Investigations On The Tissue-based Chemiluminescence Biosensor

The biosensor is a special chemical sensor. It uses the biologically sensitive components as active units (such as enzyme, antibody, nucleic acid, cell, tissue) and promises highly selective to the analyte. The analyte was sensed through various kinds of physical and chemical signal transducer. The analytical methods based on biosensor are characterized by high selectivity, easy realization for on-line continuous monitoring, fast response and low cost.This paper mainly concerns the application of tissue-based chemiluminescence biosensors in the field of bio-chemical analysis. The paper consists of two parts.The first part is an introduction dealing with the categorization, work principle and the merits of the biosensor and the development of the application of the biosensor in the fields of environment monitoring, food analysis, bio-medicine and the military field.The second part is a research report dealing with the tissue-based chemilumiescence biosensor for etanol, uric acid, lactic acid, and pyruvic acid. 1. Plant tissue-based chemiluminescence biosensor for the determination of ethanolA novel chemiluminescence tissue sensor for the determination of ethanol is presented in this section. Homogenized mushroom tissue was chopped into small pieces and packed into a mini-glass column as reactor. When ethanol passed through the sensor, ethanol was oxidized by dissolving oxygen under catalysis of ethanol oxidase present in the mushroom tissue, then produced hydrogen peroxide could react with luminol in alkaline solution and potassium ferricyanide to produce chemiluminescence. Under the optimum conditions, CL intensity was proportional to concentration of ethanol in the range of 1-2000 μmol/L with a detection limit of 0.2μmol/L(3σ),RSD=4.14% for 50 umol/L ethanol(n=11). The whole analysis including sampling and washing could finish in less than 1.5 min. The sensor could steady over 3 hours by 200 times determination. The method has been applied successfully to the analysis of ethanol in serum, beer and wine samples.2. Pork-liver Tissue-based Chemiluminescence Biosensor for the Determination of Uric acidA new flow injection analysis method based on animal tissue as recognition element for chemiluminescence sensing of uric acid is proposed in this part. The liver of the Large White Pig was used as molecular recognition element. Uric acid is oxidized by oxygen under the catalysis of urate oxidase in the tissue column to produce hydrogen peroxide, which can react with Iuminol in the presence of potassium hexacyanoferrate(III) to generate a CL signal. The experimental results show that the CL emission intensity was linear with uric acid concentration in the range of 0.001 - 3 ug mL"1 and the detection limit (3a) for uric acid was 4x10^ ug mL"1. The biosensor detection system could be used continuously for 4 hours with less than 10% decrease in the response. More than 200 measurements were carried out during this time. A complete analysis, including sampling and washing, could be performed in 1 min with a relative standard deviation of 4.47% for 1 ug mL"1 uric acid. The biosensor detection system has been applied successfully to the analysis of uric acid in plasma and urine samples.3. Pork-kidney Tissue-based Chemiluminescence Biosensor for the Determination of Lactic acidA new method based on natural animal tissue porcine kidney as recognition element for chemiluminescence sensing of lactic acid is proposed in this part. The principle for lactic acid sensing is that lactic acid is oxidized by oxygen under the catalysis of a-hydroxy acid oxidase in the tissue column to produce hydrogen peroxide, which can react with Iuminol in the presence of potassium ferricyanide to generate a CL signal. The experimental results show that the CL emission intensity was linear with lactic acid concentration in the range of 1—1000 umol/L and the detection limit (3cr) for lactic acid was 0.2 umol/L. The biosensor could be used...