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환경 오염물질의 정량법 개발과 거동에 관한 연구 ( I ) : 미셀 역상 액체 크로마토그래피에서 페놀과 벤젠 일치환체들의 용리거동
이대운,방은정,조병연,Lee, Dai Woon,Bang, Eun Jung,Cho, Byung Yun 한국분석과학회 1993 분석과학 Vol.6 No.1
The purpose of this study is to investigate the elution behavior of monosubstituted phenols and benzenes in micellar liquid chromatographic system, $C_{18}$ column-anionic surfactant, sodium dodecyl sulfate(SDS). The partition coefficients between the micellar pseudophase-water and modified stationary phase-water are calculated by the relationship between solute retention and micellar mobile phase(SDS) composition. The free energy of transfer of solute from water to micelle is also calculated from these values. There is a direct correlation between the hydrophobicity parameters in MLC and corresponding partition data for 1-octanol-water, which indicates that the hydrophobicity of molecules plays an important role in the partition for both systems and that quantitative structure activity relationships(QSAR) are available from studies on micellar partition. The other purpose of this study is to investigate methylene selectivity of alkyl homologous series through correlation between retention and the number of carbons. The correlation between hydrophobicity parameters in MLC and 1-octanol-water partition data was also observed when n-propanol was as a modifier in the mobile phase. 음이온 계면활성제인 sodium dodecyl sulfate(SDS)를 이동상으로 한 미셀 액체 크로마토그래피(micellar liquid chromatography : MLC)에서, 시료로 22종의 페놀과 벤젠 일치환체들을 선택하여 이 시료들의 용리거동을 조사하여 MLC계에서의 소수성 효과를 알아 보았다. MLC에서의 머무름과 미셀 농도간의 상관관계를 통해 시료들이 미셀-물간, 변형된 정지상-물 사이에서 분배될 때의 분배계수를 구하였고, 또 이를 이용하여 물-미셀간의 전이 자유 에너지를 구하였다. 이러한 MLC에서 얻은 소수성 파라미터들은 옥탄올-물계와 연관시켜 본 결과 좋은 상관관계를 보였다. 따라서 이 두 계에서 시료의 소수성이 머무름에 중요한 영향을 미침을 알 수 있었다. 한편, MLC를 이용하면 quantitative structure activity relationships(QSAR)에서의 소수성 정량 연구에 적용이 가능하다. 또한 알킬 치환체들에서 탄소 수의 증가에 따른 머무름 관계를 통하여 소수성의 선택성을 알아 보았고, 유기 변형제로 n-프로판올을 첨가하였을 때에도 마찬가지로 MLC계에서의 소수성 파라미터와 옥탄올-물계 사이에 상관관계가 있음을 알았다.
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High - Performance Liquid chromatography 를 이용한 정상 성인 혈장 히스타민의 일중변화에 관한 연구
윤영섭(Young Sup Yoon),이승태(Seung Tae Lee),박중원(Joong Woon Park),홍천수(Chein Soo Hong),조병연(Byung Yun Cho) 대한내과학회 1992 대한내과학회지 Vol.43 No.3
N/A Background: Histamine plays an important role in mediating various allergic reactions, and as a neurotransmitter as well as a mediator of gastric acid secretion. The measurement of plasma histamine has a great significance in physiologic, pathophysiologic, and pharmacologic investigation. Because histamine is rapidly metabolized in the circulation and excreted into the urine, its level is very low in the blood. In order to measure histamine levels in the blood, a precise quantification technique is required. Recently, many analytical methods have been developed to measure plasma histamine levels and it has been found that plasma histamine levels. show circadian variation in healthy volunteers as well as in asthmatic patients. Methods: In this study, high-performance liquid chromatography combined with fluorometric assay, one of the most commonly adopted method, was used to measure plasma histamine levels, Plasma histamine levels were measured every hour for a day in 12male volunteers. The reference value of the plasma histamine was established and the presence of circadian variation was investigated. Results: 1) The diurnal mean of plasma histamine levels was 1.12±0.28ng/ml. 2) The means of plasma histamine levels at various sampling hours ranged between 0.40±0.48ng/ml and 2.48±1.26ng/ml. 3) The means of plasma histamine levels at each hour showed circadian variation, which takes the form of a curve with 3peaks at 6,12and 19hour. 4) After dividing 24hours into 8time intervals, statistical analysis was performed to this circadian variation. The means of plasma histamine levels at 3different time intervals, which include 4 to 6, 7 to 8 and 18 to 19hours, were significantly higher than the others. Conclusion: Diurnal mean of plasma histamine levels was 1.12±0.28ng/ml in 12healthy volunteers and plasma histamine levels showed circadian variation which revealed higher values especially in early morning hours.
