KR20140011663A - A mass extraction method of platycodin d - Google Patents

Abstract

The present invention relates to a method for mass extraction of Platycodin D and, more particularly, to a method for mass extraction of Platycodin D, comprising the steps of: reaping one to four-year-old balloon flowers on February and March; washing and selecting the reaped balloon flowers, dividing the balloon flowers into roots and leaves, and then drying and pulverizing the roots and leaves by using firepower at a temperature of 50°C or lower; and extracting Platycodin D from the pulverized powder. According to the present invention, Platycodin D contained in the balloon flowers can be simply and economically obtained for a short time. Therefore, the method of the present invention can be useful in mass extraction and production of Platycodin D. [Reference numerals] (AA) Content (%); (BB) Growth year

Description

A mass extraction method of platycodin D < RTI ID = 0.0 >

The present invention relates to a method for extracting platycodin D in a large amount, and more particularly, to a method for extracting platycodin D in a large amount, Separating the leaves or roots of the bellflower and drying and pulverizing the mixture at a temperature of 50 DEG C or less, and extracting platycodine dyes from the powdered bellflower, .

Platycodi Radix is a medicinal herb that uses the roots of perennial bellflower Platycodon grandiflorum A. de Candolle belonging to the campanulaceae as a medicinal herb. Before the medicinal herb, the roots of the bellflower are washed with water, , As it is or by removing the cork layer, the dried surface is defined as the surface. Bellflower has been widely used in Korea, China and Japan, and is also widely used as a vegetable.

The oriental medicine efficacy of bellflower is mainly mentioned in Donguibogam, Dongbangjeongbang, Maternal Maternal, Divine Materia Medica, Oriental Medicine, etc. The number of Oriental medicine containing Rhododendrons is 49 prescription drugs and 287 prescriptions This is a useful herbal medicine to be recorded.

Bellflower is a traditional Chinese medicinal herb that has been widely used in Jinhae and Gadam. In addition to these uses, Bellflower extract has been reported to work on calming, fever, analgesic action, antiinflammation, anti-ulcer, hypotensive and anti-cancer effects. In particular, Choi et al. (2001) reported that in a mouse-based animal experiment, the water extract of Bellflower extract induced nitric oxide and tumor necrosis factor from mouse macrophages, And Han et al. (2001) reported that polysaccharides isolated from bellflower selectively activate B cells and macrophages, but not T cells, and demonstrate immunostimulatory effects. On the other hand, bellflower has been reported to be effective also in obesity because it is involved in fat metabolism and prevents accumulation of liver fat.

Platycodin A, C and D, which are triterpenoid saponins, have been identified as main active ingredients of bellflower, and inulin, betulin, stigmasterol ) Are included as main components. A study on platycodon was conducted by Tsujimoto for the first time in 1940, when platycodigenin, an aglycone of saponin, was reported in 1969 by Kubota et al. And Akiyama et al in 1972 by triterpenoid. And the structure of saponins in the system. In 1984, Ishii et al. Reported the structure of 17 new saponins from bellflower.

On the other hand, as a component representing the drug efficacy of the herbal medicine, the ingredient which exists only in the herbal medicine and represents the main activity of the herbal medicine is called the "indicator ingredient ", and the Korean Food and Drug Administration In the component analysis method development study, the index component of the bellflower was set to "platycodin D".

Arai et al. (1997) have shown that Gilyeong-tang, a mixed hot-water extract of bellflower and licorice, has the effect of alleviating chronic pancreatitis, one of the most persistent diseases, which is the main active ingredient of chronic pancreatitis Platycodin D, and platycodin D functions to secrete gastrointestinal hormones such as cholecystokinin (CCK) from the duodenum. Therefore, it is reported that Gyunggyang tang promotes gastric juice secretion in the pancreas. On the other hand, Ida et al. (1998) reported that the complete undegraded whole structure of platycodin D is essential for promoting the extra secretion of pancreatic secretion.

Saeki et al. (1999) compared and analyzed the contents of platycodin A, C and D in Chinese, Korean, Japanese, and Japanese cultivars distributed in Japan. Higashi et al. (1997) And the amount of platycodin D was determined.

Recently, several foods such as beverages, tea, candy, and powder have been developed and distributed based on various physiological activities of bellflower, but they are not able to form a product group targeting consumers of various ages.

Since there is no study on the variation of content of platycodin D as an index component, it is impossible to set the optimum harvesting date of the bellflower in terms of ingredients, and it is impossible to set the optimum quality of the bellflower raw material It is difficult to equalize.

Korean Patent Publication No. 10-2010-0022883

Choi CY, Kim JY, Kim YS, Chung YC, Seo JK, and Jeong HG. (2001b). Aqueous extract isolated from Platycodon grandiflorum elicits the release of nitric oxide and tumor necrosis factor-alpha from murine macrophages. Int. Immunopharmacol. 1: 1141-1151. Park, SH, Lee, KH, Lee, SH, Kim, HC, Kim, YS, Lee, HS and Kim, HM. (2001). Polysaccharide isolated from the radix of Platycodon grandiflorum selectively activates B cells and macrophages but not T cells. Int. Immunopharmacol. 1: 1969-1978. Higashiya, Arimoto K, Iwata Y, Ujita K, Kawai T, Kobayashi Y, Sakurai K, Shimada Y, Takagi A, Taniyama T. Nakajima KI, Hisata Y, Hosoda K. Yamamoto K. Yamamoto Y. and Noguchi M. 1997). Studies on the effects of preparations of platycodon root on its quality. Natural Medicines 51 (1): 56-62.

In order to solve the problems of the prior art as described above, it is an object of the present invention to provide a method for extracting and producing platycodine dodecane in large quantities.

