• Journal of Life Science
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• v.29 no.2
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• pp.231-238
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• 2019

This study evaluated the antioxidant activity of the extract and fractions of Alnus firma. Alnus firma had the highest total phenolic content ($452.80{\pm}7.01{\mu}g$ gallic acid equivalents/mg) in a methanol (MeOH) fraction and the highest total flavonoid content ($112.29{\pm}11.14{\mu}g$ rutin equivalents/mg) and antioxidant capacity ($936.23{\pm}0.07{\mu}g$ ${\alpha}$-tocopherol equivalents/mg) in an ethylacetate (EA) fraction. The antioxidant activities of various solvent extract fractions of Alnus firma were evaluated using various antioxidant assays, including ${\beta}$-carotene-linoleate assay, reducing power assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, metal chelating activity assay, superoxide anion radical scavenging assay, and lipid peroxidation inhibition assay using the ferric thiocyanate method. These activities were compared with those of ascorbic acid, butylated hydroxyanisole (BHA), gallic acid (GA), butylated hydroxytoluene (BHT), and ${\alpha}$-tocopherol. First, at a $250{\mu}g/ml$ concentration, the EA and MeOH fractions of A. firma showed 92.43% and 89.20% DPPH radical scavenging activity, respectively. Second, $50{\mu}g/ml$ of the EA fraction exhibited 72.49% superoxide anion radical scavenging activity, a little greater than the same dose of GA (60.88%). Finally, 0.5 and 1 mg/ml of the EA fraction showed 73.45% and 73.29% inhibition of peroxidation in the ${\beta}$-carotene-linoleic acid system, respectively. The decreasing order of reducing power was EA fraction > n-butanol (BuOH) fraction > dichloromethane (DCM) fraction > n-hexane (HX) fraction. The results obtained in the present study indicated that Alnus firma can be used as an easily accessible potential source of natural antioxidants.

• Journal of the Korean Society of International Agriculture
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• v.30 no.4
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• pp.339-346
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• 2018

Cyanazine is a member of the triazine family of herbicides. Cyanazine is used as a pre- and post-emergence herbicide for the control of annual grasses and broadleaf weeds. This experiment was conducted to establish a determination method for cyanazine, as domestic unregistered pesticide, residue in major agricultural commodities using HPLC-DAD/MS. Cyanazine was extracted with acetone from representative samples of five raw products which comprised apple, green pepper, Kimchi cabbage, hulled rice and soybean. The extract was diluted with saline water and partitioned to dichloromethane for remove polar extractive in the aqueous phase. For the hulled rice and soybean samples, n-hexane/acetonitrile partition was additionally employed to remove non-polar lipids. The extract was finally purified by optimized florisil column chromatography. On a $C_{18}$ column in HPLC, cyanazine was successfully separated from co-extractives of sample, and sensitively quantitated by diode array detection at 220 nm. Accuracy and precision of the proposed method was validated by the recovery experiment on every major agricultural commodity samples fortified with cyanazine at 3 concentration levels per agricultural commodity in each triplication. Mean recoveries were ranged from 83.6 to 93.3% in five major representative agricultural commodities. The coefficients of variation were all less than 10%, irrespective of sample types and fortification levels. Limit of quantitation(LOQ) of cyanazine was 0.02 mg/kg as verified by the recovery experiment. A confirmatory method using LC/MS with selected-ion monitoring(SIM) technique was also provided to clearly identify the suspected residue.

• Analytical Science and Technology
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• v.20 no.4
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• pp.347-354
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• 2007

A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in bovine, porcine, chicken, flatfish and japanese eel muscle has been developed and validated. The separation condition for HPLC/UV was optimized with phenyl hexyl ($4.6{\times}150mm$, $5{\mu}m$) column with 10 mM monobasic sodium phosphate buffer (pH 2.5)/acetonitrile (50/50, v/v) as the mobile phase at a flow rate of 1.0 mL/min and detection wavelength was set at 254 nm. Residues were extracted from tissue by blending with methanol and lipid materials were removed with n-hexane. Then, the methanol extract was evaporated to dryness under a nitrogen stream, reconstituted in the mobile phase. Aliquot of the organic extract was decanted and filtered through $0.45{\mu}m$ syringe filter. The $20{\mu}L$ of the resulting solution was injected into the HPLC system. The calibration ranges were $0.5{\sim}5{\mu}g/g$ and calibration curves were linear with coefficients of correlation better than 0.95. The limits of quantification were $0.5{\mu}g/g$ for all muscles. The recoveries of bovine, porcine, chicken, flatfish and japaneseel muscles were 99.8%, 102.4%, 91.0%, 104.0% and 93.0%, respectively. The procedures were validated according to the CODEX guideline, determining specificity, linearity, accuracy, precision, quantitation limit and recovery.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7179-7188
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• 2015

Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

• Korean Journal of Medicinal Crop Science
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• v.11 no.3
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• pp.236-245
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• 2003

