• Title/Summary/Keyword: HPLC analysis methods

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Antioxidant Effects of Sagunja-Tang (Sijunzi-Tang) (사군자탕(四君子湯)의 항산화(抗酸化) 효과(效果))

  • Lee Yong-Tae;Cho Su-In;Kim Young-Kyun
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.2
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    • pp.170-192
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    • 2000
  • Objectives : This study was carried out to research antioxidant effects of Sagunja-Tang(SA) through in vitro and vivo experiments, and tried to investigate the relation between oxidation of tissues and deficiency of Qi. Methods and results : HPLC analysis of glycyrrhizine - known to be the main compound of Radix Glycyrrhizae - was done to certify the quality of SA. Chemiluminescence was initiated by adding tort-butyl hydroperoxide (t-BHP) to rabbit polymorphonuclear leukocytes (neutrophils), and generated reactive oxygen species (superoxide anion) decreased significantly by SA as dose dependent manner. Cell injury during 60 minutes tissue incubation was initiated by adding t-BHP, a hydrophobic hydroperoxide and $H_2O_2$, an water soluble oxidant to rat renal cortical and liver slices. Percentage cell death and lipid peroxidation were estimated by measuring lactate dehydrogenase (LDH) and malondialdehyde (MDA), a product of lipid peroxidation. t-BHP induced % cell death of renal cortical slices and lipid peroxidation of renal cortical and liver slices were decreased significantly by SA. SA decreased significantly % cell death and lipid peroxidation of renal cortical and liver slices induced by $H_2O_2$, too. Acute renal and liver injury induced by $HgCl_2\;and\;CCl_4$, which initiated from free radical, were applied to mice and metabolic data were obtained. Data showed protective effects of SA on acute renal injury caused by decrease of glomerular filtration. SA protected acute liver injury too. Conclusions Through this study, we found that SA have antioxidant effects and tissue oxidation was similar to deficiency of Qi. And further studies have to be followed to certify the mechanisms.

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Enhancement of Skin Immune Activities of Spirulina maxima by High Pressure Extraction Process (Spirulina maxima 초고압 추출물의 피부 면역 활성 증진)

  • Oh, Sung-Ho;Kang, Do-Hyung;Choi, Woon-Yong;Seo, Yong-Chang;Heo, Soo-Jin;Abu, Affan Md.;Jeong, Kyung-Hwan;Lee, Hyeon-Yong
    • Ocean and Polar Research
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    • v.32 no.2
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    • pp.157-164
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    • 2010
  • A marine alga, Spirulina maxima, was extracted under high pressure and low temperature conditions at 500 MPa and $60^{\circ}C$ for 5 and 10 min. A high pressure of 500 MPa was applied to improve process yields because of low temperature extraction. This method resulted in highest higher extraction yield of 26.1% (w/w) in comparison to those results obtained from conventional extraction methods which produced a yield of 17.6% (w/w) from water. The extracts from this process also showed 19% of low cytotoxicity against human normal fibroblast cells in adding 1.0 mg/ml of the highest concentration. The crude extract significantly reduced the production of Prostaglandin $E_2$ ($PGE_2$) from CCD-986sk cells and increased nitric oxide production by macrophages. These higher activities of enhancing skin immune functions were found to have high antioxidant extract properties, like a 98% increase in DPPH radical scavenging activity. The extracts from the high pressure process showed a higher elution of active components than other processes and generated new compounds based on HPLC analysis. This clearly indicates that the extracts from high pressure and low temperature conditions have higher skin immune activation properties that have not been previously reported.

Sonchus asper extract inhibits LPS-induced oxidative stress and pro-inflammatory cytokine production in RAW264.7 macrophages

