Process for preparing ion liquid sol-gel compound membrane buried enzyme biologic sensor
A biosensor, ionic liquid technology, applied in biochemical equipment and methods, microbial determination/inspection, biological testing, etc., can solve the problems of affecting the performance of biosensors, destroying enzyme activity, etc. Effect
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Embodiment 1
[0012] Embodiment 1 Preparation of horseradish peroxidase sensor:
[0013] First, evenly drop 7.5 mg / ml ferrocene ethanol solution on the surface of the base electrode according to the ratio of 70-140 microliters: 1 square centimeter, and let it stand for 10 minutes; then add 1 milliliter of 1-butyl, 3-methylimidazole Boron tetrafluoride, 2 ml tetraethoxysilane, 1 ml water and 0.05 ml 0.1mol L -1 hydrochloric acid at room temperature, and magnetically stirred for 3 hours to obtain a homogeneous sol A, which was then placed at room temperature for 1 hour; in addition, 1 mg of horseradish peroxidase was added to 50 microliters of 0.05mol L -1 phosphate buffer solution with a pH of 6.86 to obtain solution B; then mix sol A and solution B uniformly according to the ratio of 1:1, pipette the mixed solution with a micro-syringe and evenly drop-coat it on the surface of the above-mentioned base electrode, with a thickness of about 10 microns, and then placed at 4° C. for 24 hours to...
Embodiment 2
[0014] Example 2 Preparation of glucose oxidase sensor:
[0015] 2 mL of 1-butyl, 3-methylimidazolium boron tetrafluoride, 4 mL of tetraethoxysilane, 2 mL of water and 0.1 mL of 0.1 mol L -1 hydrochloric acid at room temperature, and magnetically stirred for 3 hours to obtain a homogeneous sol A, which was then placed at room temperature for 2 hours; in addition, 3 mg of glucose oxidase was added to 50 microliters of 0.05mol L -1 Mix uniformly in phosphate buffer solution of pH 6 to obtain solution B; then mix sol A and solution B uniformly according to the ratio of 1:2.5, pipette the mixed solution with a micro-syringe and evenly drop-coat it on the surface of the substrate electrode, with a thickness of about 100 microns Then place it at 0° C. for 36 hours to prepare the glucose oxidase sensor, which can be used to detect glucose in the water phase. The balance time of the biosensor is within 6 minutes; the response time is 15 seconds; the stability is 1.5-2 months.
Embodiment 3
[0016] Example 3 Preparation of polyphenol oxidase sensor:
[0017] Mix 1.5 mL of 1-butyl, 3-methylimidazolium boron tetrafluoride, 3 mL of tetraethoxysilane, 1.5 mL of water and 0.075 mL of 0.1 molL -1 Hydrochloric acid mixed at room temperature, magnetically stirred for 3 hours to obtain homogeneous sol A, and then placed at room temperature for 1.5 hours; in addition, 10 mg of enzymes, namely glucose oxidase, horseradish peroxidase, polyphenol oxidase One, add 500 microliters of 0.05mol L -1 phosphate buffer solution with a pH of 7 to obtain solution B; then mix sol A and solution B uniformly according to the ratio of 1:2, pipette the mixed solution with a micro-syringe and evenly drop-coat it on the surface of the substrate electrode, with a thickness of about 500 microns , and then placed at 4°C for 24 hours to prepare a polyphenol oxidase biosensor, which can be used for the determination of phenol, catechol and other substances in the aqueous phase. The balance time o...
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