• Food Science of Animal Resources
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• v.33 no.3
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• pp.417-424
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• 2013

A rapid and simple analytical method for the determination of 9 isomaltooligosaccharides (IMO) species in yoghurts was developed using dispersive solid phase extraction (dSPE) clean-up technic and high performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD). In this study, 9 IMO were extracted from samples simply with chemical reagent using ISO22662 IDF198 method and additional dSPE clean-up. The optimum instrument conditions for the determination were used carbohydrate ES $5{\mu}$ column with gradient elution of water and acetonitrile and ELS detector. The linearity of this method was expressed as the correlation coefficient ($r^2$), the results of IMO 9 species were shown in 0.9999. LOD and LOQ were respectively 7.9-22.1 mg/kg, 25.9-72.8 mg/kg. The accuracy of intra- and inter-day measurements were in the range from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$, and the preceision of the intra- and inter-day measurements were in the range from 0.8 to 7.7%. The recoveries were from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$. The determination results of IMO 9 species for the 9 yoghurts circulated in the market were in the range from $0.317{\pm}0.007$ to $1.624{\pm}0.050$ g/100 g. The newly developed method is appropriate for the determination of IMO in yoghurts, is a rapid and simple method with excellent resolution in compared with previous method.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.26-32
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• 2000

Catechol 2,3-dioxygenase was purified from recombinant strain E. coli CNU312 carrying the tomB gene which was cloned from toluene-degrading Burkholderia cepacia G4. The purification of this enzyme was performed by acetone precipitation, Sephadex G-75 chromatography, electrophoresis and electro-elution. The molecular weight of native enzyme was about 140.4 kDa and its subunit was estimated to be 35 kDa by SDS-PAGE. It means that this enzyme consists of four identical subunits. This enzyme was specifically active to catechol, and$K_(m)$ value and $V_(max)$value of this enzyme were 372.6 $\mu$M and 39.27 U/mg. This enzyme was weakly active to 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol, but rarely active to 2,3-DHBP. The optimal pH and temperature of the enzyme were pH 8.0 and $40^{\circ}C$. The enzyme was inhibited by $Co^(2+)$, $Mn^(2+)$, $Zn^(2+)$, $Fe^(2+)$, $Fe^(3+)$, and $Cu^(2+)$ ions, and was inactivated by adding the reagents such as N-bromosuccinimide, and $\rho$-diazobenzene sulfonic acid. The activity of catechol 2,3-dioxygenase was not stabilized by 10% concentration of organic solvents such as acetone, ethanol, isopropyl alcohol, ethyl acetate, and acetic acid, and by reducing agents such as 2-mercaptoethanol, dithiothreitol, and ascorbic acid. The enzyme was inactivated by the oxidizing agent $H_(2)$$O_(2)$, and by chelators such as EDTA, and ο-phenanthroline.

• Journal of Pharmaceutical Investigation
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• v.40 no.2
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• pp.101-107
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• 2010

A rapid and sensitive reversed-phase high performance liquid chromatography (HPLC) method was developed for the determination of N-(-4-Chlorophenyl)-6-hydroxy-7-methoxy-2-chromanecarboxamide (KAL-1120), a novel anti-inflammation agent, in the rat plasma. The method was applied to analyze the compound in the biological fluids such as bile, urine and tissue homogenates. After liquid-liquid extraction, the compound was analyzed on an HPLC system with ultraviolet detection at 275 nm. HPLC was carried out using reversed-phase isocratic elution with a $C_{18}$ column, a mobile phase of a mixture of acetonitril (40 v/v%) at a flow rate of 1.0 mL/min. The chromatograms showed good resolution and sensitivity and no interference of plasma. The calibration curve for the drug in plasma was linear over the concentration range of 0.05-50 ${\mu}g$/mL. The intra- and inter-day assay accuracies of this method ranged from 0.06% to 9.33% of normal values and the precision did not exceed 6.28% of relative standard deviation. The plasma concentration of KAL-1120 decreased to below the quantifiable limit at 1.5 hr after the i.v. bolus administration of 2-10 mg/kg to rats ($t_{1/2,({\alpha})}$ and $t_{1/2,({\beta})$ of 2.15 and 26.7 min at a dose of 2 mg/kg, 3.91 and 33.0 min at a dose of 10 mg/kg, respectively). The steady-state volume of distribution ($V_{dss}$) and the total body clearance ($CL_t$) were not significantly altered in rats given doses from 2 to 10 mg/kg. Of the various tissues tested, KAL-1120 was mainly distributed in the lung and heart after i.v. bolus administration. KAL-1120 was detected in the bile by 30 min after its i.v. bolus administration. However, the concentration in the urine after i.v. bolus administration became too low to measure, suggesting that KAL-1120 is mostly excreted in the bile. In conclusion, this analytical method was suitable for the preclinical pharmacokinetic studies of KAL-1120 in rats.

