• Title/Summary/Keyword: HPLC analysis

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Antioxidative Activity and Componential Analysis of Chamaecyparis obtusa Leaf Extract (편백나무 잎 추출물의 항산화 활성과 성분분석에 관한 연구)

  • Lee, Dong Sook;Lim, Myoung Sun;Kwan, Soon Sik;Kim, Sun Young;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.23 no.1
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    • pp.93-99
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    • 2012
  • In this study, the evaluation of antioxidative activity and componential analysis of C. obtusa leaf extracts was carried out. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of C. obtusa leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction ($OSC_{50}$; 0.22 ${\mu}g/mL$) and aglycone fraction of C. obtusa leaf extracts (0.20 ${\mu}g/mL$) showed about 7 times more prominent ROS scavenging activity than L-ascorbic acid (1.50 ${\mu}g/mL$). The cellular protective effects of fractions obtained from C. obtusa leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction and aglycone fraction of C. obtusa leaf extracts showed the cellular protective effects in a concentration dependent manner (5~25 ${\mu}g/mL$). The inhibitory effect ($IC_{50}$) of ethyl acetate fraction and aglycone fraction on tyrosinase exhibited 74.43 and 53.80 ${\mu}g/mL$, repectively. The aglycone fraction showed four times higher tyrosinase inhibitory effect than arbutin (226.88 ${\mu}g/mL$), known as a whitening agent. The aglycone fraction of C. obtusa leaf extracts showed three bands in TLC chromatogram and three peaks in HPLC chromatogram (360 nm). Three compounds were identified as taxifolin, quercetin and kaempferol. These results indicate that the fractions of C. obtusa leaf extracts can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against reactive oxygen species. The fractions of C. obtusa leaf extracts can be applicable to new functional cosmetics for antioxidan and whitening effects.

Comparision of Preparation Methods for Water Soluble Vitamin Analysis in Foods by Reversed-Phase High Performance Liquid Chromatography (역상 고속 액체 크로마토그래피에 의한 식품 중 수용성 비타민 분석을 위한 전처리법의 비교)

  • Kim, Hyung-Soo;Jang, Duck-Kyu;Woo, Dong-Kyun;Woo, Kang-Lyung
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.141-150
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    • 2002
  • Owing to a need for simple extraction and purification for analysis of water soluble vitamins in food samples by RP-HPLC with UV-detector, the methods of bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were employed. The recoveries of standard water soluble vitamins by the bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were significantly high compared to AOAC methods in most of vitamins. The contents of pyridoxal determined with protest in the pork was similar, but in the bromelain hydrolysis and AOAC method, was high compared to the results of reference. The niacinamide, thiamin and riboflavin determined with bromelain and protease hydrolysis showed similar values to the results of references. In the potato, pyridoxamine was detected in the AOAC method, which was not detected in the bromelain and protease hydrolysis methods. Pyridoxal contents in the protease hydrolysis and AOAC methods were very similar to the results of references. The recoveries of fortified standard vitamins in food samples were significantly high and accurate compared to those of AOAC methods. The extraction and purification with $C_{18}$ Sep-Pak solid extractor might be considered superior method for the determination of water soluble vitamins in food samples.

Effect of Phenolic Mediators and Humic Acid on the Removal of 1-Indanone Using Manganese Oxide (망간산화물(Birnessite)을 이용한 1- Indanone 제거 시 페놀계 반응매개체와 휴믹산(HA) 영향 평가)

  • Choi, Chan-Kyu;Eom, Won-Suk;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.7
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    • pp.445-453
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    • 2012
  • An investigation for removal of 1-indanone (1-ID), which were commonly produced from the biological and/or chemical treatment and natural weathering of the PAHs-contaminated soils, via oxidative transformation mediated by birnessite in the presence of various phenolic mediators is described. This study also examines the potential effect of the natural occurring substance humic acid (HA) on the oxidative transformation. The experiment was carried out in aqueous phase as a batch test (10 mg/L 1-ID, 0.3 mM phenolic mediators, $1.0g/L\;{\delta}-MnO_2$, at pH 5). All of the 11 tested phenoilic mediators belong to the group of natural occurring phenols and are widely used as model constituents of humic substances. From the results of HPLC analysis, it is demonstrated that 1-ID was not reactive to birnessite itself, but it can be effectively removed in birnessite-mediated cross coupling reactions in the presence of the phenolic mediators. The percent removals of 1-ID after 2 day incubation were ranged from 9.2 to 71.2% depending on the phenolic mediators applied. The initial rate constant ($K_{int}$, $hr^{-1}$) values for the 1-ID removals obtained from the pseudo-first-order kinetic plots also widely ranged from 0.18 to 15.0. Results of the correlative analysis between the removal efficiencies and structural characteristics of phenolic mediators indicate that the transformation of the 1-ID was considerably enhanced by the addition of electron-donating substituents (e.g., -OH, $-OCH_3$) at the benzne ring, and much less enhanced by the addition of electron-withdrawing substituents (e.g., -COOH, -CHO). The presence of HA showed that removal efficiencies of 1-ID in the birnessite-phenolic mediator systems decreased with increasing HA concentrations. However at low concentration of HA (< 2 mg/L), it caused some enhancement in the removals of 1-ID as compared to the control.