It is another object of the present invention to provide a method for obtaining platycodine from platycodon in a simple and economical manner in a short time.

In order to accomplish the above object, the present invention provides a method for producing a rice seedling, comprising: harvesting 1 to 4 years old rhododendron around 2 to 3 months; Washing and harvesting the harvested bellflower, separating the leaves or roots of the bellflower, drying and pulverizing the blend at a temperature of 50 ° C or lower; And extracting platycodin D from the pulverized platycodon. The present invention also provides a method for extracting platycodin D in a large amount.

It is more preferable that the platycodine di is extracted from the crothelial layer formed of the cork layer of the rootstock.

It is further preferable that the thickness of the bellflower roots is 6 mm at the maximum.

Preferably, the drying is performed for 72 to 80 hours, and the drying rate of the bellflower is preferably 23 to 25%.

The step of extracting the platycodiandi is performed by using distilled water, ethanol, methanol, butanol, acetone, ethyl acetate, dichloromethane or the like at 4 to 80 ° C for 10 minutes to 48 hours . In particular, it is more preferable that the extraction is performed by an ultrasonic extraction method.

In addition, the present invention provides a pharmaceutical composition, a health supplement, and a cosmetic composition comprising platycodine di

According to the present invention, it is possible to obtain platycodine dianhydride contained in the bellflower in a simple and economical manner in a short time, and thus the method of the present invention is useful as a method of extracting and producing a large amount of high purity platocidyldi Can be used.

1 is a graph showing a building rate and a shrinkage rate according to an embodiment of the present invention.
FIG. 2 is a graph showing changes in the content of platycodin D according to the growth period according to an embodiment of the present invention.
FIG. 3 is a graph showing an HPLC chromatogram of an analytical sample at each growth stage according to an embodiment of the present invention.
FIG. 4 is a graph showing the building rates by the number of growing years according to an embodiment of the present invention.
FIG. 5 is a graph showing the shrinkage rate according to the number of years of growth according to an embodiment of the present invention.
FIG. 6 is a graph showing changes in the content of platycodin D according to the number of growing years according to an embodiment of the present invention.
FIG. 7 is a graph showing an HPLC chromatogram of an analysis sample according to the number of growing years according to an embodiment of the present invention.
Figure 8 is a graph showing the result of comparing the content of the Platycodin D (platycodin D) contained in the leaves, stems and roots of bellflower according to an embodiment of the present invention.
FIG. 9 is a graph showing the results of comparing the contents of platycodin D contained in the roots of the rootstock, the core and the cortex according to an embodiment of the present invention.
FIG. 10 is a graph showing a chromatogram of HPLC analysis of Broodstock root, referees and cortex according to an embodiment of the present invention.
11 is a graph showing the results of comparing the content of platycodin D according to the roots of 10-year-old rhizome according to an embodiment of the present invention.
FIG. 12 is a graph showing the results of comparison of drying time during drying at room temperature shade and 50 ° C thermal power according to an embodiment of the present invention.
FIG. 13 is a graph showing a variation in content of platycodin D according to a drying method after drying in different drying methods according to an embodiment of the present invention. FIG.
FIG. 14 is a graph showing HPLC / UVD chromatogram according to the drying method after different drying methods according to an embodiment of the present invention. FIG.

Hereinafter, the present invention will be described in detail.

The present invention aims to elucidate the optimal method for extracting platycodine dianhydride in large quantities by establishing a component optimum harvesting time by examining the content variation of platycodine diatoms according to the growth period and the number of growing years of the platycodon, We intend to contribute to the increase of farm income, the development of excellent processed products and the quality control through the production of good raw materials of the blooming cultivator.

The method of extracting platycodin D of the present invention in a large amount comprises: a step of harvesting the bellflower of about 1 to 4 years old at about 2 to 3 months, a step of washing and selecting the bellflower, Drying and pulverizing at a temperature of 50 ° C or less, and extracting platocidyldi from the powdered broth.

Hereinafter, a method for extracting platycodin D of the present invention in a large amount will be described in detail.

First, bellflowers with a growth age of 1 to 4 years are selected, and they are harvested around February to March and used as bellflower feedstock.

In the present invention, in order to establish an optimum harvesting period in which a large amount of platycodine dyes can be extracted from the bellflower, 1 to 10 years of bellflower seedlings having the same genetic background grown under the same cultivation conditions are harvested on the same date, , And the variation of platocidyldi content according to the years of growth was measured to determine the optimum growth period of platycodine digestion. In addition, 6-year-old bellflower seedlings grown on the same cultivation conditions with the same genetic background were harvested on the same date every month (January to December) and selected as a sample. The content of platcordine di And the optimum growth period and growth period for platycodine diatomic extracts were investigated.

That is, in the present invention, the optimum growth period and the growth period, that is, the optimal harvesting period, in which platycodine is highly pure and can be obtained in large quantities, are established, and the platycodon seeds harvested at this optimum harvesting period are selected, It is characterized by being used as a sample for extraction.

Therefore, in order to extract a large amount of platycodine dyes, it is preferable to use the bellflower harvested in about 2 to 3 months of the 1 to 4 years old bellflower.

The selected platycodon is washed and cleaned, and then only leaves or roots of the platycodon are separated from each other.

In general, platycodine diatoms of bellflower are evaluated as the main components of roots. In the present invention, in order to confirm whether platocidyldi exists in other parts than the bellflower roots, the leaves, stems and roots underground are used as samples As a result, it was confirmed that the content of platocidyldi was distributed in the leaf or root of the plant part of the bellflower. Therefore, in the present invention, it is preferable to use a leaf or a root of a plant part of a bellflower as a sample in order to extract a large amount of platycodine.

