KR20130061046A - Preparation method for fermented rhodiola rosea using absorbents - Google Patents

Abstract

PURPOSE: A rhodiola sachalinensis extract making method is provided to make rhodiola sachalinensis extracts high in salidroside aglycone which are easily absorbed in the human body. CONSTITUTION: Rhodiola sachalinenesis extract is treated with more than one among acid clay, bentonite, diatomite, activated charcoal and a weak base anion exchange resin as an adsorbent. A remaining solution which is not absorbed by the adsorbent is vaccinated with lactobacillus at a temperature of 25-40°C for 24-96 hours. The adsorbent is acid clay or bentonite. The weak base anion exchange resin is Dowex MWA-1. For 100 parts by weight of rhodiola sachalinenesis extracts, 5-60 parts by weight of adsorbent are mixed. The solid content of the rhodiola sachalinensis extract is 0.1-10 parts by weight %. The lactobacillus is chosen from lactobacillus acidophilus [KCTC 2182], bifidobacterium longum [ATCC 15707], bifidobacterium brevis [ATCC 15700] and pediococcus pentosaceus [KCCM 11088P].

Description

Preparation method for fermented rhodiola rosea using absorbents}

The present invention relates to a method for producing a Honggyeongcheon fermented product. In particular, the present invention relates to a method for preparing fermented red ginseng using fermentation agent, and the fermented red ginseng can be used as a material for general food, functional food, medicine or cosmetics.

Rhodiola rosea ( Rhodiola rosea ) is a plant widely distributed in the high mountains of Europe and Asia. It is a traditional medicine that is effective in stimulating the nervous system, reducing depression, improving work ability, recovering from fatigue, and preventing altitude sickness. It has been used for a long time. Especially, it is a perennial herbaceous plant that grows a lot in birch forests and mossy rocks in the mossy long-growth forest between 1,800m ~ 2,300m above sea level. It is able to survive in high altitude environment with large crossroads, dry oxygen, thin oxygen and strong UV. Due to its special adaptability, it is also known as the Gosan Gyeongcheon or Gosan Honggyeongcheon.

The main components of Rhodiola saline include Salidroside, Tyrosol and Glycoside, as well as starch, protein, fat, sugar, flavone compounds, amino acids, negligible elements and trace essential oils. Hong's modern research began in the 1950s. It is known to have a pharmacological action that surpasses some ginseng, and has recently been attracting great attention at home and abroad as it has been proved to be a new drug with no side effects.

The main effects of honggyeongcheon are reported to be effective in restoring energy, longevity, work force prevention, and fatigue recovery. The effects of hong kyung cheon have been actively researched in recent years, and the product development has been recognized as the value of 30 kinds of health functional foods or medicines such as hong kyung cheon liquor, old medicine, capsule, tea, gum, candy, etc. This is increasing even more.

Looking at the patent technology registered in relation to the conventional honggyeongcheon extract, a composition for reducing blood alcohol concentration containing honggyeongcheon extract (Korean Patent No. 10-0179087), a composition for preventing and treating diabetes containing honggyeongcheon extract (Korean Patent No. 10 -0179088), honggyeongcheon extract (Korea Patent No. 10-0265385) having a preventive and therapeutic effect of circulatory diseases, liver fibrosis inhibitory composition (Korea Patent No. 10-0316790), containing a high acid honggyeongcheon extract, Hongkyeongcheon root extract Immunostimulating agent composition (Korean Patent No. 10-0386809), whitening composition containing Honggyeongcheon extract (Korean Patent No. 10-0445404), composition for the prevention and treatment of viral diseases containing Honggyeongcheon extract (Korean Patent No. 10-999872). However, all of the above patents only confirm the efficacy of the Rhodiola sac extract itself or the fractions fractionated therefrom, and there is no description of the method of bioconversion from glycoside to non-glycoside to facilitate the absorption of salidroside, which is known as one of the active ingredients of Rhodiola saline, in the human body. None known.

The present inventors are studying the preparation method of fermentation products of Honggyeongcheon lactic acid bacteria for bioconversion of salidroside to non-glycoside which is advantageous to be absorbed by the human body. The fermentation proceeded to realize that the ratio of the non-glycoside content of the salidroside was completed the present invention.

An object of the present invention is to provide a fermentation product of the high glycosides of salidroside which is advantageous to be absorbed by the human body by providing a fermentation method of the Honggyeongcheon difficult to ferment alone.

The present invention comprises the steps of treating the Honggyeongcheon extract with at least one adsorbent selected from acidic clay, bentonite, diatomaceous earth, activated carbon and weakly basic anion exchange resin; And it provides a method for producing a fermentation product of Honggyeongcheon comprising inoculating fermentation with lactic acid bacteria in the filtrate not adsorbed to the adsorbent.

The adsorbent in the method of preparing the Honggyeongcheon fermentation product of the present invention is characterized in that acidic clay or bentonite.

The weakly basic anion exchange resin in the manufacturing method of the fermentation product of Honggyeongcheon is characterized in that the Dowex MWA-1.

In the manufacturing method of the fermentation product of Honggyeongcheon of the present invention, it is characterized in that 5 to 60 parts by weight of the adsorbent is mixed with respect to 100 parts by weight of the extract.

The solid content of the honggyeongcheon extract in the method of manufacturing the honggyeongcheon fermentation product of the present invention is characterized in that 0.1 to 10% by weight.

