CN106841187B - A kind of quick liquid oxygen indicator and its preparation method and application - Google Patents
A kind of quick liquid oxygen indicator and its preparation method and application Download PDFInfo
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- CN106841187B CN106841187B CN201710096169.5A CN201710096169A CN106841187B CN 106841187 B CN106841187 B CN 106841187B CN 201710096169 A CN201710096169 A CN 201710096169A CN 106841187 B CN106841187 B CN 106841187B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
- G01N21/783—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour for analysing gases
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N2021/775—Indicator and selective membrane
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Abstract
The present invention relates to a kind of quick liquid oxygen indicator and its preparation method and application, the composition of the oxygen indicator includes: reactive black 5, Friedlander's bacillus strain KL1, fluid nutrient medium and distilled water.Preparation method includes: that the Friedlander's bacillus strain KL1 of 24~48h of constant temperature stationary culture is linked into the fluid nutrient medium by sterilizing containing reactive black 5, it mixes, in 30~40 DEG C of constant temperature 24~48h of stationary culture under air-proof condition, liquid oxygen indicator is obtained.Under test gas is acquired with gas collecting syringe, is then injected into liquid oxygen indicator, is turned upside down concussion 3~10 seconds, can develop the color, colorimetric, quantitative oxygen concentration are then carried out with standard oxygen concentration colorimetric card.The present invention can be used for oxygen detection in seal food preservation environment, and detection oxygen whether there is and determine oxygen concentration in medical oxygen-free environment, anaerobic bacteria culture environment and other oxygen free operation environment.
Description
Technical field
The invention belongs to oxygen content measure field, in particular to a kind of quick liquid oxygen indicator and preparation method thereof and
Using.
Background technique
Oxygen is all essential for many chemistry and biology, and the reason for causing most foods rotten, because
This needs removes in packaged food.It can be by adding nitrogen, carbon dioxide etc. into food packaging for the removal of oxygen
Gas, or removed by oxygen scavenger.However, increase over time, due to encapsulating material leakproofness not
Persistently, or during transport, storage packaging is damaged, and as a result food is caused to be gone bad by oxygen gas pollution.In addition, oxygen is also
It is important, in extensive industry, including but not limited to food packaging, steel production, sewage treatment, PETROLEUM PROCESSING and glass,
The production process of paper, beer.Therefore, the measurement of oxygen is an important feature in these areas.
The method for the detection oxygen having built up at present, including clark's electrode and gas chromatography.However, these detection systems
It unites usually expensive, either original capital expenditure or the cost of operation.And visual oxygen indicator is honest and clean
Valence, consumer can be allow to detect the presence of oxygen by naked eyes.It is some successfully to realize quotient than colour pattern oxygen indicator
Industry, such as the Ageless Eye product of Mitsubishi Gas Chemical company production.In recent years, there are many more pass through UV activation
Be reported than colour pattern oxygen indicator.Their most of principles all be using reducing substances occur in conjunction with oxygen it is reversible
Redox answer, and along with the variation of color, indicate the presence or absence of oxygen in this way.But there are face
The disadvantages of color change is unobvious, does not adapt to the biggish environment of humidity, unstable under light action, easily loses effectiveness, furthermore greatly
Partial oxidation indicator cannot be recycled, and the environmental issues such as result in waste of resources.
Andrew Mills and Katherine Lawrie develop novel oxygen indicator Pt-TiO2/MB/
Glycerol/SPS controls the reaction time with platinum, by improving the load capacity of platinum, can delay recovery time, when bearing for platinum
When carrying capacity is 1.52%, recovery time is up to 12 hours.This oxygen indicator can indicate that food is opened in the case where refrigeration
Sealed how long.Experiment shows that such indicator may be reused, and in use to the 5th, declines without manifest function.But
It is that the material cost that this technology uses is expensive, market price is high.In addition, the current domestic product in terms of oxygen indicator is very
It is few, and have the shortcomings that not reproducible use, expensive, complicated for operation, detection is insensitive, without standard measure mostly.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of quick liquid oxygen indicator and preparation method thereof and answer
With the liquid oxygen indicator can quickly indicate that oxygen exists and can quantify oxygen concentration；Preparation method is simple, by liquid when use
Color reaction can occur for injection air to be measured in indicator, concussion several seconds, when anaerobic show it is light yellow, it is dense according to oxygen when aerobic
Spend the deep mixed blue-green of different displays；It can be used for oxygen detection in seal food preservation environment, medical oxygen-free environment, anaerobism
Oxygen is detected in bacterium culture environment and other oxygen free operation environment whether there is and determine oxygen concentration.
