• Title/Summary/Keyword: HPLC screening

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Determination and survey of fluoroquinolones in meats and eggs (II) (식육 및 계란에서 플루오로퀴놀론계 항균물질 정량분석 및 잔류조사 (II))

  • Choi, Yoon-Hwa;Kim, Yeon-Ju;Lee, Kyung-Hye;Kang, Young-Il;Lee, Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.32 no.3
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    • pp.281-286
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    • 2009
  • Fluoroquinolones in muscle and egg were separated by liquid extraction and determined. The analysis was carried out using following conditions; C18 column ($150{\times}4.6mm$, $5{\mu}m$), mobile phase composed of D.W. (containing 0.4% triethylamine and phosphoric acid) : methanol : acetonitrile (780:100:120, v/v/v), quarternary pump at a flow rate of 0.9ml/min and $20{\mu}l$ of injection volume, fluorescence detector with EX 278nm/Em 456nm. The calibration range of seven fluoroquinolones showed linearity ($r^2{\geq}0.999$) at concentration range of $0.025{\sim}0.8{\mu}g/ml$. The recoveries in fortified muscle and egg represented more than 81.3%. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, enrofloxacin, danofloxacin, saraloxacin and orbifloxacin were 3.1, 2.5, 3.6, 1.7, 0.9, 2.5 and $2.1{\mu}g/kg$, respectively. We also monitored fluoroquinolones residue in the sample (chicken muscle 182, cattle muscle 140, pig muscle 139, egg 212) using EEC-plate (E. coli ATCC 11303) screening and HPLC confirmation methods. The screening test results, fluoroquinolones, antibacterial substances were all negative.

Determination and survey of tetracyclines residue in honey by high performance liquid chromatography (액체크로마토그래피를 이용한 벌꿀 중 테트라싸이클린계 항생물질의 정량분석 및 잔류조사)

  • Lee Sung-Mo;Park Eun-Jeong;Hong Jee-Young;Kim Jung-Im;Lee Jung-Goo;Hwang Hyun-Soon;Kim Yong-Hee
    • Korean Journal of Veterinary Service
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    • v.28 no.3
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    • pp.203-213
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    • 2005
  • Oxytetracycline, tetracycline, chlortetracycline and doxycycline in honey were separated by solid phase extraction (SPE) and determined with high performance liquid chromatography (HPLC) with UV/Visible detector. Analysis was carried out using following conditions: XTerra $C_8$ column $(3.9\times150mm\;i.d. 5{\mu}m)$, mobile phase composed of 0.01M oxalic acid : methanol : acetonitrile (820 : 80 : 100, v/v/v), isocratic pump at a flow rate of 0.9 ml/min. and $50{\mu}l$ of injection volume, UV/Visible detector with wavelength of 360nm. The calibration curves of four tetracyclines showed linearity $(\gamma^2>0.999)$ at concentration range of $100\~1,000 ng/ml$. The recoveries in fortified honey represented more than $70\%$ with low coefficient of variation $(<10\%)$ for concentration range of four tetracyclines. The detection limits for oxytetracycline, tetracycline, chlortetracycline and doxycycline were 13.8, 14.6, 26.2 and 24.9ng/g in acacia honey. respectively. We also monitored tetracyclines residue in domestic honey [n : 38, acacia (20), wild flower (18) ] and foreign honey [n=22, legally distributed (13), illegally distributed (9)] using modified Charm II screening and HPLC confirmation methods. Seven of the 60 samples $(11.7\%)$ were suspect positive using modified Charm II screening test. Chlortetracycline residue was found in one foreign honey (illegally distributed) tested at concentrations of 0.22 ppm. Conclusively, for more effective control of tetracyclines used in beekeeping should be further survey for residues in honey and also national guidelines (maximum residue limit : MRL) and methods should be obligatory.