The lactic acid bacteria in the production method of the fermentation product of Honggyeongcheon is characterized in that any one or more lactic acid bacteria selected from the genus Lactobacillus, Bifidobacterium genus and Pediococcus genus.

Lactic acid bacteria in the preparation method of the fermentation product of the present invention, Lactobacillus ashdophyllus [KCTC 2182], Bifidobacterium long gum [ATCC 15707], Bifidobacterium brevis [ATCC 15700] and Pediococcus pentosaceus It is characterized in that any one or more lactic acid bacteria selected from [KCCM 11088P].

The step of fermentation in the method of manufacturing the Honggyeongcheon fermentation product of the present invention is characterized in that it is carried out at 25 to 40 ℃ 24 to 96 hours.

The present invention is a fermentation product obtained by inoculating fermentation of lactic acid bacteria into a filtrate treated with Honggyeongcheon extract from any one selected from acidic clay, bentonite, diatomaceous earth, activated carbon, and weakly basic anion exchange resin. It provides a honggyeongcheon fermentation, characterized in that the content of the non-glycoside in the total 100 parts by weight of 30 to 100 parts by weight.

The present invention provides a food composition comprising the honggyeongcheon fermented product.

The present invention provides a fermentation method of Honggyeongcheon, which is difficult to ferment alone, and furthermore, since the fermentation product of Honggyeongcheon produced through the method of the present invention has a high content of non-glycosides of salidroside, which is advantageous to be absorbed by the human body, new general food and health It can be used as a material for nutraceutical, medicine or cosmetics.

1 is a photograph of Honggyeongcheon used in Preparation Examples, Examples and Comparative Examples of the present invention.
Figures 2a, 2b, 2c and 2d is a graph showing the pH change according to the type and incubation time of lactic acid bacteria when the solid content of Rhodiola cheoncheon extract in Experimental Example 1 is 0.5% by weight, 1% by weight, 3% by weight and 5% by weight, respectively. to be.
Figure 3 is a graph showing the pH of the filtrate measured after 48 hours of adsorbed Honggyeongcheon extract according to the acidic clay or bentonite throughput in Experimental Example 2.
Figure 4 is lactic acid bacteria in the honggyeongcheon extract (●), honggyeongcheon extract filtrate (○), Dowex MWA-1 adsorption filtrate (▼) of the honggyeongcheon extract, the bentonite adsorption filtrate (△) of the honggyeongcheon extract without adsorbents After inoculation and fermentation, it is a graph confirming the bacterial count according to fermentation time.
Figure 5 shows the salicide side and its non-glycoside standard in Experimental Example 3, Hong Kyungcheon extract (Con), acid clay adsorbed filtrate (1B), fermented product of acid clay adsorbed filtrate (1A), Dowex MWA-1 adsorbed filtrate (2B) , Fermentation of Dowex MWA-1 adsorption filtrate (2A), Diaion HP-2MG adsorption filtrate (3B), fermentation of Diaion HP-2MG acid clay adsorption filtrate (3A), bentonite adsorption filtrate (4B), bentonite adsorption filtrate It is a photograph of the result of TLC analysis of the fermented product (4A).
6A, 6B, 6C, and 6D are HPLC chromatograms of salidroside and its non-glycoside standard in Experimental Example 3, chromatograms of Honggyeongcheon extract before adsorptive treatment with acidic clay, and Hongkwangcheon extract filtrate adsorbed with acidic clay, respectively. Chromatogram, a chromatogram of fermentation obtained by fermenting the filtrate of Hongkyungcheon extract treated with acidic clay for 48 hours.
FIG. 7 shows fermentation of Honggyeongcheon extract (Con), acid clay adsorption filtrate (1B), fermentation product of acid clay adsorption filtrate (1A), Dowex MWA-1 adsorption filtrate (2B), and Dowex MWA-1 adsorption filtrate in Experimental Example 3 Salidroside of water (2A), Diaion HP-2MG adsorption filtrate (3B), fermentation product of Diaion HP-2MG acid clay adsorbent filtrate (3A), bentonite adsorption filtrate (4B), and fermentation product of bentonite adsorption filtrate (4A) And a graph showing a comparison of the content of salicide side glycosides.

The present invention comprises the steps of treating with at least one adsorbent selected from acidic clay, bentonite, diatomaceous earth, activated carbon and weakly basic anion exchange resin; And inoculating the fermented lactic acid bacteria into the filtrate not adsorbed to the adsorbent.

Fuller's Earth (Acid clay) in the present invention is a montmorillonite clay mineral, colloidal hydrous aluminum silicate, pale yellow fine powder. The acidic clay of the present invention is suspended by adding 1 g of 50 g of water, and then the pH of the filtrate is 3 to 5.

Bentonite is a montmorillonite clay mineral, like acidic clay, and is colloidal hydrous aluminum silicate.

The acidic clay and bentonite are preferable in that the yield is high after the adsorbent treatment, and the fermentation of Honggyeongcheon is performed by adsorbing a substance that inhibits the growth of lactic acid bacteria. Fermentation properties of these extracts are not improved in all adsorbents, but fermentation properties are not improved at all in activated clay, zeolite and aluminum oxide.

In addition, diatomite is a substance composed of a protective film (diatom shell) of diatoms or single cell plants. Diatomaceous earth has a very high yield after adsorbent treatment, but the fermentation characteristics of fermenting red gland are slightly lower than acidic clay or bentonite.

