• Title/Summary/Keyword: HPLC Chromatography

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Action Patterns of Chitinase and Separations of Chitooligosaccharides Produced by Chitinolytic Hydrolysis (키티나제에 의한 키토올리고당의 생성활성 규명과 올리고당의 당별 분리 생산)

  • Kim, Kwang
    • KSBB Journal
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    • v.17 no.1
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    • pp.100-105
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    • 2002
  • N-acetyl-D-glucosamine oligosaccharides [(GlcNAc)n] whose degree of polymer-ization is from one to ten (n=1-10) were fractionated by column chromatography on CM-Sephadex. Electro dialysis from a partially deacetylated chitosan hydrolysate prepared crudely with the N-acetyl-D-glucosaminidase(chitinase) and exo-N, N'-diacetylchito-biohydrolase(chitobiase) of Serratia marcescens QM B1466. Reducing sugar compositions and sequences of the N-acetyl-glucosamine oligosaccharides were identified by N-acetylation, randomly cleavage with chitinase and ego-splitting with chitobiase. N-acetyl-glucosamine heterochitooligosaccharides with glucosamine oligosaccharides, (GlcN)n at the reducing end residues together with $(GlcN)_1\sim(GlcN)_4$ were detected. Separation was accomplished by prefractionation with election by 0 to 1.0 M NaCl gradient solution. $(GlcNAc)_1 =4.25%,\; (GlcNAc)_2=4.49%,; (GlcNAc)_3=11.1%,\; (GlcNAc)_4=2.5%,$$ $(GlcNAc)_{5}$=0.64%, $(GlcNAc)_{6}$=2.12% and $(GlcNAc)_{7}$=1.21%, respectively, were crystallized after electrodialysis and lyophilization Each N-acetyl-D-glucosamine oligosaccharides content were detected by HPLC.

The Possibility of Environmental Paraquat Exposure (파라콰트의 환경성 노출 가능성)

  • Oh, Se-Hyun;Choi, Hong-Soon;You, Ho-Young;Park, Jun-Ho;Song, Jae-Seok
    • Journal of agricultural medicine and community health
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    • v.36 no.4
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    • pp.218-226
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    • 2011
  • Objectives: Paraquat (PQ) is a widely used ionic pesticide that is fatal when ingested accidentally or for suicidal purposes. It is thought that chronic exposure of PQ is related with the development of Parkinson's disease, but epidemiological studies have not yet confirmed that theory. This study attempted to estimate the possibility of environmental PQ exposure through soil and water. Materials and Methods: We analyzed the amount of decomposed PQ solution in wet soil after exposure to ultraviolet light. An artificial rainfall condition was simulated over soil sprayed with PQ to measure the amount of eluted PQ. In addition, PQ was diluted in water from three differently rated rivers and the changes in PQ concentration were measured after ultraviolet exposure over one month. High performance liquid chromatography/ultra violet detection was used to analyze the concentrations of PQ. Results: In the method we used, the recovery rate of PQ showed a precision rate less than 5%, an accuracy greater than 88%, and the calibration equation was y=5538.8x-440.01($R^2$=0.9985). There were no significant differences in the concentrations of PQ obtained from the three specimens over a 1-week period. From the PQ-sprayed soil, the artificial rainfall conditions showed no PQ elution over a 1-month period, and there was no significant differences in PQ concentrations according to ultraviolet exposure among the three samples. Conclusions: PQ remains well adsorbed naturally in soil. However, it may still exist in an integrated state for a long time in the hydrosphere, so the possibility of PQ exposure through drinking water cannot be disqualified.

Quantification and Varietal Variation of Rutin in Mulberry Fruits (뽕나무 계통별 오디함유 rutin 분석 및 품종변이)

  • Kim Hyun-bok;Kim Sun-lim
    • Journal of Sericultural and Entomological Science
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    • v.46 no.1
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    • pp.1-5
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    • 2004
  • Rutin has many beneficial effects on human health. It was established that rutin antagonizes the increase of capillary fragility associated with hemorrhagic disease, reduces high blood pressure, decreases the permeability of the vessels, and has an antiedema effect, reduces the risk of arteriosclerosis, and shows antioxidant activity. Rutin analysis with mulberry fruits was carried out by high-performance liquid chromatography (HPLC) to reveal the possibility of source of functional food. As a result, mean content of fifty accessions was 0.14±0.050% DW. Among the tested fifty accessions, 'Sabangso' was showed the highest rutin in 0.29%DW, whereas 'Simseol' was the lowest content in 0.05% DW. Also, we researched the change of rutin content according to harvested date. The rutin contents of earlier harvested groups were higher than later harvested groups. Simultaneously, fruity characteristics as well as rutin content were researched and analyzed to select the functional mulberry varieties for the production of fruit. From the six accessions which were contained high rutin content, we selected three suitable varieties such as 'Ficus', 'Kangsun', and 'Palcheongsipyung'.

