• Title/Summary/Keyword: HPLC screening

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Improvement of Decarboxylating Agar Medium for Screening Biogenic Amine-Producting Bacteria in Kimchi

  • Mah, Jae-Hyung;Shin, Soon-Young;Lee, Heung-Shick;Cho, Hong-Yon;Hwang, Han-Joon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.491-496
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    • 2001
  • A modification of decarboxylating agar medium as described by niven was performed to improve the detection method of biogenic amine-producing bacteria and to eliminate the false-positive. A total of 120 bacterial strains isolated from kimchi were used to evaluate different dicarboxylating agar media and for screening biogenic amines. Potential false-positives ranged from approximately 66 to 79% of the strains tested in the already well-known media. In our improved medium, none of the 120 strains showed the potential false-positives. There was a good agreement (81.7%-87.5%) between the results obtained by the improved medium and by HPLC analysis. Consequently, this medium was greatly improved in screening biogenic amine-producing bacteria and discarding false-positives. Of the 120 kimchi isolates, 14.2, 18.3, 37.5, and 0.8% were found by HPLC to be the producers of histamine, tyramine, putrescine (as a form of spermine), and cadaverine, respectively. The proportion of biogenic amine producer during kimchi fermentation increased to a maximum at an immature period and decreased thereafter.

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Isolation and Identification of an Unauthorized Tadalafil Analogue in a Commercial Functional Food (시판 기능성식품으로부터의 타다라필 유도체 부정첨가물질의 분리 및 구조규명)

  • Baek, Du-Jong
    • YAKHAK HOEJI
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    • v.54 no.4
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    • pp.240-243
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    • 2010
  • High-performance liquid chromatography (HPLC) screening method revealed that a propolis product marketed as a functional food contained an undeclared substance similar to tadalafil, the active ingredient of the prescription drug Cialis$^{(R)}$ approved for the treatment of male erectile dysfunction. In order to identify the illegal additive, the propolis product was extracted with methylene chloride, and the extract was purified further using semipreparative HPLC. The chemical structure of the isolated substance was elucidated based on IR, LC/MS-ESI, and $^1H$- and $^{13}C$-NMR spectroscopy, which showed the characteristics similar to tadalafil. The only difference was the substitution of the methyl group at the piperazinedione ring of tadalafil to the amino group of the identified illegal additive.

The Use of the Online Two-dimensional Liquid Chromatography Coupled with a Universal Detector for the Screening of Non-volatile Potential Migrants in Food Packaging Materials (식품포장재내 비휘발성 잠재 이행물질들의 스크리닝을 위한 이차원크로마토 그래피와 범용검출기의 이용)

  • Yoon, Chan-Suk;Lee, Keun-Taik
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.16 no.1
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    • pp.9-18
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    • 2010
  • For screening test of the non-volatile compounds which migrate from food packaging materials into foodstuffs, the traditional high performance liquid chromatography (HPLC) systems suffer from the lack of universal detector with high sensitivity and universality and high efficiency HPLC separation column which provides complete separation of complex mixtures into all individual substances. In this work, the use possibility of online two-dimensional liquid chromatography (2D-LC) system coupled with a charged aerosol detector (CAD), a universal detector, was reviewed. 2D-LC system permits to improve peak capacity and resolving power for complex mixtures. Charged aerosol detector (CAD) offers a new feasibility for detection of any non-volatile compounds with high sensitivity and constant response factor in a calibration range. The combination of size exclusion chromatography (SEC) and normal phase HPLC (NP-HPLC) is most frequently used for the separation of the natural and synthetic polymers which are mainly used as raw materials for the manufacture of food packaging materials. However, there is no commercial software available for data acquisition and handling and therefore the quantification in 2D-LC analysis is still rare.

