嗎啡免疫生化感測器研究與應用

Preparation and application of morphine Immunosensors

張宏維(Hung-wai Chang)

指導教授 ： 施正雄

國立臺灣師範大學/理學院/化學系/碩士(2001年)

摘要

目前在檢測血液或尿液中嗎啡量常用的方法是利用管柱層析如氣象層析儀和高效能液相層析儀兩種方法。但是這樣的方法中往往需要事前的樣品處理需要相當多的時間與人力，免疫方法可以解決管柱耗時的問題。本篇所用的方法是利用抗嗎啡抗體與標幟上嗎啡的G6PDH(Glucose-6-phosphate dehydrogenase labeled with morphine, G6PDH-morphine)在光學與石英壓電免疫感測器上來進行偵測。在免疫感測器的部分，有很好的線性關係，而且偵測下限是113 ng/ml，這樣的偵測下限遠低於目前法律規定的300ng/ml，因此可以應用在實際的檢驗上，且光學免疫感測器的相對標準偏差值(R.S.D.) 3%左右，這是不錯的再線性的檢測方法。並且本篇也探討體內血清與尿液中主要的成分對於這個感測方法的影響。在石英壓電免疫感測器上，首先製備C60-G6PDH並且把C60-G6PDH固定在石英壓電晶體上作為感測膜來對於溶液中抗嗎啡抗體，當C60-G6PDH的吸附溶液中抗嗎啡抗體後會造成頻率的下降來作為偵測的標準，利用固定C60-G6PDH的石英壓電晶體來偵測溶液中的抗嗎啡抗體有相當不錯的再現性且對於抗嗎啡抗體有不錯的選擇性。

關鍵字

嗎啡

並列摘要

Current technology for determination of morphine in blood or urine is often to use the chromatographic methods such as GC and HPLC, but most of these methods are time consuming. To solve this problem, a morphine optical immuno-sensor and a quartz piezoelectric crystal sensor based on anti-morphine antibody and Glucose-6-phosphate dehydrogenase labeled with morphine (G6PDH-morphine) for opiate solutions were designed and prepared in this study. The optical morphine sensor exhibited good sensitivity with a detection limit of 113 ng/mL which was far below the allowed concentration (300 ng/mL) in law for morphine in biological solutions. The optical sensor also showed a high reproducibility with a relative standard deviation (RSD) of approximately 3 %. The interference of major biological species in simulated serum and urine solutions for the determination of morphine with the optical immuno-sensor was investigated and discussed. In the preparation of the quartz piezoelectric sensor, immobilized fullerene C60 - G6PDH-morphine was synthesized and applied as a coating material on quartz piezoelectric crystals for the immunoassay of anti-morphine antibody in aqueous solutions. The oscillating frequency of the G6PDH-morphine coated-piezoelectric crystal of the piezoelectric sensor decreased by adsorption of anti-morphine antibody in solutions. The piezoelectric crystal sensor showed nearly no interference from various biological species in simulated serum. The piezoelectric crystal sensor based on immobilized C60-G6PDH-morphine for anti-morphine antibody also exhibited good sensitivity and good reproducibility.

並列關鍵字

morphine