KR100394577B1 - Crude extracts from Schizandra chinensis RUPRECHT seed and process for preparation thereof - Google Patents

Abstract

The present invention relates to Schisandra chinensis seed functional extract and its manufacturing method. Schisandra chinensis seed extract and its fractions extracted with various organic solvents show high antioxidant activity against lard and soybean oil, and have excellent DPPH free radical scavenging activity and antimicrobial against food decay microorganisms. It has an excellent effect of showing activity and an excellent effect of showing high nitrite scavenging ability.

Description

Schizandra chinensis RUPRECHT seed and process for preparation according to Schizandra chinensis extract

The present invention relates to Schisandra chinensis seed functional extract and a method for producing the same. More specifically, the present invention relates to a schizandra seed extract with antioxidant, antimicrobial and nitrite scavenging ability extracted using an organic solvent from Schizandra seeds and a method for producing the same.

Schizandra chinensis RUPRECHT is a Magnoliaceae plant that blossoms in June-July and matures in September-October and becomes magenta. Omija grows in colonies in Odaesan, Jirisan, and Balwangsan areas, and is cultivated in Hwacheon, Inje, Pyeongchang, Gyeongsangbuk-do, Bonghwa, Jeollabuk-do, Muju, Jinan, Jangsu, and Gyeongsangnam-do. The yield is 141 M / T.

Schisandra chinensis has been used in traditional medicine to improve systemic weakness, mental and physical fatigue, bronchitis, bronchial asthma, nervous breakdown, hypotension, cardiac dysfunction, malnutrition ulcers and wounds, and to improve visual acuity in tea or summer. It is processed and used mainly in starch foods and Schisandra chinensis using pigments.

In the study of Schizandra chinensis, Kim et al. Reported on Schizandra chinensis, organic acid and anthocyanin pigment [Kim, K.I., Nam, J.H. and Kwon, T.W. On the proximate composition, organic acid and anthocyanins of Omija,Schizandra chinensisBaillon. Korean J. Food Sci. Technol. 5: 178-182 (1973)], Yang et al. Reported the stability of anthocyanin pigments [Yang, H.C., Lee, J.M. and Song, K.B. Anthocyanins in cultured OmijaSchizandra chinensisBaillon) and its stabililty. Korean Soc. Agri. Chem. Biotechnol. 25: 35-43 (1982). In addition, Lee et al. Conducted a series of studies on the composition of free sugars, lipids and nonvolatile organic acids by sites of Schizandra chinensis [Lee, J.S. and Lee, S.W. The studies of composition of free sugar, fatty acid and nonvolatilitic organic acid in part of Omija (Schizandra chinensisBaillon). Korean J. Dietary culture 4: 177-181 (1989). As a study on the pharmacological action of Schizandra chinensis, Hikino et al reported on hepatoprotective action [Hikino, H., Kios, Y., Takuchi, H. and Ikeya, Y. Validity of the oriental medicines 60. Liver-protective drugs . II Antihepatotoxicaction of lignoids fromS. chinensisfruits. Planta Med. 50: 213-216 (1984)] and the like have been described in alcohol detoxification [Lee, J.S. and Lee, S.W. Effect of water extract in fruits of Omija (Schizandra chinensisBaillon) on alcohol metabolism. Korean J. Dietary culture 5: 259-262 (1990)] reported on antidiabetic activity [Sheo, H.J., Lee, M.Y. and Hwang, J.S. Effect ofSchizandrae fructusextract on blood constituents of alloxan induced diabetic rabbits. J. Korean Soc. Food Sci. Nutri. 16: 262-268 (1987). Schisandra chinensis research has been conducted by Kang et al. [Kang, K.C., Park, J. H., Beak, S. B., Jhin, H.S. and Rhee, K.S. Optimization of beverage preparation formSchizandra chinensisBaillon by response surface methodlogy. Korean J. Food Sci. Technol. 24: 74-81 (1992)] and dried Schisandra chinensis fruit juiceSaccharomyces cerevisiae ellipsoideusWowSaccharomyces coreanusA study on the quality of fermented rice by fermentation [Chang, E.J. Studies on the production of Omija Wine. M.S. Thesis, Korea Univ., Korea (1985)] and studies on the drying and storage of Schizandra chinensis [Jung, G. T., Ju, I.O. and choi, J.S. Studies on drying and preservation of OmijaSchizandra chinensis). Korean Postharvest Sci. Technol. 5: 217-223 (1998). The antimicrobial activity of Schizandra chinensis against pathogenic microorganisms such as Lee, S.H. and Lim, Y. S. Antimicrobial effects ofSchizandra chinesisextract on pathogenic microorganism. J. Korean Soc. Food Sci. Nutri. 27: 239-243 (1998)], lactic acid bacteria isolated from kimchi [Lee, S.H. and Lim, Y. S. Effect of Omija (Schizandra chinesis) extract on the growth of lactic acid bacteria isolated from Kimchi. Korean J. applied Microbiol. Biotechnol. 25: 224-228 (1997); andListeria monocytogenesThe antimicrobial effect of this has been reported [Lee, S.H. and Lim, Y. S. Antimicrobial effects ofSchizandra chinesisextract againstListeria monocytogenes. Korean J. applied Microbiol. Biotechnol. 25: 442-447 (1997)], but studies on the physiological activities such as antioxidant, antibacterial and nitrite scavenging of Schizandra chinensis seeds are still insufficient.