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Rapid Screening Method of Peroxidase by Colorimetric Assay and Screening of 2, 4-DCP Degradable Strains (발색법에 의한 Peroxidase의 신속한 스크리닝법과 2, 4-DCP 분해균주의 스크리닝)

  • Ryu, Kang;Lee, Eun-Kyu
    • KSBB Journal
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    • v.23 no.6
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    • pp.484-488
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    • 2008
  • Chlorinated phenols are widely used by the chemical industry as intermediate products in synthesis and previously were frequently applied to various industry fields. Peroxidases catalyze the peroxide-dependent oxidation of a range of inorganic and organic compounds. Peroxidase was shown to mineralize a variety of recalcitrant aromatic compounds and to oxidize a number of polycyclic aromatic and phenolic compounds. Among monomeric phenolic and nonphenolic compounds, peroxidase is known to oxidize its compounds. In this study, a colorimetric assay was developed to quantitatively evaluate the peroxidase activity for rapid screening. Color products of different intensity were developed proportionally to the peroxidase activity on agar plate and 96-well plate. This method correlates well with the RP-HPLC result. Using this screening method, 12 colonies of strain was screened which survived at high concentration of 2,4-DCP (1000 ppm) and with peroxidase activity for the $7^{th}$ round screening step on agar plate. These strains were utilized 2,4-DCP as a sole carbon source and produced peroxidase. After the screening test, four of the bacteria have significant better effect of COD removal on dye waste-water. COD removal of these was from 44% to 61%, respectively.

Enhanced effect extraction of Antioxidant substance Homoorientin from Pseudosasa japonica and Phyllostachys bambusoides leaves using Ultrasonic wave system (초음파를 이용한 이대와 왕대 잎으로부터 항산화물질 Homoorientin의 추출효율 향상)

  • 이광진;신용국;김영식
    • KSBB Journal
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    • v.24 no.2
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    • pp.189-194
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    • 2009
  • In this work, antioxidant substance homoorientin from Pseudosasa japonica and Phyllostachys bambusoides leaves wereextracted using ultrasonic system, and analyzed by reversed-phase high performance liquid chromatography (RP-HPLC) on-line ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) antioxidant screening method. Also, the various experimental variables were the frequency and time of ultrasonic system. From the results, homoorientin was high extracted at the experimental condition of low frequency 35 kHz and time 60 min. And the content of homoorientin in Phyllostachys bambusoides was remarkably higher than that in the Pseudosasa japonica extract.

Comparison of EEC-4 Plate Test and Charm II Test in Residual drug Screening Methods on Meats (식육 중 잔류물질 스크리닝법인 EEC-4 Plate Test와 Charm II Test 비교)

  • Ham Hee-jin;Kim Doo-whan;Kweon Taek-boo;Lee Jung-hark
    • Journal of Food Hygiene and Safety
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    • v.19 no.4
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    • pp.198-202
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    • 2004
  • It was carried out to compare the relationship of EEC 4-plate test method and Charm II test method by HPLC. 46 only tetracyclines, 4 only sulfonamides, 4 sulfonamides+tetracyclines, one amphenicols+tetracyclines, and one amphenicols+sulfonarnides+tetracyclines, it was detected on 71 porks of EEC 4-plate positive or Charm II positive by HPLC. In case of sulfonamides and chloramphenicols, two test methods had related with each other, but had no relation in tetracyclines. Also, it was $10.5\%(6/57)$ tetracyclines, $81.8\%(9/11)$ chloramphenicols that were EEC 4 plate positive or Charm II positive, but not detectrd by HPLC. Consequently, EEC 4-plate test method and Charm II test method were suitable by screening test on antibiotics and sulfonamides etc.

Determination of tylosin in edible meats by high-performance liquid chromatography (HPLC를 이용한 식육내 타이로신의 잔류분석법)

  • Kim, Gon-sup;Shin, Sun-hye;Kim, Jong-su;Ra, Do-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.13-19
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    • 2001
  • A simple and rapid analytical method for the determination of tylosin in chicken, pork and muscle was established by High-Performance Liquid Chromatography(HPLC). Chicken, pork and beef muscle(5 g) were fortified by adding the $0.2{\mu}g/ml$ of standard tylosin and the drug was extracted from meats with 70% acetonitrile(ACN) and followed by liquid-liquid partition for clean-up procedure. Then $20{\mu}l$ portion of ACN elution was directly analyzed by HPLC with spectra 100 variable wavelength detector, and unfortified blank control were treated similarly. The average recovery rate of tylosin added to chicken, pork and beef muscle were $83{\pm}2.3$, $96{\pm}3.3$ and $92{\pm}1.6$(%) at the level 0.2 ppm, respectively. No tylosin residues in marketing meats. These results suggested that HPLC methodology could be acceptable for the extraction, determination and screening of tylosin residues in edible meats.

