KR101067294B1 - A method for the preparation of high quality corn bran by extrusion and ultrahigh pressure treatments - Google Patents

Abstract

The present invention relates to a method for producing high quality corn bran extract by ultra-high pressure treatment after extrusion molding, in particular the corn bran extract prepared using corn bran obtained in large quantities as a processed corn starch by-products, a large amount of antioxidant and anti-skin aging polyamine conjugate It can be widely used as a functional food or whitening cosmetic raw material for preventing adult diseases and skin aging.

Corn Bran, Extrusion, Ultra High Pressure, Polyamine Conjugate, Antioxidant, Antityrosinase, Whitening Cosmetic

Description

A method for the preparation of high quality corn bran by extrusion and ultrahigh pressure treatments}

The present invention relates to a method for producing high quality corn bran extract by ultra high pressure treatment after extrusion molding. More specifically, the present invention relates to an antioxidant and anti-skin ageing polyamine conjugate [ N, N' -dicumaroylputrisin (DCP), Np -coumaroyl- N' -ferrule by ultra-high pressure treatment after extrusion molding. It is related to the manufacture of high quality corn bran extract containing loyl putricin (CFP) and N, N' -diferuloyl putricin (DFP)] as a functional whitening cosmetic material.

Recently, with the improvement of living standards and rising interest in health, the demand for functional cosmetics for anti-aging is increasing rapidly. The domestic cosmetics market is worth more than W5trn in 2005, of which the cosmetics market is worth W600bn. Since 2000, with the development of IT / BT technology, the past cosmetics technology has been developed from usability to safety and effectiveness, and in the past, the concept of “treatment” has recently been introduced. In particular, anti-aging cosmetics, including wrinkle-improving functional cosmetics, account for 35-40% of domestic functional cosmetics sales of about W550bn as of 2005. News, Korean Cosmetics Sales Status, 2008; Lee Ok-seop, Cosmetic Trends and Forecasts in Korea, Fragrance J. 2, 59-63, 2009).

The market for imported cosmetics in Korea is about 2 trillion won, despite the active development and sales of the cosmetics industry. In 2004, there was a report that cosmetics exports increased, but the total import and export performance of domestic cosmetics until 2000 shows that the proportion of imports is increasing year by year. It is very low at 48.7% compared to developed countries (Health & Industrial News, Korea Cosmetic Sales Status, 2008; Lee Ok-seop, Cosmetic Trends and Forecasts in Korea, Fragrance J. 2, 59-63, 2009). Therefore, the development of new functional cosmetics is expected to contribute greatly to not only import substitution but also export promotion.

Currently, cosmetics classified as functional cosmetics include wrinkle improvement products, whitening products, and sunscreen products. Currently, most of the KFDA's functional ingredients are not used in various forms of cosmetics because they are very unstable raw materials. Therefore, there is a need for developing a new functional cosmetic material derived from a natural cosmetic that has higher efficacy and safety on the skin than the existing functional ingredients and ensures stability in the formulation.

As a raw material for whitening cosmetics, vitamins of tyrosinase inhibitor arbutin, tyrosinase inhibitor, free radical scavenging useful oil licorice extract, and melanin inhibitor vitamin-C derivatives are designated as KFDA's notification ingredients. Many of the ingredients have low stability and are not scientifically confirmed in terms of their efficacy, and more powerful whitening agents such as kojic acid and hydroquinone are problematic. Therefore, there is a need to develop new materials of functional whitening cosmetics derived from plants that are more safe and effective.

Of naturally occurring polyphenolic compound hydroxycinnamic acid derivative, i.e., para-Kumar acid (p -coumaric acid, CA), ferulic acid (ferulic acid, FA), caffeic acid (caffeic acid) is hanggan toxic anti-cancer, anti-inflammatory, It has various biological activities such as antibacterial, antimutagenic and antioxidant activity. In particular, N, N' -dicumaroylputrycin (DCP), Np -coumaroyl- N' -feruloylputrycin (CFP) and N, N'- diferulo among some hydroxycinnamic acid derivatives Polyamine conjugates, such as ylputrisin (DFP), have recently been shown to have tyrosinase inhibitory activity as well as strong antioxidant activity as a radical scavenger against reactive oxygen species (Choi et al., J Agric Food Chem). 23. 1090-1092, 2007). Therefore, there is a need for the search for plants containing a large amount of these polyamine conjugates and the development of technology capable of extracting them effectively.