Physical Characteristic and In vitro Transdermal Delivery of PCL-b-PEG Micelles Containing Quercetin and Rutin (Quercetin과 Rutin을 함유하는 PCL-b-PEG 고분자 미셀의 특성 및 피부 흡수에 관한 In vitro 연구)

  • Lim, Gyu-Nam;Kim, Sun-Young;Kim, Min-Ji;Park, Soo-Nam
    • Polymer(Korea)
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    • v.36 no.4
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    • pp.420-426
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    • 2012
  • In this study, we prepared polymer micelles containing quercetin and rutin, known as antioxidants, using poly(${\varepsilon}$-caprolactone)-b-poly(ethylene glycol), and evaluated in vitro skin permeation of the active materials. Quercetin and rutin loaded micelles were characterized by DSC (differential scanning calorimetry), HPLC (high performance liquid chromatography) and DLS (dynamic light scattering) measurements. The particle size of the polymer micelles increased in a concentration dependent manner (0.5~2.0% PCL-b-PEG). The Zeta potential of quercetin and rutin loaded micelles remained constant. To evaluate the skin penetration of PCL-b-PEG micelles, Franz diffusion cell experiment was performed. The aqueous solutions of quercetin and rutin were used as the control groups. Quercetin and rutin loaded PCL-b-PEG micelles showed more efficient skin permeation than the control groups. Safety assessment (patch test) of quercetin and rutin loaded PCL-b-PEG micelles on skin was performed to test application possibility of the polymer micelles to cosmetics. Any adverse symptoms were not observed.

Inhibitory Effects of Carex humilis Extract on Elastase Activity and Matrix Metalloproteinase-1 Expression (산거울 추출물의 Elastase 활성 저해 및 Matrix Metalloproteinase-1 발현 억제 효과)

  • Park, Sun-Hee;Lee, Kang-Hyuk;Han, Chang-Sung;Kim, Ki-Ho;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.129-136
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    • 2010
  • In order to evaluate anti-wrinkle activity of Carex humilis extract, free radical scavenging activity, elastase inhibitory activity and reduction of expression Matrix metalloproteinase-1 (MMP-1) mRNA and MMP-1 protein were investigated. The roots of Carex humilis were extracted with 95 % ethanol and successively partitioned with organic solvents with increasing polarity of the solvents. Each fraction of organic solvent were investigated by using free radical scavenging activity and elastase inhibitory activity test. Among them, EtOAc fraction showed antioxidant activity ($SC_{50}$=4.89 ${\mu}g/mL$) and elastase inhibitory activity ($IC_{50}$=23.5 ${\mu}g/mL$). EtOAc fraction was developed on silica gel by open-column chromatography and consecutively re-developed on C18 resin by prep-HPLC to give ${\alpha}$-viniferin as a major component, which was confirmed by spectrometric analysis. In the assay on expression of MMP-1 mRNA by RT-PCR and protein by western-blot, EtOAc layer (10 ~ 100 ${\mu}g/mL$) was reduced about 50 ~ 60 %, 50 ~ 65 % respectively and ${\alpha}$-viniferin (0.5 ~ 2 ${\mu}g/mL$) was inhibited about 60 ~ 75 %, 55 ~ 65 % respectively in human fibroblast. Therefore, our findings suggest that EtOAc layer of Carex humilis containing ${\alpha}$-viniferin can be useful as an active ingredient for cosmeceuticals of anti-wrinkle effects.

Application and Antimicrobial Activities of Casein Hydrolysates Treated with Asp.oryzae Protease (Casein 효소 가수분해물의 항균 활성과 그 응용)

  • Lee Hye-Jin;Yi Sang-Duk;Oh Man-Jin
    • Food Science and Preservation
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    • v.13 no.1
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    • pp.88-94
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    • 2006
  • This study was carried out to produce antimicrobial peptides from casein using various proteases. To examine whether the hydrolysis of casein would produce antimicrobial substance and the application as natural antimicrobial material, casein was hydrolyzed by five different proteases. The casein hydrolysate was fractionated with regenerated membrane filter (molecular weight cut-off 30,000 10,000 and 3,000) and antimicrobial activity was measured for each fraction. Antimicrobial activity appeared great in the fraction below 3,000 molecular weight The fraction was re-fractionated by high performance liquid chromatography and substance of main peak (retention time: 13.2 min) collected was used as a sample to measure antimicrobial activity. Among the casein hydrolysates produced by protease, antimicrobial activity was observed the greatest in hydrolysate treated with Aspergillus oryzae protease. The minimum inhibition concentrations of the Asp. oryzae protease hydrolysate were 1.0-1.5 mg/mL. This hydrolysate was a heat stable peptide since antimicrobial activity was maintained after treating with heat for 20 min at $121^{\circ}C$.