Selection of High Tobramycin-Producing Mutants (Tobramycin 고생산성 변이주의 분리)

  • 나규흠;김학주;김기태;양중익;김계원
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.343-347
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    • 1991
  • An improved method for the selection of high tobramycin-producing mutants of Streptomyces tenebrarius ATCC 17920 was investigated. By the use of apramycin-containing media, low nebramycin-producing mutants were eliminated. Strain No. 23, resistant to apramycin and kanamycin B and sensitive to tobramycin, was isolated from soils, identified as Pseudomonas paucimobilis and used as a test organism for overlaying the mutants on agar plate. While inhibition zones were not shown when the parent strains were overlaid with soft agar containing the strain No. 23, clear zones were shown when high tobramycin-producing mutants were overlaid. Using this screening strategy, 58 mutants showing clear zones had been isolated. antibiotic activities of which were incresed to 3~8 fold compared to that of parent strain.

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The Rapid Detection of Antioxidants from Safflower Seeds (Carthamus tinctorius L.) Using Hyphenated-HPLC Techniques (Hyphenated-HPLC 기술을 활용한 홍화씨의 항산화 성분 분석)

  • Kim, Su-Jin;Kim, Sang-Min;Kang, Suk-Woo;Um, Byung-Hun
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.414-419
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    • 2010
  • Hyphenated-HPLC techniques combine the separation power of HPLC with the structural and bioactivity information provided by NMR, ESI/MS, and an on-line antioxidant screening system. The major advantages over the traditional off-line techniques are rapidity and efficiency. In this study, we used hyphenated HPLC techniques including online HPLC-ABTS, LC-NMR, and LC-MS todirectly identify the major antioxidants of safflower (Carthamus tinctorius L.) seeds. The results demonstrated that the major antioxidant compounds from on-line HPLC-ABTS analysis were identified as 8'-hydroxyarcgenin-4'-O-$\beta$-D-glucoside, N-(p-coumaroyl) serotonin, and N-feruloylserotonin. Among them, N-feruloylserotonin accounted for almost 50% of the ABTS radical scavenging activity of the total extract. The results demonstrate that HPLC hyphenated techniques can be used to rapidly screen and structurally identify antioxidants from crude plant extracts.

Efficient Target-Site Assay of Chemicals for Melanin Biosynthesis Inhibition of Magnaporthe grisea

  • Kim, Jin-Cheol;Son, Mi-Jung;Kim, Heung-Tae;Park, Gyung-Ja;Hahn, Hoh-Gyu;Nam, Kee-Dal;Cho, Kwang-Yun
    • The Plant Pathology Journal
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    • v.16 no.3
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    • pp.125-129
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    • 2000
  • A rapid and efficient assay to determine melanin biosynthesis inhibition of Magnaporthe grisea, a causal agent of the rice blast, by chemicals was developed. Wells in 24-well plates were loaded with spore suspension of the fungus and three known melanin biosynthesis inhibitors of KC10017, tricyclazole, and carpropamid. Subsequent color changes of mycelia and culture media in the wells were observed 7 days after incubation. The wells treated with KC10017 (an inhibitor of polyketide synthesis step and/or pentaketide cyclization step) became colorless, whereas tricyclazole (an inhibitor of 1, 3, 8-trihydroxynaphthalene reductase) or carpropamid (an inhibitor of scytalone dehydratase)-treated wells exhibited red color. They did not show any inhibitory effect on fungal growth. The inhibition of reaction steps prior to 1, 3, 6, 8-tetrahydroxynaphthalene formation was easily determined by colorless medium and mycelia. However, it was impossible to distinguish between inhibition of reduction steps and inhibition of dehydration steps by colors of the cultures. It was accomplished through HPLC analysis of the melanin biosynthesis-involving pentaketide metabolites accumulated by the inhibitors. Through screening of a number of synthetic chemicals using the in vitro assay, we could find a novel chemical group of melanin biosynthesis inhibitor.