In view of the above, the present inventors extracted and fractionated Schisandra chinensis seeds with various organic solvents, and then, each of the extracts was subjected to antioxidant, food decay microorganisms by POV (peroxide value), TBA (thiobarbituric acid), and electron donor measurement methods. The present invention was completed by investigating nitrite scavenging ability, which is an antimicrobial and carcinogenic substance, against Schisandra chinensis seed extract having excellent antioxidant and antibacterial properties and nitrite scavenging ability.

Accordingly, an object of the present invention is to provide an extract with antioxidant, antimicrobial and nitrite scavenging ability extracted from Schisandra chinensis seed.

It is another object of the present invention to provide a method for preparing the Schisandra chinensis extract.

Still another object of the present invention is to provide a fraction obtained by dividing Schisandra chinensis seed extract for each solvent.

The object of the present invention is to crush the Schisandra chinensis seed, adding water, methanol, ethyl acetate, tet, chloroform and extract the extract extracted into chloroform, ethyl acetate, butanol and the like again, POV, TBA of the schisandra seed extract The antioxidant activity was investigated by measuring DPPH free radical scavenging activity by DPPH method. Subsequently, the extract of Schizandra chinensis seed per extractant was achieved by investigating the antimicrobial activity of inhibiting the growth of various microorganisms by measuring the diameter of the clear zone to confirm that it had nitrite scavenging ability.

Next, the structure of this invention is demonstrated.

1 is a photograph showing the results of observing microorganisms (right) and untreated microorganisms (right) treated with butanol fraction (50 μg / mL) of the Schizandra chinensis seed methanol extract of the present invention under a scanning electron microscope.

The present invention is prepared by grinding the Schisandra chinensis seed, degreasing it, adding 3 to 8 times the amount of an organic solvent, extracting 1 to 3 times at a temperature of 70 to 95 ° C., filtration, concentrating with a rotary vacuum evaporator, and lyophilizing. , Scavenging DPPH free radicals, Lactobacillus plantarum (KCCM 11322), Bacillus subtilis (KCCM 11314), Staphylococcus aureus (KCCM 12103) , Salmonella typhimurium (KCCM 11806), Escherichia coli (ATCC 10536), Penicillium citrinum (KCCM 11663)] inhibit the growth and inhibit nitrite The present invention extracts Schisandra chinensis seed. The present invention is pulverized Schisandra chinensis seed, and extracted with water, methanol, ethanol, ethyl acetate and chloroform, respectively. The method comprising in sequence fractions Tate, butanol, and water; Adding the Schisandra chinensis seed extract extracted for each solvent to lard and soybean oil and measuring the peroxide value to investigate the antioxidant power of the Schisandra chinensis extract; Adding the Schisandra chinensis seed extract extracted for each of the solvents to linoleic acid and measuring the TBA value to investigate the antioxidant activity of the Schisandra chinensis extract; Investigating the antioxidant power of the Schisandra chinensis seed extract by mixing the methanol dissolving Schisandra chinensis seed extract extracted for each solvent into a DPPH solution and measuring the absorbance; Drop the culture medium of each of the test strains on a plate and lay it thinly, and then put on a filter paper disk which absorbed the Schisandra chinensis extract extracted for each solvent, incubate it, and measure the diameter of the clear zone to investigate the antimicrobial activity of the Schisandra chinensis extract. ; Treating the butanol fraction of Schizandra chinensis extract in a medium cultured with food rot bacteria for 48 hours and observing the morphological changes of the cells under a scanning electron microscope; NaNO ₂ solution was added to the extract of Schizandra chinensis seed extracted by solvent, and acetic acid solution and Griess reagent were added to the reaction solution by adjusting the pH to 1.2, 3.0, and 6.0. do.

