• Title/Summary/Keyword: o--Phenylenediamine

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Competitive Enzyme-Linked Immunosorbent Assay for Detection of Gentamicin Residues in Edible Animal products (축산식품 중에 잔류하는 Gentamicin 검사를 위한 ELISA 개발에 관한 연구)

  • Kim, Jae-Myung;Lee, Mun-Han;Lee, Hang;Ryu, Pan-Dong;Cho, Myung-Haing;Park, Jong-Myung
    • Journal of Food Hygiene and Safety
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    • v.9 no.3
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    • pp.123-131
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    • 1994
  • An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

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Analytical Method of L-Ascorbic Acid Content in Green Tea (녹차중(綠茶中)의 L-Ascorbic Acid의 정량법(定量法)에 관한 연구(硏究))

  • Shin, Mee-Gyung;Nam, Chang-Woo
    • Korean Journal of Food Science and Technology
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    • v.11 no.2
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    • pp.77-80
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    • 1979
  • Effects of interfering substances on the determination of L-ascorbic acid (AsA) in green tea and it's extracts by 2,4-dinitrophenylhydrazine (DNP) method was studied. and the removal of these interfering substances was also investigated. Under the condition prescribed for DNP method, AsA content of green tea are effected by some sugar, reductones, dicarbonyl compounds, organic acids, amino acids and others. All interfering substances except amino acids were eliminated by the chloroform extraction after adding o-phenylendiamine to sample solution. and remaining amino acids were eliminated almost completely by the treatment with ion exchange resin$(Amberlite{\;}IR-120H^{+})$. After removing the interfering substances by the above mentioned procedure, total AsA in green tea was determined by DNP method. The values obtained by this method were in good agreement with those by thin layer chromatography (TLC) method. and the method was more rapid and simple than TLC method.

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Hydroxyl Radical-Mediated Commitment of HL-60 Cells to Differentiation: Modulation of Differentiation Process by Phosphodiesterase Inhibitors

  • Cho, Young-Jin;Ahn, Woong-Shick;Cha, Seok-Ho;Lee, Kweon-Haeng;Kim, Won-Il;Chung, Myung-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.3
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    • pp.369-376
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    • 1998
  • This report shows that hydroxyl radical, generated by a Fenton reaction involving adenosine $5'-diphosphate/Fe^{2+}$ complex ($5-15\;{\mu}M$) and $H_2O_2$ ($2\;{\mu}M$), induced differentiation of HL-60 cells in a dose- and time-dependent manner. This is evidenced by the increases in 12-O-tetradecanoylphorbol 13-acetate- and fMLP-stimulated superoxide production capability. The cells exposed to hydroxyl radical for defined periods (24∼96 hr) continued to differentiate even after the hydroxyl radical generating system had been removed. The differentiated cells displayed fMLP-stimulated calcium mobilization and increased expression of myeloid-specific antigen CD11b and CD14. The extent of the differentiation was markedly reduced by desferrioxamine ($100\;{\mu}M$), dimethylthiourea (5 mM), N,N'-diphenyl-1,4-phenylenediamine ($2\;{\mu}M$), and N-acetyl-L-cysteine (5 mM). The induction of differentiation by hydroxyl radical was enhanced by 3-isobutyl-1-methylxanthine ($200\;{\mu}M$) and Ro-20-1724 ($8\;{\mu}M$), and inhibited by dipyridamole (2 ${\mu}M$). These results suggest that hydroxyl radicals may induce commitment of HL-60 cells to differentiate into more mature cells of myelomonocytic lineage through specific signal-transduction pathway that is modulated by phosphodiesterase inhibitors.

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Chemopreventive Activity of Turmeric Essential Oil and Possible Mechanisms of Action

  • Liju, Vijayasteltar Belsamma;Jeena, Kottarapat;Kuttan, Ramadasan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6575-6580
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    • 2014
  • This study aimed to evaluate the antimutagenic and anticarcinogenic activity of turmeric essential oil as well as to establish biochemical mechanisms of action. Antimutagenicity testing was accomplished using strains and known mutagens with and without microsomal activation. Anticarcinogenic activity was assessed by topical application of 7, 12 - dimethylbenz[a]anthracene (DMBA) as initiator and 1% croton oil as promoter for the induction of skin papillomas in mice. Inhibition of p450 enzymes by TEO was studied using various resorufins and aminopyrene as substrate. Turmeric essential oil (TEO) showed significant antimutagenic activity (p<0.001) against direct acting mutagens such as sodium azide ($NaN_3$), 4-nitro-O-phenylenediamine (NPD) and N-methyl-N-nitro N'nitrosoguanine (MNNG). TEO was found to have significant antimutagenic effect (>90%) against mutagen needing metabolic activation such as 2-acetamidoflourene (2-AAF). The study also revealed that TEO significantly inhibited (p<0.001) the mutagenicity induced by tobacco extract to Salmonella TA 102 strain. DMBA and croton oil induced papilloma development in mice was found to be delayed and prevented significantly by TEO application. Moreover TEO significantly (P<0.001) inhibited isoforms of cytochrome p450 (CYP1A1, CYP1A2, CYP2B1/2, CYP2A, CYP2B and CYP3A) enzymes in vitro, which are involved in the activation of carcinogens. Results indicated that TEO is antimutagenic and anticarcinogenic and inhibition of enzymes (p450) involved in the activation of carcinogen is one of its mechanisms of action.

