• Title/Summary/Keyword: phenol detection

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Analysis of Mitochondrial DNA in Patients with Essential Tremor (본태성 수전증 환자의 미토콘드리아 DNA 분석)

  • Lee, Uhn;Yoo, Young Mi;Yoo, Chan Jong
    • Journal of Korean Neurosurgical Society
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    • v.29 no.2
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    • pp.188-195
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    • 2000
  • Objective : Essential tremor(ET) is the most common movement disorder, however, there has been little agreement in the neurologic literature regarding diagnostic criteria for ET. Familial ET is an autosomal dominant disorder presenting as an isolated postural tremor. The main feature of ET is postural tremor of the arms with later involvement of the head, voice, or legs. In previous studies, it was reported that ET susceptibility was inherited in an autosomal dominant inheritance. As previous results, it would suggest that ET might be associated with defect of mitochondrial or nuclear DNA. Recent studies are focusing on molecular genetic detection of movement disorders, such as essential tremor and restless legs syndrome. Moreover, authors have analysed mitochondrial DNA(mtDNA) from the blood cell of positive control(PC) and ET patients via long and accurate polymerase chain reaction(LA PCR). Materials & Methods : Blood samples were collected from PC and 9 ET patients. Total DNA was extracted twice with phenol followed by chloroform : isoamylalcohol. For the analysis of mtDNA, LA PCR was performed by mitochondrial specific primers. Results : With this technique, deletions of large quantities were detected within several regions of mtDNA in ET patients except for D-loop and CO I regions. Conclusion : The authors believe that ET is a genentic disorder with deficiency of mitochondrial DNA multicomplexes and mitochondiral dysfunction could be one of major causative factors of ET. Mitochondrial dysfunction may play an important role in the pathogenesis and possibility of disease progression among familial group with ET patients.

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Identification of the impurities in the technical product of Atonic (Atonic 원제의 부성분 구조 확인)

  • Kyung, Kee-Sung;Chung, Chang-Kook;Lee, Jae-Koo
    • The Korean Journal of Pesticide Science
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    • v.8 no.2
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    • pp.129-136
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    • 2004
  • In order to determine the amounts of impurities and to identify the chemical structures of the impurities in the technical product of the plant growth regulator Atonic, the extracts of diethyl ether and dichloromethane were analyzed with GC-FID and GC-MSD. resulting in detection of five impurities and identification of their chemical structures. The amount of the active ingredient atonic in the technical product was about 84% and those of the impurities ranged from 0.24 to 10.74%. The identified impurities in this technical product are 2-methoxyphenol (guaiacol, m/z 124), 2-chloro-6-methoxyphenol and/or 4-chloro-6-methoxyphenol (m/z 158), 1,2-dimethoxy-4-nitrobenzene (m/z 183), and 2,6-bis(1,1-dimethylethyl)-4-methylphenol (m/z 220), suggesting that they are not hazardous impurities.

Establishing Quantitative Standards for Residual Alkaline Phosphatase in Pasteurized Milk

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Lim, Jong-Soo;Kim, Hong-Seok;Kang, Il-Byeong;Jeong, Dana;Song, Kwang-Young;Kim, Hyunsook;Kim, Kwang-Yup;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.36 no.2
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    • pp.194-197
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    • 2016
  • The alkaline phosphatase (ALP) assay is a rapid and convenient method for verifying milk pasteurization. Since colorimetric ALP assays rely on subjective visual assessments, their results are especially unreliable near the detection limits. In this study, we attempted to establish quantitative criteria for residual ALP in milk by using a more objective method based on spectrophotometric measurements. Raw milk was heat-treated for 0, 10, 20, 30, and 40 min and then subjected to ALP assays. The quantitative criteria for residual ALP in the milk was determined as 2 μg phenol/mL of milk, which is just above the ALP value of milk samples heat-treated for 30 min. These newly proposed methodology and criteria could facilitate the microbiological quality control of milk.

Enzymatic Spectrophotometric Determinations of Acetylcholine and Choline in the Biological Samples (생체 시료 중 아세틸콜린 및 콜린에 대한 효소-분광학적 정량분석)