Activated carbon is also activated by carbonizing carbonaceous materials such as vegetable fiber such as sawdust, wood chips and palm bark, or attan or petroleum. Activated carbon has a low yield of acidic clay or bentonite and adsorbent treatment, and the fermentation properties of fermenting Honggyeongcheon are somewhat lower than acidic clay or bentonite.

The weakly basic anion exchange resin is an ion exchange resin that exchanges anions and combines primary to tertiary amines with a polymer matrix condensed in three dimensions. The weakly basic anion exchange resin has excellent fermentation characteristics for fermenting Honggyeongcheon, but the yield of acidic clay, bentonite and adsorbent is low. As the weakly basic anion exchange resin, for example, Dowex MWA-1 or Diaion HP-2MG can be used. On the other hand, even adsorbing honggyeongcheon extract with a cation exchange resin such as Amberlite IRC-50 or Amberlite XAD-4 does not improve the fermentation properties of honggyeongcheon extract.

In the present invention, 5 to 60 parts by weight of the adsorbent, preferably 20 to 50 parts by weight, and more preferably 25 to 40 parts by weight, based on 100 parts by weight of the rhodiola saline extract when treated with the adsorbent. . When the content of the adsorbent exceeds the upper limit, the amount of the adsorbent is large, so that the Hong Kyung-cheon extract and the adsorbent are not evenly mixed, it is not easy to separate the filtrate of the Hong Kyung-cheon extract, and the yield is also lowered. In addition, when the ratio of the adsorbent is less than the lower limit, the adsorption of the substance inhibiting the lactic acid bacteria fermentation is not enough, so that the fermentation is not sufficiently made.

In the present invention, the adsorbent treatment time and temperature need not be particularly limited, but the treatment time is 12 hours or more, preferably 24 hours or more, and more preferably 36 to 54 hours. The treatment of the adsorbent above the upper limit does not improve the fermentation anymore, and below the lower limit, the adsorption of the substance inhibiting the lactic acid bacteria fermentation is not sufficient, so that the fermentation is not sufficiently performed. The treatment temperature is possible at 5 to 40 ° C., and treatment can be performed at higher temperatures, but the adsorption efficiency is not increased through heating.

In the mixed extract of the red ginseng and red ginseng of the present invention, the solid content of the red ginseng is 0.1 to 10% by weight, preferably 0.2 to 5% by weight, more preferably 0.3 to 1% by weight, when the upper limit is exceeded. Even if it passes through, the growth and fermentation of lactic acid bacteria is not sufficiently made, below the lower limit, the content of the active ingredient derived from Honggyeongcheon is reduced and the production yield is inefficient.

Lactic acid bacteria selected from Lactobacillus genus, Bifidobacterium genus and Pediococcus genus Lactobacillus may be used in the preparation of the fermentation product of Honggyeongcheon. Preferably, the lactic acid bacteria are Lactobacillus ash also filler's, Bifidobacterium ronggeom, Bifidobacterium Brenna non Scotland and Peddie Oh Caucus pen and quarrying three-house, more preferably Lactobacillus ash also filler being held in the Korea Food Research Institute One or more lactic acid bacteria selected from among [KCTC 2182], Bifidobacterium long gum [ATCC 15707], Bifidobacterium brevis [ATCC 15700] and Pediococcus pentosaceus [KCCM 11088P], and most preferably Lactobacillus ashdophyllus [KCTC 2182], Bifidobacterium longgum [ATCC 15707].

The fermentation step of the method for producing a fermentation product of Honggyeongcheon is carried out at 25 to 40 ° C. for 24 to 96 hours.

Hong Kyungcheon fermented product prepared by the above method has a content of non-glycoside in a total of 100 parts by weight of the salidroside glycoside and non-glycoside 30 to 100 parts by weight, preferably 50 to 100 parts by weight, more preferably 70 to 100 parts by weight As a part, since it contains a lot of non-glycosides that are easily absorbed by the human body as compared to the extract, it is possible to further enhance the inherent efficacy.

For example, the fermented red ginseng of the present invention exhibits an antioxidant effect, inhibits oxidative stress of muscle cells, and has an activity of improving exercise ability, and thus fermented by inoculating fermented lactic acid bacteria into a mixed extract of red ginseng and red ginseng. The food composition which uses water as an active ingredient can be manufactured. The food composition may be a health functional food for fatigue recovery and athletic performance including the honggyeongcheon fermented product as an active ingredient.

In the present invention, the health functional food is a food prepared by adding the fermentation product of Honggyeongcheon to food materials such as beverages, teas, spices, gums, confections, or the like, encapsulated, powdered, suspensions, and the like. Means to bring the effect, unlike the general medicine has the advantage that there are no side effects that can occur when taking a long-term use of the drug as a raw material. The health functional food of the present invention thus obtained is very useful because it can be consumed on a daily basis. The amount of fermentation products of Honggyeongcheon fermentation products in such health functional foods cannot be defined uniformly depending on the type of the health functional foods to be applied, but may be added within a range that does not impair the original taste of the food, and is usually 0.01 to the target foods. To 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of a health functional food in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the dietary supplement of the present invention may be in the form of pills, tablets, capsules or beverages.

Health functional food of the present invention as an active ingredient, as well as the fermentation of Honggyeongcheon, may include ingredients that are commonly added during food production, for example, include proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. do. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As flavoring agents, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the health functional food of the present invention is prepared with a drink and a beverage, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be further included in addition to the fermented product of the red ginseng of the present invention. have.

In addition, the anti-fatigue and movement disorder treatment and prevention of the pharmaceutical composition of the present invention using the honggyeongcheon fermentation may be prepared.