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HPLC에 의한 혈청 Vitamin A, D 및 E의 동시 정량에 관한 연구
조병연,허갑범,이현철,임승길,유경자 대한내분비학회 1987 Endocrinology and metabolism Vol.2 No.2
Clinical interest in evaluating concentration of fatsoluble vitamins in serum has increased in recent years, owing to their variables roles in human. The purpose of this study was, therefore, to determine vitamin A, D and E simultaneously in human serum by high-performance liquid chromatography. The results obtained are as follows; 1) Serum proteins were precipitated by the addition of ethanol and vitamins were subsequently extracted with n-hexane. Recoveries of vitamin A, D and E were 75.48, 99.57 and 92.91%, respectively. 2) For the optimum analysis, C_18 reversed-phase column was employed with methanol as an eluent. The detection of the vitamins was monitered with UV detection at 280nm. Vitamin A acetate was used as an internal standard to correct for variable recoveries. 3) The mean concentrations were 92.79+-6.91ng/100 ul for vitamin A, 9.42+-0.61 ng/100 ul for vitamin D and 799.03+-50.85 ng/ 100 ul for vitamin E. The detection limits were 2.18, 16.73 and 27.87 ng, respectively. This method permits simple, rapid, selective and reproducible determination of vitamin A, D and E. So this assay system might be adequate for the clinical routine work as well as research purpose.
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HPCL에 의한 Urinary Hydroxyproline의 정량에 관한 연구
조병연,허갑범,이현철,임승길 대한내분비학회 1988 Endocrinology and metabolism Vol.3 No.2
Hydroxyproline is liberated during the intracellular maturation of collagen. Furthermore, degradation of the N-terminal peptides which contain 4-hydroxyproline in type I procollagen is probably also a source of urinary hydroxyproline. Therefore, increased collagen synthesis as well as breakdown will result in elevated hydroxyproline levels. In fasting individuals, urinary hydroxyproline may therefore be used as a specific index of collagen metabolism. Our objective was to develop a reliable, optimized and more rapid HPLC method for urinary hydroxyproline analysis. Total hydroxyproline in urine is derivatized with 4-chloro-7-nitrobenzofurazan after reaction of the urine with o-phthalaldehyde. The highly fluorescent adducts of hydroxyproline is separated on a -Bondapak C18 reversed-phase column using acetonitrile-0.01 M sodium phsphate buffer, pH 7.0 (10:90, v/v) as mobile phase, followed by fluorometric betection.
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모세관 전기 영동법에 의한 분석에서 피크 모양 및 검출 향상을 위한 단백질의 유도체화
조병연 경희대학교 자연과학종합연구원 2000 자연과학논문집 Vol.6 No.-
단백질 및 항체를 6-aminoquinolyl-N-hydroxysuccinimidyl (6-AQC) 시약을 사용하여 단일 피크가 생성되는 조건으로 유도체화 하였다. 유도체화를 통하여, 전기 영동법에 의한 분석에서 피크 모양 및 검출 한계 등이 향상되었으며, pI 값 등의 변화 등이 관측되었다. 이러한 실험 결과 단백질의 전기 영동법에 의한 분석에서 더 좋은 분리도 및 피크 확인이 가능하였다. 유도체화된 단백질은 항원-항체 반응을 통하여 활성도를 확인하였으며, 단백질에 따라 활성도를 유지하는 단백질이 관측되었다. Proteins and antibodies can be derivatized with 6-ammoqumoly l-N- hydroxysuccinimidyl (6-AQC) reagent, leading to a single HPCE peak. These species have greatly improved peak shape and performance, improved detectabilities, altered pI values, and in general, improved identification properties in various HPCE modes. The confirmation of the activity of the derivatized proteins through antibody-antigen recognition was also performed, and the activity of some tagged proteins was observed.
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