reproduce as fruits with less arine flavor.

The color of potato tubers is red or purple. It is an anthocyanin which is a water-soluble pigment. It is contained in fruits, vegetables, flowers and leaves, and has various physiological activities (Jang HL, Yoon KY ), Biological activities and total phenolic content of ethanol extracts of white and flesh-colored Solanum tubersum L. potatoes. J Korean Soc Food Sci Nutr 41 (8): 1035-1040). The antioxidant and anticancer mechanisms of anthocyanin are in vivo ( in vivo ) and in vitro ( in vitro) have also been demonstrated experiments have been made many studies based on it (Choi, HD, Lee HC, Kim SS, Kim YS, Lom HT, Ryu GH (2008). Nutrient components and physicochemical propertiesof new domestic potato cultivars. Kor J Food Sci. Technol 40 (4): 382-388, Jeong JC, Chang DG, Yoon YH, Park CS, Kim SY (2006). Effects of Cultural Environments and Nitrogen Fertilization Levels on the Anthocyanin Accumulation of Purple-fleshed Potato. J Bio-Environ 15 (2): 201-210).

On the other hand, in the extraction of a material composed of a structure in which the surface is covered with a sheath such as a color potato, the simple hydrothermal extraction process has difficulty in effectively penetrating the solvent into the texture of the material, However, in this case, the elution of an undesired component is limited in its use as a food or medicinal product. Therefore, there is a need to develop a technology capable of overcoming the disadvantages of the conventional extraction process as described above and enabling efficient dissolution of active ingredients.

Korean Patent Registration No. 10-1162511 (Registration Date: Jun. 28, 2012) discloses a process for extracting colored potato with an alcohol containing acid and concentrating under reduced pressure to obtain an alcohol extract; Concentrating the alcoholic extract, dissolving the alcoholic extract in a mixed solvent of an acid-containing substance and an alcohol, conducting chromatography, and concentrating each solvent fraction to obtain a concentrated liquid; Dissolving an active fraction concentrate in which the active fraction is concentrated in the concentrate into an acid-containing alcohol, and collecting the active fraction by performing chromatography; And isolating the active fraction by chromatography, removing the solvent, and freeze drying to obtain the desired compound. The novel anthocyanin compound and its preparation method are disclosed.

The present invention is to develop and provide a novel method for preparing an antioxidant-containing colorant potato extract capable of overcoming the above-mentioned problems.

In order to accomplish the above object, the present invention provides a method for producing an extract, comprising the steps of: (A) adding a colorant potato to an ultra-high pressure extraction apparatus and extracting it at a pressure of 200 to 400 MPa to obtain an ultra-high pressure extract; And (B) extracting the ultra-high pressure extract with hot water. The present invention also provides a method for preparing an extract of a color potato with antioxidant.

Hereinafter, the method for preparing the colorant potato extract containing the antioxidant of the present invention will be described in detail.

≪ Step (A): Carla little potatoes  Put in an ultra-high pressure extraction device and 200 ~ 400 MPa Pressure extract to obtain an ultra-high pressure extract, < RTI ID = 0.0 >

In this step, the colored potatoes are put into an ultra-high pressure extraction apparatus and extracted at a pressure of 200 to 400 MPa to obtain an ultra-high pressure extract. Preferably, the colored potatoes are placed in an ultra-high pressure extraction apparatus and extracted at a pressure of 200 to 400 MPa for 15 to 30 minutes to obtain an ultra-high pressure extract. When the extract is prepared under the above-described conditions, the antioxidant substance can be further concentrated and extracted from the colored potatoes.

Conventional natural material extraction methods have drawbacks such as low extraction efficiency, high energy consumption, destruction of many useful components due to heat, denaturation of proteins, loss of components, extraction of soluble fractions, and instability of heat. This problem can be solved by using extraction.

Ultra high pressure extraction is a technique for extracting useful components by applying ultra high pressure technology to medicinal crops. It can extract useful components of medicinal crops in a short time, and can obtain a single component with high purity and an extract with almost no impurities. This is because cell membranes are destroyed under ultrahigh pressure and the solvent can penetrate into the cells. As a result, more components are easily eluted out of the cells.

In addition, ultra-high pressure extraction is a non-heating treatment method which does not denature major components in the food to maintain the freshness and can overcome the texture and flavor of the food by the conventional heat treatment.

In addition, ultra-high pressure extraction promotes destruction of hydrophobic bonds or ionic bonds, and acts selectively on macromolecules including hydrophobic bonds rather than substances with small molecular weights, and thus can be used as a process for increasing the yield during extraction.

