KR100552991B1 - Extraction and Purification of Active Active Ingredients from Jinkyo and Herbal Medicine Compositions Containing the Extracts - Google Patents

Abstract

The present invention relates to a method for extracting and purifying an active ingredient from orthodontics, and to a herbal composition containing the extract. More specifically, the active active ingredient can be efficiently extracted from orthodontics, which is a herbal medicine, using an aqueous alcohol solution and a nonpolar solvent. The present invention relates to herbal compositions such as anti-inflammatory analgesics and methods containing the extracts.

Description

Extraction and Purification Method of Active Active Ingredients from Jinkyo Bridge

The present invention relates to a method for extracting and purifying an active ingredient from orthodontics, and to a herbal composition containing the extract. More specifically, the active active ingredient can be efficiently extracted from orthodontics, which is a herbal medicine, using an aqueous alcohol solution and a nonpolar solvent. The present invention relates to herbal compositions such as anti-inflammatory analgesics and methods containing the extracts.

Jinkyo refers to the roots of perennial herbaceous plants of the cognate plant of the genus Ranunculus, which grows in the dense ground in the bushes all over Korea. It is a toxic herbal medicine that has been cut and dried in the sun and used as a medicine. In oriental medicine, the same plant has been used for the treatment of analgesic, dysmenorrhea, customs pain, arthritis, yellowing, and urinary bulge. In recent years, in addition to the above-mentioned effects, it is known to have an effect on lowering blood pressure, antibacterial action against various infectious diseases, analgesic and inflammation due to increased secretion of corticosteroids. Lycoctonine, abadharidine, septinriodine are known as active ingredients contained in the orthodontic and cognate root plant, and other than gentianine and gentoianicine. Alkaloids, such as, and aromatic compounds, such as methyl-n- (3-carbamoyl propionyl) anthranilate and methyl-n- (2-acetaminobenzoyl) anthranilate, are known. , p477-478 (1990); Chinese Medicine Dictionary, Book Publishing Decision, p5171-5177 (1998)].

Regarding the medicinal properties of jinkyo extract, the traditional herbal medicines such as Dongbobogam, medicinal herb and medicinal herb, and related literature only describe some of the herbal medicinal medicinal effects of jinkyo extract prepared by the traditional method of medicinal herbs. Analytical studies on the active ingredients and studies on efficient extraction methods of the active ingredients are very small, and there have been no studies published on the efficient extraction methods of fractions containing a large amount of the active ingredient.

Therefore, the present inventors separated and prepared the shingles fraction by a simple manufacturing method in order to use more scientifically known anti-inflammatory effect not only to have anti-inflammatory analgesic effect, but also to improve nasal sensation due to wind, cold, and wet, and the fraction has excellent anti-inflammatory analgesic activity. It was found that the present invention has been completed. The present invention has a purpose of providing a simple method for producing an excellent anti-inflammatory analgesic activity from Jingyo and its use as an anti-inflammatory analgesic agent using this fraction, and the present invention has been used only in the form of medicinal herbs, etc. The present invention has been completed by developing the extract of jinhwa into a formulation of tablets and injections for easy administration.

The present invention in the preparation of Chinese herbal medicines by extracting the Chinese onions with water or alcoholic aqueous solution,

Extracting and filtering the fibrils with water;

Separating the filtrate obtained in the extraction filtration step with 0.7 to 1.2 parts by weight of a saturated lower alcohol with respect to 1 part by weight of the filtrate;

Concentrating the filtrate under reduced pressure at a temperature of 60 to 70 ° C. by adding 3 to 5 parts by weight of ethyl alcohol to 1 part by weight of the residual aqueous solution to the remaining aqueous solution obtained in the layer separation step;

It characterized by consisting of a step of producing a powder extract by freeze drying the extract obtained in the vacuum concentration step.

In addition, the present invention is another feature of the anti-inflammatory analgesic containing a powder extract for the extract, as an active ingredient, and an anti-inflammatory analgesic tablet and injection using the same.

The present invention will be described in more detail as follows.

The present invention is characterized by an extraction method capable of extracting the active active ingredient from the nebula with high efficiency and an anti-inflammatory analgesic agent containing the active active ingredient, as well as tablets and injections using the same.

First, the above-mentioned ointment is extracted and filtered with water.