Antioxidative Activity and Component Analysis of Prunella vulgaris L. Extract/Fractions (하고초 추출물의 항산화 활성 및 성분 분석)

  • Suh, Ji Young;Seong, Joon Seob;Yun, Mid Eum;Lee, Ye Seul;Ha, Ji Hoon;Park, Dong Soon;Park, Soo Nam
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.647-657
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    • 2016
  • In this study, the antioxidative effects and active component analysis of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Prunella vulgaris L. were investigated. The free radical scavenging activities ($FSC_{50}$) was investigated at 50% ethanol extract ($15.25{\mu}g/mL$), ethyl acetate fraction ($8.68{\mu}g/mL$), and aglycone fraction ($8.25{\mu}g/mL$) respectively. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay was investigated at 50% ethanol extract ($4.68{\mu}g/mL$), ethyl acetate fraction ($1.00{\mu}g/mL$), and aglycone fraction($1.02{\mu}g/mL$) respectively. In the cellular protective effect against $^1O_2$ induced cellular damage of human erythrocytes, extract/fractions of P. vulgaris L. were increased in a concentration dependent manner($1{\sim}25{\mu}g/mL$). Especially, ${\tau}_{50}$ of aglycone fraction at concentrations of $25{\mu}g/mL$ showed the most protective effects at 337.9 min. It's showed nine times higher (+)-${\alpha}$-tocopherol (${\tau}_{50}=38.7min$) as typical antioxidant in the $^1O_2$-induced photohemolysis of human erythrocytes. TLC and HPLC were used to analyse active components in the ethyl acetate fraction and aglycone fraction of P. vulgaris L. In ethyl acetate fraction, caffeic acid, rosmarinic acid, quercetin 3-${\beta}$-D-glucoside, rutin, kaempferol-3-O-rutinoside, astragalin (kaempferol-3-O-glucoside) were identified. In aglycone fraction, caffeic acid, rosmarinic acid, quercetin, kaempferol were identified. These results indicated that extract/fraction of P. vulgaris L. is may be used in cosmetics industry as natural antioxidants by quenching and/or scavenging $^1O_2$ and other ROS, and protecting cellular membranes.

Antioxidative Effect and Active Component Analysis of Quercus salicina Blume Extracts (참가시나무의 항산화 효과 및 활성 성분 분석)

  • Lee, Hye-Jin;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.2
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    • pp.143-152
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    • 2011
  • In this study, the antioxidative activity, inhibitory effects on tyrosinase and elastase, and active components of Quercus salicina Blume extracts were investigated. Q. salicina Blume was extracted using 50 % ethanol, from which ethyl acetate and aglycone fractions were prepared. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity ($FSC_{50}$) of Q. salicina Blume aglycone fraction was the highest ($8.25\;{\mu}g$/mL). The luminol-dependent chemiluminescence assay showed that reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of Q. salicina Blume aglycone fraction on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system was the most prominent at $0.039\;{\mu}g$/mL. The protective effects of extract/fractions of Q. salicina Blume against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim} 25\;{\mu}g$/mL). Especially, ${\tau}_{50}$ of aglycone fraction in $10 \;{\mu}g$/mL concentration showed the most protective effect at 259.9 min. The inhibitory effects ($IC_{50}$) on tyrosinase and elastase of Q. salicina Blume extracts were higher at aglycone fraction (respectively, $21.82 \;{\mu}g$/mL, $41.18\;{\mu}g$/mL). Active component analysis by TLC and HPLC showed quercetin, keampferol, catechin, quercitrin, isoquercitrin, hyperoside, and etc. These results indicate that Q. salicina Blume extract has strong antioxidative activity and can be used as antioxidant. Particularly, aglycone fraction of Q. salicina Blume showed superior antioxdative activity and high inhibitory effect on tyrosinase and elastase. Therefore, aglycone fraction of Q. salicina Blume could be applicable to new functional cosmetics.