A kind of quick liquid oxygen indicator of the invention, the composition of the oxygen indicator includes: reactive black 5
(Reactive Black 5, Sigma-Aldrich company), Friedlander's bacillus strain KL1, fluid nutrient medium and distilled water.
The ingredient of the fluid nutrient medium includes: potassium dihydrogen phosphate 2.66g/L, ammonium chloride 0.2g/L, anhydrous sodium sulfate
0.5g/L, yeast extract 3g/L.
The method that the liquid oxygen indicator is lived again are as follows: the oxygen indicator after dioxygen oxidation develops the color is again quiet
Set, in 30~40 DEG C constant temperature incubation 8~10 hours, the liquid oxygen indicator lived again.
The liquid oxygen indicator, which is lived again, is recycled number more than or equal to 18 times.
The reaction of the liquid oxygen indicator and storage container are 4mL chromatography column feed materials bottle (CNW Technologies
GmbH)。
The preparation method of the quick liquid oxygen indicator, comprising:
The Friedlander's bacillus strain KL1 of 24~48h of constant temperature stationary culture is linked into and passes through sterilizing containing reactive black 5
Fluid nutrient medium in, mix, (every 4mL is dispensed into chromatography column feed materials bottle, covers threaded cap) is in 30~40 DEG C under air-proof condition
Constant temperature 24~48h of stationary culture, (reactive black 5 color fade being waited until, when thoroughly becoming light yellow) obtain liquid oxygen indicator.
Friedlander's bacillus strain KL1 in the present invention, depositary institution's title: Chinese microorganism strain preservation conservator
Meeting common micro-organisms center (CGMCC), preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microorganism
Research institute, deposit number CGMCC No.13552, the deposit date is on 01 09th, 2017, it is proposed that classification naming are as follows: pneumonia gram
The primary Salmonella of thunder (Klebsiella pneumoniae).
The temperature of the Friedlander's bacillus strain KL1 constant temperature stationary culture is 30~40 DEG C, and pH is 4.0~9.0.
The inoculum concentration of the Friedlander's bacillus strain KL1 is 1%~10% (V/V).
The concentration of reactive black 5 is 100~1000mg/L in the fluid nutrient medium by sterilizing containing reactive black 5.
The present invention also provides a kind of application of quick liquid oxygen indicator, the methods of the application are as follows:
Under test gas is acquired with gas collecting syringe, is then all injected into liquid oxygen indicator, turns upside down
Concussion 3~10 seconds, can develop the color, and colorimetric, quantitative oxygen concentration are then carried out with standard oxygen concentration colorimetric card；It is particularly applicable to eat
Product pack interior oxygen content detection or other need to monitor the instruction field of ambient oxygen content.
Oxygen concentration of the present invention refers both to the percentage by volume that oxygen accounts under test gas.
The liquid oxygen indicator anaerobic reduction-state becomes oxidation state in bluish-green in light yellow after dioxygen oxidation reacts
Color.
The gas collecting syringe be 100 μ L microsyringes (Shanghai Gao Ge Biotechnology Co., Ltd), acquisition to
It surveys gas and is injected into chromatography column feed materials bottle.
The preparation method of the standard oxygen concentration colorimetric card includes: that an access liquid oxygen indicator is arranged in a row, the
One injection of oxygen, the last one injection excessive oxygen do not generate column oxysome product during which according to oxygen concentration gradient injection of oxygen
The paint color card that concentration is 0%~100%.
The solution of the present invention for being used to prepare indicator substance are as follows: potassium dihydrogen phosphate 2.66g/L, ammonium chloride 0.2g/L,
Anhydrous sodium sulfate 0.5g/L, yeast extract 3g/L, 0.1~1g/L of reactive black 5 dye.