On the other hand, the term 'comprising as an active ingredient' in the present specification means that it contains an amount sufficient to achieve the efficacy or activity of the Honggyeongcheon fermentation product of the present invention. In one embodiment of the present invention, the red gyeongcheon fermentation product in the composition of the present invention is, for example, at least 0.001 mg / kg, preferably at least 0.1 mg / kg, more preferably at least 10 mg / kg, even more preferred Preferably at least 100 mg / kg, even more preferably at least 250 mg / kg, most preferably at least 0.1 g / kg. Since the fermented red ginseng has no adverse effects on the human body even when it is excessively administered as a natural product, the upper limit of the amount of the red ginseng ferment contained in the composition of the present invention can be carried out by those skilled in the art.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, lubricants. Agents, flavors and the like can be used.

The pharmaceutical composition may be formulated to include one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.

Formulation forms of the compositions may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions and the like. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include but are not limited to natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. One or more components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like, and preferably, oral administration.

Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to reaction, and usually The skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g / kg.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or by incorporating into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following examples are provided only to aid understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Manufacturing example  One: Honggyeongcheon  Exclusive Fermented  Produce

end. material

The main raw material, Rhodiola rosea ( Rhodiola rosea , Fig. 1) was purchased in 2010 in China, was pulverized and used by pulverization (Cyclotec ^™ 1093, FOSS Co., Denmark). Red ginseng used commercially available red ginseng powder (NH Hansamin, 6 years old).

The strains used in this experiment are Lactobacillus and Pediococcus stored in the Korea Food Research Institute as shown in Table 1. And Ten lactic acid bacteria of Bifidobacterium spp. Were used. The strain was cultured in sterile 50% glycerol and stored for a long time in a -70 ℃ deep freezer.

division Strain name Used badge L1 Lactobacillus acidophillus 128 MRS L2 Lactobacillus plantarum 144 MRS L3 Lactobacillus fermentum 164 MRS L4 Lactobacillus curvatus 166 MRS L5 Lactobacillus gasseri 3162 MRS L6 Lactobacillus gasseri 3163 MRS L7 Lactobacillus acidophillus K-59 MRS B1 Bifidobacterium longum RCM  Of GAM B2 Bifidobacterium breve RCM  Of GAM P0 Pediococcus pentosaceus MRS

I. Preparation of Honggyeongcheon Extract

Hongkyeongcheon extract was added to the reflux extractor and distilled water at 90 ℃, the extract was heated for 3 hours was separated from the centrifuge (Mega 17R, Hanil Science Indusrrial Co. Ltd., Korea) adjusted to 4 ℃. The separated supernatant was filtered through a filter paper (Watman No. 2), and the filtrate was used as an extract of Honggyeongcheon.

All. Preparation of fermented red pepper

After diluting so that the solid content of the honggyeongcheon extract is 0.5, 1, 3 and 5% by weight, the diluted honggyeongcheon extract was sterilized at 121 ° C., 1.5 atm for 15 minutes.

The lactic acid bacteria of Table 1 were activated by culturing at 37 ° C. for 20 hours using MRS broth and GAM broth, respectively, and then lactic acid bacteria were inoculated with 1 wt% (10 ⁶ CFU / mL) of the culture broth, respectively. Cultured at 48 ℃ for 48 hours to prepare a fermentation of Honggyeongcheon alone.

Experimental Example  One: Honggyeongcheon  Exclusive Fermented  Fermentation Characteristics Check

In order to confirm the fermentation characteristics of the fermentation product of Honggyeongcheon extract of Preparation Example 1, the pH and viable cell number of fermented products were checked at 24 and 48 hours after inoculation.

The number of viable cells is 1 ml of the sample in 9 ml of sterile physiological saline and mixed well. After diluting step by step, 0.1 ml is taken. Smear agar and store it as anaerobic as possible using anaerobic jar (BBL Gas Pak Anaerobic System), incubate at 37 ℃ for 24 ~ 48 hours, measure the number of colonies produced, multiply the average number of colonies by dilution factor Colony per culture was calculated.

end. PH Change According to Solid Contents and Types of Lactic Acid Bacteria

The changes in pH according to the type and incubation time of the lactic acid bacteria when the content of the honggyeongcheon extract solid content is 0.5% by weight, 1% by weight, 3% by weight and 5% by weight are shown in FIGS. 2A, 2B, 2C and 2D, respectively.

In FIG. 2a, the initial pH was 6.25 when the solid content of the Honggyeongcheon extract was 0.5% by weight, and after 48 hours, the final pH value was decreased from 5.25 to 5.50.

In FIG. 2B, the initial pH was 6.21 when the solid content of the honggyeongcheon extract was 1% by weight, and after 48 hours, the final pH value was 5.61 to 5.70, which was slightly smaller than 0.5% by weight, but decreased.

However, as shown in Figures 2c and 2d when the honggyeongcheon concentration is increased to 3% by weight and 5% by weight, after 48 hours at the initial pH 6.15 and 6.13, but slightly different depending on the type of lactic acid bacteria to 5.95 to 6.00, 6.04 to 6.11, It can be seen that there is no difference in pH before and after fermentation.

From the above results, it was confirmed that at 3 wt% or more of Rhodiola savona, the pH was not decreased even when the incubation time was increased, and even at 1 wt% or less, there was no decrease in pH observed in conventional lactic acid bacteria culture.

