KR100815476B1 - Kenaf extract and novel kaempferol compound isolated them, and its use - Google Patents

Abstract

The present invention relates to a kenaf extract and a novel camperol compound isolated therefrom and its use, and more particularly, to a kenaf extract ( Hibiscus cannabbinus L., Malvaceae) extract and a new camperol compound separated therefrom are antibacterial, antioxidant The present invention relates to a new use that can be used as a preparation method and a method for obtaining the same from Kenaf, and the pharmaceutical composition and the kenaf extract containing the camphorol compound of the present invention are excellent in the above-mentioned effects and are expected to be greatly used in the future.

Kenaf (Hibiscus cannabbinus L., Malvaceae), camphorol compound

Description

KENAF EXTRACT AND NOVEL KAEMPFEROL COMPOUND ISOLATED THEM, AND ITS USE}

1 is a ¹ H NMR spectrum of a compound of Formula 1. FIG.

2 is a ¹³ C NMR spectrum for a compound of Formula 1. FIG.

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The present invention relates to a kenaf extract and a novel camperol compound isolated therefrom and its use, and more particularly, to the kenaf ( Hibiscus cannabbinus L., Malvaceae) extract and the new camperol compound separated therefrom are antibacterial and antioxidant By showing activity, the present invention relates to a food and pharmaceutical composition containing Kenaf extract and a new camperol compound isolated therefrom as an active ingredient.

Camperol (kaempferol) is a flavonoid-based component with antioxidant, antimicrobial and anti-aging effects.

Camperol isolated from acid garlic showed cholesterol-lowering activity, and camperol isolated from bouillon native to Africa was found to have significant cosmetic and whitening effects. Camperol in wormwood is known to have a good antitussive and expectorant effect. Camphorol, a flavonoid component of vegetables, has been studied to have anti-cancer and anti-obesity effects. However, the amount of camphorol produced from each of these plants is known to be very small.

In addition, since camphorol compounds derived from plants have lower side effects than camphorol compounds obtained by organic synthesis, they can be directly used in the human body, and since the development potential as environmentally friendly compounds is high, a wide range of studies are being conducted.

In order to meet the demands of the times, the inventors of Hibiscus cannabbinus L. (Malvaceae) in the process of obtaining a large amount of a camphorol compound exhibiting antioxidant, anti-aging, anti-obesity, etc. activity from natural resources The extract was selected and found to contain a large amount of camphorol while expressing various functionalities such as antioxidant, antibacterial and anti-aging. Kenaf is a fiber crop belonging to the family Malvaceae. The stem is used as paper and pulp, but the biological activity of the plant's leaves has not been identified.

Therefore, the inventors of the present invention not only investigated the antimicrobial and antioxidant activity of the kenaf extract and its derived compounds at the laboratory level, but also studied the verification of the availability of kenaf to the pharmaceutical, cosmetics industry, and functional foods. The present invention was completed by confirming the excellent antioxidant effect.

The present invention is a Kenaf leaf extract having an antioxidant, antimicrobial, and anti-aging effect and a novel compound isolated therefrom. Kaempferol-3,7-Oa-dirhamnoside (kaempferitrin) And as an invention for providing the use, Kenaf contains a large amount of the above-mentioned new camphorol compound in about 5 to 8% by weight, such as pharmaceuticals, quasi-drugs, cosmetic additives, food additives, beverage additives and functional health supplements, etc. It can be usefully used.

Accordingly, an object of the present invention is to provide a kenaf extract and a novel kempferol compound derived therefrom and their use which exhibit antibacterial and antioxidant activity.

The present invention is characterized by a pharmaceutical composition for antimicrobial agents comprising Kenip (Hibiscus cannabbinus L., Malvaceae) containing the compound represented by the following formula (1) as an active ingredient.
[Formula 1]

In addition, the present invention is characterized by another novel compound of formula 1, which is isolated from the extract.

