CN109207465A - Embedding medium, preparation method and its measuring method for BOD sensor - Google Patents
Embedding medium, preparation method and its measuring method for BOD sensor Download PDFInfo
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- CN109207465A CN109207465A CN201811114303.0A CN201811114303A CN109207465A CN 109207465 A CN109207465 A CN 109207465A CN 201811114303 A CN201811114303 A CN 201811114303A CN 109207465 A CN109207465 A CN 109207465A
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- embedding medium
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- polyvinyl alcohol
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
Abstract
The present invention proposes a kind of embedding medium, preparation method and its measuring method for BOD sensor, belongs to bioengineering field, is able to solve the technical problems such as existing embedding medium compound molding effect is bad, mechanical performance is poor, easy swelling.The embedding medium for being used for BOD sensor is made of polyvinyl alcohol, sodium alginate and xanthan gum.Embedding medium provided by the invention can embed enough strains, in the measurement that can be applied to BOD content.
Description
Technical field
The invention belongs to bioengineering field more particularly to it is a kind of for the embedding medium of BOD sensor, preparation method and
Its measuring method.
Background technique
BOD sensor has many advantages, such as that simple structure, high sensitivity, response time are short, is widely studied and is applied.
For BOD sensor, biosensor is its core, selected in the manufacturing process of biosensor
Bacterial strain, experiment condition of culture and its process for fixation have an impact to the characteristic of sensor and service life.
Wherein, the research of process for fixation becomes big hot topic, uses micro- life under normal conditions either physically or chemically
Object is limited in certain space, so that microorganism is kept activity and has good gas permeability and mechanical strength.Both at home and abroad often
Microorganism immobilization method mainly has: investment, absorption method, sandwich method and cross-linking method, and the application then mainly studies
Investment therein.All having with comparison embedding effect for core of the embedding medium as investment, selection and ratio is great
Meaning.
Compared to previous biofilm by the preparation of biomembrane and being affected for strain, the production of embedding medium method is more simple
Single, cost is lower, while can embed enough microorganisms, reduces error, keeps experimental result more accurate.Therefore it studies
Embedding medium method is imperative as biological sensor sensing element.It is very few to the research of embedding medium both at home and abroad at present, and grind
That studies carefully mainly uses single agents as embedding medium, such as polyvinyl alcohol (PVA), polyacrylamide etc., though have good
Strong bond, resistance to biological degradability and mechanical performance, but and do not have good molding effect.Therefore research mixing examination
Agent as embedding medium, allow its embedded with good molding effect and physical and chemical stability while it is enough
Microorganism, this will be significant the further research of embedding medium method.
Summary of the invention
The present invention proposes a kind of embedding medium, preparation method and its measuring method for BOD sensor, embedding medium tool
There is good molding effect, and there is good elasticity after forming, simultaneously as its stickiness reduces and water-retaining property is good, therefore
Enough strains can be embedded.
In order to achieve the above object, the present invention provides a kind of embedding medium for BOD sensor, the embedding medium is by gathering
Vinyl alcohol, sodium alginate and xanthan gum composition.
Preferably, the mass ratio of the polyvinyl alcohol, sodium alginate and xanthan gum is 1:1:2-1:2:3.
Preferably, the mass ratio of the polyvinyl alcohol, sodium alginate and xanthan gum is 1:1:2.
The present invention also provides a kind of preparation methods of embedding medium as described in the above technical scheme, include the following steps:
The each component in weighed embedding medium in proportion is put at room temperature and is dissolved in the reaction vessel for filling 1L deionized water
In, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands, is cooled to 30-50 DEG C, obtains
To the embedding medium.
The present invention also provides a kind of methods using the measurement BOD content of embedding medium described in above-mentioned technical proposal, including
Following steps:
Will be surrounded by strain embedding medium investment testing liquid in, by measure respectively system initial current value and micro- life
The final current value of system, is calculated the BOD content in testing liquid after material consumption oxygen solution Organic substance in water.
Preferably, initial current value and final current value pass through the electrode probe detection in BOD sensor.
Preferably, the strain that can be embedded includes facultative aerobe and aerobic bacteria.
