衍生化液相層析螢光法對Perhexiline之分析研究

Derivatization Liquid Chromatographic Analysis of Perhexiline as a Fluorescent Derivative

黃培羿(Pei-Yi Huang)

指導教授 ： 吳信隆

高雄醫學大學/藥學院/藥學研究所/碩士(2003年)

摘要

本研究利用衍生化高效液相層析法，對血漿中治療濃度狹窄的藥物perhexiline建立簡單靈敏之分析方法。本法利用（2 - naphthoxy）acetyl chloride（NAC）為衍生試劑，以三乙基胺（TEA）為催化劑，於甲苯溶媒下，將perhexiline 加以衍生，反應所得衍生物，以2 - naphthyl laurate（NTL）為內部標準，利用逆向層析分離管柱C-18為固定相，甲醇水溶液（95：5，v/v）為移動相進行層析；分離所得衍生物以螢光檢出器（激發波長230 nm；放射波長347 nm）加以偵測。所建立之分析方法定量範圍為0.1 ~ 20.0 mM，偵測極限為0.03 mM（S / N = 3；檢品注入量10 mL）。
本研究對影響perhexiline衍生反應之重要因素，包括反應溶媒、試劑需要量、鹼性催化劑及反應時間等，皆詳加探討。
本法對血漿中perhexiline之應用分析，經甲苯適當萃取並結合perhexiline之基本分析；本法對血漿中添加perhexiline之含量分析具有可行性，其血漿定量範圍為0.3 ~ 10.0 mM，偵測極限為0.03 mM。

關鍵字

高效液相層析法 ； 衍生 ； perhexiline

並列摘要

Analytical derivatization coupled with high performace liquid chromatography is used for the analysis of perhexiline, a prophylactic antianginal drug. The method is based on the dervatization of perhexiline with a fluorescent derivatizing reagent, ( 2 - naphthoxy ) acetyl chloride ( NAC ) in toluene, using triethylamine ( TEA ) as a base activator. After derivatization, methanol was added to the reaction mixture to inactivate the excess derivatizing reagent. The resulting derivative was separated on a C-18 column, using a mixed solvent of methanol：water ( 95：5, v/v) as the mobile phase, and monitored with a fluorometric detector at excitation 230 nm and emission 347 nm. 2 - Naphthyl laurate（NTL）was used as an internal standard ( I.S.), and the peak - area ratios of perhexiline derivative to I.S. were used for quantitative analysis.
The linear range of the method for the determination of perhexiline was 0.1 ~ 20.0 mM with a detection limit of 0.03 mM (S / N = 3；per 10mL injection). Several parameters affecting the derivatization of perhexiline were discussed including reaction solvent, amount of derivatizing reagent, base catalyst, and reaction time.
Application of the method to the analysis of perhexiline spiked in plasma was studied. After suitable toluene extraction of perhexiline from plasma, the toluene extract was directly subjected to be derivatized with NAC. The lower quantitation limit of the method is 0.3 mM for perhexiline in plasma with a detection limit of 0.03 mM.

並列關鍵字

high performace liquid chromatography ； derivatization ； perhexiline

參考文獻

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