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효소반응법을 이용한 우황 및 우황함유 액상 제제 중 총담즙산의 정량
하인식,김승환,차봉진,권종원,양중익,민신홍,Ha, In-Sik,Kim, Seung-Hwan,Cha, Bong-Jin,Kwon, Jong-Won,Yang, Joong-Ik,Min, Shin-Hong 한국약제학회 1991 Journal of Pharmaceutical Investigation Vol.21 No.2
A simple and sensitive method was developed for the quantification of free and conjugated bile acids in bezoar without prior hydrolysis. $3{\alpha}-Hydroxy$ bile acids are first oxidized to 3-keto bile acids in the reaction catalyzed by $3{\alpha}-hydroxysteroid$ $dehydrogenase(3{\alpha}-HSD)$. During this oxidative reaction, an equimolar quantity of nicotinamide adenine dinucleotide(NAD) is reduced to NADH and subsequently oxidized to NAD with concomitant reduction of nitrotetrazolium blue(NTB) to diformazan by the catalytic action of diaphorase. The diformazan has an absorbance maximum at 540 nm. The intensity of the color produced is directly proportional to bile acids concentration in the bezoar extracts. The optimum conditions for the enzymatic reaction such as effects of reaction time, reaction temperature and pH, and stability were investigated. Calibration plots for the sodium chelate observed to be linear and intra-, inter-assay analytical recovery of bile acids averaged $97.65{\pm}3.4%(S.D.)$. Therefore, it is considered that the quality control of total bile acids from bezoar or bezoar-containing liquid preparation using this simple and sensitive assay system will be acceptable. Also current bezoars and bezoar-containing liauid preparations were examined their total bile acids from this method.
2-Bromoacetylnaphthalene을 螢光誘導體化劑로 利用한 Alclofenac의 HPLC 分析에 관한 硏究
李允中,曺正吉,河仁植,金容熙 成均館大學校 科學技術硏究所 1990 論文集 Vol.41 No.1
A simple, rapid and high sensitive method for determination of alclofenac is described. 2-Bromoacetylnaphthalene was used as the pre-column fluorescent derivatizing reagent for high performance liquid chromatography. Alclofenac was derivatized quantitatively into fluorescent compound by reacting with 2-bromoacetylnaphthalene in the presence of 18-crown-6 ether in acetonitrile. The optimum conditions for the derivatization such as concentration of KOH, 18-crown-6 and 2-bromoacetylnaphthalene, reaction temperature and reaction time were investigated. The structure of alcolofenac derivative was confirmed from IR, NMR and mass spectra. The fluorescence properties of alclofenac derivative were examined. The derivative was separated on a reverse phase column (Lichrosorb RP-8) in isocratid elution mode using the secondary mixture of acetonitrile and water as the mobile phase. The effluent was monitored by fluorometer.(Ex. 303, Em. 418 nm). The calibration plots for the peak area versus concentration of alclofenac observed to be linear(r =0.999).
효소반응법을 이용한 우황 및 우황함유 액상 제제 중 총담즙산의 정량
김승환,민신홍,권종원,양중익,차봉진,하인식 한국약제학회 1991 Journal of Pharmaceutical Investigation Vol.21 No.2
A simple and sensitive method was developed for the quantification of free and conjugated bile acids in bezoar without prior hydrolysis. 3α-Hydroxy bile acids are first oxidized to 3-keto bile acids in the reaction catalyzed by 3α-hydroxysteroid dehydrogenase(3α-HSD). During this oxidative reaction, an equimolar quantity of nicotinamide adenine dinucleotide(NAD) is reduced to NADH and subsequently oxidized to NAD with concomitant reduction of nitrotetrazolium blue(NTB) to diformazan by the catalytic action of diaphorase. The diformazan has an absorbance maximum at 540 ㎚. The intensity of the color produced is directly proportional to bile acids concentration in the bezoar extracts. The optimum conditions for the enzymatic reaction such as effects of reaction time, reaction temperature and pH, and stability were investigated. Calibration plots for the sodium cholate observed to be linear and intra-, inter-assay analytical recovery of bile acids averaged 97.65±3.4%(S.D.). Therefore, it is considered that the quality control of total bile acids from bezoar or bezoar-containing liquid preparation using this simple and sensitive assay system will be acceptable. Also current bezoars and bezoar-containing liquid preparations were examined their total bile acids from this method.