CN102353798A - Biological fermentation process-oriented olfactory analog instrument and online analysis method - Google Patents
Biological fermentation process-oriented olfactory analog instrument and online analysis method Download PDFInfo
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Abstract
The invention discloses a biological fermentation process-oriented olfactory analog instrument and a biological fermentation process-oriented online analysis method. The biological fermentation process-oriented instrument is characterized by consisting of a test box and a computer; a gas-sensitive sensor array is arranged in a constant-temperature box; in the constant-temperature process, the constant-temperature box is heated only, but not refrigerated; under the control of the computer, fermentation tail gas is precisely and automatically sampled by a miniature vacuum pump, an electromagnetic valve, a flowmeter, a throttle valve and a tail gas bottle, so as to realize the online detection of the biological fermentation process; a modularized time-varying neural network and a modularized non-time-varying neural network form a cascade model to realize the online analysis of the biological fermentation process; the online analysis method comprises judgment whether the fermentation process is normal or not; recognition of tail gas of O2, NH3 and CO2, alcohols, acids, esters and other components and concentration evaluation; prediction of oxygen dissolved in a fermentation liquor, a pH value and a respiratory quotient, evaluation on concentrations of a substrate, a thallus and a product; and optimization of a material supplementing amount, a gas inflating amount and a stirring speed. By the online analysis method, the real-time dynamic display of the biological fermentation process and Internet network remote monitoring are realized.
Description
Technical field
The present invention-a kind of olfactory analog instrument and on-line analysis method towards biological fermentation process relates to computing machine, precision measurement, control automatically, precision optical machinery, bio-reactor, Fermentation Engineering, analytical chemistry field.The present invention mainly solve the accurate sampling automatically of gas sensor array operating room exact constant temperature, fermentation tail gas, olfactory analog instruments design and with being connected and communication, the online detection of biological fermentation process and remote transmission and visual, biological fermentation process status predication, control and optimization problem of fermentation tank.
Background technology
Modern Fermentation Engineering is the bridge of genetic engineering, enzyme engineering, cell engineering realization industrialization, is to improve the Fermentation Engineering gordian technique of target product concentration, production efficiency and feed stock conversion based on the online detection of the biological fermentation process of computing machine and modern control theory, control and optimisation technique.
The dreamboat of biological fermentation process optimal control is in the hope of reaching the productive capacity maximization through process optimizing.The biological fermentation process influence factor is intricate, and experimental data repeatability is relatively poor, be difficult to describe with the mathematical model of confirming, and be a non-linear stochastic dynamic process.Really, the data storage of sweat is important with showing, yet, the online detection of what is more important sweat, Real-time and Dynamic control, prediction and optimization.For example; Influence on the basis of glutamic acid fermentation principal element in deep understanding; In conjunction with empirical equation; We can set up all kinds of mathematical models of glutamic acid fermentation process; Generate rule to describe sweat thalli growth and product; Design complete process and detect and control system, according to the fermentation liquor and the fermentation tail gas data of direct or indirect test, to dissolved oxygen DO (DO) concentration, pH value, throughput, fermentation temperature, tank pressure, tail gas O
2And CO
2Parameters such as concentration, feed supplement mode and opportunity detect and predict that analysis-by-synthesis growth of microorganism and metabolism situation in time change control strategy, and the adjustment external environment condition is beneficial to the optimization of thalli growth and product building-up process.
The online detection of sweat, control and the purpose of optimizing are to utilize microorganism to obtain high value-added product with comparatively economic mode.Excellent species seed selection and improvement, nutrient culture media and supplying technics optimization, working condition control are aspects that reaches this target with optimizing; In recent years, people more with target tightening the level of the online detection of biological fermentation process technical parameter, modeling and height such as robotization control, biological fermentation process instrumentation and aspect.Wherein, Computer control and optimization system towards saccharomycete, amino acid, citric acid, beer, penicillin fermentation process; Especially sweat temperature, the control of glucose flow feeding have obtained practical application with optimization system, have obtained good economic benefit.
In saccharomycete was cultivated, people kept concentration of glucose suitable in the nutrient culture media through control stream rate of acceleration, have reached the effect that improves yeast thalline output.In penicillin fermentation, some researchers have made up the computer control system of flow feeding technology, have obtained effect preferably.Professor Zhang Siliang of East China University of Science proposes biological respinse engineering analysis and the control method based on cellular metabolism stream; Invented that sweat is multilevel, the multi-parameters optimization control technology; Designed a kind of new ideas bio-reactor; Through streams detection and process optimization and amplification, increased substantially the fermentation level of products such as penicillin, erythromycin, aureomycin, inosine, guanosine, recombination human serum albumin.