The study was performed for elucidating angiotensin converting enzyme (ACE) inhibitory activity and comparing antioxidative activity of Panax ginseng extracts prepared at different conditions. Total phenolic content, inhibitory activity on ACE and antioxidative effects were tested on 10 ethanolic extracts and correlation coefficient between total phenolic content and physiological activity was calculated. Yield and total phenolic content of 50% ethanolic extract prepared at $85^{\circ}C$ exhibited the highest value as 42.52% and 0.82%, respectively. Among the fractions obtained from 50% ethanolic extract prepared at room temperature, water fraction showed the highest value in yield as 72.08% and ethyl acetate fraction did in total phenolic content as 6.59%. In the test on ACE inhibitory activity, 50% ethanolic extract obtained at room temperature indicated the strongest effect of 93.8% which was higher than 85.2% of commercialized ACE inhibitor and solvent fractions showed potent inhibitory activity in order of hexane fraction, diethyl ether fraction, ethyl acetate fraction, butanol fraction and water fraction at concentration of $4000{\mu}g/ml$. 50% Ethanolic extract prepared at $85^{\circ}C$ had the most potent inhibition effect on human LDL oxidation as 78.2% at $200{\mu}g/ml$ and the other extracts also did above 60%. Diethyl ether fraction and ethyl acetate fraction showed strong inhibition activity $(34.38%{\sim}78.13%)$ on LDL oxidation at concentration of $10{\sim}200\;{\mu}g/ml$. From the statistical analysis via SAS program, correlation coefficient between total phenolic content and ACE inhibitory effect was 0.6353 at P<0.05. Conclusively, this report showed that the most efficient extraction condition for elevating inhibitory activity on ACE and LDL oxidation, phenolic content and yield from Panax ginseng was 50% ethanol extraction at room temperature or high temperature condition. And Panax ginseng would be used for preventing hypertension or atheroscrelosis for man via inhibitory action on ACE and LDL oxidation.

• Journal of the Korean Applied Science and Technology
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• v.29 no.3
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• pp.486-494
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• 2012

Nano-emulsion with phase inversion temperature (PIT) emulsifying system was prepared to use rapeseed oil from originating Jeju in order to apply various cosmetic applications. Natural rape seed oil (NRSO) extraction was extracted using n-hexane as a solvent. NRSO extract showed a light yellowish color of viscous liquid as well as yield was $43{\pm}2.5%$. Acid value was $2.76{\pm}0.5$ and gravity was $0.89{\pm}0.05$. Droplet size of PIT-Yuche-NE with 20wt% of rapeseed oil was 50-120nm (average: $82{\pm}5.8nm$) and zeta potential was -29.5mV. It was thermodynamically good stable emulsion due to $(PEG)_{5-30}$fattyacidether. Some conclusions from the result of characteristic experiment were obtained as follows. First, the anti-oxidative activity was measured by free radical scavenging activity using DPPH (1,1-diphenyl-2-picrylhydrazyl radical). Anti-oxidative activity of PIT-Yuche-NE was $37.2{\pm}6.7%$ on 10mg/mL compared with PIT-Toco-NE (Natural tocopherol nano-emulsion, $28.8{\pm}6.5%$ on 10 mg/mL) and PIT-Nokcha-NE (Green tea extract nano-emulsion, $29.6{\pm}7.2%$ on 10mg/mL). Second, the collagen synthesis activity of PIT-Yuche-NE was $148{\pm}15.2%$ compared with PIT-Toco-NE (Natural tocopherol nano-emulsion, $121{\pm}13.5%$ on 10mg/mL) and PIT-Nokcha-NE (Green tea extract nano-emulsion, $95{\pm}12.7%$ on 10mg/mL). Third, the effectiveness of moisturizing activity of Yuche-CRM with Aramo-TS after 6 hours increase $47{\pm}3.9%$ (*p-value￡0.05, n=7) whereas Both Toco-CRM was $30{\pm}5.2%$ (*p-value￡0.05, n=7) and Nokcha-CRM was $35{\pm}4.5%$. Therefore, Yuche-CRM has higher moisturizing effect than other two creams. Finally, Nano-emulsion stabilizing rapeseed oil using PIT emulsifying system of this study can be used to apply cosmetics industry and pharmaceutical industry.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.3
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• pp.384-394
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• 2009

