CN102279118A - Concentration gradient diffusion sampler for bioreactor on-line testing system - Google Patents
Concentration gradient diffusion sampler for bioreactor on-line testing system Download PDFInfo
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- CN102279118A CN102279118A CN2011101148888A CN201110114888A CN102279118A CN 102279118 A CN102279118 A CN 102279118A CN 2011101148888 A CN2011101148888 A CN 2011101148888A CN 201110114888 A CN201110114888 A CN 201110114888A CN 102279118 A CN102279118 A CN 102279118A
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Abstract
The invention relates to a sampler which is suitable for bioreactors of microbial fermentation, cell culture and the like, in particular to a concentration gradient diffusion sampler. The concentration gradient diffusion sampler is characterized by that: according to the diffusion principle due to concentration difference of ingredients of the liquid in the solutions in the two sides of a diffusion barrier, the concentration gradient distribution of the ingredients (test sample) is formed in a sampling pipe, and the flux or rate of the diffusion obeys Fick's law. Using the device for sampling the sample in the bioreactor can effectively prevent sampling pollution and has good security. The sampling has no filtering process of solutions, thus avoiding blocking of membrane holes and having high stability of continuous sampling. The ingredients of the sample form the gradient distribution, so that the detection can be carried out without cumbersome dilution, thus making the on-line testing system simple and quick and making the linear range large. The device is particularly applicable to the on-line testing of various flow injection analytical systems.
Description
Technical field
The present invention relates to a kind of sampler that is applicable to bio-reactor on-line detecting systems such as microbial fermentation, cellular incubation.
Background technology
Bio-reactor is the biological function that utilizes enzyme or biosome (as microorganism) to be had, at the external apparatus system that carries out biochemical reaction, it is a kind of biological function analog machine (container), as fermentation tank, enzyme or cell reactor etc., wait production and scientific research field widespread use at food and fermentation (as drinks, amino acid, organic acid), medicine (microbiotic, interferon), environment (biodegradation), bioenergy (cellulosic ethanol, the little algae of produce oil, biogas).
Keep the efficient operation of bio-reactor, promptly obtain maximum biological transformation ratio and minimum cost, need control the bioprocess in the reactor, the basis of its control is detected various parameters in the reactor exactly.
The detection mode of bio-reactor has two kinds of offline inspection and online detections.Offline inspection is by hand sampling, then, sample is delivered to laboratory or instrument room, measures by instrumentation.Online detection be detecting instrument directly with reactor links together or detecting sensor probe insertion reactor in detect.Advantage be sampling and testing process can be continuously, rapidly, accurately finish, the dynamic change of the reflection reactor biochemical process that energy is timely, accurate, continuous helps the optimization and the control of bioprocesses.
On-line detecting system generally comprises 2 parts: (1) analysis and detection technology and equipment (pick-up unit); (2) device (sampler) that pick-up unit and inside reactor medium can be combined.The present invention is exactly a kind of sampler of on-line detecting system.
Relevant both at home and abroad sampler patent situation analysis is as follows:
(1) domestic patent
1. the Zhu Xiang of Jiangsu University faces invention " the automatic sample valve of a kind of novel fermentation " (number of applying for a patent: 200520073106.0).
2. surplus few literary composition invention a kind of " the special-purpose sample valve of fermentation tank " (application number/patent No.: 201020102452).
More than the common trait of two kinds of patents all be the sampler that constitutes by pipeline and valve member, can automatically, directly take out the sample (former state) of inside reactor, steam sterilization and packing washer are arranged, prevent that sampling from polluting.
3. invention such as the Liu Fei of Southern Yangtze University a kind of " system of detecting concentration of sugar in fermentation process on line " (application number/patent No.: 200910034515).
The feature of this patent is that then, sample is sent on the detecting instrument and measures after mixing (dilution) in proportion with pure water by directly sampling (former state) from fermentation tank of constant flow pump.With the difference of 1,2 patents be to have adopted constant flow pump, be not the sampling that opens or closes by valve.By the uniflux of liquid, avoid pollution to reactor (fermentation tank).
4. invention " a kind of sampling air mix facilities of bioprocesses the on-line detecting system " (number of applying for a patent: CN02138376.6) such as Southern Yangtze University poplar Hai Lin.