The quality of biological fermentation process depends on creation, control and the optimization of good production environment fully, mainly is the online detection and the adjustment of the various parameters of sweat.At present, biological fermentation process mainly detects with controlled variable and comprises DO value, pH value, temperature, stir speed (S.S.), air mass flow, tank pressure, liquid level, viscosity, dissolving CO
2, feed supplement speed, feed supplement amount, tail gas O
2, tail gas CO
2, respiratory quotient (RQ), cell concentration etc.Wherein, these 2 parameters of DO value and pH value possibly be most important and the most widely used.Theoretical analysis is pointed out, can regulate the pH value through the stream rate of acceleration and the stream dosage of control soda acid or carbon source material; And the adjusting of dissolved oxygen DO mainly realizes through changing parameters such as throughput, stirring rate, tank pressure, ventilation composition (pure oxygen or oxygen enrichment), carbon and nitrogen (mainly being glucose and ammoniacal liquor) additional amount.
The online detection of fermentation tail gas is very important with analysis.Tail gas O
2And CO
2Content paramagnetic oxygen analyzer commonly used and infrared CO
2Analyser is measured respectively.Fast development along with computer technology and sensor technology; Laser nephelometer, flow injection type analyser, biology sensor, high performance liquid chromatography etc. can be realized substrate concentration, thalline activity and concentration, the isoparametric online detection of production concentration; Some devices and instrument have been realized commercially producing; But problems such as existing can't withstand high temperatures, high pressure, reliability are low, poor stability only are applied to laboratory and pilot-scale at present.
Mass spectrometer is a kind of typical tails assay instrument, is widely used in quick measurement tail gas various ingredients in recent years.On principle, mass spectrometer can online detection tail gas O
2, CO
2With some other composition.The mass spectrometer of being furnished with kapillary and film device is connected with computer data processing system, through the online detection to multiple gases, can carry out online detection and analysis to a plurality of fermentation tanks simultaneously.The mass spectrometric analysis method major defect is that (1) instrument price is very expensive.For example, the U.S. produces MAX300-LG mass spectrometer unit price more than 1,000,000 yuan, is difficult to large-scale promotion; (2) high to the volatile ingredient insufficient sensitivity, be difficult to realize the on-line analysis simultaneously of multiple composition.In fact, mass spectrometer is mainly used in tail gas O at present
2And CO
2Online detection.
Olfactory analog method and instrument utilize the overlapping a plurality of gas sensors of performance to form array, can carry out qualitative and quantitative analysis to the smell quality fast, cause that people pay much attention to.For example, we can confirm aroma-producing substance classification, intensity, quality grade, true and false, freshness, control production run, adjustment formula for a product and technology to the measurement data of smell according to the olfactory analog instrument, or the like.Before 2000, olfactory analog research mainly concentrates in simple composition gas identification and the concentration estimation; Today, research object is extended on the complicated smell global feature qualitative and quantitative analysis, for example, and the grade estimation of spices and essence, tea, wine; Fruit maturity is judged; Aquatic products and meat products freshness are confirmed; Surrounding air and water pollution monitoring; War gas detects to be surveyed; Medical diagnosis on disease; And even bacterium and blood smell; Or the like.Even to this day, can we find out the material that does not definitely have smell?
Along with the progress of material science and accurate manufacturing process, with SnO
2For the sensitivity of MOS type gas sensor of representative has reached 1.0 * 10
-9V/V (1.0ppb), usual range is 1.0～10000ppm.A development trend of olfactory analog research is; Has necessary sensitivity and optionally under the prerequisite at single gas sensitive device; Form array with responding range broad and the overlapping a plurality of gas sensors of performance; Utilize data analysing method to improve the selectivity of instrument emphatically; Control repeatability and the long-time stability that realize instrument with the precision of internal work condition, and the multidimensional response signal that gas produces is obtained analysis results such as gas classification, intensity, key component concentration according to gas sensor array.These research contents relate to concrete application such as precision optical machinery, precision measurement and control, computing machine, applied mathematics, analytical chemistry and Fermentation Engineering, have important significance for theories and practical value.
