氧化銅/氧化亞銅/聚吡咯複合材料於生化分析應用

Abstract

本研究利用麻布紋紙 (LT paper) 為基材，再以氧化銅/氧化亞銅/聚吡咯複合材料功能化，分別用於葡萄糖、β-半乳糖苷酶與大腸桿菌之定量檢測上。以兩步合成步驟將氧化銅/氧化亞銅/聚吡咯修飾於麻布紋紙上:吸附於麻布紋紙上之吡咯單體在抗壞血酸和果糖存下，與銅離子反應形成氧化亞銅/聚吡咯 (Cu2O/Ppy)；之後在溴化十六烷基三甲銨下和銅離子反應，完成功能化紙電極 (CuO/Cu2O/Ppy LT paper electrode) 之製備。氧化銅/氧化亞銅/聚吡咯紙電極具類似葡萄糖氧化酶之催化活性，對葡萄糖之最低偵測極限可達0.04 mM。將此電極之面積縮小至0.12 cm2，製作成微型化紙電極。微電極可有效提升質量傳輸效率並減少樣品需求體積；利用此電極對葡萄糖專一的催化活性，可偵測β-半乳糖苷酶催化水解乳糖產生之葡萄糖，進一步定量β-半乳糖苷酶；亦可檢測能產生β-半乳糖苷酶之大腸桿菌，偵測極限分別可達0.034 U mL-1與120 CFU mL-1。活化劑和抑制劑對β-半乳糖苷酶之作用、誘導劑異丙基-β-D-硫代半乳糖苷 (IPTG) 誘導β-半乳糖苷酶表達，與抗生素對大腸桿菌之生長影響亦能由此檢測法進行篩檢。本研究所合成之氧化銅/氧化亞銅/聚吡咯 (微) 紙電極具可拋性、易處理、重量輕與低成本等特點；不僅對葡萄糖檢測具有靈敏性、選擇性與穩定性之優勢，可用於血糖監測應用上；另外對 β-半乳糖苷酶與大腸桿菌之檢測都極具發展應用潛力。

Linen texture (LT) paper functionalized with CuO/Cu2O/Ppy composite has been prepared and used as an electrode for quantitation of glucose, β-galactosidase, and Escherichia coli (E. coli). The low-cost CuO/Cu2O/Ppy LT paper is prepared through a two-step process; pyrrole adsorbed on an LT paper reacts with Cu2+ in the presence of ascorbic acid and fructose to form Cu2O/Ppy that then reacts with Cu2+ in the presence of cetyltrimethylammonium bromide. The CuO/Cu2O/Ppy LT paper electrode possessing glucose oxidase-like activity catalyzes the oxidation of glucose, allowing quantitation of glucose down to 0.04 mM. Furthermore, functionalized paper size was decreased to 0.12 cm2 to fabricate the CuO/Cu2O/Ppy LT paper microelectrodes. The microelectrode enhances the mass transfer efficiency and reduces the sample volume. By taking the specificity of β-galactosidase to hydrolyze lactose to form glucose, the functionalized paper microelectrode has been employed for the quantitation of β-galactosidase, and detection of E. coli that express β-galactosidase as low as to 0.034 U mL-1 and 120 CFU mL-1, respectively. In addition, this assay is powerful for screening activators/inhibitors of β-galactosidase, and the antibiotic activities toward E. coli. Our results clearly show the merits such as low-costs, high sensitivity, specificity, and superior stability of CuO/Cu2O/Ppy LT paper (micro) electrodes for blood glucose monitor, as well as for quantitation of β-galactosidase activity and E. coli.