KR20130097589A - Extraction of natural pigment using sorghum by-product and there of composition - Google Patents

Abstract

PURPOSE: A natural pigment using a sorghum byproduct contains a functional active component such as anti-oxidation, anti-inflammation, and antibacterial and maintain inherent colors for a long time and having excellent storage stability according to pH conditions. CONSTITUTION: A natural pigment extract of a sorghum byproduct with anti-oxidation and antibacterial activity is obtained by pulverizing the sorghum byproduct and extracting the pulverized material of the sorghum byproduct by a lower alcohol under the presence of an acid. The lower alcohol is methanol or ethanol. The byproduct is a leaf or a stem of sorghum. The acid is selected from hydrochloric acid, the citric acid, and the acetic acid, and malic acid. An anti-oxidation and antibiotic cosmetic composition, a pharmaceutical composition, and food additives contain the natural pigment extract as an active ingredient. [Reference numerals] (AA) Extract amount mg/100g; (BB) Natural pigment content [mg/100g]; (CC,DD) Ethanol ratio

Description

Extraction of natural pigment using sorghum by-products and its composition {Extraction of natural pigment using sorghum by-product and there of composition}

The present invention relates to a method for extracting natural pigments from stems and leaves which are sorghum by-products, and to a health functional composition including a composition extracted from sorghum by-products, optimal pigment extraction conditions, a method of stabilizing pigments, and a food additive.

Sorghum bicolor Moench is a major grain food crop. Sorghum is rich in plant compounds such as polyphenols, tannins, phenols, anthocyanins, phytosterols, and policosanols, which are known to significantly affect human health (Awika & Rooney, 2004, Phytochemistry, 65, 1199-1221). ). Recent studies have suggested that antioxidant activity of sorghum (Choi et al., 2006, Food Chemistry, 103, 130-138), anticancer effect (Kwak et al., 2004, Journal of Korean Society Food Science Nutrition, 33, 921-929) , And has a hypocholesterolemic effect (Ha et al., 1998, Korean Journal of Food Science and Technology, 30, 224-229) and have been reported to reduce cardiovascular disease (Cho et al., 2000, Journal of the Federation of American Societies for Experimental Biology, 14, A249). Moreover, sorghum has been reported to have DPPH radical scavenging activity and antimutagenic effect (Kwak et al., 2004, Journal of Korean Society Food Science Nutrition, 33, 921-929). Flavonoids, a polyphenolic compound extracted from sorghum, are found in most of the flavonoids in higher plants and vascular plants and are found in all fruits, vegetables, and wines, especially red wines and teas. Flavonoids are naturally present primarily in glycosidic form, but may be present in phenol and sulfate free form, so-called aglycon form, or in bound form to carbohydrates or proteins. In most cases flavonoids have a chemical structure of polyphenolic compounds. More than 8,000 flavonoids have been identified in plants, and they have many functions.

High amount (sorghum) pigment, which is a non-tartar-based natural coloring material, is a pigment obtained by extracting fruit of sorghum with water or ethanol. The main pigment is apigenin of flavonoids (Apigenin, C15H10O5).

High pigments are soluble in water, hydrous ethanol, propylene glycol and alkaline solutions but not in fats and oils. May not dissolve in acid and pigment aggregation occurs around pH 4.0. It is stable to heat and light and has excellent dyeing resistance to proteins. Liquid high pigments contain ethanol and the like for stabilization. It is used as a coloring material for livestock processed products, plant proteins, and fish processed products by using excellent dyeing property. Diluent or stabilizer and solvent can be added for color value adjustment and quality preservation. It is also used in beverages, confectionery, baking, and livestock products.

Korean Patent Laid-Open Publication No. 10-2010-0122543 discloses a volume of water that is about 2 to 15 times, preferably about 5 to 10 times the weight of a sorghum sample, after grinding the seeds of sorghum, such as methanol, ethanol, butanol, and the like. A polar solvent of C1 to C4 lower alcohol or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably about 70 to 100% of methanol for about 1 hour to 1 day, more preferably 100 By using extraction methods such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for about 1 day with% methanol, preferably hot water extraction, followed by filtration and concentration to obtain a sorghum extract.