Particularly, among the root portions of the bellflower, it is more preferable to use a dripped portion formed of a dripped portion, that is, a corked layer, except for the main portion of the original root. It is better to extract a greater amount of platycodine dyes when using the crotch region formed from the cork layer rather than the lignified central region of the root of the bellflower.

Further, it is more preferable that the root of the bellflower has a smaller root diameter, preferably a root thickness of 6 mm or less, more preferably 2 to 6 mm, because it contains a large amount of platycodine.

After the leaves and roots of the bellflower are separated, drying and pulverization are carried out.

Generally, medicinal crops used as medicinal herbs are described to be dried at a temperature of 60 ° C or less, unless otherwise specified. In the case of herbal medicines, it is customary to use a room temperature shade at 25 ° C or lower have.

However, in the present invention, it is preferable to perform thermal power drying at a temperature of 50 DEG C or less so as to maximize shortening of the drying time without loss of useful components contained in the platycodon. At this time, when the temperature is higher than 50 ° C. during the thermal power drying, the content of the platycodine which can be obtained by the loss of the useful component contained in the bellflower, ie, platocidyldi, can be reduced. Inappropriate.

The dried dodder is then pulverized, wherein the dodder powder is preferably 20 to 300 mesh. When the size of the bellflower powder is within the above range, the extraction efficiency is improved and the extraction filtrate removal process is better.

Thereafter, platocidyldi is extracted from the powdered broth.

Specifically, the platycodine di is dissolved in distilled water, ethanol, methanol, butanol, acetone, ethyl acetate, dichloromethane and the like in an amount of 10 to 50% by weight (w / v) An intermediate polar organic solvent or the like is used as an extraction solvent and subjected to a conventional extraction method such as a low temperature extraction method, a room temperature extraction method, an ultrasonic extraction method or a reflux extraction method at 4 to 80 ° C for 10 minutes to 48 hours, After filtration, the filtrate can be extracted by freeze-drying, room-temperature drying or hot-air drying.

The extraction solvent may be distilled water, a lower alcohol and an intermediate polar organic solvent, or a mixture of two or more thereof. Especially, distilled water, ethanol, or a mixture thereof is preferably used.

The platycodyldi that has been extracted as described above can be isolated in high purity through various conventional separation methods as necessary. Separation methods such as solvent distribution, precipitation separation, recrystallization, and column chromatography can be used as separation methods usable at this time.

Particularly, it is more preferable to extract for 2 hours by ultrasonic extraction method, specifically ultrasonic extraction method, from pulverized platycodon in view of optimum extraction efficiency.

Generally, even if the same extraction solvent is used under the same conditions, or when the same extraction solvent is used and the same extraction method is used and the extraction is performed according to time, the extraction efficiency varies depending on each component to be extracted. In the present invention, The optimum conditions for the extraction of platycodine dianhydrides from platycodon have not been investigated until now. Especially, the optimal harvesting time and the freshly harvested platycodon from this period It is obvious that the present invention is a completely unexpected technique because it is a technique that can not be found anywhere about a method of obtaining platcordine dian in large quantities.

In the present invention, the content of platycodine di-extracted from the bellflower was evaluated by HPLC / UVD / MS or ELSD analysis. At this time, the HPLC column for the analysis of the platelet-containing platycodine diad may use a reversed phase (C18) chromatography column, and acetonitrile (ACN) and formic acid may be used as the mobile phase.

Therefore, according to the extraction method of the present invention as described above, it is possible to extract a large amount of platocidyldi from bellflower, and also to efficiently extract and separate high-purity platocidyldi by a simple and economical method in a short time have.

The present invention provides a pharmaceutical composition comprising platycodine diese extracted from platycodine as an active ingredient as described above, wherein the pharmaceutical composition comprising platycodine dyes as an active ingredient is particularly useful as an antioxidant, Obesity and anti-hyperlipemia, and anticancer activity.

The platycodiamine is preferably contained in the pharmaceutical composition in an amount of 0.05 to 20% by weight, more preferably 0.05 to 10% by weight. If the content is less than 0.05% by weight, the effect of the pharmaceutical composition may be insignificant. If the content is more than 20% by weight, the pharmaceutical effect may be relatively low compared to the usage amount.

The pharmaceutical composition containing platycodine dyes extracted from the phloem of the present invention as an active ingredient can be administered orally or parenterally. The route of administration of the pharmaceutical compositions of the present invention may be, but is not limited to, oral, intravenous, intramuscular, intraarterial, gingiva, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, , Sublingually or topically.

For such clinical administration, the pharmaceutical composition of the present invention may be formulated into a suitable formulation, such as oral administration, parenteral administration, or injection, using known techniques. For example, upon oral administration, it may be admixed with an inert diluent or edible carrier, sealed in a hard or soft gelatin capsule, or pressed into tablets. For oral administration, the active compound may be formulated as an ingestible tablet, buccal tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, suspensions, Hard or soft capsules, elixirs, suspensions, syrups, wafers, and the like. At this time, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin for formulation into tablets, capsules and the like; Excipients such as dicalcium phosphate; Disintegrants such as corn starch or sweet potato starch; Magnesium stearate, calcium stearate, sodium stearyl fumarate, or a lubricant such as polyethylene glycol wax may be mixed. In the case of a capsule formulation, a liquid carrier such as fatty oil may be mixed in addition to the above-mentioned substances.