The difference of this patent and 1,2,3 patents is: place a ceramic membrane filter spare at sampling probe end (being connected with fermentation tank), the continuous filtration in film spare of liquid in the reactor, the filtered fluid of trace is drawn (not being former state) by pipeline, behind an air mix facilities, enter detector and measure.The ceramic membrane aperture is between 0.1-0.2 μ m, and microorganism can not pass through, and can prevent effectively that reactor (fermentation tank) from polluting.
(2) external (U.S.) patent
1.Seifert Deng (1990) " a kind of sterile sampling device " (4,942,770).Be connected with reactor by a T-valve, finish sampling by the conversion valve passage, promptly connect with fermentation tank earlier, (forward) flows into sampling pipe, the conversion valve (is closed the UNICOM with fermentation tank inside then, the UNICOM of unlatching and external pipe), the sample in the sampling pipe (former state) is oppositely sent into next step air mix facilities or detecting instrument by the liquid pump.Because inside reactor and extraneous direct the contact have been avoided in the switching of T-valve, thereby play the effect of pollution-free sampling.
2.Chow Deng (1995) inventions a kind of " sampler of ultraviolet disinfection and method " (5,409,841).Adopt uviol lamp that sampling pipe and container are sterilized, do not adopt steam sterilizing.
3.Samhaber Deng (1987), " bio-reactor sampler " (4,695,551).Adopt the semipermeable membrane filtering and sampling.Semi-permeable diaphragm can be isolated microorganism and be passed through, and avoids polluting, and increases a semi-permeable diaphragm shaking device simultaneously, prevents the obstruction of fenestra.
4.Moon Deng (1999), " fermentation tank sample cock steam sterilizer " (5,914,092).Relate to a kind of about improving the apparatus and method of steam to sample cock and pipeline sterilizing efficiency.
5.Jaeger (2005) invention a kind of " liquid sampling and method " (6,860,162).When wine fermentation tank is taken a sample, designed a settler, sampling probe can be avoided sediment in the fermentation liquor, prevents to block.
In sum, existing invention mainly contains two types, and a class is made up of pipeline and valve, and a class is that diffusion barrier filters.The sampler weak point that pipeline valve is formed is: taking-up be former state in the reactor, contain thalline, the thick slag of raw material etc., often need just can enter Instrument measuring through after the filtration treatment; For avoiding the pollution of sampling process, must need the configuration bactericidal unit, as steam, uviol lamp etc.For the diffusion barrier filtering and sampling, because the aperture of film has stoped passing through of thalline, avoided the external source pollution, weak point is the obstruction that the impurity in the liquid causes the diffusion barrier mesh easily, filters poor stability, needs often to clean or change.In addition, the common drawback of these two types of sampling modes is: because the variation of sample concentration is bigger in the reactor, the sample that takes out need could be measured through after the dilution of different proportion, this just needs the complicated air mix facilities of configuration, both increase the cost of sampling system, also caused the error (dilution error) of measurement result easily.
The present invention adopts diffusion barrier (element) sampling method, but not conventional membrane filtration mode, but the diffusion principle that in the solution of diffusion barrier both sides, causes according to each component in the liquid by concentration difference, the concentration gradient that forms component (sample) in sampling pipe distributes, and the flux of its diffusion or speed are followed Fei Ke (Fick) law.
The online detection that utilizes device of the present invention to carry out bio-reactor has following advantage:
1. diffusion barrier (element) is porous ceramics or organic high molecular layer, the ability high-temperature sterilization, and the aperture is between 0.1-0.2 μ m, and microorganism can not pass through, and can effectively prevent the pollution of sampling process, and security is good.
2. do not have pressure differential between the solution of diffusion barrier (element) both sides, molecule in the solution or ion by the low side migration of the high side direction concentration of concentration, do not have the filter process of solution by simple diffusion, and fenestra should not stop up, and the stability of serial sampling is higher.
3. the sample of Qu Chuing (sample) does not have impurity, does not need filtration treatment just to be applicable to that multiple analytical instrument directly measures.