Comparing the change of pH by lactic acid bacteria Lactobacillus acidophilus 128 (L1) , Bifidiobacterium longum (B1) , Bifidiobacterium It was confirmed that the pH of breve (B2) and Pediococcus pentosaceus (P0) was reduced significantly compared to other lactic acid strains, and was selected as a suitable strain for the preparation of fermented red pepper.

I. Changes in the Number of Viable Cells According to Solid Contents and Types of Lactic Acid Bacteria

Four kinds of lactic acid bacteria ( L. acidophillus 128 , B. longum , B. breve , P. pentosaceus ) , which showed relatively large pH change among 10 kinds of lactic acid bacteria, were pre-incubated again and the content of 0.5 wt. 1% by weight of lactic acid bacteria were inoculated into% and 1% by weight of Honggyeongcheon extract, and fermented at 37 ° C. for 24 hours. The number of viable cells was measured and the results are shown in Table 2.

division Viable cell count ( CFU Of ml ) Before fermentation After fermentation (0.5 wt%) After fermentation (1% by weight) L1 7.10 * 10 ⁶ 6.10 * 10 ³ 2.25 * 10 ² B1 2.10 * 10 ⁶ 2.59 * 10 ³ 1.32 * 10 ² B2 1.64 * 10 ⁶ 4.40 * 10 ² 1.95 * 10 P0 1.50 * 10 ⁶ 2.01 * 10 ² 1.29 * 10

As shown in Table 1, the result of culturing Rhodiola sachalinensis using L. acidophillus 128 was 1.2 * 10 ³ CFU / mL at 0.5 wt% at 7.10 * 10 ⁶ CFU / mL at the first inoculation, and 1.3 * 10 at 1 wt%. ^The number of viable cells was rather reduced to ² CFU / ml. This is a relatively high number of bacteria compared to other microorganisms, but it can be seen that the higher the concentration of Hongkyeongcheon extract, the number of viable bacteria is significantly reduced.

B. longum results first live cells were cultured using the 2.10 * 10 ^6, 0.5% by weight in ^{CFU / ㎖ 2.59 * 10 3 CFU} / ㎖, the 1% by weight was 1.32 * 10 ² CFU / ㎖.

The viable cell count was 1.64 * 10 ⁶ CFU / mL at 4.64 * 10 ² CFU / mL at 0.5% by weight and 8.95 * 10 CFU / mL at 1% by weight after incubation with breve . It was.

Results number of viable cells by fermentation by using a P. pentosaceus is in a concentration of 1.50 * 10 ⁶ in CFU / ㎖ 0.5% by weight of all the lactic acid bacteria to 1.29 * 10 CFU / ㎖ the ^{2.01 * 10 2 CFU / ㎖,} 1% by weight honggyeongcheon It was found to decrease dependently.

In order to compare the lactic acid bacteria fermentation characteristics of red ginseng and red ginseng, the red ginseng extract was prepared in the same manner as the preparation method of red ginseng extract of Preparation Example 1, and the pH and viable cell number of the microorganisms were inoculated and fermented at 37 ° C. for 24 hours. The change is shown in Table 3.

division pH Viable cell count ( CFU Of ml ) Before fermentation After fermentation Before fermentation After fermentation L1 6.22 4.02 7.10 * 10 ⁶ 7.50 * 10 ⁸ B1 6.22 3.62 2.10 * 10 ⁶ 8.95 * 10 ⁸ B2 6.22 3.92 1.64 * 10 ⁶ 2.88 * 10 ⁸ P0 6.22 3.89 1.50 * 10 ⁶ 3.20 * 10 ⁸

 According to the results of Table 3, when inoculated with each lactic acid bacteria in 0.5% by weight of red ginseng extract and fermented at 37 ° C. for 24 hours, the pH was significantly decreased by about 2 or more to less than 4.2, and the viable cell count of lactic acid bacteria was rather 2 log cycles. Proliferation was confirmed. Compared with lactic acid bacteria fermentation of red ginseng extract, fermentation of red ginseng extract is greatly inhibited the growth of lactic acid bacteria due to the antimicrobial activity of the red ginseng, thereby the bioconversion effect of the useful components of red ginseng by enzymes secreted from lactic acid bacteria or lactic acid bacteria In single fermentation it was expected to be minimal.

However, in the results of Table 2, Lactobacillus higher residual viable cell number after 24 hours culture than the remaining microorganisms acidophilus 128 was selected and used in the next experiment.

Manufacturing example  2: Honggyeongcheon Fermented  Produce

end. Adsorbent Treatment of Rhodiola sachalinensis Extract

Table 4 summarizes the characteristics and application fields of the 11 types of adsorbents used in the adsorbent treatment of Hong Kyungcheon extract.

Sorbent type characteristic Applications Active carbon
(Activated carbon) Decolorant, Adsorbent Recovery of solvents, removal of organic impurities Clay activated
(Active clay) Acidic clay is treated with acid to adsorb and decolorize to increase the catalytic action Dehydration, Purification, Decolorization, Adsorbent of Petroleum Products Acid clay
(Acidic clay) Almost similar in appearance to acidic clay and bentonite, but rich in silicic acid. Stronger adsorption than bentonite Decolorization of petroleum, desiccant, catalyst, dehydrating agent Dowex MWA-1 Weakly basic anion exchange resin - Aluminum oxide Oxide of aluminum - Diaion HP-2MG Weak Base Ion Exchange Resin Zeolite
(Zeolite) Synthetic mineral, also called zeolite, used as cation exchange resin, desiccant, adsorptive separator Cleaning aids for synthetic detergents, radioactive treatments, water repellents, municipal wastewater treatment agents Amberlite IRC-50 Cation exchange resin - Diatom earth
(Diatomaceous earth) Sediments made from the remains of diatoms Adsorbents, Filters, Insulating Materials, Abrasives, Filters for Purification of Industrial Water, Water, Pharmaceuticals, Food Bentonite
(Bentonite) Clay minerals resulting from alteration of tuff and rhyolite particles Structural viscosity and protective colloid action, foam stabilization, detergency improvement, cosmetics, medicine, food, paint, etc. Amberlite XAD-4 Non Polymeric Adsorbent -