In the present invention, about 3 to 8 times the amount of the organic solvent is added to the schizandra seed, which is ground.

In the present invention, Schisandra chinensis seed extraction is carried out 1-3 times at 70-95 ° C., preferably twice at 85 ° C.

Schizandra chinensis RUPRECHT in Schizandra chinensis RUPRECHT was soaked in water overnight to remove pulp and used hot-dried seeds at 60 ° C.

The fats and oils used in the present invention used soybean oil and lard without any antioxidants.

The reagent for extracting and fractionating Schisandra chinensis seed used in the present invention was used as the first grade, and the rest of the reagent was used as an express.

In the present invention, the strains used for the investigation of antimicrobial activity were Lactobacillus plantarum (KCCM 11322), Bacillus subtilis (KCCM 11314), Staphylococcus aureus (KCCM 12103). ]), Salmonella typhimurium (KCCM 11806), Escherichia coli (ATCC 10536) and Penicillium citrinum (KCCM 11663) were used.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1 Preparation of Schisandra chinensis Seed Extract and Fraction

A sample of Schisandra chinensis seed pulverized (CEMOTEC 1090) was degreased with hexane, and 5 times the amount of the sample was extracted using water, methanol, ethanol, ethyl acetate, ethyl acetate, Chloroform was added to the flask to which the reflux condenser was attached, followed by extracting twice in an 85 ° C. water bath for 3 hours, filtered, concentrated in a rotary vacuum evaporator, and lyophilized to obtain an extract. . Among the various extracts, the most active extract was sequentially fractionated with a solvent such as chloroform, ethyl acetate, butanol, and water, concentrated and lyophilized to obtain a fraction.

As a result, as shown in Table 1, the extraction yield by solvent showed the highest extraction yield with ethyl acetate (12.0%), the chloroform extraction yield was 11.7%, the water extraction yield was 8.2%, the methanol extraction yield was 6.4%, and the ethanol extraction. The yield was the lowest as 5.0%.

Yield of Schisandra chinensis Extract Extracted with Various Extraction Solvents menstruum water Methanol ethanol Ethyl acetate chloroform Extraction yield (%) 8.2 6.4 5.3 12.0 11.7

Experimental Example 1: POV Measurement

In this Experimental Example, 100 g of lard and soybean oil were added to 500 ppm of Schisandra chinensis seed extract according to the extraction solvent obtained in Example 1 dissolved in DMSO (dimethyl sulfoxide) solution, and stored at 60 ° C. in a constant temperature flask for 1 g each. Was analyzed by leveling up. The sample in chloroform 10 mL, was added to 15 mL and KI saturated solution of 1 mL of acetic acid was shaken for 1 minutes then allowed to stand for 5 minutes cow, was shaken by the addition of distilled water to 75 mL and then titrated with 0.01 N Na ₂ S ₂ O ₃ solution It was set as POV (peroxide value).

As a result, as shown in Table 2, the peroxide value of the piglet before the test was 1.23 meq / kg and the peroxide value increased to 22.6 meq / kg after 16 days without addition. Antioxidant activity of Schisandra chinensis seed extract was in order of methanol extract> ethanol extract> water extract> chloroform extract> ethyl acetate extract, and all solvent extracts had lower antioxidant effect than δ-tocopherol, a natural antioxidant. appear. In addition, Table 3 shows the peroxide value of soybean oil, and the antioxidative power of the extract was relatively low because soybean oil rancidated faster than that of lard with peroxide value of 1.11 meq / kg. The natural antioxidant δ-tocopherol had little effect, but it was effective in Schisandra chinensis extract, of which ethanol extract was the most effective at 44.58 meq / kg, followed by methanol extract (46.71 meq / kg).