Korean Patent No. 10-0813638 discloses a concentrate of ethyl acetate layer having the highest activity among extracts extracted from Artemisia Herba of Kang Hwa methanol and extracted in the order of hexene, ethyl acetate, butanol and water in order to reverse phase silica. After filling, the mixture was eluted with chloroform / methanol = 50/1 to 1/1 (v / v) to extract an apigenine compound.

Korean Patent No. 10-0625469 is a step of preparing a sweet potato stem sample, the reaction of the sample and sulfuric acid solution to collect the filtrate by filtering with a glass filter, the filtrate is concentrated and the concentrated filtrate Separation of phenolic compounds by reaction with three times more ethanol and centrifugation, and separation and concentration of ethanol solubles in the upper part of the centrifugation, mixing the separated phenolic compounds consisting of hexane, ethyl acetate, and formic acid After dissolving in a solvent, the solvent soluble portion was extracted with a fraction containing a ferric acid (ferulic acid) using a silica gel column to extract the perric acid.

Most natural pigments extracted from sorghum grains have physiological activities, including antioxidant activity, and also contain large amounts of health functional ingredients. On the other hand, in the case of pigments extracted from sorghum grains, the pigment content is relatively low, and the pigment tends to lose its own color gradually according to the storage period, which is a significant limiting factor in the use of natural pigments. Natural pigments were separated from stems and leaves, which are by-products of sorghum, which have a much higher yield per unit area than sorghum grains.

Accordingly, an object of the present invention is to optimize the extraction of natural pigments from sorghum by-products, and to stabilize the extracted natural pigments and to test the active ingredients, thereby making it possible to use them in a wide range of fields such as health functional compositions and food additives.

 The present invention features a method for extracting natural pigments containing a large amount from leaves and stems, which have not been tried at all, and a functional composition containing the extract. In particular, a method for extracting a phenolic compound containing a pigment from the by-products of golden wax sorghum and a functional composition derived therefrom is characterized by the above-mentioned.

In addition, the present invention is characterized by a health functional composition containing an extract extracted from sorghum by-products, food additives, natural pigments and natural pigment stabilization method.

The present invention relates to a method of extracting natural pigments from sorghum by-products and stabilization of the extracted natural pigments, and extracting a large amount of natural pigments and phenolic compounds from the by-products of golden wax or myelo sorghum by the extraction method of By providing a functional composition containing health functional active ingredients such as anti-inflammatory and antimicrobial properties, and maintaining the storage stability of the extracted natural pigments, the color stability according to the pH conditions in various fields such as functional foods, cosmetics, food additives It can be used.

Figure 1 shows a comparison of the solvent and solvent content for the maximum extraction of sorghum pigment.
Figure 2 shows the analysis of extraction amount of sorghum pigment according to the type and concentration of acid.
Figure 3 shows the analysis of the amount of hydrogen pigment extraction according to the extraction time.
Figure 4 shows the pigment comparison according to the millet crushed particle size.
Figure 5 shows the cytotoxicity evaluation of sorghum by-product extract.
Figure 6 shows the antioxidant (NBT) evaluation of sorghum by-product extract.
Figure 7 shows the anti-inflammatory evaluation of sorghum by-product extract.

The present invention relates to an extraction method and a composition containing a maximum amount of natural pigments from stems and leaves which are by-products of sorghum, specifically, the conditions under which the polyphenol component containing the natural pigments is produced at the maximum when the extraction conditions are different. To identify and to identify the pigments, and to compare and analyze the health functional physiological activity of the pigments, to efficiently extract extracts showing superior activity compared to the crude extract containing the conventional sorghum-derived pigments.