Phenolic acid in corn is present in free, ester, water-soluble polyamine and non-aqueous forms, and almost all of it is occupied by water-insoluble phenolic acid combined with dietary fiber in cell walls. (Sosulski F et al., J Agric Food Chem 30, 337- 340, 1982.) Ferulic acid in corn polyphenols is not only a bioactive substance with anticancer, antihypertensive, antidiabetic, anti-inflammatory and antioxidant activities, but also a natural flavor component of vanillin biosynthesis. It is widely known as a precursor (Graf, E. Free Rad. Biol. Med . 13 , 435-448, 1992; Saulnier L et al., Carbohydrate Polymers, 45, 269-275, 2001). In addition to the development of several processing technologies for the conversion of ferulic acid combined with arybinoxylan dietary fiber components to the free form, research on the development of enzyme treatment methods has been actively conducted. While mass production of ferulic acid has been actively carried out along with the development of functional materials using the same, the development of polyamine conjugates of corn, which has recently been spotlighted as an antioxidant and anti-aging bioactive component, along with the development of mass production technology of corn There is little development of cosmetics.

On the other hand, corn bran obtained as a by-product from the wet-milling corn starch processing process is known to contain a large amount of polyamine conjugates (Moreau RA et al., Lipids 36 , 839-844). , 2002). However, these polyamine conjugate components are not easy to extract because they are combined with a large amount of dietary fiber present in corn bran, and new extraction technology that can replace the heat extraction method because the stability of heat is different depending on the type of ingredients. Need development

Is accompanied by physical and chemical changes in the food component by the heater energy supplied extrusion process (extrusion process) is from a heater mounted separately from the motor energy which is supplied by a screw rotation of the extruder (Chung MY, Lee SJ. Korean J Food Sci Technol 29: 65-71, 19978). Extrusion molding technology is an efficient and economical process compared to other heat treatment processes because the unit operation such as mixing, grinding, heating, molding and drying occurs in a short time during the food processing process (Eise K. Plastic Compounding 9: 44- 47, 1986). In addition, the extrusion molding process can produce a processed product having a variety of properties because it can adjust the properties of the desired product according to the raw material input speed, moisture content, screw rotation speed, structure of the injection port, screw arrangement (Harper JM. Food extruders and their application.In: Extrusion cooking.Merier C, Kinko P, Harper JM. (Eds.), AACC St. Paul MN, USA pp. 1-18, 1989; Linko P. Extrusion cooking in bioconversion. Extrusion cooking.mercier C, Linko P, Harper JM, eds.American Association of Cereal Chemists, Inc., St. Paul.p 235-245, 1989).

Up to now, many studies on the improvement of the processing process have been reported along with the development of high quality raw materials and various processed foods using the extrusion molding process. Studies on the liquefaction of corn starch with high efficiency using the extruder as an enzyme reactor (Roussel et al., Lebensm-Wiss u.-Technol 24: 449-458, 1991) and the effect of ethanol fermentation of sorghum using extruder (Zhan et al., Industrial Crops Products 23: 304-310, 2006). In addition, mechanical and thermal energy inputs during extrusion are known to change the physical properties of the extruded food to increase water solubility or to cause hydrolysis of starch during extrusion of cereal starches or grits (Chiang BY). & Johnson JA. Cereal Chem 54: 436-443, 1977).