[Formula 1]

Referring to the present invention in more detail as follows.

The present invention contains the Kenaf extract and the new camperol compound isolated from it by revealing that Kenaf ( Hibiscus cannabbinus L., Malvaceae) extract and the new camperol compound isolated therefrom exhibit antibacterial and antioxidant activity. Health functional food, cosmetics industry and pharmaceutical compositions.

In the present invention, in order to investigate the antimicrobial and antioxidant activity of Kenaf ( Hibiscus cannabbinus L., Malvaceae) extract and the novel camphorol compound isolated therefrom, the antimicrobial test of Kobayashi et al. ; Kobayashi, et al., Biosci.Biotech.Biochem., 58: 133-134, 1994) showed antimicrobial and antioxidant activity and can be provided as pharmaceutical compositions such as food preservatives and antibacterial agents.

In the present invention, various methods may be proposed to prepare the kenaf extract, but in the present invention, any method capable of extracting from a simple extraction method to a fat-soluble component may be applied, and the extraction method may exhibit antibacterial and antioxidant activity. Any method for preparing the kenaf extract is included in the present invention. As an example of the extraction method of the kenaf extract according to the present invention, distilled water bath extraction method, an organic solvent extraction method may be applied, and as an organic solvent, an aliphatic alcohol or an aqueous alcohol solution containing methanol, ethanol, propanol, butanol, hexane, ethyl acetate, etc. This can be used.

In addition, the separation and purification method of the novel camphorol compound represented by Formula 1 from the kenaf extract is as follows, but is not limited thereto.

1) extracting Kenap ( Hibiscus cannabbinus L., Malvaceae) leaf area with methanol to obtain a crude methanol extract;

2) extracting the crude methanol extract with a solvent selected from water, aliphatic alcohol, aqueous alcohol solution, hexane, chloroform, ethyl acetate, benzene and butanol to obtain a water layer and an organic layer;

3) After adsorbing the water layer and the organic layer on a normal silica chromatography column, fractions of toluene, acetone and methanol were eluted while changing the ratio of toluene, acetone, acetone and methanol, and the antibacterial and antioxidant fractions were confirmed. Obtaining (toluene: acetone acetone and methanol);

4) adsorbing the fraction on a second normal phase silica chromatography column, eluting with varying the ratio of water and methanol (10:10 to 5: 5) to obtain an active fraction;

5) purifying the fraction by performing gel filter chromatography (Sephadex LH-20) using 40% methanol in a solvent;

6) finally purifying the purified fraction using a recrystallization method and high performance liquid chromatography (HPLC) to purify a single active compound characterized in that the formula (1) from Hibiscus cannabbinus L., Malvaceae It includes a method for separating the compound represented by.

First, the dried kenaf is extracted and then filtered to obtain a liquid extract. At this time, the extraction solvent may be an organic solvent selected from aliphatic alcohol or alcohol aqueous solution, hexane and ethyl acetate, the aliphatic alcohol includes methanol, ethanol, propanol, butanol, preferably methanol. The extract was concentrated under reduced pressure, dissolved in distilled water, and then partitioned with hexane solvent. The mixture was gradually partitioned with ethyl acetate and butanol, and finally the aqueous layer was separated. The ethyl acetate fraction, which has been identified as antimicrobial and antioxidant activity, was separated by performing several steps of column chromatography (silica gel and Sephadex LH-20) in stages.In the final stage, compounds with high antioxidant activity were recrystallized and high-performance liquid chromatography. Purify the single active compound using (HPLC).

Thus obtained compound was analyzed by structural analysis using ¹ H-NMR, ¹³ C-NMR, FAB-MS, etc. Finally, as a yellow oily compound, the molecular formula is C ₂₇ H ₃₁ O ₁₅ , the molecular weight is 579 [ MH] ⁺ , a novel camphorol compound represented by the formula (1) showed excellent activity against antibacterial and antioxidant.

