D-乳酸螢光生物感測器薄膜化之相關研究

D-乳酸已在研究中證實與糖尿病、腎損傷、高血壓及癌症具有相關性，目前D-乳酸的檢測法是使用高效液相層析儀(HPLC)，此方法的缺點為耗時過長及操作複雜故本研究藉由偵測菸鹼醯胺腺嘌呤二核苷酸之還原態(NADH)的濃度建立了一種新型薄膜式D-乳酸螢光生物感測器。薄膜以V-group結構設計並使用聚二甲基矽氧烷(polydimethylsiloxane, PDMS)製作，將薄膜作為基板並將酵素(D-Lactate dehydrogenase)鍍在上面後形成酵素薄膜，將待測檢品及輔酶菸鹼醯胺腺嘌呤二核苷酸之氧化態(NAD+)滴上反應後，反應產物NADH以340 nm波長的UV-LED激發出490 nm波長之螢光強度並成正比，由NADH即可推算出老鼠血清中D-乳酸的濃度。此生物感測器有良好的線性程度(R2=0.9934)及偵測極限12.4 μmol/L，測定之精密度範圍在1.9-11.79%皆低於15%，準確度範圍在102.8-110.95%，大鼠血清檢測結果與HPLC比較平均誤差在0.34-11.87%。相較於高效液相層析檢測法，本薄膜式D-乳酸螢光生物感測器能夠提供一個快速、精確、成本低、且方便的檢測生物樣品中D-乳酸的新選擇。

關鍵字

薄膜；螢光； D-乳酸；生物感測器

並列摘要

In this study, we constructed a film-based D-lactate biosensor by detecting NADH. The base of film is made by polydimethylsiloxane (PDMS) coated with D-lactate dehydrogenase (D-LDH). The nicotinamide adenine dinucleotide (NAD+) is put in rat serum or urine to catalyze D-lactate to pyruvate with D-LDH while the coenzyme NAD+ was reduced to NADH. The concentration of NADH can be calculated by the fluorescence intensity and then infer the content of D-lactate. The results have good linearity (R2 = 0.9934) and the limit of detection (LOD) is 12.4 μmol/L. The intra-day accuracy and precision ranged from 102.8 to 110.95% and 2.78 to 8.85%, respectively, while inter-day accuracy and precision ranged from 101.75 to 108.04% and 1.53 to 9.05%, respectively. In addition, the film-based biosensor detection time required is about 20 times faster than HPLC method. Therefore, this new biosensor provides a rapid, accurate and convenient choice for D-lactate determination in biological sample.

並列關鍵字

Film ； Fluorescence ； D-lactate ； Biosensor

參考文獻

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D-乳酸螢光生物感測器薄膜化之相關研究

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