• Korean Journal of Food Science and Technology
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• v.42 no.5
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• pp.593-597
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• 2010

This study investigated the properties and antioxidative activities of phenolic acid concentrates of rice bran. Rice bran contains bioactive substances such as phenolic compounds, which can provide health benefits as natural antioxidants. This study examined how levels of phenolic acids can be obtained efficiently through various extraction methods. The extractions of defatted rice bran were followed by using ethylacetate (RBE-I), ethylacetate after alkaline hydrolysis (RBE-II), and 80% methanol (RBE-III). For all extracts, yields (%), total polyphenol contents (TPC), various phenolic acids and antioxidative activities were estimated. RBE-II had the highest total polyphenol contents (526.72 mg/100 g rice bran) and showed high antioxidative activity (74.7%). To concentrate the phenolic acids, RBE-II was passed through Sep-pak $C_{18}$ Vac cartridge and F1-RBE-II was collected by the elution of 50% methanol. The total phenolic content of F1-RBE-II (736.8 mg/100 g rice bran) was higher than that of RBE-II (367.1 mg/100 g rice bran), and the ratios of ferulic acid (73%) and sinapic acid (14%) increased. As RBE-II was analysed by HPLC, 6 different phenolic acids were found via chromatography, whereas F1-RBE-II showed 5 different peaks and the major phenolic acid was identified as ferulic acid. The ABTS radical scavenging activity of F1-RBE-II was the highest among the rice bran extracts. In a ${\beta}$-carotene-linoleic acid model system, linoleic acid oxidation was reduced by F1-RBE-II (73%) and RBE-II (35%).

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.2
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• pp.171-178
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• 2008

In the methanol extract from safflower seeds, two kinds of antioxidant were detected by preparative HPLC [$\mu$-Bondapak $C_{18}$ column ($7.8{\times}300\;mm$)]. Two unknown compounds were defined as CA and CB which had peaks at 22.1 min and 24.5 min, respectively. Antioxidant activity was measured by their scavenging ability on the stable tree radical of 1,1-diphenly-2-picrylhydrazyl (DPPH). For bulk extraction of antioxidants, the methanol extract was fractionated with hexan, chloroform, ethyl acetate and butanol. The ethyl acetate traction showing the highest DPPH radical scavenging activity was further purified by silica gel column chromatography to CA and CB. By NMR analysis, CA and CB were identified as N-(p-Coumaroyl)serotonin and N-feruoylserotonin, respectively. The content of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were analyzed by reverse phase HPLC using a $\mu$-Bondapak $C_{18}$ column ($3.9{\times}300\;mm$) with linear gradient elution from 10% acetonitrile to 50% acetonitrile for 30min on UV detector at 300 nm. The contents of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were 4.11 mg/g DW and 7.29 mg/g DW, respectively, and these two DPPH radical scavengers were detected only in the hull of seeds.

• Journal of Life Science
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• v.21 no.7
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• pp.976-984
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• 2011

Total phenolic content was the highest in 60% ethanol extracts at 21.91 mg/g, and inhibitory activity against tyrosinase of 60% ethanol extracts was higher than ethanol extracts of other concentration. The inhibitory compounds against tyrosinase from Persimmon leaves were purified using Sephadex LH-20, MCI-gel CHP-20 column chromatography with gradient elution. Two purified compounds were isolated as a result. The chemical structures of each compound were determined and identified using $^1H$-NMR and $^{13}C$-NMR, FAB-Mass. The compounds were confirmed as (+)-gallocatechin and prodelphinidin B-3. The tyrosinase inhibitory activities of purified (+)-gallocatechin and prodelphinidin B-3 were 29.5 and 40.2%, respectively. The inhibitory activities of (+)-gallocatechin and prodelphinidin B-3 against melanin biosynthesis in melanoma cell were 32.5 and 46.7%. The safety of essence with tyrosinase inhibitory compounds from persimmon leaves was also assessed by various safety profiles. First, changes in pH (4.90~4.95) and viscosity (23,000~26,000 cP) was not detected for 60 days. Essence also showed stability against temperature and light for 60 days. All these findings suggest that extracts from persimmon leaves have a great potential as a cosmetical ingredient with a potent whitening effect.