Compared with prior art, the advantages and positive effects of the present invention are:
Embedding medium provided by the present application perfect can be formed at 30 DEG C or more, and have good elasticity after forming, meanwhile,
Due to the reduction of its stickiness and water-retaining property is good, more good living environment can be provided for strain, therefore can embed enough
Strain.Influenced using the method for the embedding strain of the embedding medium by external condition smaller, and it is good to have the following advantages that (1) has
Good biocompatibility is metabolized without influencing or influencing very little microorganism normal life；(2) physical stability has enough
Mechanical strength；(3) good resistance to biological degradability and chemical stability；(4) enough microorganisms can be embedded；(5) honest and clean
Valence is easy to get.
Specific embodiment
The technical scheme in the embodiments of the invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is common
Technical staff's every other embodiment obtained without making creative work belongs to the model that the present invention protects
It encloses.
The embodiment of the invention provides a kind of embedding mediums for BOD sensor, and the embedding medium is by polyvinyl alcohol, seaweed
Sour sodium and xanthan gum composition.
Though the existing for example resistance to biological degradability of embedding medium chemical property etc. being compounded to form by polyvinyl alcohol and sodium alginate
It is good, but physical property is poor, it is found through experiment that, it is fixed-type relatively slow, also, since polyvinyl alcohol fusing point is higher, dissolution compared with
Difficulty, needs high-temperature process, but to its dissolution after, with the decline of temperature, which is also not easy to condense, and is squeezed after condensation with hand
Pressure does not have good elasticity yet.In order to overcome the drawbacks described above of existing embedding medium, the present embodiment improves the formula of embedding medium,
Improved embedding medium is made of polyvinyl alcohol, sodium alginate and xanthan gum, and the embedding medium formed by the formula is at 30 DEG C or more
It perfect can form, after xanthan gum especially is added in formula, the more good elasticity of embedding medium after molding can be made, meanwhile,
Stickiness reduction and water-retaining property due to the embedding medium is good, more good living environment can be provided for strain, therefore can embed
Enough strains.
In an alternative embodiment, the mass ratio of the polyvinyl alcohol, sodium alginate and xanthan gum is 1:1:2-1:2:4.
The present embodiment is further defined the mass ratio of 3 kinds of constituents in formula, in the range, can make to be prepared
Embedding medium have it is good elasticity, water content is higher, water-retaining property is preferable, by molding solid colloid exposure in air, pass through
Keep original state substantially for 24 hours；Resistance to biological degradability is good, and finished product is placed in sewage and is stirred, and will not be resolved.It is preferably implemented one
In example, the mass ratio of the polyvinyl alcohol, sodium alginate and xanthan gum is 1:1:2.
The embodiment of the invention also provides a kind of preparation methods of embedding medium, include the following steps:
The each component in weighed embedding medium in proportion is put at room temperature and is dissolved in the reaction vessel for filling 1L deionized water
In, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands, is cooled to 30-50 DEG C, obtains
To the embedding medium.
It is in above-described embodiment, embedding agent solution is cooling in 30 DEG C or more after heating for dissolving, it can slowly be frozen at this time
Type can not only make strain survive by the way that required strain package is nuzzled before embedding medium solidifies in this way, but also strain can be with
Carry out normal metabolic activity.Specifically, embedded strain includes facultative aerobe and aerobic bacteria, wherein described facultative
Aerobic bacteria is selected from and falls sour bacterium, Escherichia coli etc.；Aerobic bacteria such as bacillus subtilis and bacillus amyloliquefaciens etc..
The embodiment of the invention also provides a kind of methods using above-mentioned embedding medium measurement BOD content, include the following steps:
Will be surrounded by strain embedding medium investment testing liquid in, by measure respectively system initial current value and micro- life
The final current value of system, is calculated the BOD content in testing liquid after material consumption oxygen solution Organic substance in water.Wherein, initially
Current value and final current value pass through the electrode probe detection in BOD sensor.