Changes in the Peroxide Values Measured over Time with Storage of Pork Containing Schisandra chinensis Seed at 60 ° C (Unit: meq / kg) menstruum Save time (days) 0 8 16 water 1.23 ¹⁾ 10.91 13.76 Methanol 1.23 8.77 13.43 ethanol 1.23 9.82 13.83 Ethyl acetate 1.23 14.94 17.32 chloroform 1.23 11.22 14.05 δ-tocopherol 1.23 13.84 18.80 Control 1.23 12.46 22.60 NOTE 1) is the average value repeated three times.Unit: meq / kg

Changes of Peroxides Measured over Time with Soybean Oil Containing Schisandra chinensis Seed Extract at 60 ° C menstruum Save time (days) 0 8 16 water 1.11 ¹⁾ 24.88 53.46 Methanol 1.11 23.79 46.71 ethanol 1.11 15.27 44.58 Ethyl acetate 1.11 28.97 56.89 chloroform 1.11 28.15 58.40 δ-tocopherol 1.11 32.64 60.12 Control 1.11 30.87 62.98 NOTE 1) is the average value repeated three times.Unit: meq / kg

Experimental Example 2: TBA Measurement

In this experiment example, the substrate solution was added so that linoleic acid became 0.03 M in a solvent in which 0.1 M phosphate buffer (pH 7.0) and ethanol were mixed 4: 1. To 20 mL of this substrate solution, 19.2 mL of 0.1 M phosphate buffer (pH 7.0) and 0.8 mL of Schizandra seed extract for each extraction solvent obtained in Example 1 were added to 500 ppm, and then stored at 40 ° C in a shaker. 2.0 mL sample solution was taken and analyzed.

1.0 mL of 35% trichloroacetic acid and 2.0 mL of 0.75% TBA reagent were added to 2.0 mL of the sample solution, followed by shaking for 30 seconds, followed by reaction for 40 minutes on a 95 ° C. water bath. The reaction solution was cooled to room temperature, shaken by adding 1.0 mL of acetic acid and 2.0 mL of chloroform, followed by centrifugation at 3,000 rpm for 5 minutes to measure the absorbance of the supernatant at 532 nm, which was determined as a TBA (thiobarbituric acid) value.

As a result, as shown in Table 4, the control was 1.557 after 6 days and methanol extract was 0.065, and the TBA value was very low, which was the best antioxidant activity, which was higher than δ-tocopherol (0.229) and was similar to BHA (0.014). Therefore, the antioxidant effect of the extract on linoleic acid was methanol extract> chloroform extract> ethyl acetate extract> ethanol extract> water extract.

Changes in TBA Values of Linoleic Acid Containing Schisandra chinensis Seed Extracts at 40 ° C Storage menstruum Save time (days) One 2 4 6 water 0.119 ¹⁾ 0.353 0.537 1.262 Methanol 0.022 0.085 0.108 0.065 ethanol 0.023 0.146 0.223 0.369 Ethyl acetate 0.036 0.181 0.206 0.298 chloroform 0.004 0.133 0.192 0.157 BHA 0.002 0.065 0.059 0.014 δ-tocopherol 0.132 0.213 0.245 0.229 Control 0.294 0.569 1.474 1.557 NOTE 1) is the average value repeated three times.

Experimental Example 3: Investigation of Antioxidant Capacity by DPPH Method

This Experimental Example is an antioxidant activity assay using DPPH (1,1-diphenyl-2-picrylhydrazyl), and the sample of Schisandra chinensis seed extract extracted in Example 1 was dissolved in 4 mL of methanol and 1.5 × 10 ^-4 M DPPH methanol solution. After adding 1 mL, the solution was left at room temperature for 30 minutes and the absorbance was measured at 517 nm. The amount (μg) of the sample required to reduce the absorbance of the control group without adding the sample by 1/2 was expressed as RC ₅₀ , and compared with the existing antioxidants δ-tocopherol and BHA.

As a result, as shown in Table 5, the concentration of extract (RC ₅₀ ) required for 50% reduction of DPPH was 19.8 ㎍ / mL and 18.7 ㎍ / mL for commercial BHA and δ-tocopherol, respectively. Many. Among the methanol extracts, the highest activity was 33.2 ㎍ / mL, and the ethyl acetate and chloroform extracts were 465 ㎍ / mL.