  • Wang, Lan;Xu, Ming Lu;Liu, Jie;Wang, You;Hu, Jian He;Wang, Myeong-Hyeon
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.579-585
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    • 2015
  • BACKGROUND/OBJECTIVES: Sonchus asper is used extensively as an herbal anti-inflammatory for treatment of bronchitis, asthma, wounds, burns, and cough; however, further investigation is needed in order to understand the underlying mechanism. To determine its mechanism of action, we examined the effects of an ethyl acetate fraction (EAF) of S. asper on nitric oxide (NO) production and prostaglandin-E2 levels in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. MATERIALS/METHODS: An in vitro culture of RAW264.7 macrophages was treated with LPS to induce inflammation. RESULTS: Treatment with EAF resulted in significant suppression of oxidative stress in RAW264.7 macrophages as demonstrated by increased endogenous superoxide dismutase (SOD) activity and intracellular glutathione levels, decreased generation of reactive oxygen species and lipid peroxidation, and restoration of the mitochondrial membrane potential. To confirm its anti-inflammatory effects, analysis of expression of inducible NO synthase, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and the anti-inflammatory cytokines IL-$1{\beta}$ and IL-6 was performed using semi-quantitative RT-PCR. EAF treatment resulted in significantly reduced dose-dependent expression of all of these factors, and enhanced expression of the antioxidants MnSOD and heme oxygenase-1. In addition, HPLC fingerprint results suggest that rutin, caffeic acid, and quercetin may be the active ingredients in EAF. CONCLUSIONS: Taken together, findings of this study imply that the anti-inflammatory effect of EAF on LPS-stimulated RAW264.7 cells is mediated by suppression of oxidative stress.

Rhus verniciflua Stokes extract suppresses migration and invasion in human gastric adenocarcinoma AGS cells

  • Lee, Hyun Sook;Jung, Jae In;Kim, Kyeong-Hee;Park, Sang Jae;Kim, Eun Ji
    • Nutrition Research and Practice
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    • v.14 no.5
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    • pp.463-477
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    • 2020
  • BACKGROUND/OBJECTIVES: Many studies have suggested that Rhus verniciflua Stokes (RVS) and its extract are anticancer agents. However, RVS had limited use because it contains urushiol, an allergenic toxin. By improving an existing allergen-removal extraction method, we developed a new allergen-free Rhus verniciflua Stokes extract (RVSE) with higher flavonoid content. In this study, we examined whether RVSE inhibits the ability of AGS gastric cancer cells to migrate and invade. MATERIALS/METHODS: The flavonoids content of RVSE was analyzed by HPLC. The effects of RVSE on migration and invasion in AGS cells were analyzed by each assay kit. Matrix metalloproteinase (MMP)-9, plasminogen activator inhibitor-1 (PAI-1) and urokinase-type plasminogen activator (uPA) protein expression was analyzed by protein antibody array. The Phosphorylation of signal transducer and activator of transcription (STAT) 3 were assayed by Western blot analysis. RESULTS: RVSE treatment with 0-100 ㎍/mL dose-dependently reduced the ability of AGS cells to migrate and invade. Notably, treatment with RVSE strongly inhibited the expression of MMP-9 and uPA and the phosphorylation of STAT3. In contrast, RVSE treatment dramatically increased the expression of PAI-1. These results indicate that the inhibition of MMP-9 and uPA expression and STAT3 phosphorylation and the stimulation of PAI-1 expression contributed to the decreased migration and invasion of AGS cells treated with RVSE. CONCLUSIONS: These results suggest that RVSE may be used as a natural herbal agent to reduce gastric cancer metastasis.

GTPase Activity Analysis of eRF3 in Euplotes octocarinatus

  • Song, Li;Dong, Jun-Li;Zhao, Ya-Qin;Chai, Bao-Feng;Liang, Ai-Hua
    • Journal of Microbiology and Biotechnology
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    • v.20 no.9
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    • pp.1283-1287
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    • 2010
  • In eukaryotes, eRF3 participates in translation termination and belongs to the superfamily of GTPases. In this work, the dissociation constants for nucleosides bound to Euplotes octocarinatus eRF3 in the presence and absence of eRF1a were determined using fluorescence spectra methods. Furthermore, a GTP hydrolyzing assay of eRF3 was carried out using an HPLC method, and the kinetic parameters for GTP hydrolysis by eRF3 were determined. Consistent with data from humans, the results showed that eRF1a promoted the binding of GTP to eRF3 and the GTP hydrolyzing activity of eRF3. However, in contrast to the lack of GTP binding in the absence of eRF1 in human eRF3, the E. octocarinatus eRF3 was able to bind GTP by itself. The nucleotide binding affinity of the E. octocarinatus eRF3 also differed from the human data. A structure model and amino acid sequence alignment of potential G domains indicated that these differences may be due to valine 317 and glutamate 452 displacing the conserved glycine and lysine involved in GTP binding.