In above-described embodiment, embedding medium embedding strain mainly passes through and manually chooses transfer, specifically, by extemporaneous preparation from
The heart is good or strain is transferred in embedding medium by the strain through preparing by liquid-transfering gun, glass bar etc. already, is stirred
To uniform.It is understood that for which kind of instrument specifically used in above-described embodiment depending on the number of strain amount, simultaneously to this
It is not specifically limited.
It is introduced provided by the embodiment of the present invention in detail to become apparent from for the embedding medium of BOD sensor, its preparation
Method and its measuring method, are described below in conjunction with specific embodiment.
Embodiment 1
At room temperature polyvinyl alcohol, sodium alginate and xanthan gum that mass ratio is 1:1:2 are sequentially placed into be dissolved in and fill 1L
In the reaction vessel of ionized water, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands,
It is cooled to 30 DEG C.
Embodiment 2
At room temperature polyvinyl alcohol, sodium alginate and xanthan gum that mass ratio is 1:1:3 are sequentially placed into be dissolved in and fill 1L
In the reaction vessel of ionized water, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands,
It is cooled to 40 DEG C.
Embodiment 3
At room temperature polyvinyl alcohol, sodium alginate and xanthan gum that mass ratio is 1:2:3 are sequentially placed into be dissolved in and fill 1L
In the reaction vessel of ionized water, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands,
It is cooled to 45 DEG C.
Embodiment 4
At room temperature polyvinyl alcohol, sodium alginate and xanthan gum that mass ratio is 1:2:4 are sequentially placed into be dissolved in and fill 1L
In the reaction vessel of ionized water, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands,
It is cooled to 50 DEG C.
Comparative example 1 (biofilm)
Polyvinyl alcohol and sodium alginate that mass ratio is 1:1 are sequentially placed into be dissolved in and fill the anti-of 1L deionized water at room temperature
It answers in container, in being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands, is cooled to 30
℃。
Comparative example 2 (single embedding medium)
The polyvinyl alcohol that mass ratio is 1:1 is sequentially placed into the reaction vessel for being dissolved in and filling 1L deionized water at room temperature,
In being heated to 100 DEG C or more on heating device, while quickly stirring makes it completely dissolved, and stands, is cooled to 30 DEG C.
Performance test
1, it evaluates in terms of viscosity, hardness, saturation degree, elasticity, resistance to biological degradability to embodiment 1-4 and comparative example 1-
The molding effect of 2 embedding mediums provided, the specific test method is as follows, and molding effect is as shown in table 1:
The measuring method of hardness: being squeezed with glass bar, eye-observation
Water-retaining property measuring method: in air by embedding medium exposure, its substantially saturation state is observed by 12-24h
Elasticity measurement method: being squeezed with glass plate, is stood and is seen that it can or can not restore to the original state for a period of time.
Resistance to biological degradability measuring method: finished product is placed in water or sewage, and observation can or can not be by after stirring or concussion
Resolution.
When testing embedding medium degradation resistance, morphologic observation are as follows: three times with brine by molded embedding medium
It is put into the conical flask for filling sterile water later, in the item that temperature is 20 DEG C, revolving speed is 150 turns after being placed on constant temperature oscillator
48h is shaken under part.
1 molding effect explanation of table
It, will not be extensive slightly after modification by 1 effect of table it is found that comparative example 1 is general using the molding effect of biofilm embedding
Shape is restored, and because does not have good water-retaining property, so that finished product is more shrivelled after placing in air；Comparative example 2 uses
Single embedding medium is carried on as before after embedding for flowable slime body, therefore does not have molding possibility.Compared to comparative example 1 and 2,
The preparation-obtained embedding medium of the embodiment of the present application 1-4 has good elasticity；Water content is higher, water-retaining property is preferable, will form
Solid colloid exposure in air, process for 24 hours substantially keep original state；Resistance to biological degradability is good, and finished product is placed in sewage
Stirring, will not be resolved.