The present invention provides a standard oxygen concentration colorimetric card, using fast oxygen gas indicator provided by the invention and by being somebody's turn to do
Colorimetric card can oxygen concentration in quantitative detection low-oxygen environment.
Fast oxygen gas indicator is lived again:
Fast oxygen gas indicator becomes blue-green after, is placed on 30~40 DEG C of environment, it is small that constant temperature stands 8~10
Shi Hou, blue-green are thoroughly taken off, and present again light yellow, and oxygen indicator is lived again completion at this time.It is recyclable to make using the method
With same branch fast oxygen gas indicator 18 times or more.
Fast oxygen gas indicator provided by the invention is the biochemistry by Friedlander's bacillus strain KL1 to reactive black 5 dye
Effect preparation, and the reducing power provided by Friedlander's bacillus strain KL1 realizes that the multiple circulation of oxygen indicator makes
Use function.
The key of fast oxygen gas indicator provided by the invention is the preparation of indicator substance, and indicator substance is through analytical chemistry reality
Authenticate and be set to TAHNDS, reduction-state be it is light yellow, and become blue-green oxidation state after oxygen catalytic oxidation, as shown in Figure 1.
Beneficial effect
The present invention can quickly indicate that oxygen exists and can quantify oxygen concentration；Preparation method is simple, and when use indicates liquid
Color reaction can occur for injection air to be measured in agent, concussion several seconds, when anaerobic show it is light yellow, when aerobic not according to oxygen concentration
With the deep mixed blue-green of display；It can be used for oxygen detection in seal food preservation environment, medical oxygen-free environment, anaerobic bacteria training
Supporting detection oxygen in environment and other oxygen free operation environment whether there is and determine oxygen concentration；The present invention is with color change indicating oxygen
Gas has and is swift in response, and color change is obvious, easily observes；It can be recycled 18 times or more；It is environmentally friendly, it is pollution-free.
Detailed description of the invention
Fig. 1 is indicator substance TAHNDS color change molecule mechanism in the present invention；
Fig. 2 is that fast oxygen gas indicator uses demonstration graph in embodiment 2；Wherein, 2b is the partial enlarged view of 2a；
Fig. 3 is 0% to 100% standard oxygen concentration oxygen indicator in embodiment 3；
Fig. 4 is 0% to 100% standard oxygen concentration colorimetric card in embodiment 3.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, those skilled in the art
Member can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Range.
Embodiment 1
The preparation of fast oxygen gas indicator:
The Friedlander's bacillus strain KL1 bacterium solution of constant temperature (37 DEG C) stationary culture for 24 hours is taken, with 10% inoculum concentration (V/V)
It accesses in the fluid nutrient medium of the reactive black 5 containing 500mg/L, is sub-packed in the 4mL color of sterilizing after mixing bacterium solution by every 4mL solution
Compose in sample injection bottle, be subsequently placed in 37 DEG C of constant temperature incubation 48h, until the color of reactive black 5 is taken off completely, solution become it is light yellow i.e.
It is completed for preparation.
Aforesaid liquid medium component: potassium dihydrogen phosphate 2.66g/L, ammonium chloride 0.2g/L, sodium sulphate 0.5g/L, yeast mention
Take object 3g/L.
Aforesaid liquid culture medium and chromatography column feed materials bottle pass through 121 DEG C, and 100kPa sterilizing uses after twenty minutes.
Embodiment 2
The application method of fast oxygen gas indicator:
100 μ L of under test gas is acquired with gas collecting syringe, is then injected into fast oxygen gas indicator, injection is extracted
After device, chromatography column feed materials bottle is turned upside down concussion 3~10 seconds, observes color change.Light yellow, oxygen is presented in anaerobic state color
Blue-green is presented after oxidation, that is, oxygen presence or absence can be observed.By the oxygen indicator of discoloration and standard oxygen concentration colorimetric card
Comparison can further determine that oxygen concentration, as shown in figures 2 a and 2b.