4. in diffusion process, have liquid part (sample) the device scanning after testing of concentration gradient, can form a scanning peak, component concentrations has correlativity in peak height or peak width and the reactor.Sample for low concentration can calculate sample size in the reactor by peak height, and enriched sample can peak width be calculated content.Also but the response curve that concentration of component changes in the METHOD FOR CONTINUOUS DETERMINATION reactor is determined components contents in the reactor.Therefore, on-line detecting system does not need to dispose complicated air mix facilities, makes on-line detecting system simpler, and finding speed is faster, and error at measurment is littler.
Summary of the invention
The present invention relates to a kind of simple, fast, the sampler of stable and safe bio-reactor on-line detecting system, be specifically related to a kind of concentration gradient diffusive sampling device, form by right cylinder, diffusion barrier (element), diffusion admittance, U type diffusion pipeline, damping fluid (reagent) pipeline, diffusion component mixed liquor outflow pipeline and liquid pump.
Agent set is the cylindrical structure that the organic material rod processes, and diameter is 1.0-2.5cm.There is fluid pipeline right cylinder inside, is the U type, i.e. U type diffusion pipeline, and internal diameter of the pipeline is 0.5-2.0mm.The interlude of U type pipeline is positioned at the right cylinder front end, and two side shoot pipelines extend to the right cylinder rear end, i.e. damping fluid (reagent) conduit entrance and a liquid sample pipe outlet.There is an opening in the central authorities of U type diffusion sampling pipe interlude, are diffusion admittance, and diameter is 0.5-2.0mm.Two side shoots of U type diffusion sampling pipe flow out pipeline with damping fluid (reagent) flow ipe and diffusion component mixed liquor respectively in the right cylinder rear end to be connected, and a liquid pump controlling plumbing fixtures liquid flow is arranged.Diffusion barrier (element) can be selected plane (diaphragm or garden post) and cast for use, separates the aperture and is 0.1-0.2 μ m.Sheet diffusion barrier (element) is circular, and diameter 0.5-1.5cm is encapsulated in the right cylinder front end and gets in the circular groove, and is corresponding with the diffusion admittance position.The column resolution element is a column type, and column diameter is 0.5-2.0mm, directly is encapsulated on the diffusion admittance.The tubulose resolution element is encapsulated in U type interlude position, and bore is 0.5-2.0mm, directly is communicated with U type diffusion sampling pipe.
This device can be connected with the fluid pipeline on bio-reactor top, sidewall or the reactor, Yi Bian promptly be solution in the reactor, Yi Bian be the damping fluid (reagent) in the U type pipeline, is isolated by resolution element between the two-phase.Molecule in the solution or ion are through diffusion admittance, see through resolution element, to the low side of concentration (U type pipeline) migration, within a certain period of time, the solution on U type diffusion sampling pipe interlude and two the side shoot directions forms the Gradient distribution of sample by the high side (reactor) of concentration.According to the diffusion time of setting, the start liquid pump is sent the gradient composition mixed liquor (sample) that diffuses to form in the U type pipeline into detecting instrument and is measured, and is full of new damping fluid (reagent) in the U type pipeline simultaneously, enters sampling process next time.
Description of drawings
Accompanying drawing 1 is sampler (sheet resolution element) structure and molecular diffusion schematic diagram of mechanism.
Accompanying drawing 2 is sampler (tubulose resolution element) structure and molecular diffusion schematic diagram of mechanism.
Accompanying drawing 3 is sampler (column resolution element) structure and molecular diffusion schematic diagram of mechanism.
Accompanying drawing 4 is sampler and bio-reactor connection diagram.