After all, the present invention is derived from the sorghum by-products to extract the maximum amount of natural pigments, characterized in that the utilization of the extract and the light and heat stable pigment composition to the health functional foods and cosmetics can be efficiently developed.

After milling the sorghum by-products can be extracted by conventional methods using common extraction solvents well known in the art. As the extraction solvent, water, C ₁ ~ C ₄ alcohol, ethyl acetate, acetone or a mixed solvent thereof may be used, and preferably water is used, but is not limited thereto. The amount of the extraction solvent is preferably 1 to 50 times by weight of the by-product, more preferably 5 to 30 times, but is not limited thereto. The extraction method may be an extraction method such as room temperature extraction, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction, preferably ultrasonic extraction is not limited thereto.

The extract may include an acceptable carrier, excipient or diluent for the development of health functional foods and cosmetics, food additives, and products using a stable purple pigment.

In particular, the composition of the present invention can be ingested in the form of the extracted powder and processed products using the extract, the extract can be added as it is or used as a health food with other foods or food ingredients, and can be suitably used according to a conventional method. Can be. The blending amount of the active ingredient may be suitably determined according to the purpose of use, namely beauty, prevention, health, additives or other pigment utilization products.

Pharmaceutical compositions comprising natural pigment extracts extracted from sorghum by-products according to the present invention, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc. It may be used in the form of a sterile injectable solution, but is not limited thereto and may be appropriately adjusted depending on the method of use and use. Carriers, excipients and diluents that may be included in the pharmaceutical composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate , Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

Foods to which the natural pigment extract extracted from the sorghum dispersion can be added include drink, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, alcoholic beverage and Vitamin complexes, and the like, but is not limited thereto. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5g, preferably 0.3 to 1g based on 100ml It is good to adjust according to the type of food and the method of use.

You may mix | blend the cosmetics of this invention with the component of the natural pigment extract extracted from the said aqueous dispersion, the other component normally mix | blended with cosmetics as needed. The composition of the present invention comprises the extract and fractions in an amount of 0.01 to 10% by weight, preferably 0.1 to 8% by weight, most preferably 0.5 to 5% by weight, based on the total weight of the composition.

Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cooling agents, restriction agents, purified water and the like. The functional cosmetic composition for whitening of the present invention may be prepared in any formulation commonly prepared in the art, and includes, for example, emulsion, cream, lotion, pack, foundation, lotion, essence, hair cosmetic, and the like.

Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited thereto.

Example  1 Extraction of natural pigments from sorghum by-products

As a result of measuring the content of the total phenolic compound containing the pigment from the sorghum by-products used in the present invention, as shown in Table 1, the content of the phenolic compound of the seed was 15.0 mg / g Although it contained less than 19.1mg, the stem contained 25.2mg of more than 20.6mg of myo sorghum, and 23.3mg of leaf also contained more than 13.1mg of myosu. In the color comparison of the extract, as shown in Table 2, the color of the pigment extracted from the golden waxy corn leaf was the darkest, followed by the stem and the seed.

Golden Swordsman Milosu Kinds Phenolic Compound (mg / g) Kinds Phenolic Compound (mg / g) strain 15.9 strain 19.1 stem 25.2 stem 20.6 leaf 23.3 leaf 13.1

kind Golden Swordsman Milosu Colors of seeds, stems and leaves

As shown in Table 3, in the measurement of the color change according to the pH of the by-product extract, the color was gradually increased according to the increase in pH (alkaline) in the case of the pigment extracted from the seeds of golden ryegrass and milky lotus seeds, No significant color change was observed at the deepest point, but the stems showed the above phenomenon.

pH Sorghum by-product Golden Swordsman Milosu pH 3

pH 7

pH 10

In experiments to determine the degree of destruction of the pigments due to long-term storage at room temperature, the sorghum pigments were confirmed to have excellent stability of the pigments compared to the anthocyanin pigments of colored rice and red beans, as shown in Table 4. In the comparison after three months, pigmentation of pigment and red bean showed destruction of pigment, and intrinsic color was changed into brown system. In case of sorghum, however, no noticeable change in color was observed to a slight degree.

