An optical fiber bienzyme sensor based on the luminol chemiluminescent reaction was developed and demonstrated to be sensitive to glucose. Glucose oxidase(GOD) and horseradish peroxidase(HRP) were co-immobilized by mi...An optical fiber bienzyme sensor based on the luminol chemiluminescent reaction was developed and demonstrated to be sensitive to glucose. Glucose oxidase(GOD) and horseradish peroxidase(HRP) were co-immobilized by microencapsulation in a sol-gel film derived from tetraethyl orthosilicate(TEOS). The calibration plots for glucose were established by the optical fiber glucose sensor fabricated by attaching the bienzyme silica gel onto the glass window of the fiber bundle. The linear range was 0 2-2 mmol/L and the detection limit was approximately 0 12 mmol/L. The relative standard deviation was 5.3% ( n =6). The proposed biosensor was applied to glucose assay in ofloxacin injection successfully.展开更多
Bienzymatic biosensor for the determination of glucose by flow injection chemiluminescence(CL) de-tection was proposed.Hybrids of gold nanoparticles(GNPs) and chitosan were chosen as the immobi-lization matrix of gluc...Bienzymatic biosensor for the determination of glucose by flow injection chemiluminescence(CL) de-tection was proposed.Hybrids of gold nanoparticles(GNPs) and chitosan were chosen as the immobi-lization matrix of glucose oxidase(GOD) and horseradish peroxidase(HRP) to fabricate the biosensors with silane-pretreated glass microbeads.After the enzyme catalyzing oxidation of glucose in GOD biosensor,the produced H2O2 flowed into HRP biosensor to react with luminol.The doped GNPs in chitosan were found to enhance the classical CL reaction of luminol-H2O2-HRP.The CL enhancement was investigated in detail by CL and UV-visible spectrum.Under the optimized experimental conditions,glucose could be determined in a linear range from 0.01 to 6.0 mmol/L with a detection limit of 5.0 μmol/L at 3σ.The accuracy of the proposed method was examined by detecting the glucose level in four clinical serum samples from hospital.The proposed method provides a new alternative to deter-mine glucose.展开更多
采用分步法将仿生硅化与脂质体技术相结合,模拟细胞微结构,以卵磷脂为原料,制备脂质体微囊,以胺类为诱导剂,在其表面硅化形成氧化硅壳层,实现对葡萄糖氧化酶(GOx)与辣根过氧化酶(HRP)双酶固定。研究了超声时间、诱导剂PDADMA用量、硅前...采用分步法将仿生硅化与脂质体技术相结合,模拟细胞微结构,以卵磷脂为原料,制备脂质体微囊,以胺类为诱导剂,在其表面硅化形成氧化硅壳层,实现对葡萄糖氧化酶(GOx)与辣根过氧化酶(HRP)双酶固定。研究了超声时间、诱导剂PDADMA用量、硅前驱体水解液(TMOS)用量、Triton X-100浓度等因素对固定化酶活性影响。结果表明,在200μL PDADMA、0.7 m L TMOS、超声50 min制得的固定化酶活性最高,能迅速硅化形成直径200 nm左右的球形微囊。在此优化条件下制备的固定化酶经重复使用7次,依然达到初始催化酶活的61.46%,经一个月冷藏后酶活仍能达到最初的74.1%。可见,经过固定化,双酶系统的稳定性得到了显著提高。展开更多
文摘An optical fiber bienzyme sensor based on the luminol chemiluminescent reaction was developed and demonstrated to be sensitive to glucose. Glucose oxidase(GOD) and horseradish peroxidase(HRP) were co-immobilized by microencapsulation in a sol-gel film derived from tetraethyl orthosilicate(TEOS). The calibration plots for glucose were established by the optical fiber glucose sensor fabricated by attaching the bienzyme silica gel onto the glass window of the fiber bundle. The linear range was 0 2-2 mmol/L and the detection limit was approximately 0 12 mmol/L. The relative standard deviation was 5.3% ( n =6). The proposed biosensor was applied to glucose assay in ofloxacin injection successfully.
基金Supported by the Natural Science Foundation of Shandong Province (Grant No. Q2007B03)the Doctoral Fund of Qingdao University of Science and Technology (Grant No. 0022141)the National Natural Science Foundation of China (Grant No. 20775038)
文摘Bienzymatic biosensor for the determination of glucose by flow injection chemiluminescence(CL) de-tection was proposed.Hybrids of gold nanoparticles(GNPs) and chitosan were chosen as the immobi-lization matrix of glucose oxidase(GOD) and horseradish peroxidase(HRP) to fabricate the biosensors with silane-pretreated glass microbeads.After the enzyme catalyzing oxidation of glucose in GOD biosensor,the produced H2O2 flowed into HRP biosensor to react with luminol.The doped GNPs in chitosan were found to enhance the classical CL reaction of luminol-H2O2-HRP.The CL enhancement was investigated in detail by CL and UV-visible spectrum.Under the optimized experimental conditions,glucose could be determined in a linear range from 0.01 to 6.0 mmol/L with a detection limit of 5.0 μmol/L at 3σ.The accuracy of the proposed method was examined by detecting the glucose level in four clinical serum samples from hospital.The proposed method provides a new alternative to deter-mine glucose.
文摘采用分步法将仿生硅化与脂质体技术相结合,模拟细胞微结构,以卵磷脂为原料,制备脂质体微囊,以胺类为诱导剂,在其表面硅化形成氧化硅壳层,实现对葡萄糖氧化酶(GOx)与辣根过氧化酶(HRP)双酶固定。研究了超声时间、诱导剂PDADMA用量、硅前驱体水解液(TMOS)用量、Triton X-100浓度等因素对固定化酶活性影响。结果表明,在200μL PDADMA、0.7 m L TMOS、超声50 min制得的固定化酶活性最高,能迅速硅化形成直径200 nm左右的球形微囊。在此优化条件下制备的固定化酶经重复使用7次,依然达到初始催化酶活的61.46%,经一个月冷藏后酶活仍能达到最初的74.1%。可见,经过固定化,双酶系统的稳定性得到了显著提高。
