CN105866275A - Method and device for quantitative analysis on nitrogen-oxygen absorption capacity of nitrogen-fixing bacteria in sealed container - Google Patents
Method and device for quantitative analysis on nitrogen-oxygen absorption capacity of nitrogen-fixing bacteria in sealed container Download PDFInfo
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- CN105866275A CN105866275A CN201610204200.8A CN201610204200A CN105866275A CN 105866275 A CN105866275 A CN 105866275A CN 201610204200 A CN201610204200 A CN 201610204200A CN 105866275 A CN105866275 A CN 105866275A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/025—Gas chromatography
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Abstract
The invention discloses a method and a device for quantitative analysis on the nitrogen-oxygen absorption capacity of nitrogen-fixing bacteria in a sealed container. The method comprises the following steps: putting a nitrogen-fixing bacterium group solution of 1/10 to 1/2 in volume into a rigid container in an atmospheric pressure environment; sealing the opening of the rigid container with a rubber plug, and performing shake culture for 1 hour to 10 days; introducing an inert gas which does not react with nitrogen or oxygen into the rigid container through a gas supplementing bag so as to recover one atmospheric pressure inside the rigid container; and sampling from the rigid container, performing testing analysis by using a gaschromatograph, and comparing the testing analysis result with a testing result of common air. The method and the device disclosed by the invention are simple in principle, a new mixed gas in which the inert gas is introduced is tested and analyzed by using the gaschromatograph, and the result is compared with that of common air tested and analyzed by using the gaschromatograph, so that the amount of oxygen and nitrogen absorbed by the nitrogen-fixing bacteria can be simply, visibly and accurately measured.
Description
Technical field
The invention belongs to gas composition analysis technical field, fixed nitrogen in more specifically a kind of quantitative analysis hermetic container
Bacterium inhales the method and device of nitrogen oxygen amount.
Background technology
During cultivating nitrogen-fixing bacteria, the oxygen consumption of antibacterial and nitrogen consuming amount are the important evidence of process control, are final
The important indicator of product quality.There is presently no quickly and accurately assay method, topmost problem is, if without interruption
Fresh air, the oxygen of consumption and nitrogen are difficult to quantitative determination；If employing closed system, treat that antibacterial absorbs oxygen and nitrogen
After, it being negative pressure in system, be equivalent to gas and be diluted, in vial, the measurement result of residual gas must be with the survey of normal air
Determine result to make with reference to (standard), that is the analysis result of sample in bottle is analyzed gained knot with normal air sample same method
Fruit compares, thus the remaining nitrogen of gas and the content of oxygen in calculating bottle outlet；But, usually can only be with solid during gas sample
The method determining volume, little easily fixation pressure, in the case of normal air standard specimen is different from gas sample pressure in bottle, take
Drawing gas absolute magnitude (Molecules) that same volume obtains during sample would not equally, and analysis result is the most comparable
Property, it is impossible to it is used for making quantitative analysis.
The patent " method of a kind of biological nitrogen fixation detection " of Authorization Notice No. CN1074547C, discloses " by test sample
It is placed in the culture bottle of suitable volume, adds corresponding culture medium in right amount, carry out with rubber capsule or other closed materials
Close, after evacuation, inject 20% oxygen and 80% nitrogen, form standard pressure gas, then when being suitable for cultivating corresponding at temperature
Between, detect the change of nitrogen percentage amounts with standard sampling container sampling, thus obtain the decrement of nitrogen ", this way will be based on nitrogen
Gas, oxygen absorption ratio are different, and when nitrogen, oxygen absorption ratio are identical with original scale, this method will lose its usefulness.
Summary of the invention
For solving the problems referred to above, no matter which kind of nitrogen-fixing bacteria compare with to overcome the deficiencies in the prior art, the present invention to intend providing one
Example absorbs oxygen and nitrogen, can simply, nitrogen-fixing bacteria inhale the method and device of nitrogen oxygen amount in accurate quantitative analysis hermetic container.
For achieving the above object, in the quantitative analysis hermetic container that the present invention provides, nitrogen-fixing bacteria inhale the method for nitrogen oxygen amount such as
Under:
First: the nitrogen fixation flora solution of 1/10 to 1/2 rigid container volume is put into rigid container under atmospheric pressure environment；
Second: rigid container opening part rubber closure seals, and concussion is cultivated 1 hour to 240 hours；
3rd: in rigid container, be filled with the noble gas that can not react with nitrogen and oxygen by supplementary airbag so that
Air pressure in rigid container returns to an atmospheric pressure；
4th: sample from rigid container, use gas chromatograph to carry out test analysis, and by Measurement results and common sky
The measurement result comparison of gas.