50 g of 11 adsorbents were respectively prepared in 100 ml of hot water extract (1% solids content, pH6.46) of Preparation Example 1, and the pH and solids content of the filtrate were measured and shown in Table 5.

division pH Soluble Solids Content (%) absorbent Before processing Honggyeongcheon  extract( Control ^* ) 4.10 1.0






absorbent Active carbon
(Activated carbon) 3.55 0.5 Clay activated
(Active clay) 3.01 0.8 Acid clay
(Acidic clay) 3.96 0.8 Dowex MWA-1 7.30 0.6 Aluminum oxide 5.83 1.0 Diaion HP-2MG 4.46 0.6 Zeolite
(Zeolite) 9.31 1.0 Amberlite IRC-50 3.03 1.0 Diatom earth
(Diatomaceous earth) 7.00 1.4 Bentonite
(Bentonite) 6.04 1.0 Amberlite XAD-4 7.82 2.0

As can be seen from Table 5, the initial 1% solids content of Honggyeongcheon extract showed a pH of 4.10, but the pH of the filtrate filtered after adsorbent treatment was activated carbon, activated clay, acidic clay, Dowex MWA-1, Amberlite IRC. When -50 and the like were lowered to 3.01 ~ 3.96, they increased to 4.46 ~ 9.31 when aluminum oxide, Diaion HP-2MG, Zeolite, diatomaceous earth, bentonite and Amberlite XAD-4 were treated. In particular, zeolite showed a sharp pH change.

In the case of solids, there was a big change after adsorbent treatment depending on the type of adsorbent, which was about 2 times increased when treated with Amberlite XAD-4, whereas about 40-50% when activated carbon, Dowex MWA-1, and Diaion HP-2MG were treated. Decreased. Such a decrease in solid content may lead to a decrease in yield or main active ingredients in the future, which is considered to be an undesirable result.

I. Fermentation of Rhodiola saline Extract Filtrate

Adsorption filtrate obtained in A. was first adjusted to 6.5, the initial pH for lactic acid bacteria fermentation, and Lactobacillus in the same manner as in Preparation Example 1. acidophilus Table 6 shows the result of fermentation by inoculation.

division pH Soluble solids
content (%) Fermented without adsorbent treatment Honggyeongcheon  extract( Control ^* ) 6.5 0.9





Fermented Honggyeongcheon Extract after Treatment with Adsorbent Active carbon
(Activated carbon) 4.9 0.6 Clay activated
(Active clay) 5.8 0.8 Acid clay
(Acidic clay) 4.6 0.8 Dowex MWA-1 3.4 0.6 Aluminum oxide 5.4 1.0 Diaion HP-2MG 3.7 0.6 Zeolite
(Zeolite) 5.3 0.8 Amberlite IRC-50 5.3 1.0 Diatom earth
(Diatomaceous earth) 4.9 0.8 Bentonite
(Bentonite) 4.3 0.8 Amberlite XAD-4 7.1 1.0

The pH of honggyeongcheon extract filtered after treatment with each adsorbent was adjusted to 6.5, which is the initial pH for lactic acid bacteria fermentation, and the solids content was adjusted to 1% level, and then inoculated with lactic acid bacteria and fermented at 37 ° C for 48 hours. The results are shown in Table 6.

Looking at the pH change after 48 hours of fermentation, it was found that the fermentation process without the adsorbent exhibited a pH of 6.5, inoculated with lactic acid bacteria and fermented for 48 hours.

Compared with activated carbon, acid clay, Dowex MWA-1, Diaion HP-2MG, diatomaceous earth, and bentonite, the pH was lowered to below 5.0 by fermentation, and activated clay, aluminum oxide, zeolite, Amberlite IRC-50 and XAD. -4 was confirmed that the fermentation was not sufficiently made to pH 5.0 or more.

Therefore, activated carbon, acidic clay, Dowex MWA-1, Diaion HP-2MG, diatomaceous earth and bentonite treatment were found to help fermentation of lactobacillus. However, activated carbon, Dowex MWA-1 and Diaion HP-2MG treatment resulted in the solid content of Honggyeongcheon extract. Due to the significant reduction, acidic clay or bentonite was assumed to be the preferred adsorbent for the preparation of fermentation products.

Experimental Example  2: Confirmation of proper throughput of adsorbent

In order to examine the effect of the treatment of acidic clay and bentonite selected in Preparation Example 2 on the fermentation of lactobacillus of Honggyeongcheon extract, the adsorbent first washed with 70% MeOH and distilled water 1, 5 per 100 ml of Honggyeongcheon extract (0.8%) , 10, 15, 20, 25, 30, 35 g each treatment by adsorbing at 500rpm for one day and then inoculated lactic acid bacteria into the filtered culture as in Preparation Example 2 and fermentation for 48 hours while showing the results of the pH change of the fermentation product 3 is shown.

