• Title/Summary/Keyword: HPLC Chromatography

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Antimicrobial Effects of Ethanol Extracts from Korean and Indonesian Plants (국내 및 인도네시아산 식물의 에탄올 추출물의 항균효과)

  • Kim, Moo-Sung;Lee, Dong-Cheol;Hong, Jong-Eun;Chang, Ih-Seop;Cho, Hong-Yon;Kwon, Yong-Kwan;Kim, Hee-Yun
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.949-958
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    • 2000
  • Antimicrobial effects of 150 kinds of Korean and 82 kinds of Indonesian plants were investigated to develope natural food preservatives. Extracts of the plants with 70% ethanol were tested their antimicrobial effects against several food spoilage microorganisms, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Seventeen kinds of Korean and eighteen kinds of Indonesian plants were found relatively effective, of which Myristica fragrans and Melaleuca leucadendra were the most effective, respectively. The major fractions of the two plant extracts showing antimicrobial activity were further purified by solvent fractionation, silicagel column chromatography and preparative HPLC. The purified substances were identified as limonene and caprylic acid in M. fragrans, and ${\alpha}-terpineol$ in M. leucadendra, respectively.

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Biotransformation of Free Isoflavones by Bacillus Species Isolated from Traditional Cheonggukjang

  • Lim, Ji-Sun;Jang, Chan-Ho;Lee, In-Ae;Kim, Hyo-Jung;Lee, Choong-Hwan;Kim, Jeong-Hwan;Park, Chun-Seok;Kwon, Dae-Young;Lim, Jin-Kyu;Hwang, Young-Hyun;Kim, Jong-Sang
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.1046-1050
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    • 2009
  • Our previous study showed that isoflavone profile of soybean undergoes a significant change during cheonggukjang preparation. In particular, the content of metabolite(s) with similar retention time to glycitein under the high performance liquid chromatography (HPLC) condition was significantly increased while the levels of genistein and its derivatives were notably lowered. Therefore, we hypothesized that genistein and its derivatives might be converted to genistein glucosides with similar elution time to glycitein. Our current data suggest that genistein and its derivatives are extensively metabolized into various compounds including genistein glycosides, but not glycitein or its derivatives, by Bacillus species isolated from traditional cheonggukjang. Some of daidzein was also converted into a derivative with shorter retention time by Bacillus amyloliquefaciens 51 and 86-1 but not by Bacillus subtilis 3-5 and 3-17. As metabolism of soy isoflavones, major health-promoting components in soy products, is widely variable depending upon Bacillus species, it is essential to select microorganism that minimizes the breakdown or modification of soy isoflavones in the process of fermented soy product manufacture.

Changes of Isoflavone Distribution in Soybeans Using Almond Powder (아몬드 첨가에 의한 대두의 이소플라본 특성 변화 연구)

  • Yang, Seung-Ok;Chang, Pahn-Shick;Baek, Bong-Kwon;Hong, Sung-Dae;Lee, Jae-Hwan
    • Korean Journal of Food Science and Technology
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    • v.39 no.3
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    • pp.231-236
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    • 2007
  • The isoflavone distributions of soy samples treated with soaking, oven-drying, and almond additions were determined by high performance liquid chromatography (HPLC). Oven-drying was performed to increase the amount of ${\beta}-glucoside$ isoflavones, and almonds were added to convert the ${\beta}-glucosides$ into their corresponding aglycones. Oven-drying at $100^{\circ}C$ for 4 hr significantly increased ${\beta}-glucoside$ levels and decreased $malonyl-{\beta}-glucosides$, while almond additions of 2.5% and 5.0% (w/w) significantly increased aglycone contents (p<0.05) for samples with 12 hr of drying. The rate of increase for genistein from genistin was faster than that of daidzein from daidzin with almond additions. The ${\beta}-glucosidase$ activity in the 5.0% added almond soybean samples was significantly higher than in the samples without added almond (p<0.05). The aglycone content increased from 1.62% in the raw soybeans to 61.55% in the 2.5% added almond soybean samples for 12 hr of incubation. The information from this study could be used to increase the isoflavone aglycone contents of soybeans by using natural products such as almonds, without organic solvent additions or microorganism fermentation.