≪ Step (B): Extracting the ultra-high pressure extract with hot water>

This step is a step of extracting the ultra-high pressure extract obtained in the above step by hot water extraction. Preferably, the ultra-high pressure extract obtained above is subjected to hot water extraction at 50 to 70 DEG C for 20 to 30 hours.

According to the following Experimental Examples, it was found that when the colorant extract of the present invention was prepared in parallel with the ultra-high pressure process, the content of total polyphenols and total flavonoids was high, the extraction yield of useful components was high and the antioxidative effect Electron donating ability, reducing power, and ABTS radical scavenging ability).

According to the present invention, a colorized potato extract with an increased antioxidant can be prepared from a colored potato.

FIG. 1 shows the results of measurement of the electron donating ability of DPPH radicals of small-sized potatoes. 'HPE 15' was extracted by hot water extraction at 60 ° C, 'HPE 15' was extracted by hot water extraction at 60 ° C for 15 minutes, 'HPE30' was extracted by ultra-high pressure extraction for 30 minutes And then extracted with hot water at 60 ° C.
FIG. 2 shows the results of measuring the ABTS radical scavenging ability of the potatoes of the present invention. 'HPE 15' was extracted by hot water extraction at 60 ° C, 'HPE 15' was extracted by hot water extraction at 60 ° C for 15 minutes, 'HPE30' was extracted by ultra-high pressure extraction for 30 minutes And then extracted with hot water at 60 ° C.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Example 1: Preparation of antioxidant-containing colorant potato extract]

The tiny potatoes were supplied from Ryoya Co., Ltd., an agricultural company, and immediately washed and used. 50 g of colored potatoes were placed in a plastic bag together with distilled water so as to prevent air from entering them. Then, an ultra-high pressure extraction apparatus (Ilshin autoclave, Daejeon, Korea), and the pressure of 300 MPa was set to 15 minutes and 30 minutes, respectively, to obtain the respective ultra-high pressure extracts.

Each of the ultra-high pressure extracts obtained above was placed in an extraction flask attached to a vertical reflux condenser, and distilled water (500 ml) corresponding to 10 times the weight of the kidney potato weight was added to the obtained ultra-high pressure extract as an extraction solvent at 60 ° C For 24 hours to obtain an antioxidant-containing colorant potato extract.

Each of the obtained extracts was filtered through a rotary vacuum evaporator N-N series (Berlin, EYELA, Germany), concentrated, and lyophilized to use in the following Experimental Example.

On the other hand, after performing the ultrahigh pressure treatment for 15 minutes, the extract obtained by hot water extraction at 60 ° C was subjected to high pressure extraction (HPE 15) and ultrahigh pressure treatment for 30 minutes, followed by hot water extraction at 60 ° C Was used in the following experiment referred to as " HPE30 (high pressure extraction for 30 min) ".

[ Comparative Example  1: normal A little kid potato Hot water extract  Produce]

50 g of the colored potatoes were placed in an extraction flask attached to a vertical reflux condenser and repeatedly extracted twice at 60 캜 for 12 hours using distilled water (500 ml) 10 times the weight of the sample as an extraction solvent, A potato hot water extract was prepared. The extract was filtered through a rotary vacuum evaporator N-N series (Berlin, EYELA, Germany), concentrated and freeze-dried to be used in the following experiment.

On the other hand, the hot-water extract obtained by hot water extraction at 60 ° C was called "WE (water extraction)" and used in the following experiment.

[ Experimental Example  1: Confirm extraction yield]

In this experimental example, the extraction yield (water soluble solid content versus raw material content) of potato extract was investigated according to the treatment time and treatment time. The same procedure as in Example 1 and Comparative Example 1 was repeated three times, and the average value was shown in the following Table 1 (mean + standard deviation).

Sample menstruum Temperature Ultra high pressure treatment
time Extraction yield
(%, w / w) WE
water

60 ° C - 1.73 0.30 HPE15 15 min 2.10 ± 0.51 HPE30 30 min 2.41 ± 0.22

As a result of the experiment, the yields of the ultra high pressure treated extracts were 2.10% and 2.41%, respectively, and the extraction yields were about 1.4 times higher than those of the hot water extracts. From the above results, it can be confirmed that the ultrahigh pressure treatment increases the extraction yield of color potato.

[ Experimental Example  2: Determination of total phenol and total flavonoid content]

In this experiment, total phenol contents and total flavonoid contents of potato extracts were investigated according to the treatment time and treatment time.