On the other hand, in Korea, a report on the molecular structure change and alcohol fermentation efficiency of flour starch through extrusion molding (Lee et al., Korean J Food Sci Technol 23: 683-688, 1991) and recently, the functionalities of the extruded ginseng It was reported that the content of ingredients increased (Ha et al., Food Sci Biotechnol 14: 363-367, 2005). Also, when the extruding process was used for pretreatment of ginseng starch, the liquefaction and saccharification rate of starch by enzyme treatment increased. (Han et al., J Food Sci Nutr 11: 318-322, 2006). In addition, extrusion treatment increases the solubility of skim bean powder by enzymatic hydrolysis (Cha et al., Food Sci Biotechnol 16: 543-548, 2007) and increases the yield of levulinic acid from starch. Sikkim was reported (Cha et al., Ind Crop Prod 16: 109-118, 2002). In this way, the extrusion molding process can improve the solubility, digestibility, functionality, and palatability of food extracts along with the physicochemical changes of the food, thereby developing various food products as well as developing superior food materials. However, there have been few studies on the extraction of bioactive substances from corn bran using extrusion molding.

Ultrahigh pressure extraction technology is an extraction method that promotes decomposition of plant cell walls and membranes and induces modification of useful functional materials under ultra-high pressure (500-1,000 atm). It is done. In particular, the ultra-high pressure extraction technology facilitates the extraction of heat-stable bioactive substances present in plants, and minimizes the destruction of nutrients, tastes, and flavors of food during extraction, increases palatability, as well as new functional oligosaccharides, peptides, amino acids and fats. Is a new food processing technology that can prevent the production, discoloration and discoloration of food under oxygen and high temperature (Kim Do-yeon et al., Korean Society of Food Science and Technology, poster presentation, 2008). However, studies on the production of useful physiologically active substances from plants using ultra-high pressure extractors have been insufficient. By performing the ultra-high pressure extraction at the same time after the extrusion molding process, it is thought that the production of high quality plant extracts containing the plant-derived bioactive substances is possible.

Accordingly, an object of the present invention is to provide a method for producing high quality corn bran extract containing antioxidant and anti-skin aging polyamine conjugate by ultra high pressure extraction after extrusion molding treatment.

Another object of the present invention is to measure the scavenging activity and the radical scavenging activity and the whitening activity of polyamine conjugate of corn bran extract obtained by ultra high pressure extraction after extrusion molding treatment.

Another object of the present invention is to provide a use of corn bran extract.

The purpose of the present invention as described above is to develop a new functional material from corn bran obtained in large quantities as a corn starch processing by-product by wet-milling, anti-oxidation and anti-skin aging three polyamine conjugates by ultra-high pressure extraction after extrusion molding High-quality corn bran extract containing gates (DCP, CFP, DFP) was prepared by solvent fractionation and adsorption resin chromatography, and the antioxidant activity was measured by measuring the scavenging activity against DPPH radicals and inhibition of tyrosinase. This was accomplished by checking the activity to confirm the whitening activity.

One feature of the invention is characterized by providing a method for producing a high quality corn bran extract containing polyamine conjugates by ultra high pressure extraction after extrusion molding comprising the following steps:

First step of obtaining corn ethanol extract by putting corn bran into an extruder, and extruding and drying it, followed by adding 80% ethanol aqueous solution to the extractor, extracting continuously in an ultra-high pressure extractor, and filtering and concentrating.

A second step of dissolving the crude ethanol extract in an aqueous 80% ethanol solution and then overnight in a refrigerator and then centrifuging the supernatant obtained by partial concentration under reduced pressure, suspending with water, and then dichloromethane to fractionate it;

The dichloromethane fraction was again dissolved in dichloromethane (see FIG. 1) or in normal-hexane-ethylacetate-methanol-water (200: 200: 100: 1, v / v) mixed solvent and then overnight in the refrigerator. A third step of preparing a white powder by collecting and drying the lower layer which is left to settle separately;

The white powder obtained in the third step was again dissolved in 80% ethanol, diluted with water and previously equilibrated with 40% ethanol. Diaion HP-20 adsorption resin (Mitsubishi, Japan) cartridge (20 cm x 65 cm ) Was washed with 40% ethanol using a large purification system (Biotage, USA) to elute and remove the pigment and low molecular weight para-coumaric acid and ferulic acid, and then remove 60% ethanol solution. It consists of a fourth step of producing a polyamine conjugate-containing corn extract by eluting three polyamine conjugates (DCP, CFP, DFP) adsorbed by passing through under reduced pressure.