Parenteral administration is also applied to subcutaneous injections, intravenous injections, intramuscular injections, intra-thoracic injection injections, mucous membranes, or topical application, including dispersing agents, suppositories, powders, aerosols (nasal sprays or inhalants), gels, , Or a non-aqueous liquid suspension, an oil-in-water emulsion or a water-in-oil emulsion), a solution, and the like. For formulation into a parenteral dosage form, the composition may be formulated into a solution by mixing with a stabilizer or buffer in water to form a unit dosage form of ampoule or vial.

The effective dose of the pharmaceutical composition of the present invention as described above can be adjusted to a necessary range according to the clinical judgment in consideration of the patient's age, physical condition, and weight. Generally, the effective dose of the pharmaceutical composition is 1 to 20 mg / day, preferably 5 to 10 mg / day, per 1 kg body weight of an adult patient, Preferably two to five times a day.

The present invention also provides a health supplement containing, as an active ingredient, platycodine dianhydride extracted from the bellflower, wherein the health supplement contains the active ingredient platycodine extracted from the widely used edible bellflower Therefore, it is safe for human body and is suitable for use in food.

The health supplements containing the platycodine di-compound as an active ingredient have antioxidant, anti-inflammatory, anti-obesity and anti-hyperlipidemic activity, anti-cancer activity, Is particularly preferable.

The amount of the platycodi di is preferably 0.05 to 10% by weight, more preferably 0.05 to 5% by weight, in the health supplement. If the content is less than 0.05% by weight, the effect as a health supplement may be insignificant. If the content is more than 10% by weight, the health promotion effect as a health supplement food may be relatively low compared to the addition concentration.

The kind of the health supplements is not limited. For example, the health supplements may be added to conventional palatable foods such as noodles such as ramen noodles, raw noodles, tofu, cereals, breads, chewing gum, candies, And can be used as a food coloring matter. In addition, it can be formulated into a common formulation such as tablets, granules, pills, hard capsules, soft capsules or liquid formulations, and can be prepared as a juice, a pouch, a drink, or the like. It goes without saying that components other than the above-mentioned components can be appropriately selected and blended by those skilled in the art according to the formulations.

In addition, the platycodine dianthate contained in the health supplement food of the present invention can effectively prevent the change of the smell and flavor of the food caused by oxidation, the rancidity of the fattening, and the discoloration of the food, It can also be used to preserve foodstuffs or to maintain freshness and quality of food over a long period of time.

Such foods include not only typical foods but also beverages (including alcoholic beverages), fruit and processed foods thereof (e.g., canned fruits, jars, jam, maare marlade, etc.), fish, meat and processed foods Ham, sausage, corn oil, etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, Vegetable protein, retort food, frozen food, various seasonings (e.g., soybean paste, soy sauce, soy sauce, etc.), and the like.

In addition, the present invention provides a cosmetic composition comprising platycodine diese extracted from bellflower as an active ingredient.

The amount of the platycodine is preferably 0.01 to 5% by weight, more preferably 0.01 to 2% by weight, based on the cosmetic composition. When the content is less than 0.01% by weight, the effect as a cosmetic may be insignificant, and if the content is more than 5% by weight, the effect as a cosmetic relative to the concentration of the additive may be relatively low.

In accordance with the present invention, platycodiamine extracted from the bellflower has antioxidant, antiinflammatory, anti-obesity and anti-hyperlipemia and anticancer activity. Especially when it is applied to a cosmetic composition, it is used for antioxidant, whitening, antiallergic, .

The cosmetic composition of the present invention can be used as it is or as diluted if necessary. The platycodine dianhydride may be prepared in liquid or solid form using bases, adjuvants and additives commonly used in the cosmetics field. The liquid or solid form of cosmetics may include, for example, but not limited to, lotions, creams, lotions, bath salts, and the like.

The base, adjuvant and additives commonly used in the cosmetics field are not particularly limited, and examples thereof include water, alcohol, propylene glycol, stearic acid, glycerol, cetyl alcohol, liquid paraffin and the like.

The cosmetic composition of the present invention as described above is very useful for preventing damage due to oxidation of the skin, for example, spots (brown half), freckles, skin cracks, and ultraviolet damage (sunburn) It is very useful to maintain the quality of cosmetics.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example 1. Measurement of variation of platycodine di content according to growing time

(1) Building rate and contraction rate

Gangwon-do The gypsies of Samcheok City were harvested on the 20th day of each month from January to December 2011 and used as the samples.

The prepared sample was washed and cleaned, and thermal power was dried at 50 DEG C or less, and the building ratio and shrinkage ratio were measured. The results are shown in Fig. 1 and Table 1 below.

Growing season Building Rate (%) Shrinkage (%) January 22.9 31.1 February 20.3 31.6 In March 19.1 35.4 April 15.7 36.8 In May 18.3 34.4 June 19.3 33.8 In July 22.9 33.0 August 22.7 32.9 September 22.1 32.0 October 22.0 32.5 November 23.6 32.1 December 22.2 31.0

As shown in Fig. 1 and Table 1, it can be seen that the storage rate and shrinkage percentage of the storage of the platycodon grandiflorum harvested at different growing periods from January to December are significantly different depending on the growing time.

First, the building rate is about 22 ~ 23% from the beginning of July, which is the period when the flowering period of the top part of the bellflower, that is, the growth of the terrestrial nutrition, , But after the emergence of the buds on the ground, the overgrowth of the ground is developed by the consumption of the nutrients accumulated in the underground reservoir until the development of the overgrown growth and before the independent nutrient growth is satisfied, that is, before the complete development and formation of the leaves, By April, the storage rate of the storage tank showed a rapid decrease, which was the lowest at 15.7% in April. Since then, as the photosynthetic ability of the above-ground part has been gradually increased, the amount of nutrients required for nutrient growth has been gradually increased and the building rate has been gradually increased. 18.3% in May, 19.3% in June and 22.9% in July , And it was confirmed that the building rate was almost constant during the following period. This phenomenon is due to the fact that the top part of the bellflower is transferred to the reproductive growth, and the leaves and the top part are damaged. Therefore, it is impossible to synthesize new nutrients, and even if it is synthesized, the nutrients required for life maintenance such as breathing are consumed. It was predicted that it would be impossible.

