• Korean Journal of Food Science and Technology
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• v.53 no.1
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• pp.34-39
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• 2021

This study investigated the anti-cancer effects of Salicornia herbacea L. fractions in human ovarian cancer cells (OVCAR-3). S. herbacea powder was extracted with 95% EtOH and sequentially fractionated with hexane, dichloromethane (DCM), ethyl acetate, butanol, and H2O. Further, the growth inhibitory effects of the six fractions were determined using the MTS assay. The DCM fraction dramatically decreased cell viability. Similarly, the cell cycle was arrested at the subG1 phase in DCM-treated cells. To confirm apoptosis, the cells were stained with annexin V/FITC-PI solution. Total, early, and late apoptotic cells were significantly increased in the DCM fraction. The mRNA expression of Bcl-2 was reduced, whereas the pro-apoptotic factors Bax and Bak were increased in DCM fraction-treated cells. These results indicated that the DCM fraction in S. herbacea exhibited strong apoptotic effects through the p53-dependent signaling pathway.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.46 no.4
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• pp.339-348
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• 2020

The purpose of this study was to investigate the possible use of the Boehmeria nivea var. nipononivea extract and fractions for the development of natural cosmetic ingredients. The leaves of B. nivea var. nipononivea, extracted by 70% ethanol, were sequentially fractionated with n-hexane, dichloromethane, ethylacetate, and n-butanol. As a result of DPPH and ABTS test, ethyl acetate fractionation was shown to be excellent in radical scavenging activity. For the antimicrobial activities against Staphylococcus aureus, Staphylococcus epidermidis, Cutibacterium acnes and antibiotic resistant strains, MIC and birth control rate were observed by paper disc method. In the antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and C. acnes, the dichloromethane and ethylacetate fraction showed stronger antibacterial activity than the other fractions and extract. Moreover, the ethylacetate fraction showed strong nitric oxide (NO) production inhibitory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell. In conclusion, we found that B. nivea var. nipononivea extract was not cytotoxic and showed antioxidant, antimicrobial and anti-inflammatory effects. These results suggest that the Boehmeria nivea var. nipononivea extract and fractions could be applied as an effective cosmetic material with antioxidant activity.

• The Korea Journal of Herbology
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• v.32 no.2
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• pp.57-64
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• 2017

Objectives : Yam bean (Pachyrhizus erosus) possess various nutrients, it has been widely used as traditional cosmetic material in Indonesia. The aim of this study was to investigate the anti-oxidant activity and the anti-melanogenic effect of Yambean (Pachyrhizus erosus) extract and its fractions. Methods : The anti-oxidant activity of yam bean extract assessed based on total polyphenol, flavonoid contents, DPPH and ABTS radical scavenging assay. To evaluate anti-melanogenic effects and cytotoxicity of Yambean extract and its fractions, B16F10 melanoma cell was used. Results : In results, total polyphenol content of yam bean water extract (YW) and Yambean 70% ethanol extract (YE) were $1.18{\pm}0.03mg/g$ (mg of gallic acid/g of sample), $1.16{\pm}0.01mg/g$. Total flavonoid contents of YW, YE were $3.55{\pm}0.06mg/g$ (mg of naringin/g of sample), $1.78{\pm}0.03mg/g$. Moreover, YE scavenged DPPH and ABTS effectively in $4mg/m{\ell}$ compared to YW. Cytotoxicity of YE and its fractions in B16F10 melanoma cell was measured using MTT assays. It had no cytotoxicity up to $500{\mu}g/m{\ell}$. Melanin accumulation in B16F10 melanoma cell was induced using alpha-melanocyte stimulating hormone (${\alpha}-MSH$) and 3-isobutyl-1-methylxanthine (IBMX). B16F10 melanoma cell treated with $10-500{\mu}g/m{\ell}$ YE and hexane, ethyl acetate, butanol, $H_2O$ fractions for 24h. Non treated B16F10 melanoma cell (Control) markedly increased melanin contents. In contrast, YE ethylacetate fraction effectively suppressed melanin accumulation in a dose-dependent manner. Conclusion : In conclusion, these results suggest that Yambean extract has the potential as a cosmetic material which possess anti-oxidant and anti-melanogenic activities.

