• Title/Summary/Keyword: microbiological assay

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Microbiological Assay of Vitamin B group in Panax Ginseng Roots II. Assay of Pantothenic Acid and Biotin (인삼중 Vitamin B군의 미생물학적검정 II Pantothenic acid 및 Biotin 의 검정)

  • 김영은;허무언
    • YAKHAK HOEJI
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    • v.8 no.3
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    • pp.85-88
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    • 1964
  • Pantothenic acid and biotin contents in Panax Ginseng roots were determined microbiologically with L. arabinosus 17-5. Detection of the vitamins was achieved by the thin-layer chromatography and PPC. Pantothenic acid and biotin were found at the Rf values of 0.42 and 0.55 respectively on the thin-layer chromatograms. In order to find out whether or not the L. arabinosus growth promoting factors contained in the respective ginseng extracts, as shown by the microbiological assay, were really the vitamins respectively, PPC was carried out. Microbiological assay of the vitamins met with results that the average pantothenic acid content in the roots was 6.6r/g, calculated as Ca-pantothenate, and the average biotin content 9.24 mr/g.

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Measurement of cellulase activities

  • 편집실
    • The Microorganisms and Industry
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    • v.11 no.1
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    • pp.21-33
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    • 1985
  • 이글은 다음과 같이 구성되어져 있다. preface 1) introduction 2) general information 3) folin protein determination 4) cellobiase assay 5) filter paper assay for saccharifying cellulase 6) carboxymethyl cellulase assay for endo-.betha.-1,4-glucanase 7) additional assay procedure for endoglucanase 8) evalutaiton of cellulase under process conditions 9) general remartks, references.

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Development of Enzyme-Linked Immunosorbent Assay for Rapid and Sensitive Analysis of Biotin (Biotin의 분석을 위한 효소면역측정법(ELISA)의 개발)

  • 이경애;손동화;고영태
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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    • pp.1152-1159
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    • 1998
  • In order to develop more rapid and reproducible analysis of biotin known as vitamin H, attempts were made to establish the condition for enzyme linked immunosorbent assay(ELISA) compared with traditional microbiological assay(MBA). Antibiotin and antiserum were obtained from the immunized rabbits injected with emulsion of biotin KLH conjugate and Freund's adjuvant. The antiserum showed cross reactivity on biocytin, a derivative of biotin, which is converted to biotin in intestine, at the rate of 177%(median inhibitory concentration(IC50)=12.58ppb), but not on other derivatives such as desthiobiotin, diaminobiotin and 2 imino biotin. Specific antibody for biotin was purified from the antiserum through protein A column and desalting column. The conditions of competitive direct ELISA (cdELISA) were established. Detection range of biotin concentration by cdELISA was 0.01∼300ng/ ml(ppb). In the spike test with milk, fruit flake and pine carrot juice, the correlation coefficient between two methods of MBA and ELISA was reliably consistent at the value of r=0.992. But detection of biotin by microbiological assay(MBA) was rather restricted in range and nonspecific. Detection range of biotin by MBA was 0.1∼0.5ng/ml(ppb). It showed cross reactivities on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. In conclusion, ELISA revealed a significant improvement compared with MBA for the biotin detection in terms of sensitivity, detection range and cross reactivity. In addition, a variety of samples could be analyzed rapidly and conveniently at one time by using ELISA. These results strongly suggest that the ELISA is very promising for the practical application to detect biotin contents in a wide range of food stuffs.

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Microbiological Assay for the Detection of Chloramphenicol in Meat and Milk (식육 및 우유 중 클로람페니콜의 미생물학적 검출법에 관한 연구)

  • 손성완;조병훈;진남섭;박종명;박근식
    • Journal of Food Hygiene and Safety
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    • v.8 no.4
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    • pp.215-223
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    • 1993
  • Chloramphenicol (CAP) is a very effective broad-spectrum antibiotic which had been widely used in animal production. However, the drug is not approved in many countries for use in food-producing animals because of its potential toxicity and the possibility of residues in food products. In this study, a modified microbiological assay was developed for the sensitive detection of CAP residues in meat and milk. The method was characterized by the extraction of CAP with ethyl acetate, addition of $0.15\;\mu\textrm{g}$ oxytetracycline/ml in the phosphate buffer diluent (pH 6.0), a luteus ATCC 9341. The lowest levels of CAP detected in muscle tissues and milk were $0.025\;\mu\textrm{g}/ml\;and\;0.05\;\mu\textrm{g}/g$, respectively. Recovery rates free CAP from milk were 68.5%, from bovine muscle 65.1%, from swine muscle 63.8%, and from chicken muscle 59.4%, respectively, and the coefficients of variation were 1.8~15.1%. The results showed that the detection limits of CAP residues in animal products could significantly be improved by the modified microbiological assay than the conventional ones.