To summarize the results of Experimental Examples 1 to 3, methanol and ethanol were effective for extracting antioxidants of Schisandra chinensis seeds. In particular, methanol extract showed high activity in various antioxidant measurement methods. Among the extracts, methanol extract with the highest activity was sequentially fractionated with chloroform, ethyl acetate, butanol and water solvent, and the extraction yield was examined. As shown in Table 6, chloroform was the highest as 48.2%, water was 25.5%, butanol 15.8%, Ethyl acetate followed by 10.6%. The results of comparing the free radical scavenging activity of each fraction are shown in Table 7. The concentration of extract (RC ₅₀ ) required to reduce DPPH by 50% in the ethyl acetate fraction was 17.9 ㎍ / mL and BHA (19.8 ㎍ / mL). mL) and δ-tocopherol (18.7 ㎍ / mL) was more antioxidant activity, but the butanol fraction showed a significant activity of 22.8 ㎍ / mL, but very little in the water fraction.

DPPH Free Radical Reduction Activities of Solvent Seed Extracts by Solvents menstruum Extract concentration (µg / mL) RC ₅₀ ¹⁾ (μg / mL) 0 25 50 100 water 0.279 ²⁾ 0.259 0.245 0.202 174.7 Methanol 0.279 0.174 0.088 0.044 33.2 ethanol 0.279 0.247 0.224 0.124 93.0 Ethyl acetate 0.279 0.236 0.257 0.240 465.0 chloroform 0.279 0.217 0.207 0.233 465.0 BHA 0.279 0.093 0.088 0.090 19.8 δ-tocopherol 0.279 0.083 0.082 0.081 18.7 NOTE 1) Amount required for 50% reduction of DPPH after 30 minutes 2) Average value repeated three times

Fraction Yield of Schisandra chinensis Seed Methanol Extract menstruum chloroform Ethyl acetate Butanol water Fraction yield (%) 48.2 10.6 15.8 25.5

DPPH Free Radical Reduction Activity of Various Solvent Fractions from Schizandra Seed Extracts Extracted with Various Solvents menstruum Extract concentration (µg / mL) RC ₅₀ ¹⁾ (μg / mL) 0 25 50 100 chloroform 0.254 ²⁾ 0.220 0.186 0.124 37.7 Ethyl acetate 0.254 0.077 0.067 0.056 17.9 Butanol 0.254 0.111 0.090 0.065 22.8 water 0.254 0.239 0.229 0.201 254.0 BHA 0.254 0.093 0.088 0.091 19.8 δ-tocopherol 0.254 0.083 0.082 0.081 18.7 NOTE 1) Amount required for 50% reduction of DPPH after 30 minutes 2) Average measured over 3 times

Experimental Example 4: Antibacterial Measurement

The tested strains were Lactobacillus plantarum , MRS broth, Bacillus subtilis and Escherichia coli (ATCC 10536), nutrient broth, Staphylo Staphylococcus aureus and Salmonella typhi murium (KCCM 11806) are tryptic soy broth, Penicillium citrinum (KCCM 11663) Inoculate broth and incubate at 30 ° C for 24 hours to activate and drop 0.3 mL of this culture onto each MRS, nutrient, tryptic soy and PD agar plate and spreader The coating was applied uniformly. Place a ø6 mm filter paper disc (Whatman No. 2) containing 50 μg of Schizandra seed extract on a plate inoculated with each test strain and incubate for 48 hours at 30 ° C. The antimicrobial activity was compared by measuring the diameter of the clear zone appearing around.