Isolation of a Nisin-Producing Lactococcus lactis Strain from Kimchi and Characterization of its nisZ Gene

  • Lee, Kwang-Hee;Moon, Gi-Seong;An, Jong-Yun;Lee, Hyong-Joo;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.389-397
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    • 2002
  • Bacteriocin-producing lactic acid bacteria were isolated from kimchi. One isolate producing the most efficient bacteriocin was identified and named Lactococcus lactis B2, based on the biochemical properties and 16S rDNA sequences. The B2 bacteriocin inhibited many different Gram positive bacteria including Lactococcus, Lactobacillus, Leuconostoc, Enterococcus, Streptococcus, and Staphylococcus, but did not inhibit Gram-negative bacteria. The bacteriocin was maximally produced at temperatures between $25^{\circ}C\;and\;30^{\circ}C$ and at the initial pH of 7.0. Ninety $\%$ of the activity remained after 10 min of heat treatment at $121^{\circ}C,\;and\;100\%$, after 1 h exposure to organic solvents. The bacteriocin was purified from culture supernatant by ammonium sulfate precipitation, CM Sepharose column chromatography, ultrafiltration, and finally, by reverse-phase HPLC. A 1.58-kb fragment was amplified from B2 chromosome by using a primer set designed from the published nisA sequence. Sequencing result showed that the fragment contained the whole nisZ and 5' portion of nisB, whose gene product was involved in postmodification of nisin. The upstream sequence, however, was completely different from those of reported nisin genes.

Exopolysaccharide Produced by Pediococcus acidilactici M76 Isolated from the Korean Traditional Rice Wine, Makgeolli

  • Song, Young-Ran;Jeong, Do-Youn;Cha, Youn-Soo;Baik, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.681-688
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    • 2013
  • This work is aimed to increase knowledge of the functional exopolysaccharide (EPS) from lactic acid bacteria (LAB) in makgeolli, a Korean fermented rice wine. Among LAB strains isolated from makgeolli, strain M76 was selected as a functional strain producing a bioactive EPS, based on its antioxidative activity on the DPPH radical. The 16S rRNA gene sequencing analysis showed a high sequence similarity (99.0%) with P. acidilactici, but had different biochemical properties with the already known P. acidilactici type strains in the aspect of carbohydrates utilization. The obtained P. acidilactici M76 produced a soluble EPS above 2 g/l. One-step chromatography using gel filtration after ethanol precipitation from the supernatant of P. acidilactici M76 was enough to obtain purified EPS with a single peak, showing a molecular mass of approximately 67 kDa. Componential and structural analyses of EPS by TLC, HPLC, and FT-IR indicated that the EPS is a glucan, consisting of glucose units. The purified EPS had antioxidant activity on the DPPH radical of 45.8% at a concentration of 1 mg/ml. The purified EPS also showed proliferative effect on the pancreatic RIN-m5F cell line and remarkable protection activity on alloxan-induced cytotoxicity. This potent antioxidant and antidiabetic EPS by LAB in makgeolli may contribute to understanding the functionality of makgeolli.

Bioequivalence of Mepiril Tablet to Amaryl Tablet (Glimepiride 2 mg) by Liquid Chromatography/Electrospray Tandem Mass Spectrometry

  • Lee, Heon-Woo;Cho, Sung-Hee;Park, Wan-Su;Im, Ho-Taek;Rew, Jae-Hwan;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.35 no.4
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    • pp.287-293
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    • 2005
  • The purpose of the present study was to evaluate the bioequivalence of two glimepiride tablets, Amaryl tablet (Handok & Aventis Korea, reference drug) and Mepiril tablet (Myungmoon Pharm. Co., Ltd., Korea, test drug), according to the guidelines of Korea Food and Drug Administration (KFDA). After adding an internal standard (glibenclamide) to human plasma, plasma samples were extracted using 1mL of methyl tertiary butyl ether. Compounds extracted were analyzed by reverse-phase HPLC with multiple reaction monitoring (MRM) mode analyte detection. This method for determination glimepiride proved accurate and reproducible, with a limit of quantitation of 2 ng/mL in human plasma. Twenty-four healthy male Korean volunteers received each medicine at the glimepiride dose of 2 mg in a $2{\times}2$ crossover study. There was a one-week washout period between the doses. Plasma concentrations of glimepiride were monitored by a LC-MS/MS for over a period of 12 hr 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 $C_{max}$ ratio for Amaryl/Mepiril were log 0.9583-log 1.1357 and log 1.0570-log 1.2376, respectively. These values were within the acceptable bioequivalence intervals of log 0.80-log 1.25. Taken together, our study demonstrated the bioequivalence of Amaryl and Mepiril with respect to the rate and extent of absorption.

Antioxidative effectiveness of ether extract in Crataegus pinnatifida Bunge and Terminalia chebula Rets (산사 및 가자 에테르 추출물의 항산화 효과)

  • Kim, Jeong-Sook;Lee, Gee-Dong;Kwon, Joong-Ho;Yoon, Hyung-Sig
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.203-207
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    • 1993
  • Free phenolic acid, soluble ester and insoluble phenolic acid were extracted from defatted Crataegus pinnatifida and Terminalia chebula. Their antioxidative effectiveness was compared with BHA and BHT for soybeen oils by measuring peroxide and thiobarbituric acid values at $45^{\circ}C$ for 25 days. The patterns of these extracts were compared by using high performance liquid chromatography. Ether extracts from Crataegus and Terminalia showed a higher antioxidative effectiveness than BHA and BHT, and the latter was more effective than the former Among phenolic extracts, free phenolic acid and soluble phenolic acid ester were found most effective in Crataegus and Terminalia, respervively, Each phenolic extract was confirmed to be composed of two or three individual compounds.