The Isoflavonoid Constituents and Biological Active of Astragalus Radix by Fermentation of β-glucosidase Strains (β-glucosidase 활성 균주 발효에 의한 황기 Isoflavonoid 성분변화 및 생리활성)

  • Kim, Chul Joong;Choi, Jae Hoo;Seong, Eun Soo;Lim, Jung Dae;Choi, Seon Kang;Yu, Chang Yeon;Lee, Jae Geun
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.5
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    • pp.371-378
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    • 2020
  • Background: In this study, the radix of Astragalus membranaceus Bunge extract fermented by Saccharomyces cerevisiae, Weissella cibaria, and Pediococcus pentosaceus to increase the levels of isoflavonoid aglycone contents. Methods and Results: In order to change the in isoflavonoids, we fermented the radix of A. membranaceus extracts with microorganisms that have β-glucosidase activity. Besed on the β-glucosidase activity, we selected three strains, Weissella cibaria, Pediococcus pentosaceus, and Saccharomyces cerevisiae. HPLC analysis revealed that the levels of isoflavonoid aglycones were increased in all fermentation cases, and the extracts fermented by S. cerevisiae showed the highest levels of isoflavonoid aglycones. We evaluated the antioxidant activity, anti-wrinkle effects and whitening effects of the S. cerevisiae-fermented extracts using the DPPH assay, tyrosinase inhibition activity assay, and collagenase inhibition activity assay. We confirmed higher activity in S. cerevisiae-fermented extracts than in control, with the half maximal inhibitory concentration (IC50) value of 565.1 ± 59.1 ㎍/㎖ in DPPH radical scavenging activity, tyrosinase inhibition rate of 78.4 ± 0.9%, and collagenase inhibition rate of 83.8 ± 1.1%. Conclusions: We selected three stains of microorganisms showing high β-glucosidase activity, W. cibaria, P. pentosaceus and S. cerevisiae. Isoflavonoid glycones in the radix of A. membranaceus were converted to isoflavonoid aglycones by fermentation. In addition, the fermented radix of A. membranaceus exhibited antioxidant activity, anti-wrinkle effect, whitening effect and radical scavenging activity.

Residue Dissipation Behavior of Bistrifluron and Cyenopyrafen in Peach for the Cultivation Periods under Field Conditions (노지재배 복숭아 중 Bistrifluron과 Cyenopyrafen의 생산단계 잔류특성에 따른 감소추이)

  • Hwang, Eun-Jin;Park, Jung-Eun;Kwon, Chan-Hyeok;Kim, Jin-Sook;Chang, Hee-Ra
    • Korean Journal of Environmental Agriculture
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    • v.37 no.1
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    • pp.41-48
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    • 2018
  • BACKGROUND: This study was performed to calculate the biological half-lives and regression coefficient of bistrifluron and cyenopyrafen dissipation in peach and to estimate the Pre-Harvest Residue Limits (PHRLs). METHODS AND RESULTS: The bistrifluron and cyenopyrafen were prepared on the basis of good agricultural practice for peach and treated with a single application. Peaches were harvested at 0, 1, 3, 5, 7, 10 and 14 days after application, prepared for analysis, and analyzed by HPLC-DAD. The limits of quantitation (LOQ) of bistifluron and cyenopyrafen were 0.02 mg/kg and 0.04 mg/kg, respectively. The recoveries of bistrifluron and cyenopyrafen were 99.5~108.7% and 88.4~98.9% at two different concentration levels. The biological half-lives of field I (Sejong) and field II (Pyeongtaek) were 6.1 and 7.0 days for bistifluron, and 6.3 and 7.0 days for cyenopyrafen, respectively. The 95% confidence intervals of dissipation rate constants of bistrifluron in peach were 0.0805~0.1457 and 0.0577~0.1417 for field I and field II, respectively, and those of cyenopyrafen were 0.0911~0.1278 and 0.0576~0.1417, respectively. CONCLUSION: Residue dissipation of bistrifluron and cyenopyrafen in peach were similar to that of correction to sample weight difference during the harvest periods. This study suggests that residue dissipation rate would be helpful to set the PHRLs that protect public health.