Another feature of the invention is characterized in that the corn bran extract is provided as a cosmetic or food composition for whitening.

The present invention relates to a method for producing an antioxidant and anti-ageing polyamine conjugate-containing corn bran extract by ultra high pressure extraction after extrusion molding, and its use, and has a strong antioxidant activity against DPPH radicals and a tyrosinase inhibitory activity, and thus whitening is strong. There is an excellent effect of providing corn bran extract that works. Therefore, the corn bran extract of the present invention contains a large amount of antioxidant and anti-ageing polyamine conjugate, so it can be utilized as a new antioxidant material in food and cosmetics is a very useful invention in the food or cosmetics industry.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example 1: Preparation of high quality corn bran extract by ultra high pressure extraction after extrusion molding

After extrusion molding, the preparation of high quality corn bran extract containing three polyamine conjugates with anti-oxidation and anti-aging properties was performed as shown in FIG. 1 by ultra high pressure extraction. The used corn bran was supplied in February 2009 from Korean grains in Icheon, Gyeonggi-do.

Specifically, corn bran (10 kg) is first put into a twin-screw extruder (FX-6, milling industry, Korea) extrusion processing (screw length 50 cm, diameter 4.5 cm, pot temperature 65, 130, and 140 ℃, Water supply rate 100 ml / min, sample feed rate 0.2 kg / min, injection diameter 5 mm), dried and then extracted repeatedly with 80% ethanol using an ultrahigh pressure extractor (ITC20, Korea), filtered and concentrated to extract corn bran ethanol extract (2.1 kg) was obtained.

The ethanol extract was again dissolved in 80% ethanol, left in the refrigerator overnight, and the supernatant obtained by centrifugation at 3,000 rpm for 3 minutes was partially concentrated under reduced pressure, suspended in water (10 L), and then diluted with dichloromethane (20 L) was added twice, and the lower layer was concentrated separately to obtain a dichloromethanol fraction (53 g).

The dichloromethanol fraction was dissolved in 80% ethanol, diluted with water and equilibrated to 40% in advance. A large purification system equipped with a DIION HP-20 adsorption resin (Mitsubishi, Japan) cartridge (20 cm x 65 cm) , USA) and washed with 40% ethanol solution (10 L) to elute and remove the pigment and low molecular weight free phenolic acid (para-coumaric acid and ferulic acid) and then 60% ethanol solution (20 L) again Three polyamine conjugates (DCP, CFP, DFP) were eluted and concentrated under reduced pressure to give a final corn bran extract (25 g, yield: 0.25%).

Example 2: Determination of Three Polyamine Conjugate Contents of Corn Bran Extract by HPLC

The content of the three polyamine conjugates of the final corn bran extract was measured as follows according to the best method (Cho et al., Food Sci & Biotechnol, 2009, submitted). 200 mL of 80% ethanol aqueous solution was added to corn bran extract powder (10 g), dissolved, filtered and concentrated under reduced pressure to remove ethanol, and then, CH ₂ Cl ₂ (400 mL) was added twice and fractionated. The lower layer was obtained and concentrated under reduced pressure to obtain a CH ₂ Cl ₂ fraction.

The extract was dissolved in 10 mL of 80% ethanol and passed through a Sep-pak C18 cartridge (Waters, USA) to remove impurities and then diluted appropriately and analyzed by HPLC. The three polyamine conjugates were identified by comparison with the holding time of standard polyamine conjugates isolated in a previous study (Choi et al., J Agric Food Chem, 23, 1090-1092, 2007), and the calibration curve of each compound ( The calibration curve) was prepared using a regression analysis graph and then the content of each polyamine conjugate was calculated.

TABLE 1

Contents of Three Polyamine Conjugates in Corn Bran Extract

Corn bran
Production time Corn bran
extract Polyamine Conjugate Content (%, Corn Bran Extract) DCP CFP DFP 2008. Sep Sample 1 1.2 ± 0.2 3.4 ± 0.4 50.6 ± 1.3 December 2008 Sample 2 1.5 ± 0.3 3.8 ± 0.5 53.5 ± 2.0 February 2009 Sample 3 1.7 ± 0.4 3.6 ± 0.4 55.3 ± 2.9 * Each value represents the mean ± standard deviation of three repeated measurements.