The pharmaceutical composition for an antimicrobial agent provided by the present invention includes a kenaf extract or a new camphorol compound derived from canap as an active ingredient, wherein the active ingredient may be a single component of each, but a mixture of two or more kinds depending on the purpose and method of use. It may be. The mixing ratio of the mixture is preferably adjusted according to the intended use, for example, the components of each mixture may be the same weight ratio.

The pharmaceutical composition for an antimicrobial agent provided by the present invention may include only an active ingredient alone, and may include 0.1 to 20% by weight of the active ingredient, preferably 1 to 5% by weight, and further include a residual amount of an excipient.

Based on the above results, it can be seen that the kenaf extract and the novel camphorol compound of the present invention have various applicability to the food industry as well as the pharmaceutical field, and particularly, antibacterial agents, antioxidants, functional foods and It can be seen that the applicability as a constraint is high. The kenaf extract and the novel camphorol compounds of the present invention can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically for clinical administration, and can be used in the form of general pharmaceuticals and health foods. have.

Kenaf extracts and novel camphorol compounds of the invention may be formulated for oral administration such as tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, hard or soft Formulated in capsules, syrups or elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin for preparation in formulations such as tablets and capsules; Excipients such as dicalcium phosphate; Disintegrants such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. Capsules contain liquid carriers, such as fatty oils, in addition to the substances mentioned above.

In addition, the kenaf extract and the novel camphorol compound of the present invention can be administered parenterally, and parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection. To formulate into parenteral formulations, the aminated β-glucans are mixed in water with stabilizers or buffers to prepare solutions or suspensions, which are formulated in unit dosage forms of ampoules or vials.

In addition, the dosage of the active ingredient is generally 1 to 50 mg / day per kg body weight of an adult patient, preferably 5 to 20 mg / day, 1 to several times at regular intervals according to the judgment of the doctor or pharmacist Preferably, it can be divided into 2-3 times a day.

In addition, the kenaf extract of the present invention and a novel adjuvant includes a health supplement containing the active ingredient. The health supplement is a food prepared by adding the kenaf extract and the novel camphorol compound to food materials such as beverages, teas, spices, gums, confectionery, or the like, encapsulated, powdered, and suspensions. Means to bring a certain effect, unlike the general medicine has the advantage that there is no side effect that can occur when using the drug as a raw material for long-term use.

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

Note

The apparatus used in the embodiment of the present invention is as follows. ¹ H-NMR and ¹³ C-NMR were measured using Varian Gemini 200 (200 MHz). MS was measured in FAB positive mode and EI mode using Micromass. TLC plate used Merck's Precoated Kieselgel 60 F _254s (art. NO. 5715) and RP-8 F _254s (Art. NO. 15424), and the column chromatography carrier was Kieselgel 60 (70-230). mesh ASTM, Merck, Art.NO.7734), Kieselgel (230-400 mesh ASTM, Merck, Art.NO.9385), YMC gel ODS-A (70-230 mesh, Art.NO.AA12SA5) and Sephadex LH- 20 (Pharmacia Biotech AB) was used. TLC developing solvents and other reagents were used as first-class and special reagents, and extraction solvents and solvents for column chromatography were re-distilled with industrial reagents.

Example  1: Preparation of Kenaf Extract

Kenaf (kenaf) used in the present invention was cultivated in Buan reclaimed land farmland collected in September 2005 and used as a sample, it is being stored in the plant biotechnology major specimen room of the Department of Biotechnology.

Kenaf leaves (133g) dried at room temperature in a shade with a homogenizer (Homogenizer, PH-91, AMT) and then triturated with methanol three times a week repeated cold extraction, the extracts were collected and concentrated under reduced pressure, methanol extract (MeOH) ext.) to give 24.6 g.

