传感器类型
其他(溶解氧电极生物传感器)
检测对象
葡萄糖(glucose/D-glucose);样品基质:PBS标准液及商业食品样品(果汁、牛奶、酸奶、奶粉、巧克力、蛋糕、饼干等)
检测原理
该传感器以蛋壳膜为生物平台,将N-乙酰-L-半胱氨酸保护的金纳米颗粒(Au NPs)经EDC/NHS活化后与葡萄糖氧化酶(GOx)共价连接,再用戊二醛交联固定于蛋壳膜表面,并置于溶解氧电极上。样品中的D-葡萄糖扩散进入膜内,被GOx特异性催化氧化为D-葡萄糖酸和过氧化氢,同时消耗溶解氧。Au NPs提高GOx负载量、稳定性和催化活性,使氧消耗速率加快。溶解氧浓度下降被O2传感器转换为电信号,信号下降量与葡萄糖浓度成正比,从而实现定量检测。
检测灵敏度
LOD: 2.5 μM (S/N = 3);线性范围: 5.0 μM–0.525 mM;灵敏度斜率: 15.78 mg/L O2 depletion/mM glucose;R^2 = 0.9998
效应效果
该传感器响应时间<60 s(含Au NPs为58 s,无Au NPs为120 s),重复性RSD=3.6% (n=6)。6个月储存后保留95%初始响应,300次测量后保留99%。果糖、乳糖、柠檬酸、苯甲酸钠、抗坏血酸、蔗糖、尿素、DL-半胱氨酸、可溶性淀粉及多种无机盐不显著干扰。食品样品加标回收率92.1–104%,RSD 0.5–5.3%。与同类O2型传感器相比,校准灵敏度15.8 mg/L O2/mM,高于8.34、9.42、6.75、6.28;LOD 2.5 μM优于3.5、8.0、9.0 μM;6个月稳定性95%优于85.2%、87.3%、86.5%、83.0%。作者认为其简单、低成本,适合食品与生物技术应用。
传感器的构成
- 换能器电极:Pasco CI-6542 O2传感器(溶解氧电极),检测溶解氧下降并输出信号
- 生物平台基底:蛋壳膜(eggshell membrane, ESM),高度交联蛋白纤维,气/水透过,承载Au NPs/GOx
- 纳米材料修饰层:N-乙酰-L-半胱氨酸保护的金纳米颗粒(Au NPs, 5000 ppm),提供生物相容界面并增强GOx活性
- 偶联/交联层:EDC/NHS(75 mM)活化Au NPs并与GOx连接;戊二醛(glutaraldehyde, 50% w/w)将Au NPs/GOx交联固定于ESM
- 识别元件:葡萄糖氧化酶(GOx, EC 1.1.3.4, 0.8% w/v),特异性催化D-葡萄糖氧化
- 反应介质:50 mM PBS(pH 7.0),维持酶活性与离子强度
- 信号输出:溶解氧(O2)消耗,由O2传感器转换为电信号
中文摘要
建立了一种简单且相对廉价的葡萄糖生物传感器,其将金纳米颗粒(Au NPs)与葡萄糖氧化酶(GOx)复合并固定于蛋壳膜生物平台上。扫描电镜证实Au NPs/GOx成功固定于蛋壳膜。详细研究了pH、磷酸盐缓冲液浓度和温度对传感器的影响。该传感器在5–525 μM葡萄糖浓度范围内呈线性响应,检出限为2.5 μM(S/N=3)。传感器重复性良好,RSD=3.6%(n=6);操作稳定性良好,可完成300次以上测量;长期储存稳定性好,保质期至少6个月。响应时间小于60 s。成功测定了商业食品样品中的葡萄糖含量。该工作展示了开发用于生物技术、生物医学和工业应用的低成本生物传感器的潜力。
英文摘要
A simple and relatively cheap glucose biosensor based on a combination of gold nanoparticles (Au NPs) and glucose oxidase (GO(x) ) immobilized on a bioplatform eggshell membrane was established. Scanning electron microscopy showed successful immobilization of Au NPs/GO(x) on the eggshell membrane. The effects of pH, phosphate buffer concentration, and temperature on the glucose biosensor were studied in detail. The biosensor shows a linear response at a glucose concentration range of 5-525 μM. The detection limit of the biosensor is 2.5 μM (S/N = 3). The biosensor exhibits good repeatability with RSD = 3.6% (n = 6), good operational stability with over 300 measurements and long-term storage stability with a shelf life of at least 6 months. The response time is less than 60 s. The glucose level in commercial food samples has been successfully determined. The proposed work shows potential to develop cost-effective biosensors for biotechnological, biomedical and industrial use.