The extraction filtration was performed by adding 8 to 12 parts by weight of water to 1 part by weight of the duct and reflux extraction for 5 hours, then filtrating to collect the filtrate, and again adding 3 to 7 parts by weight of water to 1 part by weight of the duct to the residue and filtering by heating. Then, the extraction efficiency is increased by mixing with the previous filtrate. At this time, if the amount of water is less than the above range, the stirring becomes difficult and the solubility of the extract is lowered, the extraction efficiency is lowered. If the amount of the water exceeds the above range, the amount of the saturated low alcohol solvent used in the next purification step is increased and economical. And problems with handling occur.

In the present invention, a method of re-extracting after the first extraction is adopted. This is to increase the extraction efficiency. However, when the bulky extract is produced in large quantities, even though it is effectively filtered, the loss occurs because the water content of the herbal medicine is high. Tea extraction alone is to reduce the extraction effect is to prevent this. As a result of verifying the extraction efficiency of each step, it was found that about 90% of the total extraction amount is extracted by the second extraction, and therefore, the third or more multistep extraction is not economical.

The filtrate obtained in the extraction filtration step is separated into 0.7 to 1.2 parts by weight of a saturated lower alcohol with respect to 1 part by weight of the filtrate.

As described above, the extract obtained by extraction with water in the first and second stages is filtered and purified from impurities such as unnecessary fats and low molecular weight nonpolar substances contained in the filtrate, and in the present invention, 0.7 to 1.2 parts by weight per 1 part by weight of the filtrate. The impurities are purified by performing layer separation two to three times with saturated lower alcohol to remove the solvent fraction. In the present invention, a saturated butyl alcohol or a saturated propyl alcohol can be used as the saturated lower alcohol, and it is preferable to use a saturated butyl alcohol in the present invention.

If the amount of the saturated lower alcohol is less than 0.7 parts by weight based on 1 part by weight of the filtrate, the purification efficiency is drastically reduced. When the amount of the saturated lower alcohol exceeds 1.2 parts by weight based on 1 part by weight of the filtrate, the amount of unnecessary lower alcohol is increased. In order to maintain the purification effect efficiently, it is most preferable to separate the filtrate and two times with an equivalent amount of a saturated low alcohol solvent.

The filtrate was concentrated under reduced pressure by adding 3 to 5 parts by weight of ethyl alcohol to 1 part of the residual aqueous solution to the remaining aqueous solution obtained in the layer separation step.

Ethyl alcohol was added to 3 parts by weight of 1 to 5 parts by weight of the remaining aqueous solution to the remaining aqueous solution obtained in the step of separating the layer, that is, an aqueous solution in which impurities such as fatty acids and low molecular weight nonpolar substances were purified using a saturated lower alcohol. If the amount of the ethyl alcohol is less than 3 parts by weight based on 1 part by weight of the remaining aqueous solution, some protein polysaccharides and polysaccharides that are impurities are not efficiently removed and are toxic. This increases and decreases in activity, and when the amount of the ethyl alcohol used exceeds 5 parts by weight with respect to 1 part by weight of the residual aqueous solution, some of the active ingredients are also removed by precipitation, which is undesirable since the physiological activity is reduced. The filtrate obtained by adding ethyl alcohol to the remaining aqueous solution was concentrated under reduced pressure at a temperature of 50 to 60 ° C., if the reduced concentration was outside the above range, the concentration efficiency decreased or the active active ingredient was exposed to excessive heat. It is not preferable because the activity is degraded by exposure and decomposition.

The extract obtained in the vacuum concentration step is freeze-dried to prepare a powder extract.

As a result of analyzing the powder extract extracted from orthogonal acid by HPLC using the same method as shown in FIG. 1, the orthodontic extract was treated with croton-oil-induced mouse ear edema, carrageenan. Induced rat hind edema and seconds of induced mouse writhing test results, it was confirmed that the anti-inflammatory analgesic effect on the extracts.

It contains the powder extract obtained in the present invention and is formulated by a conventional manufacturing method to prepare tablets and injections, etc. The bases used at this time, such as lactose, microcrystalline cellulose, magnesium stearate The anti-inflammatory analgesic effect can be further enhanced by using a mixture of base ingredients such as) and the powdered powder extract. In particular, when the herbal composition of the present invention is administered to the human body, since the dermis is a natural extract, as well as the side effects compared to other synthetic drugs, there are almost no side effects, even in actual toxicity test results.