Example  2: Preparation of Water and Organic Solvent Extract from Kenaf Extract

24.6 g of the methanol extract obtained in Example 1 was suspended in distilled water, fractionated with hexane (5.2 g), and the remaining aqueous layer was partitioned with ethyl acetate (EtOAc) to obtain an ethyl acetate fraction (4.4 g). The aqueous layer was partitioned with butanol (BuOH) again to obtain a butanol fraction (4.6 g), and finally 9.3 g of water fraction.

The antimicrobial and antioxidant activities were investigated using the respective fractionation layers, and the antimicrobial and antioxidant activities were confirmed in the hexane and ethyl acetate fractions in the fractionation layer.

Example  3: New from Kenaf Camperol ( kaempferol Separation of Compounds

An indicator was separated and purified in a solvent fraction obtained from the methanol extract of Example 2. 4.4 g EtOAc fraction containing indicator material (see HPLC chromatogram in FIG. 1) was dissolved in methanol and then subjected to a normal silica chromatography column using a glass column (Ø 5 x 70 cm, 7734, Merck) filled with silica gel. After adsorption, the ratio of toluene, acetone, and methanol mixture was eluted with varying the ratio of toluene, acetone, acetone, and methane to obtain 10 small fractions (toluene: acetone, acetone, methanol) in which antioxidant activity was confirmed.

Second open column chromatography was performed on small fraction 7 (toluene: acetone, 1.153 g) containing the indicator substance. ODS (YMC gel ODS-A (70-230 mesh, Art. NO. AA12SA5) was charged into a glass column (Ø 1.6 x 100 cm) and eluted with a pump for HPLC, eluting with varying the ratio of water and methanol. An active subfraction was obtained, and a small fraction 259-316 (water: methanol, 605.8 mg) with double indicators was obtained.

The obtained fractions containing the indicator material were subjected to gel filter chromatography [Sephadex LH-20, Pharmacia Biotech, glass column (Ø 1.0 x 50 cm)] using 40% methanol again as a solvent to purify a single active compound. The index compound (25 mg) was obtained. The purified fractions were finally purified using a recrystallization method and high performance liquid chromatography (HPLC) to purify a single active compound to finally obtain an indicator compound (25 mg).

Example  4: new Camperol  Structural Analysis of Compounds

In order to determine molecular weight and molecular formula of the isolated compound through this experiment, ¹ H, ¹³ C spectra were obtained using nuclear magnetic resonance ( ¹ H-NMR and ¹³ C-NMR using Varian Gemini 200 (200 MHz). Mass spectrometry was measured in FAB positive mode and EI mode using a mass spectrometer (Micromass, Hewlett packard 5989A) to determine the molecular weight and molecular formula, and the results are shown in FIGS.

IR ν _max : KBr cm ⁻¹ 3470-3353 (OH), 2927, 1727 (> C═O), 1601 (aromatic), 1458 (cycloalkane), 1189 (phenol);

¹ H-NMR (200 MHz, MeOH- d 4) ppm: δ 7.78 (2H, d, J = 8.8, H-2 '&H-6'), 6.91 (2H, d, J = 8.8, H-3 ' 6.77 (1H, d, J = 1.6, H-8), 6.44 (1H, d, J = 1.4, H-6), 5.42 (1H, brs, H-1 "), 1.18 (1H, H-6 ′), 0.79 (3H, d, J = 4.4 Hz, H-6 ″);

¹³ C-NMR (50 MHz, MeOH- d 4) ppm: 178.7 (s, C-4), 162.5 (s, C-7), 161.7 (s, C-5), 160.9 (s, C-4 ') , 158.6 (s, C-2), 156.9 (s, C-9), 135.3 (s, C-3), 131.5 (d, C-2 ', C-6'), 121.1 (s, C-1 '), 116.2 (d, C-3', C-5 '), 106.5 (s, C-10), 102.6 (s, C-1 "), 100.2 (s, C-6), 99.1 (s, C-1 "'), 95.3 (s, C-8), 72.3 (d, C-4", 4 "'), 71.8 (s, C-3"'), 71.4 (s, C-3 ") , 70.9 (d, C-2 ", 2"'), 70.8 (s, C-5 "'), 70.5 (s, C-5"), 18.5 (s, C-6 "'), 18.1 (s , C-6 "). FAB-MS: m / z 579 [M + H] ⁺