The olfactory analog instrument application has a extensive future, but present ability is also quite limited.Relevant document of olfactory analog and patent search result are following: (1) document.Nineteen ninety had only more than 60 piece in the past, was more than 500 piece by 2000, reached 4000 now surplus a piece of writing, olfactory analog research extensively expansion in recent years be described.(2) patent.100 multinomial international monopolies and more than 20 domestic patents were applied in recent years mostly or were authorized, and showed to have begun to pay attention to olfactory analog domain knowledge property right protection both at home and abroad.(3) technical standard.Through IHS international standard database is retrieved, do not find the product technology standard relevant with olfactory analog.Domestic units such as Zhejiang University, Dalian University of Technology, the Central China University of Science and Technology had carried out the research of this respect in recent years.Yet by the end of now, a piece of writing surplus the relevant paper 300 of the domestic olfactory analog of delivering wherein is published on the international well-known magazine less than 100 pieces.The The above results explanation, the olfactory analog theory and application research is demanded urgently deeply.Through to ISI and two data library inquiries of EI, do not find that as yet the olfactory analog method is applied to the document of online detection of sweat and analysis.
Core-the gas sensor array of olfactory analog instrument is to organic volatile gas such as alcohol, ester, acid, aldehyde, ketone, hydrocarbon, alkene and CO, NH
3, O
2Has very high sensitivity Deng inorganic gas.SnO
2The semi-conductor type gas sensor is to tested gas; Fermentation tail gas for example; From begin to touch reach steady state (SS) response time less than 10 seconds; Do not need secondary instrument to amplify; Directly with 0～10V or the output of 0～5V voltage signal, online detection is very attractive to biological fermentation process for this.The olfactory analog method is passed through O in the fermentation tail gas
2, NH
3, CO
2Online detection with numerous micro-organic principles; By means of computing machine and data analysing method; Confirm fermentation tail gas composition and concentration, and then parameters such as concentration of substrate, thalline activity and concentration, production concentration and fermentation liquor DO, pH value, RQ are predicted and optimize.
External causes are the conditions for change, and internal cause is the basis that changes.Sweat is controlled and optimized according to the fermentation dynamics principle, must grasp the influence of demand factor and the various fermentation condition of growth of microorganism state, matrix and oxygen thalli growth and product building-up process.Along with temperature raises, along with stir speed (S.S.), throughput, tank pressure and feed supplement amount increase, also along with sweat continues, dissolved oxygen DO, tail gas O
2, tail gas NH
3, tail gas CO
2, parameter such as cell concentration all will change, be embodied in exhaust gas component variation taken place.For example, the microorganism oxygen utilization increases, tail gas O
2Concentration will correspondingly reduce; Mend the ammonia amount and suitably increase tail gas NH
3Concentration will correspondingly increase; Mending the sugar amount suitably increases, and constituent concentrations such as tail gas ethanol, lactic acid will respective change.Say that in a sense the pH value changes and also can cause exhaust gas component to change.In a word, tail gas O
2, CO
2, NH
3Variation in inorganic gas and organic volatile gas such as alcohol, acid, ester means that variation has taken place for thalli growth state and growing environment, concentration of substrate, product building-up process; In view of the above, we can carry out online detection, prediction, control and optimization to biological fermentation process with the olfactory analog method.This is the purpose of the application's project just.
For the olfactory analog method being applied to online detection of biological fermentation process and analysis, the key technical problem that the present invention will solve is: (A) the accurate sampling of tail gas head space, gas sensor array and olfactory analog instruments design.
Not only contain O in the tail gas
2, CO
2, NH
3Etc. inorganic constituents, also contain organic metabolic products such as ethanol, lactic acid, the latter often the trace, therefore, gas sensor array should by sensitivity enough the height, selectivity preferably a plurality of gas sensor form, the single gas sensor life-span is answered long enough.The tail gas amount of 50 liters of capacity and above fermentation tank is abundant, and this is an advantage, yet a sweat continues a couple of days and even tens of days, and gas sensor Long contact time tail gas can produce " paralysis " and even " poisoning " phenomenon; Fermentation site environment air is often unholiness, and ambient temperature and humidity changes in addition; Therefore; In order to realize continuous detecting, we need study tail gas the head space accurate method of sampling and device automatically, realize the tail gas head space sample mode that a kind of macroscopic view is continuous, microcosmic is discrete; Guarantee repeatedly the repeatability of head space sampling, and guarantee the long-term consistency of gas sensor array baseline.The olfactory analog instrument should be as the special-purpose mass spectrometer of tail gas, and is easy to operate, full automation.
(B) the online detection of sweat, status predication and visual.