• The Korea Journal of Herbology
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• v.24 no.3
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• pp.39-50
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• 2009

Objectives : Smilacis glabrae rhizoma (SG) has been traditionally used as a herbal medication of musculoskeletal disorders like arthritis, pain, convulsions, and syphilis in traditional Korean medicine. This study was investigated anti-oxidative and anti-inflammatory effect of fractionated extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (HUVEC). Methods : SG extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of SG onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymethoxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of ICAM-1 and VCAM-1 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially ethyl acetate (EA) extract, significantly inhibited free radical generation, the TNF-$\alpha$-induced intracellular oxidation. Furthermore, the EA extract protected TNF-$\alpha$-induced adhesion to THP-1, expression of adhesion molecules accompanied by an attenuation of IL-6 and IL-8 formation in HUVEC. Conclusions : These results indicate that EA extract of SG have potential as an agent of atherosclerosis and other chronic inflammatory diseases including diabetes, hypertension, and arthritis.

• Clean Technology
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• v.29 no.2
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• pp.109-117
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• 2023

The purpose of this study is to develop a method for separating antioxidants and sugars from grape skin extract. The extract was first mixed with a variety of organic solvents to investigate whether the separation was feasible. When employing acetone, ethanol, dimethylsulfoxide, or dimethylformamide, the organic solvent-extract combination formed a single phase. However, when benzene, ethyl acetate, or n-hexane was added to the extract, the mixture separated into an organic and an aqueous phase and the pigments remained in the aqueous phase. On the other hand, when polyethylene glycol-2,000 (PEG-2000) and sodium citrate were added to the extract, the mixture was separated into three layers, with the majority of the flavonoids migrating to the top layer and 53% of the extract's glucose migrating to the bottom layer. The top layer had significant antioxidant activity, whereas the bottom layer showed no antioxidant activity. The glucose recovery in the bottom layer increased as the molecular weight of PEG increased and the highest recovery (67%) was observed when PEG-8,000 was added. The highest flavonoid separation was observed with PEG-2,000, followed by PEG-8,000 and PEG-400. The flavonoid separation when PEG-2,000 was added resulted in a flavonoid recovery of 48% and 0.2% from the top and bottom layers, respectively. Examining the effect of the separated solution using the agar disc diffusion method on yeast cell growth confirmed that the addition of the extract, the top, and the bottom layer did not inhibit cell growth.

• Journal of Life Science
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• v.33 no.7
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• pp.538-548
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• 2023

The research has been conducted on the isolation of antimicrobial compounds from plant natural extracts and their potential application in oral health care products. This study aimed to investigate the antimicrobial mechanism by analyzing the changes in gene expression of Streptococcus mutans, a major oral pathogen, in response to complex compounds extracted from Aralia continentalis and Arctii Semen using organic solvents. Transcriptome analysis (RNA-seq) revealed that both natural extracts commonly upregulated or downregulated the expression of various genes associated with different metabolic and physiological activities. Three genes (SMU_1584c, SMU_2133c, SMU_921), particularly SMU_921 (rcrR), known as a transcription activator of two sugar phosphotransferase systems (PTS) involved in sugar transport and biofilm formation, exhibited consistent high expression levels. Additionally, component analysis of the A. continentalis extract was performed to compare its effects on gene expression changes with the A. Semen extract, and two active compounds were identified through gas chromatography-mass spectrometry (GC-MS) analysis of the active fraction. The n-hexane fraction (ACEH) from the A. continentalis extract exhibited antibacterial specificity against S. mutans, leading to a significant reduction in the viable cell counts of Streptococcus sanguinis and Streptococcus gordonii among the tested multi-species bacterial communities. These findings suggest the broad-spectrum antibacterial activity of the A. continentalis extract and provide essential foundational data for the development of customized antimicrobial materials by elucidating the antibacterial mechanism of the identified active compounds.

