高效能液相層析法對人體血漿中inosine、hypoxanthine及adenosine 之同時分析研究

Simultaneous determination of inosine, hypoxanthine and adenosine in human plasma by high performance liquid chromatography

王筱琄(Hsiao-Chuan Wang)

指導教授 ： 柯黃盛

高雄醫學大學/藥學院/藥學研究所/碩士(2008年)

摘要

本研究建立一個快速簡單的HPLC-DAD方法，同時偵測人體血漿中adenosine、inosine和hypoxanthine的含量，並應用於缺血性心臟病病人血漿分析。本研究使用高能液相層析法，以C18層析管柱(250 × 4.6 mm I.D.; 5 µm) 做為固定相，以乙腈和醋酸鈉緩衝溶液(100 mM，pH= 3)之混和溶液(3 : 97，v/v)作為液相層析之移動相，且流速則控制在1.4 mL/min。以allopurinol為本實驗之內部標準品，待測物則以二極體陣列檢測器(DAD)在260 nm吸收波長檢測adenosine，在250 nm吸收波長檢測inosine和hypoxanthine。線性範圍分別為: adenosine(0.02 – 30 µM)；inosine(0.5-30µM)；hypoxanthine (0.5-30µM)。注入體積為100µL。層析分析可以在7分鐘以內完成。
這個方法可以用來幫助評估當病人發生發生缺血性心臟疾病時，血漿中ATP的降解產物adenosine、inosine、hypoxanthine是否有增加，以作為診斷缺血性心臟疾病診斷或治療之參考。

關鍵字

adenosine ； inosine ； hypoxanthine

並列摘要

A high performance liquid chromatographic (HPLC) method with diode array detection (DAD) is described for the simultaneous determination of adenosine, inosine and hypoxanthine in human plasma. The HPLC separation of the analyte was achieved on a C18 column (250 × 4.6 mm I.D.; 5 µm), using a mobile phase, consisting of 100 mM sodium acetate buffer, pH 3.0, containing 3% of acetonitrile at a flow rate of 1.4 mL/min. Allopurinol was used as internal standard. Diode array detector (DAD) was used, with adenosine monitored at UV 260 nm, while inosine and hypoxanthine at 250 nm. The linear range for the determination of adenosine, inosine, and hypoxanthine were 0.02 - 30 µM, 0.5 - 30µM and 0.5 - 30µM.
The method was to facilitate evaluation of a hypothesis that humans undergoing cardiac ischemia will elevated blood levels of ATP catabolic products (adenosine, inosine, and hypoxanthine).

並列關鍵字

adenosine ； inosine ； hypoxanthine

參考文獻

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