  • Nam, Myung-Hwa;Lee, Sung-Ho;Kim, Ke-Tack;Hahn, Young-Hee
    • YAKHAK HOEJI
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    • v.56 no.4
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    • pp.222-229
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    • 2012
  • In order to determine acetylcholine and choline in the biological samples, the specific enzymes of acetylcholinesterase (AChE) and choline oxidase (ChO), which utilize acetylcholine and choline as substrates, were employed to convert substrates to $H_2O_2$. The produced $H_2O_2$ was coupled to 4-aminoantipyrine/phenol with peroxidase (PO) yielding quinoneimine dye which was measured at 508 nm. In the present enzymatic spectrophotometric analysis the product at the equilibrium state was measured considering accuracy, precision, time and cost of the analysis. The developed analytical method yielded good linearity (calibration curve; $A_{508}$=9534[acetylcholine]+0.009, correlation coefficient ($R^2$); 0.999) with detection limit of $1.11{\times}10^{-7}M$, reasonable precision (relative standard deviation; 0.10~1.62% at $2.5{\times}10^{-6}M{\sim}1.25{\times}10^{-4}M$) and accuracy (relative error; -0.24~0.97% at $4.13{\times}10^{-6}M{\sim}1.01{\times}10^{-4}M$) for acetylcholine chloride standard solution. The concentrations of acetylcholine and choline in human serum were found as $3.20{\times}10^{-5}M$ and $1.14{\times}10^{-4}M$, respectively. The brain tissues of Sprague-Dawley strain rat contained 9.82${\mu}g/g$ of acetylcholine and 6.53 ${\mu}g/g$ of choline in the cerebrum, while 7.37 ${\mu}g/g$ of acetylcholine and 5.34 ${\mu}g/g$ of choline in the cerebellum.

Immobilization of Recombinant Bacterial Biosensors: a Simple Approach for the On-Site Detection of Phenolic Compounds (재조합 박테리아 바이오센서의 고정화: 페놀계 화합물의 현장 검출을 위한 간단한 접근 방법)

  • Shin, Hae-Ja
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1323-1328
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    • 2011
  • We herein report the development of an agarose-gel-immobilized recombinant bacterial biosensor simple system for the field monitoring of phenolic compounds. Escherichia coli cells harboring the pLZCapR plasmid, which was previously designed to express the ${\beta}$-galactosidase reporter gene in the presence of phenolic compounds, were co-immobilized with a substrate [chlorophenol red ${\beta}$-galactopyranoside (CPRG) in agarose gel, and dispensed to the wells of a 96-well plate. Field samples were added to the wells and color development was monitored. In the presence of 5 ${\mu}M$ to 10 mM of phenol, the biosensor developed a red (representing hydrolysis of CPRG) color. Other phenolic compounds were also detected by this immobilized system, with the pattern resembling that previously reported for the corresponding non-immobilized biosensor. The immobilized cells showed optimum activity when the gel was simultaneously supplemented with 6% dimethyl formamide (DMF), 0.1% SDS and 10 mM $CaCl_2$. The immobilized biosensor described herein does not require the addition of a substrate or the use of unwieldy instruments or sample pretreatments that could complicate field studies.

The Effect of Fermented Diet with Whole Crop Barley Silage on Fecal Shape and Odorous Compound Concentration from Feces in Pregnant Sows (청보리 사일리지 첨가 발효사료가 임신돈의 분 성상 및 악취물질 농도에 미치는 효과)

  • Cho, S.B.;Kim, C.H.;Hwang, O.H.;Park, J.C.;Kim, D.W.;Sung, H.G.;Yang, S.H.;Park, K.H.;Choi, D.Y.;Yoo, Y.H.
    • Journal of Animal Environmental Science
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    • v.17 no.3
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    • pp.145-154
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    • 2011
  • The study was conducted to investigate the effect of feeding fermented diet including whole crop barley silage on the odor reduction and microbial population change in feces, and the constipation prevention of pregnant sows. The concentration of phenol was not detected in tapioca, beet pulp, wheat bran and lupin seeds, while that of p-cresol was ranged between 9.62 and 52.11 mg/L showing that lupin was highest and tapioca was lowest. It was determined that tapioca and beet pulp were useful feed ingredients to reduce odor due to their lower contents of phenol and indole compounds. Ten pregnant sows were allocated to control group and fermented diet group in 5 sows in each group. They were fed 3.0 kg DM/d of diets for 28 days. Feces was examined and showed that the feces from the fermented diet group was observed with the higher moisture content and the lower hardness than that of the control diet group and the population of E. coli was decreased and the population of lactobacilus was higher than that of the control diet group. The concentrations of p-cresol and skatole were lower than the detection levels at 33% and 67% among the samples of feces of the control group and at 67% and 100% among the samples of the feces of fermented diet group respectively. Thus it is expected that the odor from the feces of pregnant sows fed the fermented diet could be reduced compared with that of control group. Therefore, it is suggested that feeding fermented whole barley diet to pregnant sows improve the function of intestine and reduce the rate of occurrences of constipation and odor levels.

Rapid Extraction of DNA using Ion Exchange Resin for Early Detection of Mycobacterium tuberculosis by the Polymerase Chain Reaction (결핵균 PCR에서 이온교환수지를 이용한 신속한 DNA 분리)

  • Kim, Cheol-Min;Park, Seung-Kyu;Shon, Mal-Hyun;Song, Sun-Dae;Kim, Young;Jun, Eun-Sook;Son, Han-Chul;Jung, Byung-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.30-37
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    • 1996
  • Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.