Accompanying drawing mark: 1. bio-reactor; 2. sheet resolution element (film); 3.U type diffusion pipeline (interlude); 4.U type diffusion pipeline (side shoot); 5. right cylinder; 6. damping fluid (reagent) flow ipe (import); 7. diffusion component mixed liquor flows out pipeline (outlet); 8. diffusion admittance; 9. tubulose resolution element (film); 10. column resolution element (film); 11. sampler; 12. liquid pump; 13. damping fluid (reagent) bottle; 14. detecting instrument
Embodiment
A kind of concentration gradient diffusive sampling device of bio-reactor on-line detecting system can be realized in the following way:
With reference to the accompanying drawings 1,2,3, agent set is the cylindrical structure (5) of organic material rod, can select perspex bar, tetrafluoroethene rod etc. for use, and excellent diameter is 1.0-2.5cm, and length is in 3cm.Processing U type pipeline in rod, internal diameter of the pipeline is 0.5-2.0mm.U type interlude (3) pipeline is near the right cylinder front end, and both sides side shoot pipelines (4) extend to the right cylinder rear end.There is an opening in U type pipeline interlude (3) central authorities, with right cylinder (5) front end exterior, are the passages that carries out molecular diffusion between reactor solution and U type pipeline solution, i.e. diffusion admittance (8), and channel diameter is 0.5-2.5mm.
With reference to the accompanying drawings 1,2,3, separating film element (2) is encapsulated in diffusion admittance (8) position, the outside is to contact with bio-reactor solution (1), and inner face contacts with the interior solution of U type pipeline (3), is formed on the molecular diffusion system that is isolated by microporous separation membrane between two solution phases.Diffusion barrier (element) can be selected plane (diaphragm or garden post) and cast for use, separates the aperture and is 0.1-0.2 μ m.Sheet diffusion barrier (element) is circular, and diameter 0.5-1.5cm is encapsulated in the right cylinder front end and gets in the circular groove, and is corresponding with the diffusion admittance position.The column resolution element is a column type, and column diameter is 0.5-2.0mm, directly is encapsulated on the diffusion admittance.The tubulose resolution element is encapsulated in U type interlude position, and bore is 0.5-2.0mm, directly is communicated with U type diffusion sampling pipe.
With reference to the accompanying drawings 4, be damping fluid or biochemical reagents in damping fluid (reagent) bottle (4).Damping fluid (reagent) flow ipe (import of U type diffusion sampling pipe) (6) is connected with damping fluid (reagent) bottle (4); Diffusion component mixed liquor flows out pipeline (outlet of U type diffusion sampling pipe) (7) and is connected with detecting instrument (scanner) (5).Start liquid pump (3) is full of damping fluid or reagent in the U type sampling pipe.
With reference to the accompanying drawings 4, this sampler (2) inserts from bio-reactor (1) top and connects, and also can be connected with the fluid pipeline on bio-reactor sidewall or the reactor, Yi Bian promptly be solution in the reactor, be the damping fluid (reagent) in the U type pipeline on one side, isolate by resolution element between the two-phase.
With reference to the accompanying drawings 4, the liquid part (sample) that has concentration gradient in the U type sampling pipe of sampler (2) is sent into detecting instrument (5) by liquid pump (3) and is measured.When liquid pump (3) is sent the gradient test specimens, be full of new damping fluid or reagent again in the U type sampling pipe of sampler (2).
With reference to the accompanying drawings 4, after the diffusion time of setting, the liquid part (sample) that has concentration gradient in the U type pipeline by liquid pump (3) intermittently or constant current send into detecting instrument (5) and measure.During intermittent sampling, device scanning after testing can form a scanning peak, and component concentrations has correlativity in peak height or peak width and the reactor.Sample for low concentration can calculate sample size in the reactor by peak height, and enriched sample can peak width be calculated content.During the constant current sampling, can form the response curve that changes with concentration of component in the reactor, determine components contents in the reactor.Sample for low concentration can adopt the low speed sampling and measuring, and enriched sample can adopt quick sampling and measuring.So the sample of taking-up need not just can directly be measured through dilution.Measured value is sample size measurement result in the reactor through standard correction.When liquid pump (3) is sent the gradient test specimens, be full of new damping fluid or reagent again in the U type pipeline, enter diffusion sampling next time automatically.
Claims (6)
1. the concentration gradient diffusive sampling device of a bio-reactor on-line detecting system is characterized in that: this device by right cylinder, diffusion barrier (element), diffusion admittance, U type diffusion pipeline, damping fluid (reagent) pipeline, diffusion component mixed liquor flows out pipeline and the liquid pump is formed.