The color difference of the pigment extracted from the sorghum by-products was measured by Lab color and CMYK color using the Minolta CM-3500 as shown in Table 5. The pigments extracted from the leaves showed similarity in the stem pigments and brightness, and the stems were higher in the reddish color. This phenomenon was confirmed in the case of Misusu.

Breed name part Extracted
Color of pigment KS system color Color L * a * b * C M Y K Golden Sword
Sorghum leaf

Dark brown 20.15 8.93 6.84 0 32 40 75 stem

dark
maroon 20.10 18.41 10.83 0 67 62 66 Milo leaf

Brown 40.52 28.83 35.12 0 51 75 41 stem

bright
Red 50.61 65.8 48.01 0 78 81 11

Ethanol, methanol, and water were used as solvents to extract natural pigments from sorghum to the maximum, and the extraction amounts according to the ratios are as shown in FIG. In the type of solvent, methanol extract amount was better than ethanol, and in the content of ethanol and methanol, the highest amount of pigment was extracted in 100%. In the pigment composition, the extraction of epigeninidine, which is the main component of sorghum pigment, was the highest in 100% of solvent, followed by luteolinidine, apigenin, and luteolin.

In the experiment for extracting the pigment from the sorghum to the maximum, the pigment extraction amount analysis according to the acid type such as hydrochloric acid, citric acid, acetic acid, malic acid, etc. shows that the pigment is extracted from the sorghum at maximum concentration of 0.5% or more as shown in FIG. Confirmed.

In the analysis of extraction amount of sorghum according to the extraction time, the difference in the amount of pigment extraction was not observed in more than 24 hours of extraction experiments from 12 hours to 72 hours. It was confirmed.

As a result of comparing the natural pigment content of each sorghum cultivar and each part, as shown in Table 6, 24.5 mg of white sorghum and 13.8 mg of golden wax sorghum were 34.5 mg / 100 g compared to white sorghum and milo sorghum. The total amount of natural pigments was higher than that of Milo sorghum and the major pigment component of golden wax sorghum was significantly higher in epigenidine than in the other two varieties. In the case of white waxed sorghum, epigenin was the highest. In addition, it was confirmed that the pigment content of the stem and leaves, which are by-products, was 1,881 mg / 100 g in the total amount of the stem and the golden sorghum stem was much higher than the seed, and the sorghum was 957 mg / 100 g in the total amount. It was confirmed that there is. In the case of the leaves, the golden tender sorghum tended to be similar to the stem, and the content of the half was 895mg / 100g. It was.

Breed name Natural pigment type (mg / 100g, in building) Sum Luteol
Linidine Epigae
Ninidine Luteolin Epigenin Perulic acid Qumaric acid Golden Swordsman 1.46 30.54 0.94 1.09 0.47 - 34.5 Milosu 1.34 2.63 1.46 3.51 0.80 4.04 13.78 Golden Stalker (Stem) 7.62 1356.85 31.68 355.32 10.73 119.43 1,881.63 Milosu (stem) 24.60 310.14 233.19 185.27 4.16 200.38 957.74 Golden Waxy Sorghum (Leaf) 1.68 758.31 10.07 83.21 4.72 37.44 895.43 Milo sorghum (leaves) 2.39 32.57 25.69 11.68 3.42 32.41 108.16

The pulverized particles were separated by size for extracting pigment from sorghum, and the results of pigment extraction were as follows. Among them, the most extracted was 18.1, and among the smallest particles, between 53 and 200, 17.0 mg was extracted. Comprehensive extraction amount according to the particle size was confirmed that the maximum extraction of the sorghum when pulverized to less than 300um size. Figure 4 shows the result of extracting the actual pigments for each size of the pulverized particles in a photograph.