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A Study on Optimization for Separation of Phenols and Preconcentration-Separation of Trace Phenols in Reversed-Phase Liquid Chromatography (역상 액체 크로마토그래피에서 페놀류의 분리 최적화 및 미량 페놀류의 농축-분리에 관한 연구)

  • Lee Dai Woon;Lee Sung Won;So, Min Jeong;Cho Byung Yun
    • Journal of the Korean Chemical Society
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    • v.37 no.5
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    • pp.513-522
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    • 1993
  • The purpose of this study is to optimize the selectivity of mobile phase solvents for separation of 25 phenols in reversed phase liquid chromatography and to accomplish the simultaneous preconcentration and separation of trace phenols from water samples. Phenols used in this study were classified into three groups, chloro-, methyl-, and nitrophenols. Quaternary solvent mobile phases were employed to improve the selectivity. Overlapping resolution maps(ORM) as a statistical simplex techniques was used to predict the optimum solvent system. Additional criterion such as pH and temperature were also investigated. In order to improve the resolution and decrease the analysis time, isoselective multisolvent gradient elution system was employed with ORM-Prism method. The simultaneous preconcentration and separation of trace phenols from water samples were performed by using XAD-2/Dowex 1-X8 tandem column. When the extraction efficiency was evaluated by sampling up to 1 L of distilled water, recovery of the phenols, except phenol, was above 90% and the limit of detection of the phenols was 5 ppb. The XAD-2/Dowex 1-X8 method was superior to C18 cartridge in terms of recovery and selectivity.

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Diversity of VC and incidence of hypovirulence-associated ds-RNAs in the chestnut blight fungus Cryphonectria parasitica in Korea

  • Byeongjin Cha;Jinyoung Lim;Ju, Young-Jik;Kim, Dae-Hyuk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.23-23
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    • 2003
  • Chestnut blight, caused by Cryphonectria parasitica, is the most destructive disease of American and European chestnut trees. A total of 672 C prasitica was isolated from blight lesion on chestnut twigs, which were collected from major chestnut plantations all over Korea in 1999. Isolation rates were over 30% in Kyunggj-, Kyongnam-, and Chonnam-do. The highest isolation rate was 37.4% and recorded in Kyongnam-do. On the other hand, Chonbuk-do had the lowest isolation rate as 13.5%. In grouping of C parasitica by colony shape and color, yellow colony with irregular margin were the most dominant colony type with a frequency of 65.2%. When the 672 isolates were inoculated on the chestnut twigs, 380 isolates (56.5%) caused lesions larger than the standard virulent isolate EP155-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. In Bavendamm test that determines phenol oxidase activity, 97.1% of all the isolates resulted the same or darker discoloration than EP155-2, and only 12.2% resulted the same or lighter discoloration than UEP-1. In the vegetative compatibility (VC) tests, total 670 isolates were divided into 121 VC groups (VCGs). Kyongnam-, Chonnam-, and Chungnam-do, the three principal chestnut plantation area, had 49, 33, and 27 VCGs, respectively. Among the VCGs, the biggest VCG, KR-VC104, was composed of 164 isolates and the second biggest VCG had 62 isolates. But, 64 of 121 VCGs consisted of sole member. More than 65.8% of KR-VC104, was isolated from the three provinces, Kyongnam-, Kangwon-, and Chungbuk-do. In KR-VC104, 62.8%, 59.1%, and 85.9% of the isolates looked like virulent in colony type, pathogenicity test, and Bavendamm test. In ds-RNA detection tests using cellulose chromatography, 77 of total 650 isolates were ds-RNA positive and detected ds-RNA segments were approximately 12kb, 3kb, 2.7kb, 2kb, and 1.8kb in size. Among the 77 isolates, 46 isolates had 12kb and 25 isolates had 12kb and 2.7kb. Other 6 Isolates had small ds-RNA segments. Kyongnam-, Chonnam-, and Chungnam-do had 43, 16, and 5 ds-RNA positive isolates, respectively. Among the 121 VCGs, only 29 VCGs had ds-RNA positive isolates.(중략)

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Standardization for Analysis Method of Total Polyphenol in Complex of Picao Preto (피카오프레토 등 복합물 중 총 폴리페놀 분석법 표준화)

  • Hu, Soojung;Kim, Ji-An;Moon, Myung-Hee;Lee, Sung-Hye;Yoon, Hae-Seong;Hong, Jin-Hwan
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.44-49
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    • 2018
  • As generic health functional food items have been expanded, this research project has been conducted to prepare a scientific and systematic standardized analytical method of relevant food item and examine the suitability of the method for health/functional foods on sale. Total polyphenol was necessary for development and verification of standardized analytical method. The method exhibited high linearity in the tannic acid calibration curve ($r^2$ > 0.999) over concentrations of $5-50{\mu}g/mL$. The limits of detection and quantitation for tannic acid were $5{\mu}g/mL$ and $15{\mu}g/mL$, respectively, while tannic acid recovery was 102.3-112.4% with standard deviations of 0.8-3.2%. To verify the accuracy of the analytical method, the labeled amounts of purchased health functional foods were monitored. The recovery for tannic acid was 105.6% of the labeled amounts. Thus, the new method was suitable for all cases.