• The Korea Journal of Herbology
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• v.23 no.1
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• pp.109-116
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• 2008

Objectives : This study was performed to investigate the discriminative criteria of 6 kinds of Achyranthis Radix by HPLC/DAD. Methods : 20-hydroxyecdysone is isolated by silica gel column chromatography ($CHCl_3$:MeOH, 7:1-1:1 v/v) and identified by nuclear magnetic resonance, A high-performance liquid chromatographic method with diode array detection was used to identify 20-hydroxyecdysone in A. japonica. The analysis was performed using $C_{18}$ column with isocratic elution consisted of 18% acetonitrile and 82% water and the detection was carried out by DAD at 254 nm. 6 kinds of Achyranthis Radix from different locations were extracted in MeOH. Each extracts was analyzed by HPLC in same condition as used in analysis of 20-hydroxyecdysone. The identities of each extracts were determined by comparing the retention time and UV spectrum with that of reference compound. Results : 1. A. japonica and A. bidentata showed the similar patterns of HPLC chromatogram and 20-hydroxycedysone was present in both of them because the peaks having the same retention time and UV spectrum as 20-hydroxyecdysone were shown in the HPLC chromatograms of A. japonica and A. bidentata 2. Cyathula officinalis and C. capitata showed the similar patterns of HPLC chromatogram. The peak having the same retention time and UV spectrum as 20-hydroxyecdysone was shown in the HPLC chromatogram of C. capitata but not shown in the HPLC chromatogram of C. officinalis. 3. Two species of medicinal drugs from Sacheon province showed similar patterns of HPLC chromatogram. Achyranthis Radix from Sacheon(wild) did not have 20-hydroxycedysone but Achyranthis Radix from Sacheon(cultivated) showed the peak having the same retention time as 20-hydroxyecdysone but UV spectrum of the peak was different from that of 20-hydroxyecdysone. Conclusions : These results suggested that 20-hydroxyecdysone could be the discriminative criteria for Achyranthis Radix contain 20-hydroxyecdysone though they belong to different genus and species. And the patterns of HPLC chromatogram also could be the discriminative criteria as the different species of Achyranthis Radix belonging to the same genus showed similar patterns of HPLC chromatogram.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.17 no.5
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• pp.368-372
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• 1984

This paper deals with a rapid method for determination of ATP and its related compounds in fish and shellfish products using high performance liquid chromatography(HPLC). The HPLC used is a HPLC/ALC-224 equiped with UV-spectrophotometer (254 nm) as detector and integrator (Yanagimoto system-1000). The column used is a stainless steel tubing ($30.0\;cm{\times}3.9\;mm\;i.d.$) packed with ${\mu}-Bon-dapak\;C_{18}$. A mixture of $1\%$ triethylamine-phosphoric acid(pH6.5) was used as an eluent and the flow rate of the eluent was controlled at 2 ml/min. For the separation of ATP and its related compounds, a standard mixture of ATP, ADP, AMP, IMP, inosine and hypoxanthine was subjected to HPLC under the above mentioned conditions. Six peaks were obtained with retention times within 20 min, and elution order were hypoxanthine, IMP, inosine, AMP, ADP and ATP. But 5'-IMP and 5'-GMP fractions were not separated by this method. In generally, IMP content in boiled-dried fish and shellfish products purchased from the market was comparatively higher than that of other nucleotides. Especially, boiled-dried big eye herring marked higher value in IMP content than other boiled-dried ones. Hypoxanthine and inosine were major components of ATP-related compounds in dried products and seasoned-dried ones. And IMP content in seasoned-dried products was higher than that of dried ones. This fact is suggested that a part of IMP in seasoned-dried ones was derived from flavoring matter (MSG, 5'-IMP and 5'-GMP) which is added during the seasoning treatment.

• 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.

• Food Science and Preservation
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• v.16 no.5
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• pp.754-758
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• 2009

To develop soybean cake as a functional food material, the anti-oxidative and cytotoxic activities against human colon cancer cells of crude saponins isolated from 70% (v/v) ethanol extracts of cake were investigated. The Diaion HP-20 adsorption method was used for isolation of crude saponins, which were then eluted with 100% ethanol. The non-saponin fraction was removed by elution with $H_2O$ and 20% (v/v) ethanol. The results of thin layer chromatography (TLC) analysis confirmed that crude saponins were present in the 100% ethanol extract of soybean cake. The hydrogen-donating properties of saponins were more than 60% at a concentration of $1,000\;{\mu}g/mL$. malondialdehyde(MDA) production was $1,200\;{\mu}mol\;MDA/g$ in mouse liver homogenate treated with crude saponins at the concentration of $1,000\;{\mu}g/mL$. This value was lower than that of the control, which was $3,700\;{\mu}mol\;MDA/g$. Saponins inhibited the growth of colon cancer cells in a dose- and time-dependent manner. Saponins also resulted in a decrease in the proportion of cells in the G1 phase of the cell cycle, whereas the cell proportion in G2/M phase was increased with $1,000\;{\mu}g/mL$ saponins. Thus, we conclude that saponins may induce G2/M cell cycle arrest.