The effect of improving fermentation characteristics by acidic clay or bentonite treatment is effective from 1 part by weight of the adsorbent to 100 parts by weight of honggyeongcheon extract based on honggyeongcheon extract having a solid content of 1% by weight, preferably 5 parts by weight or more, and more preferably. The addition of 20 parts by weight or more can achieve a sufficient fermentation improvement effect, even if more than 25 parts by weight did not show a fermentation improvement effect anymore.

100 ml of 1 wt% solid content of Honggyeongcheon extract was adsorbed for 24 hours on each of 25 g of acidic clay, bentonite and Dowex MWA-1, followed by Lactobacillus acidophillus in the same manner as in Preparation Example 1. The inoculation of 128 to confirm the change in the number of lactic acid bacteria for 48 hours is shown in Figure 4. As a control, the same inoculation of lactic acid bacteria was used in the Hongkyungcheon extract that was not treated with the adsorbent.

Lactic acid bacteria fermentation of Hongkyungcheon after acidic adsorption treatment at 3.1 × 10 ⁶ CFU / mL was slightly decreased to 5.14 × 10 ⁵ CFU / mL after 24 hours and 5.01 × 10 ⁵ CFU / mL after 48 hours. 1 at 24 hours after fermentation at 5.01 × 10 ⁵ CFU / mL, after 48 hours at 5.85 × 10 ⁵ CFU / mL, at 24 hours after bentonite adsorption at 8.41 × 10 ⁵ CFU / mL, after 48 hours at 8.52 × 10 ⁵ It was confirmed that the initial bacterial count was somewhat lowered by CFU / ml. On the other hand, in the case of Hong Kyung-cheon extract without the adsorbent (control), the viable cell count decreased significantly from the beginning of fermentation, and the viable cell count decreased significantly to the level of 6.10 × 10 ² CFU / ml on the second day of fermentation. It can be seen that there is an effect promoting.

Experimental Example  3: Honggyeongcheon  Before and after fermentation Salidroside  Glycosides and Non-glycoside  Content comparison

After adsorbing 25 g of acidic clay and bentonite for 24 hours to 100 ml of 1 kg by weight of red ginseng extract, each filtrate was subjected to Lactobacillus acidophillus in the same manner as in Preparation Example 1. Soluble total sugar, acidic polysaccharide and total phenolic content of the fermentation product prepared by fermentation with 128 inoculation for 48 hours are shown in Table 7 in comparison with the extract of Honggyeongcheon.

division % Per total Acid polysaccharide (%) Total phenol (%) Honggyeongcheon Extract 75.97 ± 0.92 26.16 ± 1.29 20.12 ± 0.81 Acid clay After adsorption 74.05 ± 3.17 24.51 ± 0.12 13.94 ± 0.43 After fermentation 58.30 ± 3.10 18.10 ± 0.19 15.74 ± 0.74 Bentonite After adsorption 64.81 ± 2.39 20.15 ± 0.91 12.09 ± 0.02 After fermentation 59.40 ± 2.31 19.02 ± 0.32 13.02 ± 0.30

The total sugar content was measured by phenol-sulfuric acid method using D-glucose as a standard substance, and the acidic sugar content was m-hydroxydiphenyl method using D-galacturonic acid as standard substance, and the content of polyphenol compound was phosphomolybdic by phenolic substance. According to the Folin Denis method which reacts with acid and shows blue color, gallic acid was measured by colorimetric method.

The total sugar content of the early red ginseng extract was 75.97%, the acid polysaccharide content was 26.16%, and the total phenol content was 20.12%, but the total phenolic and acidic sugars did not change significantly, but the total phenolic compound content decreased slightly. . These results were judged to be due to the adsorption and removal of some phenolic compounds and components that affect the total phenolic compound content such as pigments.

On the other hand, when the lactic acid bacteria fermented, the total sugar and acid polysaccharide contents were generally decreased and the total phenolic compound contents were slightly increased. The decrease in sugars was thought to be due to the increase in the content of soluble phenolic compounds by fermentation as in the previous studies, while sugars were consumed as a nutrient for lactic acid bacteria growth.

Experimental Example  4: Honggyeongcheon  Before and after fermentation Salidroside  Glycosides and Non-glycoside  Content comparison

100 ml of 1 wt.% Solid content of Honggyeongcheon extract was adsorbed with 25 g of acidic clay, bentonite, Dowex MWA-1 and Diaion HP-2MG for 24 hours, and then the filtrate was prepared in the same manner as in Preparation Example 1 Lactobacillus acidophilus After inoculating 128 and fermenting for 48 hours, the filtrate obtained by treatment with adsorbent before fermentation and the active ingredients of the fermentation were analyzed by TLC and HPLC.

Samples were deposited on Silica gel TLC plates (60F ₂₅₄ , Merck, Germany) for TLC analysis, developed with chloroform / methanol / H20 (70: 30: 4) and sprayed with 30% H ₂ SO ₄ reagent at 105 ° C. Color development was carried out for 30 minutes to confirm the position of each component, the results are shown in FIG. Salicide side (S1) as a standard and ρ-tyrosol (S2) were used as a non-glycoside of salicide side.

HPLC analysis was Waters Sunfire ^TM The C18 column and the mobile phase were carried out using a 20% methanol (v / v) solution. The sample injection amount was 20 µl, the solvent development rate was 1 ml / min, and measured at 278 nm using a UV detector. Salidroside (S) was used as a standard, and ρ-tyrosol (SH) was used as a non-glycoside of salidroside.