Because the sweat time is long and the multiple organic component content of tail gas is very little, from hardware aspect, data collecting card should have enough sampling precisions and resolution; From the software aspect, cause hard drive space to take too much phenomenon for avoiding a large amount of gibberish storages, need a kind of macroscopic view of research is continuous, microcosmic amplifies data recording and online display packing, realize Data Dynamic local enlargement display and remote transmission; The visual problem that needs to solve of yeastiness comprises that single gas sensor response is visual; The gas sensor array Whole Response is visual; Tail gas O
2, CO
2, NH
3, composition such as ethanol, lactic acid visual; Substrate, thalline and production concentration visual.The yeastiness prediction mainly should propose effective non-linear autoregression kinetic model, time-delay neural network forecast model, SVM prediction model towards time series data, to realize the on-line prediction to concentration of substrate, thalline activity and concentration, production concentration.
(C) sweat control and optimization.
The purpose of fermentation is to obtain high value added product with comparatively economic mode.On the basis of setting up the sweat mathematical model; Utilizing the non-linear autoregression kinetic model of time series, time-delay neural network and SVM prediction model to realize respectively with DO, pH, RQ, concentration of substrate, cell concentration, production concentration is the sweat control of target, optimizes parameters such as feed supplement speed and feed supplement amount, throughput, stirring rate, tank pressure.
The present invention is mainly with amino acid; Microbiotic; Biomaterial; Fermented products such as industrial spirit are application background; Realization is based on online detection of the biological fermentation process of olfactory analog and analytical approach; Mainly comprise the gas sensor array exact constant temperature and accurate New sample method of tail gas head space and the new equipment that detect towards the fermentation tail gas direct-on-line; The modularization that becomes neural network by towards the modularization of status predication the time and estimate towards concentration becomes integrated model and on-line learning algorithm and the popularization ability that neural network is formed when non-, is the online detection of sweat; Status predication; Control and optimization provide new analytical instrument and method.
Summary of the invention
The present invention is (referring to number of patent application: 02111046.8) in existing patent of invention " a kind of machine olfaction device and olfactory analog method of testing thereof "; " a kind of small automatic machine sense of smell instrument and smell analytical approach " (application number: 200710036260.4); " a kind of parallel-serial mode identification method and the application in machine olfaction thereof " (application number: 200710042607.6); " a kind of olfactory analog instrument and multiple smell qualitative and quantitative analysis method " (application number: 201010115026.2; Publication number: on basis CN 101806763A), for biological fermentation process is carried out online detection; The presence prediction; Control and optimization; The remote data transmission; Visual a kind of olfactory analog instrument and the on-line analysis method that proposes with remote monitoring.
To achieve these goals, the present invention-a kind of olfactory analog instrument comprises test box and computing machine two parts, and fermentation tail gas comes from the fermentation tank of production unit, and pure air is provided for oneself by the user.
Establish constant temperature oven, minipump, two-position three way magnetic valve 103, two-position two-way solenoid valve 105 in the test box, two-position two-way solenoid valve 107, two-position two-way solenoid valve 111, flowmeter, D.C. regulated power supply, driving and control circuit, throttling valve 106, throttling valve 110; Constant temperature oven comprises gas sensor array, gas sensor array annular working chamber, resistance heating wire, temperature sensor, fan, thermofin; Described gas sensor array comprises 16 gas sensors that performance overlaps each other.
The data acquisition of biological fermentation process, show with analysis and realize by main frame, the data collecting card and the display that are inserted in the main frame.
Test box is connected through cable with computing machine; Test box is connected with the pure air bottle with tested fermentation tank respectively through gas pipeline, and the tail gas bottle that it is 1500～2000ml that the gas circuit centre of test box and fermentation tank is provided with a capacity is with the savings fermentation tail gas, to avoid being extracted into vacuum; The tail gas bottle is commercial glass flask, and the user provides for oneself; Two pores on the tail gas bottle cap are connected with fermentation tank and test box respectively; Pure air in the pure air bottle makes gas sensor array before fermentation tail gas is measured, be in normal condition.
In tail gas when sampling,, under the swabbing action of minipump, the fermentation tail gas of savings in the tail gas bottle is with 600 ml/min and the above flow gas sensor array of flowing through, and gas sensor array produces i.e. 16 road analog voltage signals of responsive response thus; Data collecting card changes into 16 way word signals with this 16 road analog voltage signal, and the olfactory analog instrument carries out on-line analysis, prediction to biological fermentation process in view of the above and optimizes.
