電化學式丙胺酸轉胺酶檢測試片之開發

Development of the electrochemical strips for alanine transaminase determination

麥耀仁(Yau-Ren Mai)

指導教授 ： 吳太光 ； 蔡祐輔

中原大學/理學院/化學研究所/碩士(2006年)

https://doi.org/10.6840/cycu200600692

摘要

近年來，肝病已成為國人十大死亡原因的前幾名，因此發展一既簡便又快速的肝功能生物感測器便成為重要的研究主題。一直以來，化學網版修飾電極在電化學的偵測上就很容易受到檢體成份的干擾，而且傳統電極的造價昂貴而且容易受到污染，因此我們以化學網版修飾電極來取代一般的傳統電極。本論文是利用浸泡的方式將塗覆物質包括 Meldola’s Blue、麩胺酸去氫酶（NADH）、輔酶（NAD+）及介面活性劑（CMC與Tween 20）固定在吸附層（不織布）的表面，製備成化學網版修飾電極後再利用循環伏特安培法（cyclic voltammetry）及安培電流法（amperometry）來進行測試。
本研究以網版印刷電極針對（1）塗覆物質的各成分（2）測試轉胺酶ALT的活性（3）全血，來進行的檢測。由實驗結果顯示，自製之化學網版修飾電極的變異係數（CV）約為10 %，在測量ALT試劑的操作電位設定在0.1 V時，可以有效地偵測到電流訊號。在室溫下操作，反應時間為3分鐘，可以測得ALT的線性範圍在30∼300 U/L，R＝0.9961（n＝5）。
這些結果雖然仍不盡理想，但相信如果再進一步修正塗覆物質的種類與比例，必能提升對偵測ALT之靈敏度及線性範圍，而些部份的研究也將是我們持續努力的方向。

關鍵字

麩胺酸去氫酶 ； 網版印刷電極 ； 菸鹼醯胺核苷酸 ； 麩胺酸鹽

並列摘要

For the past few years ,liver disease has become one of the main reason to cause people’s death . It is crucial to develop a method which is cheap , fast and accurate to liver function detection. Because of some excellent characteristic of chemical modified electrodes (CMEs) , such as low contamination by the matrix , low cost , mass production, flexibility and disposable type , the traditional electrodes have been replaced by the chemical modified electrodes .
In this thesis , CMEs was produced by the method of screen printing , reagents that thesis including Meldola’s Blue , NADH , NAD+ , CMC and Tween 20 are coated on a reaction medium which is on the electrode surface . Chemical modified electrodes were tested by the methods of cyclic voltammetry and amperometry with the usage of (1) the components of the reagents that make up the
CMES ,(2) Alanine aminotransferase (ALT) and(3) whole blood . The result shows that , the precision of the CMEs is about 10% . The
dynamic range is 30~300 U/L for ALT determination , tested at the potential of 0.1V . The response time is 3 minutes , which indicated that the method is highly effective in the detection of ALT activity .
According to above results , we believe that the CMEs can be improved by varing the reagents that coated on the surface of the electrode and the preparation conditions , and the sensitivity and dynamic range of ALT determination can be improved as well .

並列關鍵字

GLDH ； Screen-Printed Electrode ； NADH ； Glutamate