Development of Analysis Method of Caffeine and Content Survey in Commercial Foods by HPLC (HPLC를 이용한 카페인의 분석법 개발 및 시판 식품중 함유량 조사)

  • Kim, Hee-Yun;Lee, Young-Ja;Hong, Ki-Hyoung;Lee, Chul-Won;Kim, Kil-Saeng;Ha, Sang-Chul
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1471-1476
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    • 1999
  • A simple and practical method for determination of caffeine in foods was developed. The analysis of caffeine was performed by reverse phase high performance liquid chromatography using a ${\mu}-Bondapak\;C_{18}$ column at isocratic condition with methanol-acetic acid-water(20 : 1 : 79) on UV detector at 280 nm. The clean-up and extraction of caffeine in samples were based on a simple pretreatment using a Sep-Pak $C_{18}$ cartridge. Recovery rates obtained with this method for cider, candy, cookie, milk, ice cream and persimmon leaf tea were 99.23%, 99.50%, 99.17%, 99.37%, 98.93% and 99.10% respectively. And the detection limit of caffeine was $0.1\;{\mu}g/mL$. With this method, the range of caffeine contents extracted from coffee, green tea, black tea, Oolong tea(tea bag), soft drinks, ice cream, milk and commercial confectionery were $3.38{\sim}37.50\;mg/g,\;16.30{\sim}26.10\;mg/g,\;10.80{\sim}16.65\;mg/g,\;11.25\;mg/g,\;0.06{\sim}0.11\;mg/g,\;0.04{\sim}0.44\;mg/g,\;0.04{\sim}0.39\;mg/g\;and\;0.10{\sim}1.80\;mg/g$, respectively. But caffeine was not detected in the other tea such as Acanthopanax sessiliflorum tea, Angelica gigas tea, Angelica tea, Arrow root tea, Duchu'ng tea, Dunggulle tea, Ganoerma lucidum tea, Ginger tea powder, Persimmon leaf tea, Ssanghwa tea and Cocoa mix powder.

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Evaluation of Biological Activity and Analysis of Functional Constituents from Different Parts of Mulberry (Morus alba L.) Tree (뽕나무(Morus alba L.) 부위별 생리활성 측정 및 기능성 물질 분석)

  • Choi, Sang Won;Lee, Yu Jin;Ha, Se Bee;Jeon, Young Hee;Lee, Dong Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.6
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    • pp.823-831
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    • 2015
  • Evaluation of biological activity and analysis of functional constituents from water and ethanol extracts of four different parts of mulberry (Morus alba L.) tree were carried out to develop functional ingredients and foods using extracts of mulberry tree. The water and ethanol extracts of four different parts of mulberry tree were prepared and their biological activities and functional constituents determined by in vitro assays and HPLC, respectively. In general, ethanol extracts showed stronger biological activities and higher functional constituents than water extracts. Ethanol extracts of mulberry fruit, root bark, and twig showed stronger antioxidant ($IC_{50}=128.4{\mu}g/mL$), ${\alpha}$-glucosidase ($IC_{50}=12.0{\mu}g/mL$), and lipoxygenase ($IC_{50}=36.3{\mu}g/mL$) and tyrosinase ($IC_{50}=410.3{\mu}g/mL$) inhibitory activities, respectively, than those of other parts. Mulberry fruit and leaf showed the highest contents of anthocyanin (cyanidin 3-glucoside: 213.20 mg/100 g) and chlorogenic acid (514.97 mg/100 g), and especially ethanol extract of mulberry leaf contained higher quercetin 3-O-(6-O-malonyl)glucoside (143.25 mg/100 g) and kaempferol 3-O-(6-O-malonyl)glucoside (30.25 mg/100 g) contents without water extract of mulberry leaf. Meanwhile, mulberry twig contained both oxyresveratrol glycoside (48.90 mg/100 g) and its aglycone (21.88 mg/100 g), whereas mulberry root bark contained mostly oxyresveratrol glycoside (724.05 mg/100 g). Additionally, mulberry root bark and leaf contained much higher ${\gamma}$-aminobutyric acid (223.90 mg/100 g) and 1-deoxynojirimycin (86.07 mg/100 g) contents, respectively, than other parts of mulberry tree. These results suggest that high quality processed foods and functional foods using mixtures of mulberry fruits, leaves, twigs, and root barks should be developed for prevention and inhibition of several pathological disorders.

Antioxidative Activity and Component Analysis of Quercus glauca Leaf Extracts (종가시나무 잎 추출물의 항산화 활성, 성분 분석)