The total phenolic content was determined according to the Folin-Denis method (Gutfinger T 1981). Folin-Ciocalteau reagent and 10% Na ₂ CO ₃ solution were added to 1 ml of each of the extracts obtained in Example 1 and Comparative Example 1 (hot water extraction, 15 minutes of ultra-high pressure treatment, and 30 minutes of ultra high pressure treatment) After 1 ml each, the mixture was allowed to stand at room temperature for 1 hour and absorbance was measured at 700 nm using a spectrophotometer (UV 1600 PC, Shimadzu, Tokyo, Japan). Thereafter, a concentration of 0 to 100 占 퐂 / ml of gallic acid (Sigma Co., St. Louis, Mo., USA) was prepared and analyzed by the same method as the above extracts. From the standard calibration curve, The phenolic content was calculated.

On the other hand, total flavonoid content was determined by adding 0.1 ml of 10% aluminum nitrate, 0.1 ml of 1 M potassium acetate, and 10 ml of ethanol (pH 7.0) to 0.5 ml of each extract (hot water extraction, ethanol), and the mixture was allowed to stand at room temperature for 40 minutes, and the absorbance was measured at 415 nm. Then, the total flavonoid content of each of the above extracts was calculated from a standard calibration curve obtained at a concentration range of 0 to 100 μg / ml using a routine (Rutin, Sigma Co., St. Louis, Mo., USA) MIN, Isla MIN, Sampietro AR, Vattuone MA (2000), Comparison of the Free Radical Scavenging Activity of Propolis from Several Regions of Argentina . J Enthropharmacology 71 (1-2): 109-114).

Total polyphenol and total flavonoid content measurements are shown in Table 2 below.

Sample Total polyphenol content (GAL ¹⁾ mg / g) Total flavonoid content (RE ²⁾ mg / g) WE 48.21 + 1.89 13.12 ± 0.73 HPE15 50.20 + - 0.14 14.35 + 0.17 HPE30 51.34 + - 0.94 15.17 ± 0.28 Values are mean SD Values are mean of triplicates
¹⁾ Gallic acid equivalent
²⁾ Routine equivalent

As a result, total polyphenol content and total flavonoid content of the two high - pressure extract samples were higher than the total polyphenol content and total flavonoid content of the ordinary hot - water extract.

[Experimental Example 3: Measurement of electron donating ability for DPPH radical]

In this experiment measuring the electron donating ability of the DPPH radical of color kid potato extract according to the concentration of the ultra-high pressure treatment or not, and extract the processing time (In vitro) was to.

The electron donating ability (EDA) was measured by the electron donating effect of DPPH (α, α-diphenyl-picrylhydrazyl). DPPH used in the determination of electron donating ability is a stable free radical, a violet compound which exhibits characteristic light absorption near 517 nm. When these radicals come into contact with antioxidant substances, radicals are extinguished by giving electrons, which change color from purple to yellow, and the absorbance decreases.

(0.2, 0.4, 0.6, 0.8, and 1 mg / ml) of each extract obtained in Example 1 and Comparative Example 1 (hot water extraction, 15 minutes of ultrahigh pressure treatment and 30 minutes of ultra high pressure treatment) , And used as a sample.

0.5 ml of 0.5 mM DPPH solution (Abs. EtOH soln.) Was added to 1 ml of ethanol, 10 L of each sample and 990 μl of 100 mM sodium acetate buffer (pH 5.5), and the mixture was stirred. After 5 min of reaction, the concentration of residual radicals was measured at 517 nm using UV spectrophotometer (Lee HH, Lee SY, 2008), Cytotoxic and antioxidant effects of Taraxacum coreanum Nakai. and T. officinale WEB. extracts. Korean J Medicinal Crop Sci 16 (2): 79-85).

 The electron donating ability (EDA) (%) was calculated using the following equation (1).

As: Absorbance of extract-added sphere

Ac: Absorbance of sphere without extract

As a result, the electron donating ability increased as the concentration of the sample increased, and the antioxidant activity of the extract treated with ultra high pressure was higher than that of the hot water extract (FIG. 1).

[ Experimental Example  4: Measurement of reducing power]

In this experiment, the reducing power of potato extract was investigated according to the treatment time, the treatment time and the concentration of extract.

