衍生化液相層析法分析葡糖醛酸內酯之研究

Derivatization Liquid Chromatographic Analysis of Glucuronolactone

葉朱娘(Chu-Niang Yeh)

指導教授 ： 溫燕霞 博士

高雄醫學大學/藥學院/藥學研究所/碩士(2003年)

摘要

很多綜合維生素製劑中含有glucuronolactone (GL)，由於GL具有促進肝臟代謝機能，通常用來治療高膽紅素血症(bilirubinemia)，最近的文獻報告也建議GL具有抗壓及保護神經免於受損的作用。
本研究嘗試以前置衍生反應系統( pre-column derivatization system ) 配合高效液相層析法，對綜合維生素製劑中glucuronolactone建立簡單且高感度之微量分析方法。本分析方法是利用2,4-dinitrophenylhydrazine ( 2,4-DNP )為衍生試劑，以氰甲烷和水為反應溶媒，在硫酸催化劑下，將GL衍生為具有紫外光吸收特性之衍生產物，其結構證實為glucuronolactone-2,4-dinitrophenyl-hydrazine ( Glu-DNP )。所得之衍生物利用逆相層析管柱COSMOSIL 5C18-AR column，以氰甲烷︰水 ( 25︰75 ；v/v )溶液為移動相，coumarin為內部標準，以紫外光偵測器在波長360 nm下進行偵測。
影響GL衍生反應之各種因素，包括催化劑之需要量、衍生試劑之濃度、反應時間及反應溫度等等，皆詳加探討。本法可測定製劑中GL之線性範圍是7.5 ~ 120 mmol/mL，而同日內及異日間分析所得的相對標準偏差 (RSD) 均小於1.3 %，說明本分析法具有良好之定量可靠性，可應用於綜合維生素製劑中GL之含量測定。

關鍵字

衍生化液相層析法 ； 葡糖醛酸內酯

並列摘要

Many of multivitamine pharmaceuticals contain glucuronolactone (GL), which was used in the treatment of bilirubinemia because it improve the liver function. In recent report suggest that GL have protective effects against neuron damages and anti-stress effect.
A pre-column derivatization system with high performance liquid chromatographic method is described for the determination of GL in multivitamine pharmaceuticals, GL was derivatized with strong chromophore reagent, 2,4-dinitrophenylhydrazine (2,4-DNP), using sulfuric acid as catalyst. The derivative obtained was separated on a COSMOSIL 5C18-AR column with acetonitrile : H2O ( 25:75, v/v ) as the mobile phase, and coumarin as the internal standard.
The parameters affecting the derivatization of GL, including catalyst concentration, the concentration of sulfuric acid, the reaction temperature and reaction time, and the amount of derivatizing agent were investigated. The linear range for the quantitative of GL in pharmaceuticals was between 7.5 ~ 120 μmol/mL. The intraday relative standard deviation ( n=5 ) and the interday relative standard deviation ( n=5 ) were all less than 1.3 %.

並列關鍵字

Derivatization Liquid Chromatographic Analysis ； Glucuronolactone

參考文獻

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