As described above, the extract extracted and purified in accordance with the present invention has a marked difference in efficacy in comparison with the extract obtained by the conventional method of extracting boiling water from Jingyo. In addition, the traditional Chinese herbal medicines can be prepared in the form of tablets and injections easy to administer the extract of Jingyo, which has been used only in the form of herbal medicines.

If the present invention will be described in detail based on the embodiment as follows, the present invention is not limited to the embodiment.

Example 1

12 l of water was added to 1 kg of commercially available oak and extracted under reflux for 5 hours, followed by filtration, and 5 liters of water was added to the residue, followed by extraction. The filtrate was mixed, and then 1.2 times the amount of saturated n-butyl alcohol in the filtrate. Layer was added three times. To the remaining aqueous solution after layer separation, ethyl alcohol four times the volume of the aqueous solution was added dropwise to remove insoluble components. The supernatant was concentrated under reduced pressure at a temperature of 55 ° C. and freeze-dried to obtain 464 g of a powder extract. In addition, the powder extract obtained was subjected to high performance liquid chromatography (HPLC) under the following conditions, and the results are attached to FIG. 1.

Column: μ-Bondapak C ₁₈ (3.9 × 300 mm)

Mobile phase: water / methanol / acetic acid = 8/2 / 0.05

Detector: UV 254 nm

Example 2

8 ℓ of water was added to 1 kg of well-dried jinchuan, which was commercially available, and refluxed for 4 hours, filtered. The residue was added again with 3 ℓ of water, and the filtrate was mixed. The filtrate was 0.7 times the weight of the filtrate. Saturated n-butyl alcohol was added and the layers were separated three times. To the remaining aqueous solution after layer separation, ethyl alcohol three times the volume of the aqueous solution was added dropwise to remove insoluble components, and the supernatant was concentrated under reduced pressure at a temperature of 53 ° C and freeze-dried to obtain 400 g of powder extract.

Example 3

1 kg of well-dried jinchuan was washed with tap water to remove contaminants. 10 l of water was added thereto, followed by extraction under reflux for 3 hours, followed by filtration. 7 L of water was added to the residue, and the filtrate was mixed. Of saturated n-butyl alcohol was added and the layers were separated three times. Elution component was removed by dropwise addition of ethyl alcohol 5 times the volume of the aqueous solution to the remaining aqueous solution after layer separation, the supernatant was concentrated under reduced pressure at a temperature of 60 ℃ and lyophilized to give 527 g of powder extract.

Comparative Example 1

Commercially purchased jinkyo was extracted with water using a standard solution solution, and the filtrate obtained by filtration was freeze-dried to obtain a powder extract.

Comparative Example 2

Water extraction was carried out in the same manner as in Comparative Example 1, and the filtrate obtained by filtration was separated three times with the same amount of saturated n-butyl alcohol, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure at a temperature of 65 ° C. The mixture was concentrated azeotropically with water, and finally, the same amount of distilled water was added and suspended, followed by freeze drying to obtain a powder extract.

Comparative Example 3

Water extraction was performed in the same manner as in Comparative Example 1, and the filtrate obtained by filtering the extract was separated three times with the same amount of saturated n-butyl alcohol, and then the precipitate formed by dropwise addition of 10 L of ethyl alcohol to the filtrate. It dried and the powder extract was obtained.

Formulation Example 1

Using the powder extract prepared in Example 1 to prepare a tablet for oral administration in the following composition.

< Furtherance >

Example 1 200 mg powder extract

Lactose 60 mg

Magnesium Stearate 40mg

Fine powder cellulose 60 mg

Corn starch 40 mg

Aerosil 30 mg

Avicel (PH-102) 70 mg

Formulation Example 2

The powder extract prepared according to Example 1 was prepared in the following composition with the following composition.

< Furtherance >

Injection ampoule: Example 1 powder extract 200 mg

                        Mannitol 360 mg

Corresponding Solvent Ampoule: Na ₂ HPO ₄ 12H ₂ O 52 mg

                        2974 mg of water for injection

Experimental Example 1

As a comparison of the acute anti-inflammatory activity of the extract prepared in Example 1 and Comparative Examples 1 to 3, the mouse ear edema test was performed as follows, and the results are shown in FIG. 2.