As shown in Table 1, the content of three polyamine conjugates of corn bran extract produced according to the corn bran produced as shown in Table 1, the corn bran extract produced from corn bran produced in September 2008 was DCP (1.2%) , CFP (3.4%), and DFP (50.6%), DCP (1.5%), CFP (3.8%), and DFP (53.5%) in December 2008, and finally DCP (1.7) in February 2009. %), CFP (3.6%), and DFP (55.3%). As such, the polyamine conjugate content of corn bran extract was slightly different according to the time of corn bran production, but generally showed the contents of DCP (1.5%), CFP (3.6%), and DFP (53%).

Example 3: Measurement of Antioxidant Activity of Corn Bran Extract

Antioxidant activity of corn bran extract was measured by DPPH radical scavenging activity, and DPPH radical scavenging activity was performed as follows according to the method of Tagashira and Ohtake. 200 ml of the sample solution (each compound dissolved in ethanol and properly diluted in 100 mg / ml) was added 4 ml of ethanol solution containing 100 μM of DPPH and mixed vigorously for 10 minutes at room temperature. The absorbance was measured to determine the decrease in absorbance due to the reduction of DPPH. DPPH radical scavenging activity was calculated according to the following equation.

DPPH radical scavenging activity (%) = (1-A / B) × 100

A: absorbance of the sample at 517 nm, B: absorbance of the control at 517 nm.

Here, the control without the sample was measured together, and the relative DPPH radical scavenging activity of the sample was measured, and then the IC ₅₀ value (concentration of the sample inhibiting 50% of the DPPH radical) was calculated by regression analysis.

% Inhibition = (1-A / B) x 100A: absorbance of the sample at 517 nm, B: control of the control at 517 nm

Absorbance

TABLE 2

Comparison of DPPH Radical Scavenging Activity between Corn Bran Extract and Other Natural Antioxidants

sample DPPH radical scavenging activity (IC ₅₀ , ㎍ / mL) Corn Bran Extract 4.5 ± 0.3 L-ascorbic acid 6.4 ± 0.7 α-tocopherol 22.5 ± 1.0 * Each value represents the mean ± standard deviation of three repeated measurements. The values in each row differ significantly at p <0.05.

As shown in Table 2, corn extract (IC ₅₀ = 4.5 μg / ml) was stronger than control L-ascorbic acid (IC ₅₀ = 6.4 μg / ml) and α-tocopherol (IC ₅₀ = 22.5 μg / ml). It showed a radical trapping activity, and thus, corn bran extract can be widely used as a functional antioxidant raw material as well as a functional food as a natural antioxidant material.

Example 4: Investigation of the whitening effect of corn bran extract

The inhibitory effect of corn bran extract on tyrosinase of mushrooms was measured using L-tyrosine and L-DOPA as substrates, and their IC ₅₀ values are shown in Table 3.

Tyrosinase activity was slightly modified by Masamoto et al. (Masamoto et al., Biosci. Biotechnol. Biochem. 67, 631-634, 2003) using L-tyrosine and L-DOPA as substrates. Used.

When using L-tyrosine as the substrate, 0.5 ml of 0.1 M phosphate buffer (pH 6.8), 0.45 ml of deionized water, 0.05 ml of DMSO concentration sample solution, and 0.05 ml of tyrosinase (2,000 U / ml) The reaction mixture (1.55 mL) was preincubated at 25 ° C. for 5 minutes, and then 0.5 mL of 2.5 mM L-tyrosine was added. The reaction mixture was preincubated in a 37 ° C. water bath for 5 minutes and then left for 5 minutes on cold ice to terminate the reaction. The reaction was monitored for 2 minutes at 475 nm. The reaction without sample was used as a control. Absorbance was measured at 475 nm with a UV spectrophotometer (Jasco, Japan). Tyrosinase inhibition was calculated as follows:

% Inhibition = (A-B) / A × 100

A and B represent the absorbances of the sample solution and the control, respectively.