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Cyanobacteria and Secondary Metabolites (시아노박테리아의 이차대사물질에 대한 연구)

  • Kim, Gi-Eun;Kwon, Jong-Hee
    • KSBB Journal
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    • v.22 no.5
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    • pp.356-361
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    • 2007
  • Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Although some cyanobacteria produce lethal toxins such as microcystins and anatoxins, some may be useful either for development into commercial drugs or as biochemical tools. Detection of unknown secondary metabolites was carried in the present study by a screening of 98 cyanobacterial strains from Cyanobiotech GmbH in order to establish a screening process, isolate pure substances and determine their bioactivities. A degenerated polymerase chain reaction technique as molecular approaches has been used for general screening of NRPS gene and PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%), respectively. A screening of interesting strains was performed by comparing PCR screening results with HPLC analyses of extracts. HPLC analysis for a detection of natural products was performed in extracts from biomass. 5 strains were screened for further scale-up processing. 7 pure substances were isolated from the scale-up cultures and tested for bioactivities under consideration to purity, amount and molecular weight of substances. One substance isolated from CBT 635 showed cytotoxic activity. This substance may be regarded as Microcystin LR.

On-line Trace Enrichment for the Determination of Insulin in Biological Samples Using Reversed-Phase High Performance Liquid Chromatography with Column Switching

  • Lee, Jung-Sook;Lee, Heeyong;Lee, Hye-Suk;Lee, Kang-Choon
    • Archives of Pharmacal Research
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    • v.17 no.5
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    • pp.360-363
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    • 1994
  • Column--swtiching technique with a reversed-phase high performance liquid chromatographic method has been developed for the routine analysis of radioiodinated insulin and its degadation products in biological fluids. The diluted biological samples were loaded onto a precolumn packed with LiChrosorb RP-8 $(25-40{\;}{\mu}m)$ using 0.1% trifuoroacetic acid (TFA) in water as a washing solvent. After valve switching, the concentrated insulins were eluted in the back-flush mode and separated by a W-Porex $C_{18}$ column with a gradient of 0.1% TFA in water and 0.1% TFA in acetonitrile as the mobile phase. The method showed good precision, accuracy and speed with the detection limit of 20 pg/ml. Total analysis time per sample was about 40 min and the coefficients of variation were less than 8, 2%.

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Enhanced extraction and Antioxidant activity analysis of Flavone C-glycosides from Black bamboo using Ultrasonic wave (초음파를 이용한 오죽으로부터 Flavone C-glycosioes의 추출 및 항산화활성분석 향상)

  • Choi, Sun-Do;Lee, Kwang-Jin
    • KSBB Journal
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    • v.23 no.4
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    • pp.297-302
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    • 2008
  • In this work, the amounts of Flavone C-glycosides homoorientin, orientin extracted from Black bamboo by various ultrasonic waves frequency (35, 72, 170 KHz, 300 Watt $\pm$1) time (15, 30, 60 min) and temperature ($25^{\circ}C$) were compared using 50% aqueous ethanol solution. And describesanalysis of the antioxidant potential of Black bamboo using an high-performance liquid chromatography (HPLC) on-line $ABTS^+$antioxidant screening method. In conjunction with the analysis of their 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid ($ABTS^+$) radical scavenging ability. The optimum operating conditions were experimentally determined to analyze the Flavone C-glycosides homoorientin, orientin in the pretreatment ultrasonic extracts. From the results, the effect on extraction yield of variations in frequency, solvent composition and extraction time was investigated. The highest yield of Black bamboo was obtained by ultrasonic waves with a frequency of 72 KHz and an extraction time of 60 min, The extraction frequency 35 KHz ($25^{\circ}C$), time 30 min was selected as an optimal antioxidant activity condition.