• 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.

• Analytical Science and Technology
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• v.22 no.5
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• pp.407-414
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• 2009

A simultaneous determination method for cholesterol, lanosterol and desmosterol was developed using gas chromatography/mass spectrometry. Urine was enzymatically hydrolyzed with ${\beta}$-glucuronidase/arylsulfatase. Samples were prepared using extractions with a mixture of ethyl acetate-hexane (2:3, v/v), followed by derivatization with a mixture of MSTFA/TMSI/TMCS (100:2:5 v/v/v). All analyses were performed using GC/MS in selective ion monitoring mode. Good linearities ($r^2=0.998{\sim}0.999$) in calibration curve and a satisfactory recovery (80.0%~113%) were achieved. Accuracy and precision values within ${\pm}15%$ in the concentration range of 5 to 200 ng/mL were also observed for all compounds. The developed method was applied to pravastatin-treated (70 and 250 mg/kg/day for 7 days, oral) hyperlipidemia rats. Those sterols were significantly lower in drug-treated rats compared to the controls, which justifies the drug efficacy. Therefore, these results indicate that the developed method was successfully applied to examine statin drug efficacy with urine sample.

• Analytical Science and Technology
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• v.22 no.6
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• pp.514-518
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• 2009

Coenzyme $Q_{10}$($CoQ_{10}$), a vitamin E-like substance, represents a components of the complex antioxidant system of the human organism. $CoQ_{10}$ levels in human plasma were determined by high performance liquid chromatography (HPLC) with UV detection. It was dissociated from lipoproteins by methanol and extracted into n-hexane with liquid-liquid extraction procedure, after centrifugation, the supernatant was dried under nitrogen gas stream. The residue was dissolved in the absolute ethanol. Determination of $CoQ_{10}$ was performed on a $C_{18}$ reversed-phase analytical column with ultraviolet detection at 275 nm and the mobile phase containing 15% (v/v) ethanol in methanol at a flow rate of 1.7 mL/min. The low limit of quantitation was 0.02 mg/L (S/N=10), the linearity between the concentration and peak height is from 0.1 to 2.0 mg/L. Twenty-four randomly selected plasma samples from apparently healthy, 27 to 44 year old individuals (males and females) were analyzed for total $CoQ_{10}$. The average level in these subjects was $0.62{\pm}0.13mg/L$ with the range of 0.41-0.98 mg/L. This method has a specific and a sufficient limit of quantitation (LOQ) for analysis of $CoQ_{10}$ in human plasma in both a clinical study and research at laboratories.

• Analytical Science and Technology
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• v.20 no.1
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• pp.84-90
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• 2007

To determine levels of 11 sulfonamides for animals in food, simultaneously, a selective method of high performance liquid chromatography with UV detector has been applied. The targets were sulfachlorpyridazine (SCP), sulfadiazine (SDZ), sulfadimethoxine (SDM), sulfisoxazole (SSX), sulfamerazine (SMZ), sulfamethazine (SMT), sulfamethoxazole (SMX), sulfamethoxypyridazine (SMP), sulfamonomethoxine (SMM), sulfaquinoxaline (SQX) and sulfathiazole (STZ). Food samples were beef, pork, chicken, milk and whole egg that were collected at the main 6 cities in Korea as Seoul, Busan, Daejon, Incheon, Mokpo and Gangneung. After homogenizing food samples with sodium phosphate solution and acetonitrile, it was extracted with n-hexane. The mobile phase gradient was a mixture of 5 mM potassium phosphate (pH 3.25) and methanol with a gradient ratio from 100:0 to 30:70. The UV wavelength was 270 nm. The overall recoveries were ranged from 75% to 95% and the limit of detection was minimum 0.004 mg/kg for SMT, and 0.007 mg/kg for STZ at signal/noise > 3, respectively. As results, sulfonamide drugs were not detected in most of the selected food samples, however, sulfamonomethoxine was detected in meat. The determined level of sulfamonomethoxine were 0.03 and 0.06 mg/kg for beef that were below the MRLs.