In other words, when considering the weight after drying in determining the harvest period of the bellflower, the yield rate was almost constant from July to February, so that there was no decrease in yield due to drying after harvest.

On the other hand, the shrinkage rate also markedly decreased from May to February of the next year, and remarkable decrease of shrinkage was observed until April (36.8%) just before the independent growth of the shoots was observed. Of the total population. Considering the fact that the underground building rate was the lowest in April, it seems that the photosynthesis anabolism product of the upper part of the ground was accumulated as a stockyard rather than the accumulation of the nutrients needed for growth of the above ground from the storehouse until April. At this time, It was judged that the shrinkage rate was greatly decreased due to the supply for growth, and the shrinkage rate decreased again after May.

(2) Platycodin D content

Dried granules were prepared in a powder state of 60 mesh. Then, 30 mL of distilled water as an extraction solvent was added to 1 g of the powder, and the mixture was extracted with an ultrasonic extractor at 40 ° C for 2 hours, followed by primary filtration with Whatman No. 2 filter paper , And secondary filtration was performed with a syringe filter (0.45 mu m) immediately before HPLC analysis. The content of the Platicodin Di was evaluated by HPLC / UVD / MS or ELSD analysis, HPLC column was used for reverse phase (C18) chromatography column, HPLC / UVD / MS and ELSD were Agillent 1200 series (USA) and 6100 Quadrupole MS (USA) was used. The flow rate was 1.0 mL / min, the column temperature was 30 ° C, and the sample injection volume was 20 μL. The mobile phase conditions used for HPLC analysis were two phase gradient and linear concentration gradient method using acetonitrile (ACN) as organic solvent. The HPLC / MS analysis conditions are shown in Table 2 below, and the experimental results are shown in FIG.

LC / MS system Agilent 6110 Quadrupole LC / MS Column Imtakt C ₁₈ (250 ㅧ 4.6 mm ID., 5 탆, Japan) Column temp. 30 ℃ Mobile phase 0.1% formate-H ₂ O: 0.1% formate-ACN Flow rate 1.0 mL / min Sample size 10 μl Ionization ESI Negative-ion mode Gas temp. 350 ℃ Drying gas 10 mL / min Capillary voltage 3.5 kV Mass range (m / z) 500 to 1,500

As shown in FIG. 2, the content of platycodine di was measured from the October of the previous year, that is, the content of platycodine, which is a main component of the platycodon, . From January to February of the next year, there was no significant change in content, and it was 0.24 ~ 0.26%. In the preparation stage of the undergrowth, , The content of platycodine di was increased rapidly to about 0.34% and showed the highest content in one year. Platycodine diatoms were also found in about 0.20% of the early leaves completely unfolded on the ground in April, probably related to the physiological plant growth associated with development of the superficial lobes from the bladder from around February after wintering I could infer that there would be.

As the growth of the above-ground growth progressed, the growth rate was 0.31% in April, 0.24% in May and June at the peak season, and 0.20% at the flowering season from July to September. Especially from July, when flowering began, to September, when the growth was advanced, and remained at 0.25% from October when flowering, which is the next year, was formed.

From the above results, there was no significant difference in the content of platycodine in stored root between October and January, which is the conventional harvest season during the year, The content of platycodine was increased rapidly, and it was considered that harvesting should be carried out around 2-3 months in terms of the content of platycodine in the absence of problems such as drying,

On the other hand, the content of platycodine di in dried platycodon was varied according to the growth stage, and the content of platycodine in 100g of platycodon root , And the results are shown in Fig. 3 and Table 3 below.

Growing season Dry muscle content (%) Building Rate (%) Crude muscle content (mg / 100g) January 0.26 22.90 59.72 February 0.34 20.27 68.56 In March 0.35 19.14 66.82 April 0.31 15.72 48.31 In May 0.24 18.32 44.47 June 0.24 19.27 45.76 In July 0.19 22.86 42.45 August 0.19 22.72 42.51 September 0.20 22.06 44.18 October 0.25 22.03 55.53 November 0.24 23.60 55.75 December 0.25 22.24 54.70

As shown in Table 3, the highest content of platycodine diad in about 100 mg of platycodine was obtained in February, and the highest content was found in platycodine di The content was decreased to 42.5mg from July to August in the flowering period. In conclusion, the dry matter distribution of dried rhododendrons shows no significant difference in the dry matter yields of dry Rhododendrons from July to January of the following year, Considering the content of platycodine which can be extracted, the highest level of about 67 mg per 100 g of raw muscle of March to March is obtained. Therefore, the optimum harvest time of raw root used for processing or platycodine is 2 ~ March was the most optimal.

As shown in FIG. 3, the HPLC chromatogram of the blooming period of the blooming period showed that the peaks of the specific components were not detected or disappeared according to the difference in growth period, but only the component content evaluation And there were no other singularities.

Example 2. Measurement of Platycodine Di Content Variation According to Growing Years

(1) Building rate and contraction rate

Gangwon-do The ginseng broodstock of Samcheok City, Gangwon-do, was harvested on October 13, 2011 for each year, and the 1 ~ 10-year-old bellflower brood grown on the same cultivation condition was used as a sample.