The separated camp Ferrol compounds have absorption band represents the aromatic (aromatic) C = C bond from C = O, 1601㎝ ^-1 in absorption by OH in the vicinity 1720㎝ ^{-1 -1} 3471㎝ For IR spectrum (spectrum) Absorption peaks at 216, 242, 300 (sh) and 326 nm of the UV spectra indicate that the compound is a flavonoid glycoside compound (Sakushima et al ., 1985).

Signals of two aromatic protons meta- coupled in the ¹ H-NMR spectrum δ 6.77 (1H, d, J = 1.6, H-8), 6.44 (1H, d, J = 1.4, H-6 Δ 7.78 (2H, d, J = 8.8, H-2 '&H-6'), 6.91 (2H, d, J = 8.8, H- derived from two aromatic protons with ortho coupling 3 '&H-5') signal was observed (Pakulski et al ., 1996).

In addition, the signals of carbons 4 and 7 were found at δ 178.7 and 162.5 of the ¹³ C-NMR spectrum, and the signals of carbons 5 and 2 were identified at δ 161.7 and 158.6. rhamnosyl) caused 5 and 6 carbon signals.

Based on the above results, various spectral data were obtained from Inigo PAR, De Iglesisa IAD and Catalan ANC (1988) Kaempferol 3-aD-glucopyranoside-7-a- rhamnopyronoside from Erythroxylon cuneifolium.Phytochemistry 27 (4): 1230-1231. 1988), this compound was identified as kaempferol-3,7-Oa-dirhamnoside (kaempferitrin).

The new purified purified camphorol compound is a yellow oil (Yellow oil), the molecular formula of C ₂₇ H ₃₁ O ₁₅ , the molecular weight was determined to be 579 [MH] ⁺ , the structural formula Indicated.

[Formula 1]

Example 5 Antioxidant Activity Assay of Kenaf Extract and New Camperol Compounds

Antioxidant Activity

Antioxidant activity was measured using free radical 1, 1-diphenyl-2-picrylhydrazyl (1, 1-diphenyl-2-picrylhydrazyl, DPPH) (Blois, Nature, 188: 1199). , 1958; Choi, et al., Kor. J. Phamacogn, 24: 299-303, 1993). 1 ml of the sample compound solution was added at each concentration (100-2000 ppm), and then 4 ml of the DPPH (0.15 mM) solution was added thereto to react at room temperature for 30 minutes, and the absorbance was measured at 517 nm. At this time, RC ₅₀ (μg / ml) represents the concentration of the compound which reduces the value of the control group without the compound by 50%.

In order to investigate the antioxidant activity of the novel camphorol compound represented by Chemical Formula 1 isolated from Kenaf, 1, 1-diphenyl-2-picrylhydrazyl (1, 1-diphenyl) which is a free radical The experiment was carried out using the antioxidant activity measurement method (Blois, Nature, 188: 1199, 1958; Choi, et al., Kor. J. Phamacogn, 24: 299-303, 1993) using -2-picrylhydrazyl (DPPH). The results are shown in Table 1 below.

Extracts and Fractions RC ₅₀ ^{2 )} (µg / ml) MeOH Extract 599 Hexane fraction 755 EtOAc fraction 294 BuOH fraction 441 Water column fraction 1992 1) 1,1-diphenyl-2-picryl-hydrazyl 2) Amount required for 50% reduction of DPPH after 30 min.