Selection of Tropane Alkaloids High-Producing Lines by Single Cell Cloning of Hyoscyamus niger L. Root Cultures

  • Min, Ji-Yun;Park, Dong-Jin;Jeong, Mi-Jin;Song, Hyun-Jin;Kang, Seung-Mi;Kang, Young-Min;Choi, Myung-Suk
    • Journal of Korean Society of Forest Science
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    • v.98 no.2
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    • pp.142-147
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    • 2009
  • Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.

A Study on Vitamin C Content of Nutrition Emphasized Products (영양강조표시제품 중 비타민 C 함량 조사)

  • Jeong, Da-un;Lee, Heon-Ok;Kim, Young-Kyoung;Om, Ae-Son
    • Korean Journal of Community Nutrition
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    • v.21 no.6
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    • pp.574-579
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    • 2016
  • Objectives: Vitamin C has various functions such as antioxidative effect and supporting absorption of iron (Fe). Aim of this present study was to provide vitamin C nutrition information and to briefly evaluate absorption interaction of vitamin C and Fe content of vitamin C emphasized products. Methods: Vitamin C emphasized foods including beverages, cereal, snacks, chocolate products, other cocoa products, and sugary products were examined by HPLC. Fe contents in samples after dry-ashing were examined by ICP. Results: Vitamin C content ranges in various products tested were the following: beverages (n=11) $20.15{\pm}0.08{\sim}845.41{\pm}6.07mg$, cereal (n=11) $52.50{\pm}0.23{\sim}262.50{\pm}0.07mg$, snacks (n=1) $50.00{\pm}0.25mg$, chocolate products (n=1) $311.73{\pm}2.44mg$, other cocoa products (n=1) $311.73{\pm}2.44mg$, other sugary products (n=2) $52.50{\pm}0.23{\sim}262.50{\pm}0.07mg$. Vitamin C (n=27) analysis values ranged from 82 to 450% of the labeled value. Vitamin C content in vitamin C emphasized food (n=6) was estimated 7.7 times~56.6 times more than Fe content. Conclusions: Analyzed samples ranged more than 80% of the labeled value in vitamin C emphasized products, which complied with food labeling regulation. But, beverages (n=3), cereal (n=4), chocolate products (n=1) were 2 times more than the labeled value. To provide accurate nutrition information, food manufactures should supervise nutrition labeling and understand the interactions between nutrients. Also, consumer should decide about the adequate amount of nutrient intake by thoroughly checking nutrition labeling.

Changes of Isoflavones and Fatty Acids in Eoyukjang, a Traditional Korean Fermented Soysauce Prepared under the Ground in a Pot or an Incubator

  • Park, Yong-Woo;Lee, Seung-Wook;Chang, Pahn-Shick;Lee, Jong-Mee;Kim, Young-Suk;Lee, Jae-Hwan
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.48-52
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    • 2009
  • Eoyukjang is a traditional Korean sauce made of cooked soypaste or meju supplemented with fish and meats at least 1 year of fermentation period. Eoyukjang was recovered according to the traditional method and stored under the ground in a pot without plastic packaging (13G: 13 month fermentation under the ground) or in an incubator wrapped with plastic packaging (6I, 12I, and 18I: 6, 12, and 18 month fermentation, respectively). Distribution of isoflavones and fatty acids were monitored by high performance liquid chromatography (HPLC) and gas chromatography (GC), respectively. Total isoflavones in 13G, 6I, 12I, and 18I were 3.792, 0.387, 0.460, and $0.510{\mu}mol/g$, respectively. Samples of 13G had at least 8.24 times higher isoflavone contents than samples from 12I. Aglycones were the major isoflavones in eoyukjang and were found more than 92% in 13G and 39-63% in incubated samples. In fatty acid analysis, the ratios of unsaturated to saturated fatty acids from 13G were higher than those from 6I, 12I, and 18I. Traditional fermentation methods using a pot may allow more migration of air and moisture than samples wrapped with plastic packaging, which caused the difference in the distribution of isoflavones and fatty acids.