Process for preparing membrane used to immobilize microbes to biosense biochemical oxygen demand

A technology of microbial immobilization and biochemical oxygen demand, applied in biochemical equipment and methods, microbial determination/inspection, biological material pretreatment, etc., can solve problems such as poor stability of BOD microbial membranes, and maintain activity and stability. , good adhesion, small diffusion resistance effect

Inactive Publication Date: 2003-02-12
XIAMEN UNIV
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Problems solved by technology

[0004] Some existing patents on BOD biosensors mainly focus on the design of the overall measuring instrument and the improvement of the measuring device. Most of them use the electrochemical method of the three-electrode system to measure dissolved oxygen. Solve the shortcomings of poor stability of BOD microbial film

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Example 1: Add tetramethoxysilane (TMOS) and dimethyldimethoxysilane (DiMe-DMOS) into a vial at a volume ratio of 1:1.2, shake fully and evenly, then stir and add the same solution as TMOS dropwise. The volume of 0.01mol / L HCl aqueous solution, the pH is 3, the mixed solution was stirred and hydrolyzed in a 60°C water bath for 2 hours, the mixed solution was milky white, and it was allowed to stand for 30 minutes, leaving a layer of gel solution for later use; prepare 8% (W / V) PVA-124 aqueous solution, after cooling, mix the bacterial suspension preserved with 0.067mol / L phosphate buffer (pH is 7.2) by 1:1 volume ratio, then add the above-mentioned gel solution of equal volume, Mix well and evenly, and spread microbial films of different thicknesses on the surface of glass substrates, oxidation sensing probes, etc.

[0010] The above-mentioned preparation process mainly has the following characteristics: 1) No alcohol is added to the initial reaction solution, and the ...

Embodiment 2

[0011] Example 2: Using Hansennula anomala as the working strain, the above-mentioned preparation method was used to prepare a microbial immobilized membrane. TMOS:DiMe-DMOS was mixed with HCl aqueous solution (pH 6) at a volume ratio of 1:0.5. The mixed solution was stirred and hydrolyzed in a water bath at 50°C for 2.5 hours. After the mixed solution was milky white, it was allowed to stand at room temperature. The mixed solution was divided into upper and lower layers, and the supernatant was removed. Warm water can be used to prepare the PVA-124 aqueous solution, and the prepared microbial film is directly cured on the surface of the oxygen sensing film, and the oxygen sensing film is used for secondary sensing to detect the change of the dissolved oxygen concentration, so as to investigate the preparation of the method. Response properties of microbial immobilized membranes. The oxygen sensing membrane used uses 4,7-diphenyl-1,10-phenanthroline ruthenium as a fluorescent ...

Embodiment 3

[0012] Example 3: The TMOS:DiMe-DMOS is configured at a volume ratio of 1:1.8, and the resulting microbial film can be placed in a refrigerator at 4° C. for about 2 days and then dried to shape. Glucose-glutamic acid (GGA) solution with a BOD value of 100 mg / L can be added to continue storage at 4°C in the refrigerator. It is found through experiments that the microbial immobilized membrane prepared by the above method has the characteristics of sensitive and rapid response to glucose-glutamic acid (GGA) standard solution, and usually the time to complete the measurement of a GGA standard solution is within 10 minutes. By changing the amount of bacteria embedded in the microbial film, the linear range of the concentration of different BOD values ​​can be obtained. Under the same conditions, the reproducibility of the GGA solution with a BOD value of 100 mg / L was continuously measured 6 times, and the maximum change rate of the fluorescence intensity The relative deviation is 2...

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Abstract

A microbe immobilizing membrane used in the probe of microbe sensor for biochemical oxygen demand is prepared through mixing tetramethoxy silane with dimethyl dimethoxy silane, adding the aqueous solution of HCl, hydrolysis while stirring, taking gel liquid reasly for use, preparing PVA-124 aqueous solution, mixing it with the bacteria suspension and adding the above-mentioned gel liquid. Its advantages are no loss of microbes, high activity and stability of microbes and long low-temp. storge period.

Description

(1) Technical field [0001] The invention relates to a microorganism immobilized film which can be used in the sensor probe of biochemical oxygen demand microorganism sensor. (2) Background technology [0002] Biochemical Oxygen Demand (BOD) is one of the most commonly used and important water quality organic pollution indicators and monitoring parameters in the world. 5 ) standard dilution method is still a commonly used analysis and detection method at home and abroad, but this standard method requires a 5-day analysis cycle, and the operation process is cumbersome, which brings a lot of inconvenience to sewage treatment and environmental monitoring. Today, with the high development of environmental analysis, an instrument called BOD microbial sensor has been gradually developed. The determination of one sample is completed, which greatly shortens the time required for determination. [0003] The sensing probe of the BOD microbial sensor is mainly composed of an oxygen se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/02
Inventor 陈曦钟振明戴媛静王小如
Owner XIAMEN UNIV
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