The prepared sample was washed and cleaned, and thermal power was dried at 50 DEG C or less, and the building ratio and shrinkage ratio were measured. The results are shown in Figs. 4 to 5 and Table 4 below.

Growing years Fresh root (g) Dry root (g) Building Rate (%) 1 year old 3.3 0.8 24.1 2 years 7.4 1.5 20.7 3-year-old 7.0 1.6 23.4 4 years old 1.4 3.6 26.8 Five years 19.0 3.7 19.6 Six years 28.1 6.0 21.5 7 years old 35.3 8.7 24.8 8 years 48.6 10.1 20.8 9 years old 76.2 14.5 19.0 10 years 90.1 19.8 22.0

As shown in Table 4 and FIG. 4, the mean dry weight of the four-year-old specimens was the highest at 26.8% in terms of the number of years of growth, And 19.0% for 9 year olds, respectively, and the samples of other growth years ranged from 20 to 25%.

On the other hand, when we look at the mean weight change of rootstock root and dry root, we can see that the weight increases with increasing number of years of growth. In particular, the rate of dry matter growth by year is similar, And the weight of the water contained therein was found to be larger.

As shown in FIG. 5, the average shrinkage ratio of the samples was 58.5% for one year, and the samples for other growth years ranged from 60 to 71%.

(2) Platycodin D content

Dried granules were prepared in a powder state of 60 mesh. Then, 30 mL of distilled water as an extraction solvent was added to 1 g of the powder, and the mixture was extracted with an ultrasonic extractor at 40 ° C for 2 hours, followed by primary filtration with Whatman No. 2 filter paper , And secondary filtration was performed with a syringe filter (0.45 mu m) immediately before HPLC analysis. The content of the platycodine di was evaluated in the same manner as in Example 1, and the results of the experiment are shown in FIG.

As shown in FIG. 6, the content of platycodine, which is the main component of the bellflower, was quantitatively evaluated by HPLC after the dried bellflower samples were prepared in the form of powder, The content of 1 - year - old root was the highest at 0.45% and showed a declining trend with the progress of growing years. The 10 - year root was the lowest at 0.21%.

Generally, bellflower is considered to be difficult to grow for more than 3 years because the root rot disease occurs seriously as the number of years of planting after the planting increases in one area. Thus, the old bellflower is considered to be relatively precious, It is estimated that the medicinal value of the bellflower is high, so that the value of the bellflower bellflower, that is, the old bellflower, is highly appreciated.

However, in the content of platycodine dianthocyanidins, which are evaluated as main components of platycodon, the content of platycodin is increased in long - term broodstock, I could confirm.

In general, the main component of medicinal crops using roots is known to contain a major component in the cortex, ie the cork layer, than the core of the root (woody part). The ratio of the percentage of the cortex The higher the proportion of the crustacean, that is, the lower the number of years of growth, the higher the content of platycodine.

On the other hand, the content of platycodine di contained in 100 g of corn borer root was calculated by converting the platycodine di content of the borage, the planting rate of the borage, .

Growing years Dry muscle content (%) Crude muscle content (mg / 100g) 1 year old 0.45 108.89 2 years 0.43 88.00 3-year-old 0.41 96.20 4 years old 0.37 100.36 Five years 0.33 64.81 Six years 0.36 78.45 7 years old 0.36 88.39 8 years 0.30 63.46 9 years old 0.29 55.84 10 years 0.21 46.72

As shown in Table 5, the content of platycodine diad was about 109 mg in 100 g of first-year-old bellflower, and the content of platycodine in the raw soybean was significantly And the 10 year old muscle was the lowest at 46.7 mg.

In conclusion, the content of platycodine di was highest in dry root and raw root in 1 ~ 4 years.

On the other hand, the HPLC chromatogram of the seedlings analyzed for the number of years of cultivation of the platycodon was shown in Fig. 7, and it was not confirmed whether the peak of the specific component was detected or extinguished even when the growth years progressed, And only the size of the peak which is a measure of the constituent content was different, but no other singularity was found.

Example 3: Comparison of content of platycodine di

In order to confirm whether platycodine diegy which is evaluated as the main component of the bellflower root is also present in other parts except for the root, 6-year old bellflower was harvested and washed in June, and the leaves, stems and roots of the ground were dried respectively, The content of platycodine di was quantitatively evaluated, and the results are shown in Fig. At this time, the content of platycodine di was evaluated in the same manner as in Example 1 above.

As shown in FIG. 8, the content of platycodine di was highest in leaves of 0.46%, 0.04% in stem, and 0.23% in root. On the other hand, since the flower was not bloomed in June, only the flower part was further analyzed in July, so that platycodine was not detected in both purple and white flowers.

These results suggest that fliticodine diatoms are present in large amounts on the leaves of platycodon, in the stem, and in the root, relatively large amounts are detected in the platycodon, And that it is transferred to the root through the stem and is stored permanently in the root of the root.

Example 4: Content of Platycodine di

In general, it is known that the main component of medicinal crops using roots contains a major component in the cortex, that is, the cork layer, rather than the core of the root (woody part). In order to verify the difference in the content of platycodine contained in the cortex, that is, in the cortex than in the core of the root, the content of platcordine di Were analyzed by HPLC analysis chromatogram, and the results are shown in FIGS. 9 and 10. FIG.

As shown in Figs. 9 and 10, the content of platocidyldi in the raw roots, the referees and the cortex was 0.30% for the roots, 0.28% for the urn, 0.34% for the cortex It was confirmed that the content of platycodine di was higher in the cortex area formed by the cork layer than the refined core area which was clearly irradiated.

