电化学生物传感器 2011

Immobilized multi-species based biosensor for rapid biochemical oxygen demand measurement.

Biosensors & bioelectronics Liu C, Ma C, Yu D, Jia J, Liu L, Zhang B, Dong S
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组成图示

Immobilized multi-species based biose... 传感器构成示意图

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传感器类型

电化学生物传感器

检测对象

生化需氧量(BOD/BOD5,可生物降解有机物总量);样品基质:市政废水、工业废水、合成废水(GGA、OECD)

检测原理

该传感器属于呼吸型BOD传感器。废水中的可生物降解有机物经透析膜扩散至固定化BODseed微生物膜,被多物种微生物无差别氧化降解;微生物呼吸消耗样品中的溶解氧,使氧探头附近O2浓度下降。金盘工作电极在-0.7 V vs Ag/AgCl下发生O2还原反应,稳态电流/电压与溶解氧浓度相关。无BOD缓冲液的稳态电压为E0,加入样品后达到新稳态Es,响应ΔE=E0−Es随BOD升高而增大。系统采用流动注射和GGA/OECD标准曲线校准,响应时间通常小于10 min,恢复时间30–60 min。

检测灵敏度

LOD: 0.5 mg/l BOD(S/N=3);校准曲线: y = 19.0x + 19.7,R^2 = 0.9927;不同有机物响应: y = 19.704x,R^2 = 0.9441;与BOD5对比: y = 1.0645x - 1.7635,R^2 = 0.9705

效应效果

传感器活化时间约2 h,对温度、pH和PBS浓度耐受性较好:20–45 ℃下活性随温度升高,pH 4.9–9.2响应基本一致,PBS 5–50 mM响应稳定。重复性RSD为2.8%–4.3%(GGA 4–40 mg/L及活化过程),长期稳定性约50 d无明显下降,期间分析近500次。与BOD5对比,废水样品相关系数R^2=0.9705,斜率1.06,最大/最小标准偏差8.4%和0.4%(均值75.2和14.0 mg/L,n=5)。作者认为其无差别降解能力与BOD5一致,适合废水在线监测。

传感器的构成

  • 换能器电极:金盘工作电极(Au disk,0.8 mm)、Ag/AgCl参比电极(0.1 M KCl)和铂丝辅助电极(Pt wire),用于电化学检测溶解氧。
  • 气体扩散膜:Teflon膜(Orbisphere)贴附于氧探头表面,允许O2扩散并隔离电解质。
  • 载体屏障:透析膜(dialysis membrane,孔径10 μm,Oxyphen GmbH)承载生物膜并阻挡样品悬浮物。
  • 固定基质:有机–无机杂化材料,由silica sol与PVA-g-P(4-VP)接枝共聚物经溶胶–凝胶形成,多孔结构固定微生物并防止泄漏。
  • 识别元件:多物种微生物群落BODseed(Cole-Parmer E-05466-00,含7种活性污泥来源微生物),无差别降解有机物并消耗O2。
  • 信号读出:XSF-1在线BOD分析仪,在-0.7 V vs Ag/AgCl下读取稳态电压差ΔE=E0−Es。

中文摘要

为提高快速生化需氧量(BOD)检测方法的实用性,作者提出一种基于固定化多物种微生物BODseed的稳定BOD传感器,用于流动注射系统中的废水监测。采用有机–无机杂化材料固定BODseed后,生物膜的活化时间显著缩短。系统考察了温度、pH和磷酸盐缓冲液(PBS)浓度等影响因素,证实该传感器对环境具有较高耐受性,可在较宽pH和PBS浓度范围内工作。在优化固定化浓度1.0 mg/mL BODseed下,最低可检测BOD约为0.5 mg/L。所制备传感器对不同样品表现出优良稳定性和重现性,并对不同有机化合物及其混合物具有类似传统BOD5的无差别生物降解能力。传感器法结果与常规BOD5法测定废水样品的结果一致,表明该快速BOD传感器在废水监测实际应用中具有前景。

英文摘要

To improve the practicability of rapid biochemical oxygen demand (BOD) method, we proposed a stable BOD sensor based on immobilizing multi-species BODseed for wastewater monitoring in the flow system. The activation time of the biofilm was greatly shortened for the biofilm prepared by BODseed in the organic-inorganic hybrid material. Some influence factors such as temperature, pH, and concentration of phosphate buffer solution (PBS) were investigated in detail in which high tolerance to environment was validated for the BOD sensor permitted a wide pH and PBS concentration ranges. The minimum detectable BOD was around 0.5 mg/l BOD under the optimized 1.0 mg/ml BODseed immobilized concentration. The as-prepared BOD sensor exhibited excellent stability and reproducibility for different samples. Furthermore, the as-prepared BOD biosensor displayed a notable advantage in indiscriminate biodegradation to different organic compounds and their mixture, similar to the character of conventional BOD(5) results. The results of the BOD sensor method are well agreed with those obtained from conventional BOD(5) method for wastewater samples. The proposed rapid BOD sensor method should be promising in practical application of wastewater monitoring.