Concentration of Polycyclic Aromatic Hydrocarbons in Cereals, Pulses, Potatoes, and their Products (곡류, 두류, 서류 및 그 가공품 중 다환 방향족 탄화수소 (polycyclic aromatic hydrocarbons) 화합물 함량)

  • Kim, Hee-Yun;Chung, So-Young;Sho, Yo-Sub;Park, Seong-Soo;Lee, Eun-Ju;Suh, Jung-Hyuk;Lee, Yun-Dong;Choi, Woo-Jeong;Kim, Jung-Soo;Eom, Ji-Yoon;Park, Hui-Og;Jin, Myeong-Sig;Kim, Dong-Sul;Ha, Sang-Chul;Lee, Jong-Ok
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.537-541
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    • 2005
  • Estimation of concentrations of PAHs [benzo(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenzo(a,h)anthracene, benzo(g,h,i)perylene, indeno(1,2,3-c,d)pyrene] in cereals, pulses, potatoes, and their products available in Korean markets gave mean levels of 0.13, 0.08, 0.06, 0.03, 0.08, 0.15, 0.45, and $0.14{\mu}g/kg$, respectively, with recoveries between 82.6-106.6%. Methodology involved saponification and extraction with n-hexane, purification on Sep-Pak Florisil cartridges, and high performance liquid chromatography using a fluorescence detector.

Analysis of Flumeqnine in Meats (식육 중 항생제 flumequine의 분석)

  • Kuk, Ju-Hee;Ko, Yong-Seok;Kim, Yong-Hoon;Yoon, Chang-Yong;Sun, Nam-Kyu;Kim, Eun-Jung;Seo, Jee-Woo;Park, Ji-Won;Kang, Kil-Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.7-10
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    • 2009
  • An analytical method for the determination of flumequine in meats was developed and validated using high-performance liquid chromatography with fluorescence detection. The samples were mixed with sodium sulfate and extracted with ethyl acetate. After clean-up, the residues were dissolved in mobile phase. The calibration curves showed high linearity ($r^2$=0.9979) within the concentration range of 0.1-1.0 mg/kg. The limit of detection and limit of quantification were validated at 0.005 and 0.017 mg/kg, respectively. The recoveries in fortified meats ranged from 90.8 to 101.1%. The method was then validated in correspondence with the CODEX guidelines for flumequine residue in meats. Herein we monitored 150 samples of meats that were purchased in Korea (Seoul, Busan, Daegu, Daejeon, and Gwangju). Among the tested samples, flumequine was detected in 1 of beef and 1 of pork at levels in the range of 0.048-0.080 mg/kg. Overall, the flumequine residues in the tested samples were within the Maximun residue limit.

Bioequivalence of EtodolTM Tablet to KuhnillodineTM Tablet (Micronized Etodolac 200 mg) (건일로딘 정(미결정에토돌락 200 mg)에 대한 에토돌 정의 생물학적동등성)

  • Lee, Jung-Ae;Lee, Yun-Young;Cho, Tae-Seob;Park, Young-Joon;Moon, Byoung-Seok;Kim, Ho-Hyun;Lee, Ye-Rie;Lee, Hee-Joo;Lee, Kyung-Ryul
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.319-325
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    • 2004
  • A bioequivalence of $Etodol^{TM}$ tablets (Yuhan corporation) and $Kuhnillodine^{TM}$ tablets (Kuhnil Pharm. Co., Ltd.) was evaluated according to the guideline of Korea Food and Drug Administration (KFDA). Single 200 mg dose of etodolac of each medicine was administered orally to 24 healthy male volunteers. This study was performed in a $2{\times}2$ crossover design. Concentrations of etodolac in human plasma were monitored by a high-performance liquid chromatography. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 24 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was performed using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for $Etodol^{TM}/Kuhnillodine^{TM}$ were 1.01-1.10 and 0.87-1.06, respectively. This study demonstrated a bioequivalence of $Etodol^{TM}$ and $Kuhnillodine^{TM}$ with respect to the rate and extent of absorption.

Bioequivalence of BestidineTM Tablet to Dong-A GasterTM Tablet (Famotidine 20 mg) (동아가스터 정(파모티딘 20 mg)에 대한 베스티딘 정의 생물학적동등성)

  • Park, Chang-Hun;Joung, Sun-Koung;Choi, Mee-Hee;Kim, Ho-Hyun;Lee, Ye-Rie;Lee, Hee-Joo;Lee, Kyung-Ryul
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.505-511
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    • 2004
  • A bioequivalence study of $Bestidine^{TM}$ tablets (Choong Wae Pharma. Corp., Korea) to Dong-A $Gaster^{TM}$ (Dong-A Pharmaceutical Co., Ltd., Korea) tablets was conducted according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty four healthy male Korean volunteers received each medicine at the famotidine dose of 40 mg in a $2{\times}2$ crossover study. There was a one-week wash out period between the doses. Plasma concentrations of famotidine were monitored by a high-performance liquid chromatography for over a period of 12 hours after the administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 12 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the Cmax ratio for $Bestidine^{TM}/Gaster^{TM}$ were log 0.90-log 1.06 and log 0.98-log 1.20, respectively. These values were within the acceptable bioequivalence intervals of 0.80-1.25. Thus, our study demonstrated the bioequivalence of $Bestidine^{TM}$ and $Gaster^{TM}$ with respect to the rate and extent of absorption.