Experimental results, as shown in Table 8, the antimicrobial activity of Schisandra chinensis seed extract was higher than that of sorbic acid, a synthetic antimicrobial agent, Lactobacillus plantarum.,Bacillus subtilis, Salmonella typhimurium and Escherichia coli The antimicrobial activity was the same or higher, and lower for Staphylococcus aureus and penicillin citrinum. All extracts showed antimicrobial effects against the food decay strains used in the experiments: Lactobacillus plantarum, Bacillus subtilis, Escherichia coli And penicillin citrinum had the highest antibacterial activity of 12.7 mm, 14.0 mm, 11.3 mm, and 10.6 mm, respectively, and the clear zones of methanol extracts were ethyl acetate extracts from Staphylococcus aureus and Salmonella typhimurium. The clear zone was 11.6 mm and 12.0 mm, respectively. Schisandra chinensis extract showed antimicrobial activity against all test strains, especially against Bacillus subtilis, and showed higher tendency than other test strains for Lactobacillus plantarum and Salmonella typhimurium. According to the above results, the antimicrobial activity was examined by fractionation into chloroform, ethyl acetate, butanol, and water layer using methanol extract showing strong antibacterial activity on all test bacteria. The results are shown in Table 9. Antibacterial activity was generally low in the water fraction, and Staphylococcus aureus and Salmonella typhimurium showed no antimicrobial effect. Lactobacillus plantarum for butanol fraction,Antibacterial stronger than sorbic acid in Bacillus subtilis, Staphylococcus aureus, Salmonella typhimurium and Escherichia coli, with clear zone diameters of 13.1 mm, 14.2 mm, 12.4 mm, 12.4 mm and 13.0 mm, respectively The antimicrobial activity against penicillium citrineum was the best as the water fraction formed a clear zone of 11.9 mm, but it was lower than the synthetic antibacterial sorbic acid (13.8 mm).

Antimicrobial Activity of Schizandra koji Extracts Extracted with Various Solvents Strain Clear zone on plate (mm) water Methanol ethanol acetate Sorbic acid L.plantarum 11.5 ¹⁾ 12.7 12.0 12.5 12.7 B.subtilis 11.9 14.0 12.6 11.7 13.8 S.aureus 10.4 10.6 10.0 11.6 12.3 S.typhimurium 10.5 11.1 10.5 12.0 11.3 E.coli 9.1 11.3 9.7 9.7 11.3 P.citrinum 9.0 10.6 9.9 9.0 13.8 1) Average value measured 3 times

Antimicrobial Activity of Various Solvent Fractions from Schizandra chinensis Seed Extracts Strain Clear Zone on Plate (mm) chloroform Ethyl acetate Butanol water Sorbic acid L.plantarum 12.7 ¹⁾ 12.5 13.1 8.4 12.7 B.subtilis 12.8 13.1 14.2 10.2 13.8 S.aureus 12.0 12.5 12.4 0 12.3 S.typhimurium 11.5 11.7 12.4 0 11.3 E.coli 12.3 12.3 13.0 9.8 11.2 P.citrinum 9.2 9.8 10.8 11.9 13.8 1) Average value measured 3 times

Experimental Example 5: Change of Microorganism Morphology

Butanol fraction of the Schizandra chinensis extract obtained in Example 1 was added to the food decay culture incubated for 48 hours, and left for 3 hours. The cells were separated by centrifugation, immersed in 2% glutaraldehyde solution, and then heated at 4 ° C. for 2 hours. After pre-fixing for 3 hours, washed three times with 0.1 M phosphate buffer and post-fixed with 1% osmium tetroxide solution at 4 ° C. for 2 hours three times with 0.1 M phosphate buffer. Washed. The fixed sample was dehydrated sequentially for 20 minutes using 50%, 70%, 80%, 90%, 95%, and 100% of ethanol, and then immersed in isoamylacetate overnight. Thereafter, the liquid was dried in a critical point dryer using liquid nitrogen and coated with gold using an ion coater to scan a scanning electron microscope (JSM-5410LV, JEOL, Japan). ) And 1000 and 3500 times at 15-20 KV.

As a result, as shown in Figure 1, when the Schisandra chinensis fraction treatment compared to the control cells showed a morphological change swelling, cell wall collapsed, cells were destroyed.

Experimental Example 6: Measurement of nitrite scavenging ability

2 mL of 1 mM NaNO ₂ solution was added to 1 mL (1 mg) of Schisandra chinensis extract extracted with various extracting solvents in Example 1, and the pH of the reaction solution was adjusted to 1.2, 3.0, and 6.0 with 0.2 N citric acid buffer, respectively. After the volume was adjusted to 10 mL, take 1 mL of each reaction solution obtained by reacting at 37 ° C. for 1 hour, add 5 mL of 2% acetic acid solution, add 0.4 mL of Griess reagent, and mix well. After leaving for one minute, absorbance was measured at 520 nm to determine the amount of residual nitrite.

