Optical biosensor chip technology for biochemical oxygen demand monitoring in environmental samples

M. Kashem, Masayasu Suzuki
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Abstract

An optical biosensor chip technology has been studied to perform as an effect free device for rapid biochemical oxygen demand (BOD) monitoring in environmental samples. A biofilm was embedded onto a polyethylene-polypropylene (PE-PP) film shielded oxygen sensing film for developing the biosensor chip. The oxygen sensing film was prepared by coating the ruthenium complex dye solution onto the SO32- group embedded glass slide and the biofilm was prepared by immobilizing baker's yeasts (Saccharomyces cerevisiae) with polyvinyl alcohol-styrylpyridinium (PVA-SbQ) matrix. Sensor sample injection cavity was made by using silicone rubber (SR) sheet upon the embedded biofilm. An inverted microscope was used to measure the biosensor responses as the changing of fluorescence intensity (FI) due to microbial respiration in presence and absence of BOD standard solutions, glucose and glutamic acid solution (GGA) with time profiles. Taking the maximum response at 3 min of the each BOD standard solution, calibration curves were drawn as It=3 (intensity at 3 min time) divided by I0 (intensity at 0 min time) against GGA concentrations. An equal performance was obtained by the biosensor either the GGA in phosphate buffer solution (PBS) or in environmental samples such as river water (RW). A linear relationship was achieved in low GGA concentrations (up to 20 mg/L GGA having BOD 14.6 mg/L, R2=0.99). The PE-PP shielding approach has completely eliminated the effects of heterogeneous components of environmental samples onto the sensor performances.
用于环境样品生化需氧量监测的光学生物传感器芯片技术
研究了一种光学生物传感器芯片技术,用于环境样品中生化需氧量(BOD)的快速监测。将生物膜嵌入聚乙烯-聚丙烯(PE-PP)膜屏蔽的氧传感膜上,用于开发生物传感器芯片。将钌络合染料溶液包覆在SO32-基团包埋的玻片上制备氧感膜,用聚乙烯醇-苯乙烯基吡啶(PVA-SbQ)基质固定化酿酒酵母制备生物膜。在嵌入的生物膜上用硅橡胶(SR)片制作传感器进样腔。利用倒置显微镜测量微生物在存在或不存在BOD标准溶液、葡萄糖和谷氨酸溶液(GGA)的情况下,荧光强度(FI)随时间变化的生物传感器响应。取每种BOD标准溶液在3 min时的最大响应,绘制针对GGA浓度的校准曲线:It=3 (3 min时的强度)/ I0 (0 min时的强度)。该生物传感器在磷酸盐缓冲溶液(PBS)或环境样品(如河水(RW))中的GGA均获得了相同的性能。在低GGA浓度(高达20 mg/L GGA的BOD为14.6 mg/L, R2=0.99)下,二者呈线性关系。PE-PP屏蔽方法完全消除了环境样品非均质组分对传感器性能的影响。
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