Embodiment 3
The preparation of standard oxygen concentration colorimetric card:
The liquid oxygen indicator of fetching is arranged in a row, first not injection of oxygen, the last one injection excessive oxygen,
Intermediate oxygen indicator injects quantitative gradient concentration oxygen according to oxygen concentration incremental manner, correspondingly, oxygen indicator
Blue-green from the superficial to the deep will be presented from low to high according to the oxygen content of injection in color, i.e., corresponding to generate a column oxygen concentration
By 0~100% gradient.By this row's oxygen indicator shooting at photo, as shown in Figure 3.After Fig. 3 is cut, that is, it is made one
Standard oxygen concentration colorimetric card of the oxygen concentration from 0% to 100%, as shown in Figure 4.
Embodiment 4
Fast oxygen gas indicator is lived again:
Fast oxygen gas indicator becomes blue-green after, is placed on 37 DEG C of environment, bluish-green after constant temperature stands 10h
Color thoroughly takes off, and presents again light yellow, and oxygen indicator is lived again completion at this time.Using the method, same branch can be recycled
Fast oxygen gas indicator 18 times.
Claims (9)
1. a kind of quick liquid oxygen indicator, which is characterized in that the composition of the oxygen indicator includes: reactive black 5, pneumonia
Citric acid bacterial strain KL1, fluid nutrient medium and distilled water；Wherein, the ingredient of the fluid nutrient medium includes: potassium dihydrogen phosphate
2.66g/L, ammonium chloride 0.2g/L, anhydrous sodium sulfate 0.5g/L, yeast extract 3g/L.
2. a kind of quick liquid oxygen indicator according to claim 1, which is characterized in that the liquid oxygen indicator
The method lived again are as follows: the oxygen indicator after dioxygen oxidation develops the color is stood again, it is small in 30~40 DEG C of constant temperature incubations 8~10
When, the liquid oxygen indicator lived again.
3. a kind of quick liquid oxygen indicator according to claim 2, which is characterized in that the liquid oxygen indicator
It lives again and number is recycled more than or equal to 18 times.
4. a kind of preparation method of quick liquid oxygen indicator as described in claim 1, comprising:
The Friedlander's bacillus strain KL1 of 24~48h of constant temperature stationary culture is linked into the liquid by sterilizing containing reactive black 5
It in body culture medium, mixes, in 30~40 DEG C of constant temperature 24~48h of stationary culture under air-proof condition, obtains liquid oxygen indicator.
5. a kind of preparation method of quick liquid oxygen indicator according to claim 4, which is characterized in that the pneumonia
The temperature of citric acid bacterial strain KL1 constant temperature stationary culture is 30~40 DEG C, and pH is 4.0~9.0.
6. a kind of preparation method of quick liquid oxygen indicator according to claim 4, which is characterized in that the pneumonia
The inoculum concentration of citric acid bacterial strain KL1 is 1%~10%.
7. a kind of preparation method of quick liquid oxygen indicator according to claim 4, which is characterized in that described to contain
The concentration of reactive black 5 is 100~1000mg/L in the fluid nutrient medium by sterilizing of reactive black 5.
8. a kind of application of quick liquid oxygen indicator as described in claim 1, which is characterized in that the method for the application
Are as follows: under test gas is acquired with gas collecting syringe, is then all injected into liquid oxygen indicator, concussion 3 of turning upside down
It~10 seconds, can develop the color, colorimetric, quantitative oxygen concentration are then carried out with standard oxygen concentration colorimetric card.
9. a kind of application of quick liquid oxygen indicator according to claim 8, which is characterized in that the standard oxygen is dense
The preparation method of degree colorimetric card includes: that fetch a liquid oxygen indicator is arranged in a row, first not injection of oxygen, last
A injection excessive oxygen generates the face that a column oxygen volumetric concentration is 0%~100% during which according to oxygen concentration gradient injection of oxygen
Colour atla.
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| CN109856125B (en) * | 2018-12-15 | 2021-02-19 | 华南理工大学 | Oxygen-indicating type multi-tungstic acid cluster compound deoxidizer and preparation method and application thereof |
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Effective date of registration: 20200305 Address after: 315500 Dongfeng Road, Fenghua District, Ningbo, Zhejiang Province, No. 80 Patentee after: Ningbo reputation Environment Technology Co., Ltd. Address before: 201620 No. 2999 Renmin Road North, Songjiang new town, Shanghai, Songjiang District Patentee before: DONGHUA University |