Crushed particles
size
(탆) Pigment Content (mg) 4-Hydoxybenzoic acid Coumaric acid Ferulic acid Luteolinidin Apigeninidin Luteolin Apigenin system 840 or more 0.014 0.877 0.403 1.34 3.908 0.368 0.838 7.7 300 to 840 0.072 2.14 0.475 1.843 7.22 0.238 1.152 13.1 200 to 300 0.407 2.961 0.622 3.432 8.724 0.528 1.443 18.1 53 to 200 0.106 3.097 0.923 2.541 8.935 0.195 1.16 17.0

Example  2 Biological Activity Assay of Sorghum By-Product Extract

In order to evaluate the cytotoxicity during the evaluation of the physiological activity of the sorghum by-product extract used in the present invention, sorghum extract was dispersed in the medium at concentrations of 0.01%, 0.05%, 0.1%, 0.5% and 1% for fibroblast (1 × 10) for 24 hours. MTS ((3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium) assay was performed on the cells that survived the culture of ⁵ cells). As a result of the cytotoxicity evaluation, as shown in Fig. 5, the cells grew better at 0.5% concentration, and it was confirmed that there was almost no cytotoxicity at the concentration of 1%.

Antioxidative evaluation of sorghum by-product extracts was evaluated using NBT assay and DPPH assay. In the NBT assay, as the concentration of the sorghum by-product extract increased, the antioxidant activity increased, and the antioxidant power of 100% was shown in FIG. 6 at 1% concentration.

On the other hand, antioxidant activity was observed in the extracts of leaf extracts of golden waxy by DPPH assay, and no antioxidative activity was observed in the by - products of stem and white waxy wastes.

Golden Stalk Water (IC _{50 ,} ppm) White horse leaf 173.0 > 200 stem > 200 > 200

For anti-inflammatory testing of sorghum by-product extracts, they were dispersed in medium at concentrations of 0.01%, 0.05%, 0.1%, 0.5%, and 1% and cultured macrophage (RAW264.7 cell) to pro-inflammatory cytokine IL-1β, IL As a result of measuring the incidence amount of -6, TNF-α and PGE ₂ , as shown in FIG. 7, the amount of cytokine such as IL-1β which initiates inflammation, IL-6 causing excessive fibrosis, and TNF-α as shown in FIG. As the reduction was noticeable, it was confirmed that the anti-inflammatory properties were excellent. In the case of IL-1β, when the concentration of sorghum extract was more than 0.1%, the incidence was reduced by more than 50% .In the case of IL-6, cytokine production was significantly decreased from 0.05% of sorghum extract. TNF-α showed an effect at the concentration of more than 0.5% of sorghum extract.

Claims (10)

Natural pigment extract of sorghum by-products having antibacterial and antioxidant activity extracted from lower alcohol in the presence of acid after milling sorghum by-products. The natural pigment extract of sorghum by-product according to claim 1, wherein the lower alcohol is methanol or ethanol. The natural pigment extract of sorghum by-products according to claim 1, wherein the by-products are leaves or stems of sorghum. The natural pigment extract of sorghum by-products according to claim 1, wherein the sorghum by-products are leaves or stems of golden wax or sorghum. The natural pigment extract of sorghum by-product according to claim 1, wherein the acid is one selected from hydrochloric acid, citric acid, acetic acid and malic acid. After milling the sorghum by-products, a method for stabilizing the natural pigment extraction of sorghum by-products having antibacterial and antioxidant activity extracted with lower alcohol in the presence of acid at pH 10. Antioxidant and antimicrobial cosmetic composition containing the extract of any one of claims 1 to 4 as an active ingredient. A pharmaceutical composition for antioxidant and antimicrobial containing the extract of any one of claims 1 to 4 as an active ingredient. Food additive for antioxidant and antimicrobial containing the extract of any one of claims 1 to 4 as an active ingredient. A method for extracting natural pigment extract of sorghum by-products having antimicrobial and antioxidant activity extracted from low-alcohol for 12 to 36 hours after grinding sorghum by-products into 200-300 μm particles.

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