6a is an HPLC chromatogram of salidroside (S) and its non-glycoside standard (SH), FIG. 6b is a chromatogram of Honggyeongcheon extract before adsorptive treatment with acidic clay, and FIG. 6c is a Honggyeongcheon extract filtrate adsorbed with acidic clay Chromatogram of Fig. 6d is a chromatogram of a fermentation product obtained by fermenting a filtrate of Hongkyungcheon extract filtrate adsorbed with acidic clay for 48 hours. The chromatogram was shown only in the case of acidic clay treatment, and in the case of treatment with other adsorbents, the contents of the salicide side and the salicide side non-glycoside are shown in FIG. 7. In FIG. 7, Con is a Hong Kyungcheon extract, 1B is an acid clay adsorbed filtrate, 1A is an acid clay adsorbed filtrate, 2B is a Dowex MWA-1 adsorbed filtrate, 2A is a fermented product of Dowex MWA-1 adsorbed filtrate, 3B is Diaion HP-2MG adsorption filtrate, 3A represents a fermentation product of Diaion HP-2MG acid clay adsorption filtrate, 4B is a bentonite adsorption filtrate, 4A represents a fermentation product of a bentonite adsorption filtrate.

TLC and HPLC results showed that salidroside, one of the major indicators, had almost the same content as that of Hongkyungcheon extract when treated with acidic clay, Dowex MWA-1, and bentonite, whereas salidroside content of filtrate when treated with Dianion HP-20M. It was confirmed that this is reduced. In the case of salidroside aglycon type tyrosol, acidic clay, Dowex MWA-1 and bentonite were slightly increased compared to before fermentation of lactic acid bacteria. In particular, the acidic clay showed the lowest salidroside content, which was almost the same as that of non-adsorption Hongkyeongcheon extract, which showed the lowest loss of surface components due to adsorption, and also showed the tendency of conversion to tyrosol due to fermentation.

In general, when the fermentation of natural materials using lactic acid bacteria and the like to convert the glycosides to the non-glycoside form to increase the absorption and functionality in the body, these results have the effect of enhancing the functionality and absorption rate of the functional components of Honggyeongcheon It was expected to indicate.

Hereinafter, an example of the preparation of the composition containing the fermentation of Honggyeongcheon of the present invention will be described.

Formulation example  One. Sanje  Produce

20 mg of Hongkyungcheon fermented powder treated with acid clay of Experimental Example 2

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2. Preparation of tablets

10 mg of Hongkyungcheon fermented powder treated with acid clay of Experimental Example 2

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3. Preparation of capsules

10 mg of Hongkyungcheon fermented powder treated with acid clay of Experimental Example 2

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  5. Liquid  Produce

20 mg of Hongkyungcheon fermented powder treated with acid clay of Experimental Example 2

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added to purified water in accordance with the conventional liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water and adjusted to 100 as a whole. To prepare a liquid agent.

Formulation example  6. Manufacture of health functional foods

1,000 mg of Hongkyungcheon fermented powder treated with acid clay of Experimental Example 2

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 [mu] g vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is a composition that is relatively suitable for the health functional food, the composition is mixed in a preferred embodiment, but the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules may be prepared and used for preparing the nutraceutical composition according to a conventional method.

Formulation example  7. Preparation of Functional Beverages

1,000 mg of honggyeongcheon fermented product powder of Example 3

Citric acid 1,000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to the flask to obtain a total of 900 mL

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the functional beverage composition of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (10)

Treating the Hong Kyungcheon extract with at least one adsorbent selected from acidic clay, bentonite, diatomaceous earth, activated carbon and weakly basic anion exchange resin; And
Inoculating the lactic acid bacteria in the filtrate that is not adsorbed to the adsorbent and fermentation method comprising the step of fermentation.
The method of claim 1, wherein the adsorbent is acid clay or bentonite.
2. The method of claim 1, wherein the weakly basic anion exchange resin is Dowex MWA-1.
The method according to claim 1, wherein 5 to 60 parts by weight of an adsorbent is mixed with respect to 100 parts by weight of the extract of Honggyeongcheon.
The method of claim 1, wherein the solid content of the honggyeongcheon extract is 0.1 to 10% by weight.
According to claim 1, wherein the lactic acid bacteria Lactobacillus genus, Bifidobacterium genus and Pediococcus genus Lactobacillus, characterized in that any one or more lactic acid bacteria selected from lactic acid bacteria manufacturing method.
According to claim 6, The lactic acid bacteria Lactobacillus ashdophyllus [KCTC 2182], Bifidobacterium long gum [ATCC 15707], Bifidobacterium brevis [ATCC 15700] and Pediococcus pentosaceus [KCCM 11088P Method for producing a fermentation product of Honggyeongcheon, characterized in that any one or more lactic acid bacteria selected from.
The method of claim 1, wherein the fermentation is carried out at 25 to 40 ℃ 24 to 96 hours, characterized in that the manufacturing method of the fermented Honggyeongcheon.
A fermentation product obtained by inoculating fermentation of Honggyeongcheon extract with lactic acid bacteria in a filtrate treated with one or more adsorbents selected from acidic clay, bentonite, diatomaceous earth, activated carbon and weakly basic anion exchange resin, and the total weight of salidroside glycoside and nonglycoside Honggyeongcheon fermentation, characterized in that the content of non-glycosides in 30 to 100 parts by weight.
A food composition comprising the fermented red ginseng of claim 9.

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