When using L-DOPA as the substrate, a reaction mixture containing 1 ml of 1.5 mM L-DOPA solution, 0.1 ml of DMSO with or without sample and 1.8 ml of 1 / 15M phosphate buffer (pH 6.8) (3 Ml) was incubated at 25 ° C. for 10 minutes. As soon as 0.1 ml of 1,000 units / ml tyrosinase aqueous solution was added to the reaction mixture, the initial rate of linear increase in absorbance at 475 nm was measured immediately. The reaction without DMSO was used as a control. Tyrosinase inhibition was calculated as above, and L-ascorbic acid and kojic acid were used as positive control standards.

TABLE 3

Inhibitory Activity of Corn Bran Extract and Standard on Mushroom Tyrosinase (IC ₅₀ )

compound Inhibitory activity (IC ₅₀ , μg / mL) Tyrosinase
(L-Tyrosine) Tyrosinase
(L-DOPA) Corn Bran Extract 26.5 ± 1.5 2.9 ± 0.2 L-ascorbic acid 58.4 ± 2.7 - Kojisan 1.1 ± 0.2 * IC ₅₀ represents the concentration of compound required to inhibit the activity of mushroom tyrosinase by 50%.
* Experimental results were measured three times and expressed as mean ± standard deviation.
* The mean value of each sample is significantly different in the p <0.05 range.
L-ascorbic acid and kojic acid were used as positive controls.

As shown in Table 3, when L-tyrosine was used as the substrate, corn bran extract (IC ₅₀ = 26.5 μg / mL) showed stronger tyrosinase inhibition than the control L-ascovinic acid (IC ₅₀ = 58.4 μg / mL). Activity was shown.

In contrast, when L-DOPA was used as a substrate, corn bran extract (IC ₅₀ = 2.9 μg / mL) had slightly lower tyrosinase inhibitory activity than control kojic acid (IC ₅₀ = 1.1 μg / mL). These results suggest that corn bran extract strongly inhibits the pathway of L-tyrosine to dopa-quinone in tyrosinase-catalyzed reactions.

The tyrosinase inhibitory effect of corn bran extract is thought to be due to the three polyamine conjugates contained therein (Choi et al., J Agric Food Chem, 23, 1090-1092, 2007), and from the present corn bran extract The three polyamine conjugates are isolated to measure the tyrosinase inhibitory effects of these compounds at the molecular level, along with molecular modifications to increase their solubility and functionality.

These results indicate that corn bran extract prepared by ultra-high pressure extraction after extrusion molding inhibits lipid peroxidation induced by free radicals and prevents various adult diseases such as cancer, hypertension, diabetes and dementia, as well as tyrosinase. It is suggested that it will be useful as a skin whitening prevention cosmetics such as blemishes and freckles caused by.

Figure 1 shows the manufacturing process of high quality corn bran extract containing antioxidant and anti-ageing polyamine conjugate by ultra high pressure treatment after extrusion molding.

Claims (4)

Corn bran powder was extruded and dried; First step of obtaining crude ethanol extract by adding 80% ethanol, ultra high pressure extraction, and then filtrating and concentrating; A second step of dissolving the crude ethanol extract in 80% ethanol solution and then overnight in a refrigerator and then centrifuging the supernatant obtained under reduced pressure, suspending the extract with water and then fractionating it with dichloromethane; A third step of dissolving the dichloromethane fraction again with dichloromethane and leaving it in a refrigerator overnight to dry powder the precipitated lower layer; The dichloromethane extracted powder was again dissolved in 80% ethanol solution, suspended in water, equilibrated with 40% ethanol, and subjected to DIION HP-20 adsorption resin column chromatography to dissolve the pigment and paramolar acid and ferulic acid having low molecular weight. After removing, passing the 60% ethanol solution again to elute the 60% ethanol fraction containing the polyamine conjugate, and then concentrated under reduced pressure. Corn bran extract manufacturing method characterized in that consisting of. delete Cosmetic composition for antioxidant and skin aging functional whitening containing corn bran extract prepared according to the method of claim 1 as an active ingredient. Antioxidant functional food composition containing corn bran extract prepared according to the method of claim 1 as an active ingredient.

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