As a result, as shown in Table 10, all the extracts were able to decompose more than 56% of nitrite when the reaction solution was pH 1.2, and methanol and ethanol extracts showed the highest resolution of 98.6% and 91.9%, respectively. At pH 3.0, methanol extract was the highest at 94.1%, followed by water extract (51.7%)> ethanol extract (46.0%)> chloroform extract (5.9%)> ethyl acetate extract (3.9%). At pH 6.0, nitrite resolutions of 21.3% and 3.8% were observed in methanol and water extracts, respectively. The nitrite scavenging effect of methanol extract was higher than that of other extracts at all pHs. The nitrite scavenging effect of each solvent extract was similar to that of the other extracts. Showed little or very low. Since carcinogenic nitrosamines, which are combined with nitrites and amines, are easily produced under strong acid conditions, especially in pH conditions in the human or animal stomach, the fact that methanol extracts of Schisandra chinensis seeds have a high nitrite resolution under strong acid conditions as shown in this experiment. This substance inhibits the production of nitrosamines in the stomach.

In addition, the nitrite scavenging ability of the fraction obtained by distilling methanol extract having the best nitrite resolution using chloroform, ethyl acetate, butanol and water is shown in Table 11. Nitrite resolution at reaction pH of 1.2 was good in the order of butanol extract (98.9%)> ethyl acetate extract (98.4%)> chloroform extract (84.1%)> water extract (57.4%). The chloroform fraction could not degrade nitrite at all, but the butanol, ethyl acetate, and water fractions could degrade 61.8%, 48.4%, and 18.6%, respectively. Butanol fraction showed the highest nitrite resolution of 98.9%, 94.7%, 61.8% in all pH ranges, pH 1.2, pH 3.0 and pH 6.0.

According to the results of Experimental Examples 1 to 6, the organic solvent extract and the fraction having high antioxidant and antimicrobial activity showed the same tendency and the nitrite scavenging effect was high. Therefore, the organic solvent solubles of Schisandra chinensis seeds were found to be involved in antioxidant, antimicrobial and nitrite scavenging activity.

Nitrite Scavenging Ability of Schizandrae Fructus Extracts Extracted with Various Solvents pH Nitrite scavenging ability (%) water Methanol ethanol Ethyl acetate chloroform 1.2 60.2 ¹⁾ 98.6 91.9 55.9 56.5 3.0 51.7 94.1 46.0 3.9 5.9 6.0 3.8 21.3 0 0 0 NOTE 3 average value measured 3 times

Nitrite Scavenging Ability of Various Solvent Fractions from Schizandra chinensis Seed Extracts pH Nitrite scavenging ability (%) chloroform Ethyl acetate Butanol water 1.2 84.1 ¹⁾ 98.4 98.9 57.4 3.0 35.1 83.5 94.7 40.5 6.0 0 48.4 61.8 18.6 1) Average value measured 3 times

As described through the above examples and experimental examples, Schisandra chinensis seed extract and its fractions extracted with various organic solvents exhibit high antioxidant power against lard and soybean oil, have excellent DPPH free radical scavenging activity, and antibacterial against food decay microorganisms. It is a very useful invention in the health food industry because it has an excellent effect of showing activity and an excellent effect of showing a high nitrite scavenging ability.

Claims (4)

After grinding the Schisandra chinensis seed, degreasing it, adding 3 ~ 8 times of organic solvent, extracting 1 ~ 3 times at 70 ~ 95 ℃ temperature, filtering, concentrating with rotary vacuum evaporator and freeze drying, it is prepared by DPPH free Radical scavenging, Lactobacillus plantarum (KCCM 11322), Bacillus subtilis (KCCM 11314), Staphylococcus aureus (KCCM 12103), Salmonella Ty Inhibition of growth of Salmonella typhimurium (KCCM 11806), Escherichia coli (ATCC 10536), Penicillium citrinum [KCCM 11663], and scavenging of nitrite Schizandra seed functional extract, characterized in. The method of claim 1, wherein the organic solvent Schija seed functional extract, characterized in that any one selected from water (water), methanol (methanol), ethanol (ethanol), ethyl acetate, chloroform (chloroform). delete delete