Use of the enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine (ELISA를 이용한 돼지 톡소플라스마병의 조기 진단에 관한 연구)

  • Suh, Myung-deuk;Jang, Dong-hwa;Joo, Hoo-don
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.567-575
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    • 1989
  • This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

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Studies on avian infectious bronchitis: II. Standardization of an indirect enzyme-linked immunosorbent assay (ELISA) for antibody measurement (닭 전염성 기관지염에 관한 연구: II. 간접 enzyme-linked immunosorbent assay(ELISA)에 의한 항체가 측정)

  • Chang, Chong-ho;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.503-515
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    • 1989
  • Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.

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Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Dongchimi (동치미로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성)

  • 주길재;이창호;우철주
    • Journal of Life Science
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    • v.11 no.5
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    • pp.432-438
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    • 2001
  • Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

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Synthesis of the Low-Hygroscopic Polyimide for 2-Layer Flexible Copper Clad Laminate (2층 연성동박적층판용 저흡습 폴리이미드의 합성)

  • Kim, W.;Park, S.J.;Baek, J.O.;Gong, H.J.;Ahn, B.H.
    • Elastomers and Composites
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    • v.43 no.2
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    • pp.82-87
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    • 2008
  • In this study, nine kinds of polyimides were synthesized from 1,2,4,5-benzenetetracarboxylic dianhydride (PMDA), 4,4'-(4,4'-isopropylidenediphenoxy)bis(phthalic anhydride) (BPADA), m-pheny lenediamine (m-PDA) and 4,4'-oxydianiline (ODA) by controlling molar ratio of monomers. Synthesized polyimides were used as insulator films for 2-layer Flexible Copper Clad Laminate(FCCL) which were manufactured by the casting method. Glass transition temperature and thermal degradation temperature for 5% weight loss of the polyimide film were improved by increasing contents of m-PDA and PMDA, respectively. Water absorption of polyimide film was reduced by increasing contents of ODA and BPADA which have relatively long structure, respectively. Peel strength of 2-layer FCCL was improved by increasing contents of ODA and BPADA.

Studies on the Separation and Determination of Dicarbonyl Compounds by Gas Chromatography (Gas Chromatography에 의(依)한 Dicarbonyl류(類)의 분리정량(分離定量)에 관(關)한 연구(硏究))

  • Sin, Mee-Gyung;Nam, Chang-Woo
    • Korean Journal of Food Science and Technology
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    • v.8 no.2
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    • pp.85-89
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    • 1976
  • The separation and determination of dicarbonyls such as diacetyl, methylglyoxal and triose reductone in their mixed aqueous solution were carried out by means of gas chromatography with transformation of these compounds into quinoxaline derivatives with o-phenylenediamine. A column used for this experiment was consisted of Celite 545 (80-100 mesh) coated with 5% Silicon Gum SE-30. The column temperature was $180^{\circ}C$. It is desirable that this approach will be applicable to dicabonyl study in gas chromatographic determination.

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Synthesis and Characterization of a Near-Infrared Optical Materials for Shielding Infrared Rays

  • Park Su-Yeol;Sin Seung-Rim;Sin Jong-Il;O Se-Hwa;Jeon Geun
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2005.05a
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    • pp.213-215
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    • 2005
  • The metal complexes can be influenced not only by the central metal atoms and the substituent groups, but also by the native of the chelating atoms. For example, near-infrared absorbing chromophores were synthesized by the reaction of phenylenediamine derivatives with a solution of pottassium hydroxide followed by the addition of nickel(II) chloride. These dyes provide absorbing infrared light over 780-840 nm with an extinction coefficient of $2.5-6.0{\times}10^4$. By introduction of alkyl, alkoxyl, cyano, and other functional group into the parent dye, these dyes greatly improved the solubility in organic solvent. New near-infrared absorbing donor-acceptor chromophores have been investigated by varying the electron donating and accepting strength of the two halves of the molecule. The cyanine chromophores permit the simplest way of obtaining systems that absorb well into the near-infrared region of the spectrum. Cyanine dyes possess high extinction coefficients that initially increase with Increasing chain length. These chromophores could be useful in near-infrared optical materials.

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