2, BOD content in water body is measured respectively using embedding medium prepared by the embodiment of the present invention 1 and comparative example 1
By embodiment 1 (polyvinyl alcohol: ammonium alginate: xanthan gum is the combination of 1:1:2) and (polyvinyl alcohol: sea of comparative example 1
Mosanom is the combination of 1:1) it is dissolved separately in 30ml distilled water, then mixed liquor is respectively placed on heating device and is heated to
It 100 DEG C or more, is completely dissolved to its ingredient, is stood cooling on one side, the strain that will be centrifuged at 35-50 DEG C later
(microorganism) is added separately in two kinds of mixed liquors, then completely cooling to it.
After cooling, 1 gained embedding medium of embodiment is the solid of similar g., jelly-like, and 1 gained embedding medium of comparative example still can be with
Slight flow fails perfect molding.Two kinds of embedding mediums are gently squeezed with glass plate respectively, are observed after two kinds of embedding mediums are squeezed
State, 1 gained embedding medium of embodiment can restore to the original state substantially for a period of time after extrusion, and 1 gained embedding medium of comparative example exists
Irrecoverable original state after extruding.
When 1 gained embedding medium of embodiment is added in prepare liquid, normal read of popping one's head in can be passed through by 3 seconds or so
Electrical signal data, and as time increases, a kind of variation tendency of rule, rule view embedding can be also presented in electric signal therewith
The type of microorganism in agent and the quantity of microorganism and change.
1 gained embedding medium of comparative example can generate each other after being injected into prepare liquid with liquid-transfering gun coheres phenomenon, that is, unites
Phenomenon, can hinder stirring coheres phenomenon as a result, special crosslinking agent need to be configured to inhibit this, after crosslinking agent is added,
Phenomenon is cohered to be alleviated.Start just to begin with electric signal after 10 seconds after test, electric signal herein refers to stablizing
Electric signal, the electric signal less stable of 1 gained embedding medium of comparative example floats up and down more.Using 1 gained embedding medium of comparative example
Obtained result or the law curve therefrom found out also have opposite error, influence last measurement result.
Claims (7)
1. a kind of embedding medium for BOD sensor, which is characterized in that the embedding medium is by polyvinyl alcohol, sodium alginate and Huang
Virgin rubber composition.
2. embedding medium according to claim 1, which is characterized in that the matter of the polyvinyl alcohol, sodium alginate and xanthan gum
Amount is than being 1:1:2-1:2:4.
3. embedding medium according to claim 2, which is characterized in that the matter of the polyvinyl alcohol, sodium alginate and xanthan gum
Amount is than being 1:1:2.
4. the preparation method of embedding medium as described in any one of claims 1-3, which comprises the steps of:
The each component in weighed embedding medium in proportion is put into the reaction vessel for being dissolved in and filling 1L deionized water at room temperature, in
100 DEG C or more are heated on heating device, while quickly stirring makes it completely dissolved, and stands, is cooled to 30-50 DEG C, obtains institute
State embedding medium.
5. utilizing the method such as embedding medium according to claim 1-3 measurement BOD content, which is characterized in that packet
Include following steps:
It will be surrounded by the embedding medium investment testing liquid of strain, consumed by the initial current value and microorganism that measure system respectively
The final current value of system after oxygen solution Organic substance in water, is calculated the BOD content in testing liquid.
6. according to the method described in claim 5, it is characterized in that, initial current value and final current value pass through BOD sensor
In electrode probe detection.
7. according to the method described in claim 5, it is characterized in that, the strain that can be embedded includes facultative aerobe and aerobic bacteria.
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| CN101733074A (en) * | 2008-11-26 | 2010-06-16 | 北京化工大学 | Method for preparing film type biological adsorbing medium |
| CN101497880A (en) * | 2009-03-11 | 2009-08-05 | 清华大学 | Novel method for improving PVA immobilized microorganism |
| CN101639472A (en) * | 2009-08-31 | 2010-02-03 | 清华大学 | Reactor type BOD rapid measuring method and measuring instrument |
| CN102260663A (en) * | 2011-07-13 | 2011-11-30 | 清华大学 | Method for preparing magnetic immobilized particles for BOD (Biochemical Oxygen Demand) biological sensor |
| CN104894097A (en) * | 2015-05-09 | 2015-09-09 | 浙江省农业科学院 | Long-acting water-purifying bacterium capsule and preparation method thereof |
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