Reducing power refers to the ability to donate electrons to reactive oxygen species and free radicals. The antioxidant activity can be assayed by measuring the reducing power. The stronger the reducing power, the closer the color is to green. Therefore, the higher the antioxidant activity, the higher the absorbance value (Kim JH, Kim JK, Kang WW, Ha YS, Choi SW, Moon KD (2003), Chemical composition and DPPH radical scavenger activity in different sections of safflower. J Korean Soc Food Sci Nutr 32 (5): 733-738).

(0.2, 0.4, 0.6, 0.8, and 1 mg / ml) of each extract obtained in Example 1 and Comparative Example 1 (hot water extraction, 15 minutes of ultrahigh pressure treatment and 30 minutes of ultra high pressure treatment) , And used as a sample.

To 1 ml of each sample was added 200 mM phosphate buffer solution of pH 6.6 and 1% potassium ferricyanide each in the order of 1 ml, and the mixture was stirred and reacted for 20 minutes in a water bath at 50 ° C. 1 ml of 15% trichloroacetic acid (TCA) solution was added thereto, and 1 ml of the supernatant obtained by centrifugation at 12,000 × g for 15 minutes was added with 1 ml of distilled water and 1 ml of ferric chloride. 700 ml of 700 The absorbance was measured in mm. The reducing power of the sample was expressed by the value of the absorbance, and the results are shown in Table 3 below Respectively.

Sample Concentration (mg / ml) 200 400 600 800 1,000 WE 0.09 0.11 0.09 0.22 0.09 ± 0.12 0.10 0.13 0.11 + - 0.32 HPE15 0.10 0.12 0.10 ± 0.53 0.11 0.35 0.14 + 0.75 0.24 ± 0.64 HPE30 0.11 + - 0.12 0.12 + - 0.45 0.12 + - 0.37 0.13 + - 0.54 0.42 + 0.13 Values are mean S. D. Values are mean of triplicates

As a result of the experiment, the ultrahigh pressure extract showed higher reducing power than the ordinary extract. From the above results, it was confirmed that the reducing power is increased when the extract is prepared in parallel with the ultrahigh pressure extraction.

[Experimental Example 5: Measurement of ABTS radical scavenging ability]

In this experimental example, the ABTS radical scavenging ability of potato extract was investigated according to the treatment time, treatment time and extract concentration.

ABTS assay is a method of measuring antioxidant activity by using ABTS ^{+, which} is a peroxide radical produced by the reaction with potassium persulfate, by removing antioxidants and discoloring cyan. In the case of the DPPH assay, the free radicals are cleaved while the ABTS assay utilizes the cleavage of the cation radicals (Que F, Mao L, Zhu C, Xie G (2006) Antioxidant properties of Chineses yellow wine, its concentrate, and volatiles. LWT-Food Sci Technol 39 (2): 111-117).

(0.2, 0.4, 0.6, 0.8, and 1 mg / ml) of each extract obtained in Example 1 and Comparative Example 1 (hot water extraction, 15 minutes of ultrahigh pressure treatment and 30 minutes of ultra high pressure treatment) Respectively.

(ABTS), and 2.6 mM potassium persulphate were mixed and incubated in the dark for one day. The cation radicals (ABTS ⁺ ) And then diluted such that the absorbance at 734 nm was 1.5 or less. 20 μl of each sample was added to 1 ml of diluted ABTS ⁺ solution, and after 30 minutes, the change in absorbance was measured. Antioxidant activity was determined by the percentage of the radical scavenging activity of the control group using dimethyl sulfoxide (DMSO) as a control (Roberta R, Nicoletta P, Anna P, Anath P, RE (1999) Antioxidant activity using an improved ABTS radical cation decolorization assay. Free Radical Bio Med 26 (9-10): 1231-1237). The ABTS radical scavenging activity was calculated using the following equation (2).

A _test : absorbance of experimental group

A _control : absorbance of _control group

As a result, it was confirmed that the antioxidant activity of the ultra high pressure extract was higher than that of the hot water extract (FIG. 2). From the above results, it was confirmed that the antioxidant activity was increased by the combination of the ultra high pressure process.

Claims (3)

(A) a step of putting a small-sized potato into an ultra-high pressure extraction apparatus and extracting it at a pressure of 200 to 400 MPa to obtain an ultra-high pressure extract; And
(B) extracting the ultra-high-pressure extract with hot water.
The method according to claim 1,
The extraction of step (A)
Wherein the antioxidant-containing colorant extract is carried out for 15 to 30 minutes.
The method according to claim 1,
The hot water extraction in the step (B)
Wherein the extract is carried out at 50 to 70 DEG C for 20 to 30 hours.

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