Example 6 Antimicrobial Activity Assay of Kenaf Extract and New Camperol Compounds

Antimicrobial  Active investigation

Antimicrobial activity was investigated according to the method of Kobayashi (Kobayashi, et al., Biosci. Biotech. Biochem., 58: 133-134, 1994). Fungal strains include Aspergillus Staphylococcus aureus ( Staphylococus) aureus ), Salmonella typhimurium , and Kelbsiella pneumoniae , and 2 ml of mold spore germ inhibition test medium were added to these mold culture PDB slants . The mold spores were separated by addition of glass rods and filtered again with gauze. The filtrate was obtained and dispensed into a 96 well plate by 100 [mu] l and diluted until 50 spores were observed in the field while observing under a microscope. The sample is prepared at a concentration of 4 to 1000 ppm, added to the well, and after 24 hours of cancer culture at 27 ° C., spore germination is inhibited under a microscope.

In addition, Bacillus subtilis and Escherichia coli were used as bacterial strains, and these bacterial strains were transplanted into 10 ml of NB medium, followed by shaking culture for 12 hours at 27 ° C., followed by Bacillus subtilis . Tilly seuneun to be 10 ⁶ / ml, S-cyano cherry cola which 10 ⁷ / ml and diluted with NB medium. Then, 100 μl of the solution is dispensed into 96 well plates, and the sample is prepared at a concentration of 4 to 1000 ppm. The sample is added to the well and cultured at 27 ° C. for 24 hours to measure bacterial growth inhibitory activity based on suspension.

Method of Kobayashi (Kobayashi, et al., Biosci. Biotech. Biochem., 58: 133-134, 1994) to investigate the antimicrobial activity of the novel camphorol compound represented by Formula 1 isolated from Kenaf The experiment was performed as shown in Table 2 below.

Strain solvent Kenaf leaf extract (μg / μl) Staphylococus aureus E. coli Salmonella tyhimurium Klebsiella pneumoniae Bacillus subtilis MeOH Extract > 1000 500 125 500 1000 Hexane fraction 250 125 125 250 500 EtOAC Fraction 500 125 125 250 500 BuOH fraction > 1000 1000 500 500 500 Water fraction > 1000 500 125 500 500

Example 7: Acute Toxicity Test of Kenaf Extract and Camperol Compounds

① for experimental rats Parenteral  Dose Toxicity Test

To carry out acute toxicity test on Kenaf extract and camphorol compound of Formula 1, 6-week-old ICR mice (♂, Bio), weighing 25 g, were divided into 4 groups of 2 animals per group, temperature 22 ± 3 ℃, humidity 55 It was bred in an animal room of ± 10%, illumination 12L / 12D. Mice were allowed to acclimate for about a week before being used in the experiment. Feed for experimental animals (first feed, mice and rats) and drinking water were supplied after sterilization and freely ingested.

Kenaf extract and camphorol compound of formula 1 were dissolved in a small amount of dimethylsulfoxide (DMSO) and diluted with 1% carboxymethylcellulose solution to 100 mg / kg, 50 mg / kg, 10 mg / kg, 5 A single dose orally administered at a dose of mg / kg, and the control group used a 1% carboxymethyl cellulose solution. Seven days after administration, side effects or lethal were observed as follows. That is, on the day of administration, changes in general symptoms and the presence or absence of dead animals were observed at least once in the morning, at least once every afternoon from 1 hour, 4 hours, 8 hours, 12 hours, and the next day after administration.

In addition, on the 7th day of administration, animals were killed and dissected, and the internal organs were visually examined. Changes in body weight were measured at daily intervals from the day of administration, and the weight loss phenomenon of the animals by the kenaf extract and the camphorol compound of Formula 1 was observed.

As a result, there were no clinical symptoms that were related to the administration of the test substance, and it was estimated that the minimum lethal dose (LD50) of oral administration was 100 mg / kg or more because the animal did not die in the 100 mg / kg group. No specific findings related to test-related substances were observed in animals and live animals.