  • Yang, Hee-Jung;Ahn, You-Jin;Kim, Jae-Hyun;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.189-200
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on elastase, and components of Quercus glauca extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ of extract I fractions of Quercus glauca leaf was in the order: 50% ethanol extract $(12.45{\mu}g/mL)$ < ethyl acetate fraction $(10.47{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(8.57{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Quercus glauca leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was 50% ethanol extract $(OSC_{50},\;4.2{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(1.58{\mu}ug/mL)$ < ethyl acetate fraction $(0.66{\mu}g/mL)$. Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract / fractions of Quercus glauca leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Quercus glauca leaf extracts suppressed photohemolysis in a dose dependent manner, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect $({\tau}_{50}$, 398.67 min at $50{\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Quercus glauca leaf extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (360 nm) as well. Two components were identified as quercetin (55.77%), and kaempferol (44.23 %). TLC chromatogram of ethyl acetate fraction of Quercus glauca leaf extracts revealed 6 bands $(QG1{\sim}QG6)$, Among them, isoquercitrin (QG3), hyperin (QG4), and rutin (QG6) were identified. The inhibitory effect of aglycone fraction on tyrosinase $(IC_{50},\;73.5{\mu}g/mL)$ and elastase $(IC_{50},\;16.2{\mu}g/mL)$ was high. These results indicate that extract / fractions of Quercus glauca can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Quercus glauca leaf extract and inhibitory activity on tyeisinase and elastase of the aglycone fraction could be applicable to new functional cosmetics.

Simultaneous analysis for 2-thiothiazolidine-4-carboxilic acid and thiocarbamide using butanol extraction method (부타놀 추출법을 이용한 2-thiothiazolidine-4-carboxilic acid와 thiocarbamide의 동시정량에 관한 연구)

  • Lee, Sanghoi;Song, Jaesok;Yoon, youngshik;Kim, Chinyon;Won, Jonguk;Roh, Jaehoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.10 no.1
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    • pp.208-222
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    • 2000
  • This study was conducted to supplement limit of previous study, The objectives of this study were to select optimal conditions of high performance liquid chromatography(HPLC) operation for detecting urinary 2-thiothiazolicline-4-carboxylic acid(TTCA) and thiocarbamide simultaneously, and to evaluate recovery rates for various liquid-liquid extration method of these metabolites, The results are as follows : 1. The urinary TTCA and thiocarbamide were separate sharply when flow rate is $0.7m{\ell}/min$, using a series $C_8$ and $C_{18}$ column, 50 mM $KH_2PO_4$ : acetonitrile (93.5 : 6.5) and pH 3.5 as a mobile phase. The retention time was TTCA, $12.07{\pm}0.11$(mean${\pm}$SD, n=06), thiocarbamide, $7.85{\pm}0.01$ (mean${\pm}$SD, n=6), respectively. The calibration curve for TTCA and thiocarbamide was linear within the range 0.05 to $30{\mu}g/m{\ell}$. 2. By the liquid-liquid extration, butanol extration with $(NH_4)_2$ as a salting-out reagent was used as a simultaneous extration method for these metabolites in acid state, and recovery rates of this method are urinary TTCA, $49.6{\pm}17.7$ (mean${\pm}$SD, n=16), thiocarbamide, $43,9{\pm}5.50$ (mean${\pm}$SD, n=16), respectively 3. The precision(pooled coefficients of variation for 4 concentration) of the urinary thiocarbamide analysis was 0.03754 by butanol liquid-liquid extraction with $(NH_4)_2$ as a salting-out reagent, and TTCA was 0.04082 by ethyl acetate liquid-liquid extration with $(NH_4)_2$ as a salting out reagent The above results show that the butanol liquid-liquid extraction with $(NH_4)_2$ as a salting-out reagent in acid state, and using a series $C_8$ and $C_{18}$ column, 50 mM $KH_2PO_4$ : acetonitrile (93.5 : 6.5) and pH 3.5 as a mobile phase are suitable for the analysis of urinary TTCA and thiocarbamide simultaneously. The detection limit of TTCA and thiocarbamide was about $0.17{\mu}g/m{\ell}$, $0.07{\mu}g/m{\ell}$.

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Analysis of selenoaminoacids and selenoproteins in blood serum of sows fed by selenium fortified feed (셀레늄강화 사료를 먹인 모돈 혈청에서의 셀레노아미노산 및 셀레노단백질 분석)

  • Park, Myungsoon;Lee, Sung Hoon;Pak, Yong-Nam
    • Analytical Science and Technology
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    • v.28 no.3
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    • pp.196-203
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    • 2015
  • Selenium species (inorganic selenium, selenoaminoacids, and selenoproteins) were analyzed using anion exchange and affinity chromatography, which were connected to ICP/MS for the blood serum of sows fed by seleniumfortified feed. The Anion Exchange PRP X-100 column was used for the analysis of inorganic selenium (Se4+ and Se6+) and selenoaminoacids. The HEP column was used to separate SelP from GPx+SeAlb in selenoproteins. A quantitative analysis was performed using the post-column isotope dilution technique. The lactating sows were divided into three groups and fed by selenium fortified feed (organic 0.3 mg/kg, 0.6 mg/kg and inorganic 0.6 mg/kg) for four weeks. The test groups showed increases in selenoaminoacids compared with the control group, except the inorganic feed group. There was no significant difference between the organic feed groups. All test groups showed increases in selenoproteins. In particular, SelP showed a large increase that was 1.5 times higher than the other proteins.