When giving containing bacterium rigid container make-up gas, opening valve, the negative pressure in rigid container promotes to supplement in airbag
Noble gas flows into rigid container by trachea and syringe needle, and airbag deformation retract is until rigid container internal gas pressure and atmospheric pressure one
Cause.
For realizing said method, present invention also offers nitrogen-fixing bacteria in a kind of quantitative analysis hermetic container and inhale the dress of nitrogen oxygen amount
Putting, including rigid container and supplementary airbag, described rigid container can be used for containing nitrogen fixation flora solution, and sealing can use rubber closure
Seal；Described supplementary airbag is airtight and air pressure inside can be made to keep the case equal with external pressure, preferably
Lightweight is nonelastic PVC film balloon；The opening of described supplementary airbag connects one and carries valvular trachea, and the trachea other end is provided with
Syringe needle, syringe needle is connected with rigid container by piercing through rigid container sealing rubber closure.
The invention has the beneficial effects as follows: assembly of the invention structures and methods principle is simple, by being filled with noble gas, extensive
The gas pressure in rigid container after multiple nitrogen-fixing bacteria effect, draws the gas that same volume obtains absolute when solving sampling
Measuring different, analysis result does not has comparability, it is impossible to is used for obtaining problem as quantitative analysis, is passed through with gas chromatograph test analysis
Mixed gas new after noble gas, and result and the result of the normal air of gas chromatograph test analysis carried out right
Ratio, simple, intuitive is accurately measured nitrogen-fixing bacteria and is absorbed oxygen and the amount of nitrogen.
Accompanying drawing illustrates:
Accompanying drawing 1 is the structural representation of the present invention；In accompanying drawing:
1, rigid container, 2, syringe needle, 3, valve, 4, trachea, 5, supplement airbag
Detailed description of the invention:
Clearer for the purpose making the present invention implement, technical scheme and advantage, below in conjunction with the accompanying drawing 1 of the present invention, to this
Invention is further described in more detail.
No matter nitrogen-fixing bacteria absorb oxygen and nitrogen with which kind of ratio to the invention provides one, can simple, accurate must determine
In component analysis hermetic container, nitrogen-fixing bacteria inhale the method and device of nitrogen oxygen amount, in FIG, nitrogen-fixing bacteria in quantitative analysis hermetic container
Inhaling the device of nitrogen oxygen amount, including rigid container 1 and supplementary airbag 5, described rigid container 1 is used for containing nitrogen fixation flora solution, sealing
Can seal with rubber closure；Described supplementary airbag 5 is airtight and air pressure inside can be made to keep the appearance equal with external pressure
Device device, uses lightweight nonelastic PVC film balloon；The opening supplementing airbag 5 connects a trachea 4 with valve 3, trachea 4
One end is provided with syringe needle 2, and syringe needle 2 seals rubber closure be connected with rigid container 1 by piercing through rigid container 1.
Embodiment 1
In a vial, add the nitrogen fixation flora solution of 1/3 bottle of inner volume, seal with rubber closure, then concussion 24 hours,
Owing to antibacterial absorbs the oxygen in bottle and nitrogen, the air pressure inside vial is down to subatmospheric；Now open valve 3,
Negative pressure in vial promotes the noble gas supplementing in airbag to flow into rigid container 1 by trachea 4 and syringe needle 2, supplements airbag 5
Deformation retract, final airbag and vial internal pressure reach balance, and are equal to outside atmosphere due to the deformation of supplementary airbag
Pressure.At this moment the volume obtained with sample introduction needle sampling in vial and the volume equivalent directly sampled from normal air, it is possible to
The simply and accurately content of each gas componant in quantitative analysis bottle.