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Quantitative Immunoassay for Polychlorinated Biphenyl Compounds in Electrical Insulating Oils

  • Kim In Soo
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.119-127
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    • 2000
  • The development and performance of a competitive indirect immunoassay for the quantitative measurement of polychlorinated biphenyl compounds in insulating oils is presented. Reagent preparation and the assay characterisation, optimisation and validation steps are described. The dynamic range of the assay for Aroclors 1254 and 1260 in methanol was $50-800 {\mu}g\;ml^{-1}$ with $50\%$ signal inhibition values of 217 and $ 212 {\mu}g\;ml^{-1}$ respectively. Impending legislation in the UK is likely to decree that oils containing $ >50 {\mu}g\;ml^{-1}$ PCB be considered contaminated. Assay sensitivity increased with the degree of PCB chlorination. The assay of structurally related compounds of environmental concern yielded cross-reactivity values of under $0.6\%$. The immunoassay proved reliable for the analysis of transformer oils containing $>70{\mu}g\;ml^{-1}$ PCB, but over-estimated PCB levels in oils containing $<20{\mu}g\;ml^{-1}$ of the analyte with the oils requiring pre-treatment using either solid-phase extraction techniques or washing with KOH-ethanol/sulphuric acid to remove matrix interferents. The analytical performance of the assay was compared against a commercially available semi-quantitative immunoassay kit for PCBs in soil and water.

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A Modification of A Microbiological Assay of Vitamin $B_{12}$ In Fermented Vegetables

  • Lim, Sook-Ja
    • Journal of Nutrition and Health
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    • v.15 no.3
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    • pp.202-211
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    • 1982
  • A modification of a microbiological assay of vitamin $B_{12}$ was made and used to determine the vita­min levels during kimchi fermentation. A cyanide-buffer solution of pH 6.0 replaced the metabisufite-buffer specified in the A.O.A.C. method. The vitamin $B_{12}$ activity was decreased by blending kimchi samples for 5 minutes and retained the activity by steaming for 10 minutes before blending. The alkali hydrolysis of kimchi at pH 12.0 for 30 minutes at $121^{\circ}C$ was sufficient to destroy the vitamin $B_{12}$ and permit the detection of analogs with the same assay organism. Vitamin $B_{12}$ reached a maximum of 47ng/100g during the fermentation of kimchi ${15}\;4^{\circ}C.$ Inoculation of the kimchi with Propionibacterium shermanii (ATCC 13673) increased the vitamin production to a maximum of 102ng/100g at 1 week of fermentation. Soy flour (0.5%) or beef extract (0.05%), which were regarded as protein sources, added to the inoculated kimchi further increased the vitamin $B_{12}$ activity to 197 and 203ng/100g.

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A study on the residues of fluoroquinolones in hen's eggs consumed in Incheon Metropolitan city (인천광역시에서 유통되는 계란의 플루오로퀴놀론계 항균물질 잔류 실태 조사)

  • Park, Eun-Jeong;Lim, Ji-Huen;Lee, Sung-Mo;Im, Jeong-Soo;Oh, Dae-Kyu
    • Korean Journal of Veterinary Service
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    • v.31 no.3
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    • pp.385-395
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    • 2008
  • This study was carried out to investigate the residue level of fluoroquinolones in hen's general eggs and specific eggs by microbiological assay method and high performance liquid chromatography (HPLC) method. HPLC separation was carried out by reversed phase chromatography on a Symmetry $C_{18}$ (250${\times}$4.6 mm, $5{\mu}m$ particle size) with a phase composed of distilled water (containing 0.4% triethylamine and phosphoric acid) : Methanol (780 : 220, v/v), pumped isocratically at a flow rate of 1.0ml/min. A fluorescence detector was utilized with an excitation wavelength of 278nm and an emission wavelength of 456nm. The calibration curves were linear $({\gamma}^2{\geq}0.999)$ over a concentration range of $0.025{\sim}0.4{\mu}g/ml$. Average recoveries of the five fluoroquinolones in whole eggs at fortified levels of $0.05{\sim}0.2{\mu}g/g$ were ranged mean $78.1{\sim}91.7%$ and low coefficient of variation was less than 10% for all analysed samples. The limits of detection and limits of quantification for whole eggs were $1.2{\sim}6.0ng/g$ and $2.3{\sim}9.1ng/g$, respectively. Only one hen's general eggfrom chicken farm in Incheon was detected with the residual fluoroquinolones (Microbiological assay method; 1 of 47 general eggs) ; the range of residual concentration enrofloxacin was 0.12ppm. Those in food stores were detected with the residual fluoroquinolones (Microbiological assay method; 4 of 88 general eggs) ; the ranges of residual concentration enrofloxacin were $0.15{\sim}2.2 ppm$, ciprofloxacin $0.01{\sim}0.06ppm$, and hen's specific eggs (40) in food stores were not detected. For the microbiological assay method of fluoroquinolones in hen's eggs, as the results of comparative analysis, the disc diffusion method with E coli may be a little highly detected for the residual fluoroquinolones.

