Development Of Fӧrster Resonance Energy Transfer Based Homogeneous Immunoassays For Determination Of (R)-Methadone

摘要

Analisis ubatan biasanya dijalankan dengan menggunakan kaedah kromatografi yang tepat dan spesifik seperti GC-MS dan LC-MS. Kaedah tersebut diketahui mempunyai kepekaan dan spesifisiti yang tinggi. Walau bagaimanapun, kaedah tersebut memerlukan peralatan yang canggih dan kepakaran profesional untuk menjalankan analisis. Bagi menganalisis sampel yang banyak, kaedah analisis yang lebih mudah seperti imunoasai akan dijalankan sebelum pengesahan keputusan dengan menggunakan GC-MS dan LC-MS. Oleh itu, asai “high throughput screening” adalah penting untuk meningkatkan kecekapan analisis perubatan. Dalam kajian ini, ciri-ciri berpendarfluor pewarna organik, titik kuantum dan kelat lantanida akan dikenal pasti kemudian digunakan. Sebagai keputusan, imunoasai homogen berasaskan FRET dan TR-FRET yang baru telah dihasilkan dan diuji untuk mengesan (R)-metadon dalam serum manusia. Kepekatan (R)-metadon dalam 50 pesakit MMT kemudian diuji. Kepekaan dan ketepatan asai yang dihasil kemudian dikaji dan disahkan dengan menggunakan kaedah LC-MS/MS. Hasil kajian juga menunjukkan bahawa terdapat kolerasi antara ketiga-tiga imunoasai dengan kaedah kromatografi.ududDrug analysis is commonly performed using an accurate and specific chromatographic method, such as GC-MS and LC-MS. These methods are known to be highly sensitive and specific. However, they require sophisticated equipment and also skilled manpower for the analysis. For analysis of large number of samples, less descending methods like immunoassays are used prior to confirmation by GC-MS or LC-MS. Thus, high throughput screening is essential in increasing the efficiency of drug testing. In this study, the fluorescence properties from various types of fluorescence materials, such as organic dyes, quantum dot and lanthanide chelates were investigated and employed. As a consequence, new FRET and TR-FRET based homogeneous competitive immunoassays were successfully designed and developed for detection of (R)-methadone in serum sample. Then, the concentrations of (R)-methadone in the 50 MMT patient’s serums were tested. Consequently, the accuracy and sensitivity of the developed assays were studied and validated by using LC-MS/MS method. Generally, comparison of the LC-MS/MS method with other three immunoassays methods showed adequate correlation.ud