衍生化高壓液相層析法分別對Glucosamine及 Kanamycin之微量分析研究

Trace Analysis of Glucosamine and Kanamycin by Derivatization and High-performance Liquid Chromatography

梁藝倩(Yi-Chain Liang)

指導教授 ： 陳素惠

高雄醫學大學/藥學院/藥學研究所/碩士(2003年)

摘要

中文摘要
本論文嘗試以衍生化液相層析法對glucosamine及kanamycin建立簡單且靈敏度高之微量分析法。
Ⅰ. 以1-naphthyl isothiocyanate (NITC) 為衍生試劑，對glucosamine的微量分析
此方法以1-naphthyl isothiocyanate與glucosamine上的胺基反應，接上發色基團後加以偵測。衍生反應是在50°C下反應1小時，以吡啶為反應溶媒，衍生物利用逆向層析分離管柱分離，以230 nm 紫外光檢出器偵測。此方法可利用於glucosamine市售膠囊之含量分析及血液檢品中glucosamine之含量分析。
Π. 以1-naphthyl isothiocyanate (NITC) 為衍生試劑，對kanamycin的微量分析
此方法1-naphthyl isothiocyanate與kanamycin上的胺基，接上發色基團後加以偵測。衍生反應是在70°C下反應1小時，以吡啶為反應溶媒，衍生後加入methylamine的乙腈溶液消除過量衍生試劑。衍生物利用逆向層析分離管柱分離，以230 nm 紫外光檢出器偵測。此方法可利用於kanamycin市售膠囊之含量分析及血液檢品中kanamycin之含量分析。

關鍵字

衍生化高壓液相層析法

並列摘要

英文摘要
A simple and sensitive high performance liquid chromatographic method has been established for trace determination of glucosamine and kanamycin by derivatization.
Ι. Trace analysis of glucosamine with 1-naphthyl isothiocyanate (NITC) as derivatizing agent
The method is based on detection of glucosamine derivatized at amino group with 1-naphthyl isothiocyanate. The reaction was carried out in pyridine at 50°C for 1 h. The derivative was analysed by reverse-phase column and detected at 230 nm. Application of the proposed method to analysis of glucosamine in commercial sources and human plasma has proved to be satisfactory.
Π. Trace analysis of kanamycin with 1-naphthyl isothiocyanate (NITC) as derivatizing agent
The method is based on detection of kanamycin derivatized at amino groups with 1-naphthyl isothiocyanate. The reaction was carried out in pyridine at 70°C for 1 h. After derivatization, methylamine acetonitrile solution was added to the reaction mixture to eliminate the excess derivatizing agent. The derivative was analysed by reverse-phase column and detected at 230 nm. Application of the proposed method to
analysis of glucosamine in commercial sources and human plasma has proved to be satisfactory.

並列關鍵字

Glucosamine ； Kanamycin

參考文獻

參考資料

1. W. D. Wagner, A more sensitive assay discriminating galactosamine and glucosamine in mixtures, Anal. Biochem., 94, 394 (1979).

2. Setnikar, C. Giachetti and G. Zanolo, Absorption, distrubition and excretion of radioactivity after a single intravenous or oral administration of [14C] glucosamine to the rat, Pharmatherapeutica, 3, 538 (1984).

3. T. P. Mawhinney, M. S. Feather, G. J. Barbero and J. R. Martinez, The rapid, quantitative determination of natural sugars (as aldononitrile acetates) and amino sugars (as o-methyloxime acetates) in glycoproteins by gas-liquid chromatography, Anal. Biochem., 101, 112 (1980).

4. T. Shinohara, Use of a flame therminoic detector in the determination of glucosamine and galactosamine in glycoconjugates by gas chromatography, J. Chromatogr., 207, 262 (1981).