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Isolation of Erythritol Producing Microorganisms from Nature (자연계로부터 Erythritol 생산 균주의 분리)

  • 이광준;주영란;이길웅;오경수;이윤진;박상희;임재윤
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.38-42
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    • 1997
  • For the purpose of obtaining microorganisms producing high amount of erythritol, the screening test was carried out. Productivity of erythritol was analyzed by paper chromatography and HPLC' methods. Among more than two hundred isolates, one strain(KJX1) was selected as an erythritol prtducer from thc soil of corn shock. The isolated strain was identified as Pmicilliurn sp. KJ81 from the morphological and physiological characteristics. Penicillium sp. KJ81 showed white to green colony color, two- to three-stage branching conidiophcvc and flask-shaped phialides.

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Studies on Microbial Transformation of Meloxicam by Fungi

  • Shyam Prasad, G.;Girisham, S.;Reddy, S.M.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.922-931
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    • 2009
  • Screening-scale studies were performed with 26 fungal cultures for their ability to transform the anti-inflammatory drug meloxicam. Among the different fungi screened, a filamentous fungus, Cunninghamella blakesleeana NCIM 687, transformed meloxicam to three metabolites in significant quantities. The transformation of meloxicam was confirmed by high-performance liquid chromatography (HPLC). Based on the liquid chromatography-tandem mass spectrometry (LC-MS/MS) data, two metabolites were predicted to be 5-hydroxymethyl meloxicam and 5-carboxy meloxicam, the major mammalian metabolites reported previously. A new metabolite was produced, which is not detected in mammalian systems. Glucose medium, pH of 6.0, temperature of $27^{\circ}C$, 5-day incubation period, dimethylformamide as solvent, and glucose concentration of 2.0% were found to be suitable for maximum transformation of meloxicam when studied separately. It is concluded that C. blakesleeana can be employed for biotransformation of drugs for production of novel metabolites.

Characterization and Food Application of a Potentiometric Biosensor Measuring $\beta$-Lactam Antibiotics

  • Park, In-Seon;Kim, Dong-Kyung;Kim, Nam-Soo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.698-706
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    • 2004
  • $\beta$-Lactam antibiotics such as penicillin G, amoxicillin, and ampicillin were determined by a potentiometric biosensor system which exploited penicillinase immobilized on Immobilon cellulose nitrate membrane and a flat-bottomed pH electrode-as the biological component and transducer. The optimum reaction buffer for maximum sensitivity was found as 2 mM of sodium phosphate buffer (pH 7.2). The detection limit of the biosensor could be extended to 1 $\mu{M}$ of the analytes by increasing the enzyme loading for immobilization to 100 units/$m\ell$. The model samples spiked with each of the standard penicillins were measured for their biosensor responses and HPLC peak area, resulting in the relative responses of 82.1-103.5% and 79.5-106.1% for the biosensor method along with HPLC analysis, respectively. This result showed a good precision of the current biosensor method for screening the penicillin compounds.

Screening of Fungal Strains Producing Lovastatin, an Antihypercholesterolemic Agent (콜레스테롤합성저해제 lovastatin을 생산하는 곰팡이 균주의 탐색)

  • Bang, In-Young;Whang, Seung-Whan;Kim, Jung-Wan;Kim, Sang-Yong;Park, Cheon-Seok
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.442-446
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    • 2003
  • Over two hundred fungal strains from Korean soil were tested for the production of cholesterol-lowering agent, lovastatin. Each fungal strain was cultivated in the rapeseedmeal production medium (RPM). After growing for 7 days, the presence of lovastatin in the culture was examined by TLC analysis and HPLC. Nine different fungal strains were determined to produce detectable amounts of lovastatin, among which one fungal strain isolated from barnyard manure of Kanghwa island produced 25.58 mg/L of lovastatin in the production medium. The morphological observation combined with the determination of 18S-rRNA sequence suggested that the selected strain belongs to a member of Aspergillus sp..