Determination of Vitamin B12 (Cyanocobalamin) in Fortified Foods by HPLC

  • Park, Youn-Ju;Jang, Jae-Hee;Park, Hye-Kyung;Koo, Yong-Eui;Hwang, In-Kyeong;Kim, Dai-Byung
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.301-305
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    • 2003
  • This study was conducted to develop an HPLC method for determining vitamin B$_{12}$ in fortified foods which has typically been determined by microbiological assays according to AOAC and Korean Food Code approved methods. Vitamin B$_{12}$ (cyanocobalamin) was determined by reversed-phase HPLC with a triple column and UV/VIS dectector (550 nm) using the column switching technique after extraction with 5 mM potassium phosphate solution by sonication without a clean-up procedure. The recovery of spiked samples and limit of detection (LOD) by HPLC were 78.6 ∼107.5 % and 2 ppb ($\mu\textrm{g}$/kg), respectively. The LOD of the microbiological assay (MBA) was much lower than that of HPLC. The concentrations of vitamin B$_{12}$ analyzed in all tested samples (n=12) confirmed compliance with declared label claims. The range of recovery ratio by the HPLC method when compared to the microbiological assay was 76.2 ∼140.0 %. There was not significant difference between the HPLC and MBA methods (p < 0.01) with r=0.9791 and linear regression y=0.9923x-0.04. The HPLC method for determining vitamin B$_{12}$ using the column-switching technique appears to be suitable for determining vitamin B$_{12}$ concentrations above 1 $\mu\textrm{g}$/100 g in fortified foods.ied foods.

CLINICAL, MICROBIOLOGICAL, BIOCHEMICAL AND IMMUNOLOGICAL STUDY ON THE PROGRESSION OF ADOLESCENT PERIODONTITIS (청년기 치주염 진전의 임상적, 미생물학적, 생화학적 및 면역학적 연구)

  • Lee, Ju-Yeon;Kim, Sung-Jo;Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.648-658
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    • 1995
  • The present study has been performed to evaluate the clinical, microbiological, biochemical and immunological parameters associated with the periodontal disease activity in adolescent periodontitis. 21 young adolescents with evidences of periodontal attachment loss participated in the study for upto 3 years of examination. Probing pocket depths and attachment levels of whole dentitions were annually recorded and 4 deepest pockets, with initial probing depth ${\geq}$ 4mm, were selected as the representative experimental sites of a patient. Sites experiencing attachment loss ${\geq}$ 1mm during the 3-year experimental period were designated as the active sites and these sites were examined for the microbiological and biochemical profiles at the time when attachment loss occurred. Microbiological assay included cultural studies and PerioScan for monitoring BANA(+) organisms(e.g. Porphyromonas gingivalis, Treponema denticola, Bacteriodes forsythus). Biochemical assay has been performed for monitoring GCF levels of neutral protease. Serum IgG and IgG2 titers against Porphyromonas gingivalis 381 were determined of a patients at the beginning and the end of the study, respectively for patient-based analysis. The results indicated that the parameters consisting of microbiological cultures and GCF neutral protease exhibited low association with the periodontal disease activity in adolescents. However, the specificity for microbiological culture of the selected periodontopathic organisms(Aa,Pg,Pi) were considerably high. Moreover, the clinical pameters such as bleeding on probing and presence of plaque as well as IgG levels against Pg at the baseline exminations were closely associated with the subsequent evidences of attachment loss during the whole experimental period(3-year).

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