Example 5: Comparison of the content of platycodine with respect to the thickness of bellflower roots

On the other hand, in order to examine the difference in the contents of platycodine diatoms according to the difference in the root thickness of the seedlings grown in the same individual, 10-year-old seedlings were harvested in June and classified into 5 groups according to the root thickness And the content of platidomine was quantitatively analyzed. The results are shown in FIG.

As shown in FIG. 11, the content of platycodi di was lowered as the roots were thicker, and the root with the lowest root of 2 mm or less showed the highest pattern with an average of 0.49% The content of 2 to 6 mm was 0.32%, 7 to 9 mm was 0.28%, 10 to 15 mm was 0.17%, and 16 to 28 mm was 0.12%. These results were similar to those of other pharmacological effects. Platycodin was found to be more active when the growth years were lower, ie, the root thickness was smaller.

Table 6 shows the results of the comparison of dry matter rate and shrinkage ratio after drying using 10-year-old roots of each roots.

Root thickness 16 to 28 mm 10-15 mm 7 to 9 mm 2 to 6 mm 2 mm or less Average Building Rate (%) 18.3 18.0 17.2 25.7 Impossible to investigate 19.8 Shrinkage (%) 57.0 57.4 55.2 61.9 Impossible to investigate 57.9

Since it is not possible to determine the thickness of each root as an exact number of years, it is impossible to directly compare directly with the number of years of growth. However, as shown in Table 6, the result is similar to the result of a rough year of growth, When the average building rate and shrinkage rate were compared with the 10 - year - old building rate and shrinkage rate, it was found that they showed almost similar pattern.

Example 6: Comparison of the content of platycodine with respect to the drying method of bellflower roots

In general, medicinal plants used as medicinal herbs are described as drying at 60 ° C or less unless otherwise specified. In the case of herbal medicines, it is customary to use the shade at room temperature at 25 ℃ or below, and the thermal drying is often performed for the purpose of quick drying recently. Therefore, in this example, the time required for drying in a normal temperature shade and drying temperature of 50 ° C. thermal power using the October harvest sample, which is one of the samples of the growing season, was compared and the results are shown in FIG.

As shown in Fig. 12, it took 14 days to dry until the building rate reached 50% in the case of the room temperature sound, and until the drying time (building rate 22.5%) in which no further change in the building rate occurred It took about 58 days after. On the contrary, the shrinkage rate was about 55 days after drying, until the root thickness was not changed any more. The average shrinkage rate at this time was 59.4%.

On the other hand, in the case of 50 ℃ thermal drying, it takes about 19 hours after the drying until the building rate reaches 50%, and 78 hours (6 days in 3 days) . In addition, the shrinkage rate did not change due to complete drying after 54 hours (2 days and 6 hours) after drying, and the average shrinkage rate at this time was 58.0%.

As a result, it takes at least 58 days for complete drying of Rhododendron root at room temperature shade condition, but it takes 6 hours (78 hours) for 3 days at 50 ℃ when using thermal power for rapid drying, It can be confirmed that when the thermal power drying is performed at a temperature of 50 캜 as in the present invention, the drying time of about 55 days or more can be shortened.

In order to investigate the variation of content of platycodi dianthus major component depending on the drying method, freeze drying, room temperature shade, drying at 50 ° C and 80 ° C, and drying at 121 ° C were carried out using 6 - year - , The content of platycodine di contained in the bellflower roots was compared and analyzed. The results are shown in Figs. 13 and 14.

As shown in FIG. 13 and FIG. 14, the content of platycodine di was the highest at 0.23% when dried with a room temperature sound gun, followed by 50 ° C thermal drying, freeze drying, 80 ° C thermal drying, The content of platcordine di was decreased with increasing temperature at above room temperature. The content of platcordine di was decreased with increasing temperature. In the experiment for comparing the days required for drying, it was found that the thermal drying at 50 ° C can drastically shorten the drying time. In the variation of the content of platycodine according to the present drying method, Although the thermal drying showed lower contents than the normal temperature shade, statistically, no difference in contents was recognized. Therefore, when thermal drying is required during drying of raw green tea, when the thermal drying of less than 50 ° C is carried out, And it was confirmed that it would be suitable for use.

From the above results, it can be seen that the bellflower harvested from about 1 to 4 years old in the present invention was harvested from February to March, and the leaf or root portion thereof was separated and dried and pulverized at a temperature of 50 ° C or lower It has been confirmed that when platycodine dianhydride is extracted from the bellflower, it is possible to extract and produce a high-purity platycodine dianhydride in a short time in a simple and economical manner. In addition, according to the present invention, it is possible to establish an optimum harvesting period of the bellflower on the component side, which is expected to be industrially applicable.

Preparation Example 1 Cream Preparation

Three year old bellflower seedlings were cultivated in February, and the roots of the harvested bellflower seedlings were washed and dried at a temperature of 50 ° C or less for 70 hours After completely dried, it was pulverized to 60 mesh. 6 L of distilled water as an extraction solvent was added to 200 g of the above-mentioned platycodon powder, and the mixture was extracted with an ultrasonic wave extender at 40 ° C for 2 hours, and extracted three times in the same manner. 2 filter paper and syringe filter (0.45 ㎛), followed by freeze-drying of the filtrate. The dried filtrate was subjected to solvent partitioning and C18 column chromatography to obtain high purity platycodine di Respectively.

1.0% by weight of platidic anhydride, 2.2% by weight of acetic acid, 2.8% by weight of bentonite, 4.0% by weight of butylene glycol, 3.0% by weight of phenoxyethanol, 2.0% The cream was prepared by mixing 1.2 wt% of paraben, 1.2 wt% of butylparaben, 1.1 wt% of ethylparaben, 1.0 wt% of isobutylparaben, 1.0 wt% of propylparaben, and a remaining amount of purified water.