② for the rat Parenteral  Dose Toxicity Test

Using the same experimental mice as the oral administration toxicity test, 2 mg each of the camphorol compound of Formula 1 isolated according to Example 3 of the present invention was administered by the intramuscular injection method. One rat was administered twice at two week intervals, and the other rat was administered twice at two week intervals, followed by one month intervals, and then again three doses at two week intervals.

Six months after the first injection, the clinical signs, body weight, body temperature, physical examination, blood cell abnormalities (haematology) and fecal abnormalities (urinalysis) were observed, but all showed normal values.

Example 8 Formulation Example

As confirmed above, the camphorol compound of formula (1) exhibits excellent antibacterial and antioxidant activity, can be formulated into pharmaceutical compositions of food and medical products, and can also be prepared as a dietary supplement.

① Preparation of syrup

Syrup containing 2% by weight of the camphorol compound of Formula 1 as an active ingredient was prepared by the following method.

2 g of the camphorol compound of Formula 1 and 25.4 g of sugar were dissolved in 80 g of warm water. After the solution was cooled, a solution consisting of 8.0 g of glycerin, 0.8 g of saccharin, 0.04 g of flavor, 4.0 g of ethanol, 0.4 g of sorbic acid, and distilled water was prepared and mixed. Water was added to this mixture to 100 ml.

② manufacture of tablets

A tablet containing 15 mg of active ingredient was prepared by the following method.

2 g of the camphorol compound of Formula 1 were mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.

③ Preparation of injection solution

Injection solution containing 10 mg of the active ingredient was prepared by the following method.

0.2 g of the camphorol compound of Formula 1, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

④ Manufacture of health supplement drinks

After dissolving 2 g of the camphorol compound of Formula 1 in an appropriate amount of water, vitamin C as an auxiliary component, citric acid, sodium citrate, and oligosaccharides as an auxiliary component were added, and an appropriate amount of sodium benzoate was added as a preservative, followed by adding water. 100 ml was prepared to prepare a beverage composition. At this time, taurine, myo-inositol, folic acid, pantothenic acid, etc. may be added alone or together.

As described above, the kenaf extract according to the present invention and the camphorol compound of formula 1 isolated therefrom can be used in food and pharmaceuticals.

Claims (5)

Next, Kenaf (Hibiscus cannabbinus L., Malvaceae) containing the compound represented by Formula 1 as an active ingredient, the pharmaceutical composition for an antimicrobial agent. [Formula 1]

The pharmaceutical composition of claim 1, wherein the extraction solvent of the extract is selected from water, aliphatic alcohol or aqueous alcohol solution, hexane, chloroform and ethyl acetate. delete The pharmaceutical composition for an antimicrobial agent comprising the compound represented by the following Formula 1 as an active ingredient. [Formula 1]

1) extracting Kenap ( Hibiscus cannabbinus L., Malvaceae) leaf area with methanol to obtain a crude methanol extract; 2) extracting the crude methanol extract with a solvent selected from water, aliphatic alcohol, aqueous alcohol solution, hexane, chloroform and ethyl acetate to obtain a water layer and an organic layer; 3) After adsorbing the water layer and the organic layer on a normal silica chromatography column, the mixture of toluene, acetone, and methanol was eluted while changing the ratio of toluene, acetone, acetone, and methanol, and the antibacterial and antioxidant activity was confirmed. Obtaining; 4) adsorbing the fraction on a second normal phase silica chromatography column, eluting with varying the ratio of water and methanol (10:10 to 5: 5) to obtain an active fraction; 5) purifying the fraction by performing gel filter chromatography (Sephadex LH-20) with 40% methanol using a solvent; 6) Final purification of the purified fractions from the Hibiscus cannabbinus L., Malvaceae comprising the step of purifying a single active compound using a recrystallization method and high performance liquid chromatography (HPLC) Separation method of a compound represented by. [Formula 1]

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