This study was performed to determine the antioxidative and anticancer effects of extracts from Adenophora remotiflora leaves. The antioxidative effects of the extracts were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity and hemoglobin-induced linoleic acid oxidative inhibition assays. The results indicated that the extracts had stronger effects than the synthetic antioxidant BHT at the same concentration. The $SC_{50}$ values (50% radical scavenging effect on $1{\times}10^{-4}$ M DPPH) of the methanol fraction, water extract, and BHT were 47.5 ${\mu}g$/mL, 74.6 ${\mu}g$/mL and 102.2 ${\mu}g$/mL, respectively. In addition the $IC_{50}$ values (hemoglobin-induced linoleic acid oxidation inhibition) of the methanol fraction, water extract, and BHT were 120.8 ${\mu}g$/mL, 135.6 ${\mu}g$/mL, and 150.2 ${\mu}g$/mL, respectively. This research also assessed decreases in the survival of BNLcl2 cells (normal liver cells) by solvent fractions of the A. remotiflora leaf extracts at various concentrations (1, 5, 10, 25, 50, 100, 250, 500, 1,000, 2,000 ${\mu}g$/mL). The water extract did not decrease survival at any of the concentrations when compared to the control group. The hexane, ethyl acetate, and methanol fractions decreased survival as compared to the control group by inducing cell toxicity at a concentration of 1,000 ${\mu}g$/mL and above. Therefore, an anticancer activity experiment was conducted using concentrations below 500 ${\mu}g$/mL. At 500 ${\mu}g$/mL, the methanol fraction decreased A549 cell (human lung carcinoma cells) survival by 46% as compared to the control group, presenting the greatest effect against cell survival. All extracts showed greater anticancer activity in Hep G2 cells (human liver carcinoma cells) as compared to the A549 cells. For the Hep G2 cells, the methanol extract decreased survival by 28% as compared to the control group at the concentration of 500 ${\mu}g$/mL, thus restraining lung cancer cell growth.

• Korean Journal of Food Science and Technology
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• v.53 no.1
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• pp.12-18
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• 2021

The in vitro antioxidant activity of oregano seed fractions, fractionizing with 80% ethanol, n-hexane, ethyl acetate, n-butanol, and water, was evaluated, and their effects on edible oils were determined in corn oil at 180℃. The ethyl acetate fraction had the highest radical-scavenging activity. The ferric reducing antioxidant power activity and total phenol content of the ethyl acetate fraction were determined as 6,130 µmol ascorbic acid equivalents/g extract and 770 µmol tannic acid equivalents/g extract, respectively, which were significantly higher than those of the other fractions (p<0.05). Primary and secondary oxidation products in corn oil added with the ethyl acetate fraction of oregano seed significantly decreased by 1.5 and 1.26 times, respectively, compared with those in the control groups. The major volatile ingredients in the ethyl acetate fraction of oregano seeds were determined to be carvacrol, thymoquinone, and 3-cyclopentylcyl-cyclopentan-1-one. Ethyl acetate is a suitable solvent for extracting antioxidant compounds from oregano seeds and can be used as a natural antioxidant.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.10
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• pp.1295-1301
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• 2009

This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effect of Codonopsis lanceolata (CL). CL was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of CL extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. CL extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of CL (200 ${\mu}g$/plate) showed approximately 72.1% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 69.6% and 67.0% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of CL extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (HepG2), human breast adenocarcinoma (MCF-7), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL CL ethyl acetate fraction had the highest cytotoxicity of 74.5%, 70.7% and 80.3% against HeLa, MCF-7 and A549 cells, respectively. In contrast, the extract and its fractions showed only 2$\sim$31% cytotoxicity for a normal human kidney cell line (293). In vivo anticancer effect of CL extract was tested using Balb/c mice transplanted sarcoma-180 cells. CL ethyl acetate fraction showed the highest inhibition rate of 56.4% at the 50 mg/kg concentration.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.2
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• pp.228-238
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• 2007

This study was conducted to increase sweetpotato utilization and to determine the vegetative value of sweetpotato tips by investigating the phenolic compounds, antioxidative effect in oil, electron donating ability, nitrite scavenging effect and ACE inhibition activities. The phenolic compounds present in sweetpotato tips are the gallic, chlorogenic, gentisic, caffeic, couramic and ferulic acid, which are 16-122 times higher compared to other vegetables such as spinach, soybean sprout, and perilla leaves. In each solvent extract, the total phenolic compounds (175.8mg/g) was composed of 55% EtOAc extraction and 39% BuOH extract, respectively. The results of induction period using the Rancimat method showed that the antioxidant activity of SP tips was higher than the tocopherol or BHT. The relative levels of each solvent extract in SP tips were as follows: EtOAc>BHT>BuOH>Tocopherol>Water>$CHCl_3$>Hexane. The peroxide value was measured every 5 days for 25 days during storage and results showed that the peroxide value, the tips, tuberous root and tocopherol were lower compared to spinach, soybean sprout and perilla leaves. Nitrite scavenging effects were excellent in sweetpotato tips, perilla leaves and soybean sprout, especially, inhibition rate of perilla leaves (72%) were superior to the others. In process of solvent extraction, activity of BuOH and water extractions were the best. ACE inhibition activity in sweetpotato tips was 1.5 times higher than in tuberous roots and $1.9{\sim}3.7$ times higher than in spinach, soybean sprout, perilla leaves.