Preparation Example 2 Pack Preparation

0.5% by weight of platycodine, 3.0% by weight of ethyl alcohol, 0.05% by weight of EDTA-2Na, 5.0% by weight of propylene glycol, 4.5% by weight of glycerin, 1.5% %, A small amount of flavor, preservative, and a remaining amount of purified water were mixed and packed according to a conventional method.

Preparation Example 3 Preparation of Ointment

2.0% by weight of platycodine di obtained in Production Example 1, 10% by weight of wax, 5.0% by weight of Polysorbate 60, 2.0% by weight of Cigarette hardened castor oil, 0.5% by weight of sorbitan sesquioleate, % By weight, and residual vaseline were mixed to prepare an ointment agent according to a conventional method.

Preparation Example 4 Powder Preparation

5.0% by weight of platycodine di, 0.1% by weight of vitamin A and 0.1% by weight of vitamin E were obtained in the same manner as in Preparation Example 1, And the powder was mixed according to a conventional method.

Preparation Example 5 Liquid Preparation

A mixture of 5% by weight of platycodine, 25.0% by weight of glucosamine salt, 20.0% by weight of chondroitin sulfate, 10.0% by weight of collagen, 5% by weight of vitamin C, 0.5% by weight of vitamin A, 0.5% by weight of vitamin E, 1% by weight of aspartame and the remaining amount of purified water were mixed to prepare a liquid agent by a conventional method.

Production Example 6. Production of functional beverage

2.0% by weight of platycodine di obtained in Preparation Example 1, 15.33% by weight of isomerized sugar, 0.1% by weight of citric acid, 0.04% by weight of vitamin C, 0.1% by weight of perfume, 15.0% by weight of the composition of Example 2, Were mixed to prepare a functional beverage according to a conventional beverage preparation method.

Preparation Example 7 Skin Lotion

0.5% by weight of platycodine diborate obtained in Preparation Example 1, 2.5% by weight of camphor oil, 5.2% by weight of 1,3-butylene glycol, 1.5% by weight of oleyl alcohol, 3.2% by weight of ethanol and 3.2% by weight of Polysorbate 20 2.0% by weight of benzophenone-9, 1.0% by weight of a carboxyl vinyl polymer, 3.5% by weight of glycerin, and purified water of a small amount of fragrance, preservative and remaining amount were mixed to prepare a skin lotion according to a conventional method.

Preparation Example 8 Milk Lotion (Nutrition Lotion)

1.0 wt% of platycodyldi, 1.0 wt% of cetostearyl alcohol, 0.8 wt% of glyceryl monostearate, 0.3 wt% of sorbitan monostearate, 0.1 wt% of captopyl paraben obtained in Preparation Example 1, A mixture of 1.0 wt% of Bait 60, 5.0 wt% of mineral oil, 3.0 wt% of cyclomethicone, 0.5 wt% of dimethicone, 0.1 wt% of allantoin, 5.0 wt% of glycerin, 2 wt% of alcohol, 3.0 wt% of propylene glycol, Preservative, and purified water of the remaining amount were mixed to prepare a milk lotion (nutrition lotion) according to a conventional method.

Preparation Example 9 Essence

0.5% by weight of platycodine, 4.0% by weight of 1,3-butylene glycol, 10% by weight of glycerin, 0.5% by weight of allantoin, 0.1% by weight of panthenol, 0.01% by weight of EDTA-2Na, By weight of the composition, 1.5% by weight of triethanolamine, 2.0% by weight of squalane, 2.5% by weight of beeswax, 3.5% by weight of polysorbate 60, 1.0% by weight of carboxyl vinyl polymer, 2.5% by weight of sorbitan sesquioleate, The remaining amount of purified water was mixed to prepare an essence according to a conventional method.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Claims (12)

1 to 4 years old bellflower is harvested around February to March;
Washing and selecting the harvested bellflowers, separating the leaves or roots of the bellflowers, thermally drying and powdering at a temperature of 50 ° C. or lower; And
Extracting platicodine di from the powdered bellflower;
Platycodin D (platycodin D) characterized in that it comprises a method for mass extraction. The method of claim 1,
The method for extracting a large amount of placodin di, characterized in that the placodin di is extracted from the cortex formed of the cork layer of the bellflower root. The method of claim 1,
The thickness of the bellflower root is a method of extracting a large amount of placodine di, characterized in that up to 6mm. The method of claim 1,
Characterized in that the drying is carried out for 72 to 80 hours (23 to 25% of dry matter of the platycodon). The method of claim 1,
Extracting the Platicodin Di, 10 minutes to 48 hours at 4 to 80 ℃ using any one or more solvents selected from distilled water, ethanol, methanol, butanol, acetone, ethyl acetate and dichloromethane in the powdered bellflower Method for extracting a large amount of Platicodin D, characterized in that during the extraction. The method of claim 5,
Wherein said extraction is performed by ultrasonic extraction. A pharmaceutical composition comprising platicodine di extracted by the method of claim 1 as an active ingredient. The method of claim 7, wherein
The platicodine di is a pharmaceutical composition, characterized in that contained in 0.05 to 20% by weight in the pharmaceutical composition. A dietary supplement comprising the platyodidine di extracted by the method of claim 1 as an active ingredient. 10. The method of claim 9,
The platyodidine di is a dietary supplement, characterized in that contained in 0.05 to 10% by weight in the dietary supplement. A cosmetic composition comprising a platicodine di extracted by the method of claim 1 as an active ingredient. 12. The method of claim 11,
The platinum composition Di is a cosmetic composition, characterized in that contained in 0.01 to 5% by weight in the cosmetic composition.

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