When taking sample, pierce through vial rubber closure by the sample introduction needle of gas chromatogram and quantitatively draw the gas in sack, then
The gas being drawn onto is injected directly into gas chromatograph be analyzed；Then sample introduction needle is repeatedly cleaned by normal air, then general
The gas of draws equal amounts in blowing air, is expelled in gas chromatogram be analyzed by same method；Because oxygen in normal air
The content of gas is 21%, nitrogen 78%, and the oxygen peak area obtained through chromatography is if it is Ao, and nitrogen peak area is An, and
If the oxygen of sample and nitrogen peak area are Axo and Ano respectively, then:
In sample, the consumption of oxygen is exactly: %O=21* (1 Aox/Ao)
In sample, the consumption of nitrogen is exactly: %N=78* (1 Anx/An)
The present invention is to use gas chromatogram in gas analysis, it is also possible to any other detectors that gas can produce signal,
Include but not limited to that thermal capacity detector, conduction of heat, ionic flame, catalysis burning, ionic discharge, electron capture etc. detect
Device.
Embodiment 2
According to embodiment 1, save the process that wherein gas chromatogram separates, utilize the mass spectrum part in gas chromatograph-mass spectrometer directly to enter
Sample does Flow Injection Analysis；The way of this pattern is the chromatographic column in short circuit gas chromatogram, allows injection port be directly connected to mass spectrum
Capillary inlet, sample injects injection port and is later brought into mass spectrograph by carrier gas immediately and measure.Because mass spectrographic resolution is
More than 0.1%, although nitrogen and oxygen do not have any separation, it is ionized simultaneously and produces mass signal, by the ion extraction,
Nitrogen ion 28, oxygen ion 32 can differentiate the corresponding signal of extraction, and the signal extracted is integrated gained integrated value,
Can be used to accurate quantitative analysis.Owing to eliminating separation process, the method can complete to analyze test, for greatly the soonest in 20 seconds
Amount sample and instant on-line determination have particularly important practical value.
Claims (3)
1. the method that in quantitative analysis hermetic container, nitrogen-fixing bacteria inhale nitrogen oxygen amount, it is characterised in that including:
First: the nitrogen fixation flora solution of 1/10 to 1/2 rigid container volume is put into rigid container under atmospheric pressure environment；
Second: rigid container opening part rubber closure seals, and concussion is cultivated 1 hour to 240 hours；
3rd: in rigid container, be filled with the noble gas that can not react with nitrogen and oxygen by supplementary airbag so that
Air pressure in rigid container returns to an atmospheric pressure；
4th: sample from rigid container, use gas chromatograph to carry out test analysis, and Measurement results is empty with common
The measurement result comparison of gas.
2. the method that in quantitative analysis hermetic container as claimed in claim 1, nitrogen-fixing bacteria inhale nitrogen oxygen amount, it is characterised in that described
Supplementary airbag is before being filled with noble gas, first by supplementary airbag evacuation, is then charged with to occur anti-with nitrogen and oxygen
The noble gas answered, meanwhile, remains that supplementary airbag internal gas pressure is equal with external pressure.
3. in a quantitative analysis hermetic container as claimed in claim 1, nitrogen-fixing bacteria inhale the device of nitrogen oxygen amount, it is characterised in that
Including rigid container (1) and supplementary airbag (5), described rigid container (1) can be used for containing nitrogen fixation flora solution, and sealing can be used
Rubber closure seals；Described supplementary airbag (5) is airtight and air pressure inside can be made to keep the container equal with external pressure
Device；The opening of described supplementary airbag (5) connects a trachea with valve (3) (4), and trachea (4) other end is provided with pin
Head (2), syringe needle (2) is connected with rigid container (1) by piercing through rigid container (1) sealing rubber closure.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610204200.8A CN105866275A (en) | 2016-04-05 | 2016-04-05 | Method and device for quantitative analysis on nitrogen-oxygen absorption capacity of nitrogen-fixing bacteria in sealed container |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110703171A (en) * | 2019-11-11 | 2020-01-17 | 中国科学院武汉物理与数学研究所 | Multi-core magnetic resonance system gas imaging quality testing die body |
| CN113498776A (en) * | 2021-06-17 | 2021-10-15 | 深圳逗点生物技术有限公司 | Biological transport system, biological transport method, and blood cell preservation system |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110703171A (en) * | 2019-11-11 | 2020-01-17 | 中国科学院武汉物理与数学研究所 | Multi-core magnetic resonance system gas imaging quality testing die body |
| CN110703171B (en) * | 2019-11-11 | 2020-11-24 | 中国科学院武汉物理与数学研究所 | Multi-core magnetic resonance system gas imaging quality testing die body |
| CN113498776A (en) * | 2021-06-17 | 2021-10-15 | 深圳逗点生物技术有限公司 | Biological transport system, biological transport method, and blood cell preservation system |
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Application publication date: 20160817 |
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