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Effect of Sea Buckthorn Leaves on Inosine Monophosphate and Adenylosuccinatelyase Gene Expression in Broilers during Heat Stress

  • Zhao, Wei;Chen, Xin;Yan, Changjiang;Liu, Hongnan;Zhang, Zhihong;Wang, Pengzu;Su, Jie;Li, Yao
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.92-97
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    • 2012
  • The trial was conducted to evaluate the effects of sea buckthorn leaves (SBL) on meat flavor in broilers during heat stress. A total 360 one-day-old Arbor Acre (AA) broilers (male) were randomly allotted to 4 treatments with 6 replicates pens pretreatment and 15 birds per pen. The control group was fed a basal diet, the experimental group I, II and III were fed the basal diet supplemented with 0.25%, 0.5%, 1% SBL, respectively. During the 4th week, broilers were exposed to heat stress conditions ($36{\pm}2^{\circ}C$), after which, muscle and liver samples were collected. High performance liquid chromatography (HPLC) was performed to measure the content of inosine monophosphate (IMP); Real-Time PCR was performed to determine the expression of the ADSL gene. The results showed that the content of breast muscle IMP of group I, II and III was significantly increased 68%, 102% and 103% (p<0.01) compared with the control, respectively; the content of thigh muscle IMP of group II and III was significantly increased 56% and 58% (p<0.01), respectively. Additionally, ADSL mRNA expression in group I, II and III was increased significantly 80%, 65% and 49% (p<0.01) compared with the control, respectively. The content of IMP and expression of ADSL mRNA were increased by basal diet supplemented with SBL, therefore, the decrease of meat flavor caused by heat stress was relieved.

Simultaneous Determination of Prostaglandin E1 and Prostaglandin E1 Ethyl Ester in Hairless Mouse Skin Homogenate by High-Performance Liquid Chromatography

  • Choi, Han-Gon;Kim, Ji-Hyun;Li, Dong-Xun;Piao, Ming-Guan;Kwon, Tae-Hyub;Woo, Jong-Soo;Choi, Young-Wook;Yoo, Bang-Kyu;Yong, Chul-Soon
    • Journal of Pharmaceutical Investigation
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    • v.35 no.5
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    • pp.375-381
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    • 2005
  • A rapid and specific high-performance liquid chromatographic method was developed and validated for the simultaneous determination of prostaglandin $E_{1}\;(PGE_{1})$ and prostaglandin $E_{1}$ ethyl ester $(PGE_{1}-EE)$ in hairless mouse skin homogenate. The sample treatment procedure involved deproteination and precipitation by acetonitrile. $PGE_{1}$ and $PGE_{1}-EE$ in supernatant were separated in a reversed-phase C18 column without being interfered by other components present in hairless mouse skin homogenate. 9-Anthracenecarboxylic acid was used as an internal standard. The retention times of $PGE_{1}$, 9-anthracenecarboxylic acid and $PGE_{1}-EE$ were, 4.5, 9.5 and 18.0 min, respectively. The assay showed linearity from 1 to $40\;{\mu}g/ml$ for both $PGE_{1}$ and $PGE_{1}-EE$. Precision expressed as RSD ranged from 2.3 to 14.1 % for $PGE_{1}$ and 1.6 to 11.0% for $PGE_{1}-EE$. Accuracy ranged from 100.5 to 119.6 % for $PGE_{1}$ and from 98.0 to 103.7% for $PGE_{1}-EE$. This method was employed successfully to follow the time course of concentrations of $PGE_{1}$ and $PGE_{1}-EE$ in hairless mouse skin homogenate for stability study.

Study on the Components of the Anti-oxidant Activity of Panax Ginseng (인삼(人蔘)의 항산화(抗酸化) 성분(成分)에 관(關)한 연구(硏究))

  • Kim, Man-Wook;Choi, Kang-Ju;Cho, Yung-Hyun;Hong, Soon-Keun
    • Applied Biological Chemistry
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    • v.23 no.3
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    • pp.173-177
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    • 1980
  • The anti-oxidant components in Panax ginseng, which were known to have antiaging effect, were fractionated by various solvents and then isolated by the preparative thin layer chromatography. A developing system with silicagel G using toluene-chloroform-actene (20 : 13 : 13) as mobile phase was applied. The components were investigated the anti-oxidant activity by ${\alpha}$, ${\alpha}'-diphenyl-{\beta}-picrylhydrazyl$ (DPPH) at 517nm. Two effective components were obtained from red ginseng and were indophenol reducing substances.

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