2. according to claim 1, it is characterized in that: agent set adds the cylindrical structure that forms down for the organic material rod; There is fluid pipeline right cylinder inside, is the U type, i.e. U type diffusion sampling pipe; The interlude of U type sampling diffusion pipeline is positioned at the right cylinder front end, and two side shoots extend to the right cylinder rear end, connect a damping fluid (reagent) pipeline (import) and a liquid sample pipeline (outlet) respectively; There is an opening in the central authorities of U type sampling diffusion pipeline interlude, with the external communications of right cylinder front end, i.e. diffusion admittance.
3. according to claim 1, it is characterized in that: encapsulate diffusion barrier (element) on the right cylinder diffusion admittance, resolution element is plane (diaphragm or garden post) and cast; Sheet diffusion barrier (element) is circular, is encapsulated in the right cylinder front end and gets in the circular groove, and is corresponding with the diffusion admittance position; The column resolution element is a column type, directly is encapsulated on the diffusion admittance; Tubulose separates the assorted U type diffusion sampling pipe interlude position that is encapsulated in of unit, and its inner chamber directly is communicated with U type sampling pipe.
4. according to claim 1, it is characterized in that: two side shoots of U type diffusion sampling pipe flow out pipeline with damping fluid (reagent) flow ipe and diffusion component mixed liquor respectively in the right cylinder rear end to be connected.
5. according to claim 1, it is characterized in that: flowing of the liquid pump control U type interior liquid of diffusion sampling pipe arranged.
6. according to claim 1, it is characterized in that: this device can insert with the bio-reactor top and be connected, also can be connected with the fluid pipeline on sidewall of reactor or the reactor, promptly be solution in the reactor on one side, be the damping fluid (reagent) in the U type diffusion pipeline on one side, isolate by resolution element between the two-phase.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267814A (en) * | 2013-04-19 | 2013-08-28 | 南开大学 | Diffusion separator |
| CN104215782A (en) * | 2013-05-28 | 2014-12-17 | 恩德莱斯和豪瑟尔测量及调节技术分析仪表两合公司 | Method and apparatus for performing automated affinity based assays |
| CN104845879A (en) * | 2015-06-01 | 2015-08-19 | 山东省科学院生物研究所 | Automatic sampling system and control method of fermentation tank |
| CN104995509A (en) * | 2012-10-23 | 2015-10-21 | 剑桥企业有限公司 | Fluidic device |
| CN105890923A (en) * | 2014-12-11 | 2016-08-24 | 山东省科学院生物研究所 | Sterile trace on-line sampling device and sampling method |
| CN107063767A (en) * | 2017-06-14 | 2017-08-18 | 山东省科学院生物研究所 | A kind of sampler and fermentation tank sterile sampling method |
| CN110938537A (en) * | 2019-12-27 | 2020-03-31 | 乐绪枝 | Nondestructive sampling detection equipment during wine making |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104995509A (en) * | 2012-10-23 | 2015-10-21 | 剑桥企业有限公司 | Fluidic device |
| CN104995509B (en) * | 2012-10-23 | 2018-04-27 | 剑桥企业有限公司 | Fluidic device |
| US9958369B2 (en) | 2012-10-23 | 2018-05-01 | Cambridge Enterprise Limited | Fluidic device |
| US10670504B2 (en) | 2012-10-23 | 2020-06-02 | Cambridge Enterprise Limited | Fluidic device |
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| CN105890923A (en) * | 2014-12-11 | 2016-08-24 | 山东省科学院生物研究所 | Sterile trace on-line sampling device and sampling method |
| CN105890923B (en) * | 2014-12-11 | 2019-08-09 | 山东省科学院生物研究所 | A kind of sterile micro on-line period device and sampling method |
| CN104845879A (en) * | 2015-06-01 | 2015-08-19 | 山东省科学院生物研究所 | Automatic sampling system and control method of fermentation tank |
| CN107063767A (en) * | 2017-06-14 | 2017-08-18 | 山东省科学院生物研究所 | A kind of sampler and fermentation tank sterile sampling method |
| CN107063767B (en) * | 2017-06-14 | 2019-05-21 | 山东省科学院生物研究所 | A kind of sampler and fermentor sterile sampling method |
| CN110938537A (en) * | 2019-12-27 | 2020-03-31 | 乐绪枝 | Nondestructive sampling detection equipment during wine making |
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Application publication date: 20111214 |