Experimental method: Oral administration of the extract from Examples 1 and 3 to ICR (Institute of Cancer Research) mice 1 hour after applying 2.5% croton oil (right) to the right ear and after 4 hours each Ear thickness was measured with a dial thickness gauge. The control group was orally administered sterile distilled water in place of the extract.

As can be seen in Figure 2, the case of Example 1 to which the extract of the present invention is administered has the highest inhibition efficiency against inflammation.

Experimental Example 2

Rat hind paw edema test was performed as a comparison of the acute anti-inflammatory activity of the extract prepared by Example 1 and Comparative Examples 1 to 3, the results are shown in Table 1 below.

Experimental Method: After 15 minutes of intravenous injection of each of the extracts prepared by Examples 1 and 3 into SD (Spraque-Dawley) rats, 0.1 ml of 1% carrageenan solution (1% carrageenan in saline) solution) was injected subcutaneously into the rear left foot of the rat and the edema of the left hind paw was measured using a volumetric meter (UGOBASILE TYPE 7150) at a 2-hour interval for 5 hours. As a control, sterile saline was injected intravenously in place of the extract.

TABLE 1

As can be seen from the results of Table 1, the case of Example 1 to which the extract of the present invention is administered has the highest inhibition efficiency against inflammation.

Experimental Example 3

Acetic acid induced mouse twist test was performed as a comparison of the analgesic effect of the extract prepared in Example 1 and Comparative Examples 1 to 3 as follows, the results are shown in Table 2 below.

Experimental method: Orally administered to each of the extracts prepared in Example 1 and Comparative Examples 1 to 3 ICR mice, and after 1 hour 0.6% acetic acid was intraperitoneally injected at a dose of 0.1ml per 10g of mouse weight, 10 minutes after the injection, the number of times of writhing (writhing: stretching the back or stretching the hind legs completely) was observed. The control group was orally administered sterile distilled water in place of the extract.

TABLE 2

As can be seen from the results of Table 2, it can be seen that in the case of Example 1 to which the extract of the present invention is administered, the analgesic action is the best.

Experimental Example 4

When the powder extract obtained in Example 1 was administered intravenously to an ICR mouse at a dose of 6.00 g / kg and observed for two weeks, no animals were found to be dead, and other anatomical findings were not abnormal compared to the control group. Was not found, and it was found that the powder extract obtained in Example 1 was an extremely safe substance.

As described above, the extract extracted and purified according to the present invention has a marked difference in efficacy compared to the extract obtained by the conventional method of extracting boiling water from Jingyo, and it is also limited to only conventional herbal medicines and the like. The dosage has been made much simpler by preparing the powdered extracts that have been used in the formulation of tablets and injections.

1 is a result of high performance liquid chromatography (HPLC) of the extract of Jinkyo prepared by Example 1,

Figure 2 is a graph showing the percentage of inhibition of ear edema induced by croton-oil (croton-oil) of the extracts prepared by Examples 1 and Comparative Examples 1-3.

Claims (5)

In preparing the Chinese herbal medicine by extracting the Chinese herbal medicine with water or alcoholic aqueous solution, Extracting and filtering the fibrils with water; Separating the filtrate obtained in the extraction filtration step with 0.7 to 1.2 parts by weight of a saturated lower alcohol with respect to 1 part by weight of the filtrate; Concentrating the filtrate under reduced pressure at a temperature of 60 to 70 ° C. by adding 3 to 5 parts by weight of ethyl alcohol to 1 part by weight of the residual aqueous solution to the remaining aqueous solution obtained in the layer separation step; Freeze-dried extract obtained in the step of concentration under reduced pressure to prepare a powder extract. The method of claim 1, wherein the extraction filtration step is carried out by reflux extraction for about 5 hours by adding 8 to 12 parts by weight of water to 1 part by weight of pulverized, filtered to collect the filtrate, and 3 to 7 parts by weight based on 1 part by weight of pulverized Partial water, warmed and filtered, and then the mixture is carried out by mixing with the previous filtrate. 2. A process according to claim 1, wherein the saturated lower alcohol is saturated butyl alcohol or saturated propyl alcohol. Anti-inflammatory analgesic containing a powder extract of the extract of claim 1 as an active ingredient. Anti-inflammatory analgesic tablets and injections comprising the powder extract of the extract of claim 1 as an active ingredient.