Gas sensor array annular working chamber through screw retention, and forms the annular cavity of sealing with 2 O RunddichtringOs by sensor holder and sender unit cap; The diameter that sensor holder upper edge φ 120～φ is 160 millimeters uniformly-spaced is provided with 18 holes, comprises 16 7 core ceramic socket mounting holes, 1 air admission hole and 1 venthole; Air admission hole and venthole are adjacent, and the centre is separated by a dividing plate; 16 7 core ceramic sockets are glued one by one in 16 mounting holes of sensor holder with tasteless bonding agent, and are connected with control circuit with driving with D.C. regulated power supply through lead; 16 gas sensors directly are inserted on 16 7 core ceramic sockets, form gas sensor array; Fermentation tail gas gets into gas sensor array annular working chamber from air admission hole, skims over 16 gas sensors successively, discharges from venthole then.
The venthole of the air admission hole of gas sensor array annular working chamber and two-position three way magnetic valve 103, gas sensor array annular working chamber and two-position two- way solenoid valve 105 and 111, two-position two-way solenoid valve 105 and throttling valve 106 and two-position two-way solenoid valve 107, throttling valve 106 and flowmeter, minipump and flowmeter and two-position two-way solenoid valve 107, two-position two-way solenoid valve 111 and throttling valve 110, throttling valve 110 and pure air bottle 5 all are connected through gas circuit.
Constant temperature oven is heated by the resistance heating wire, and fan makes the temperature inside the box even, and thermofin prevents that heat from distributing, and the temperature sensor detected temperatures changes; Temperature regulating range is 50～60 ℃, and the constant temperature precision is ± 0.1 ℃, and thermostatic process only heats, and does not freeze.
D.C. regulated power supply comprises 4 road voltages, is respectively 5V, 10V, 12V and 36V; Wherein, 5V is the heating voltage of gas sensor array; 10V is the operating voltage of gas sensor array, and 12V is the operating voltage of 103,3 two-position two-way solenoid valves of two-position three way magnetic valve, minipump, fan, and 36V is resistance heating wire's a heating voltage; 5V and 10V are linear power supply, and 12V and 36V are Switching Power Supply.
Minipump is worked in a continuous manner; Computing machine is controlled the energising and the outage of two-position three way magnetic valve 103, two-position two-way solenoid valve 105,107 and 111 through data collecting card and cable; To control the through and off of gas sensor array annular working chamber and fermentation tail gas, pure air and surrounding air respectively; And on this basis, through the size of throttling valve 106 and 110 pilot-gas flows.
See that from the angle that gas flow changes complete gas sampling cycle comprises preliminary recovery 225 seconds successively, accurately demarcates 40 seconds, balance 5 seconds and tail gas sampling 30 seconds totally 4 stages, continues 300 seconds.
During the tail gas sampling; Fermentation tail gas is aspirated by minipump; With 600 ml/min and above flow flow through successively two-position three way magnetic valve 103, air admission hole, gas sensor array annular working chamber, venthole, two-position two-way solenoid valve 105, throttling valve 106, flowmeter, be discharged into the surrounding air from waste gas outlet by minipump then; The tail gas sampling duration is 30 seconds.
Accurate timing signal; Pure air oppositely flows into gas sensor array annular working chamber through throttling valve 110 and two-position two-way solenoid valve 111 from venthole with 600 ml/min and above flow; And finally be discharged in the surrounding air, thereby make gas sensor array when repeatedly tail gas is sampled, be in same normal condition through air admission hole and two-position three way magnetic valve 103; Accurately demarcating the duration is 40 seconds.
During balance, all solenoid valves all are in off-state, and the indoor gas of gas sensor array annular working remains static, and continue 5 seconds.
During preliminary the recovery; Under minipump suction, surrounding air with the flow of 5000～6500 ml/min flow through successively two-position three way magnetic valve 103, air admission hole, gas sensor array annular working chamber, venthole, two-position two-way solenoid valve 105, two-position two-way solenoid valve 107, be discharged into the surrounding air from waste gas outlet by minipump then; The effect of surrounding air, the one, make gas sensor array tentatively return to normal condition fast; The 2nd, the heat that accumulation produces when taking away 16 gas sensors and working simultaneously; The 3rd, wash away the scent molecule that is adsorbed on the gas sensor array annular working chamber interior walls; The preliminary recovery duration is 225 seconds.
In the gas sampling one-period, in 60 seconds of balance zero hour, gas sensor array response voltage data are by record, and promptly data acquisition time length is 60 seconds; Data collecting card converts gas sensor array 16 road analog voltage signals that perception obtains to fermentation tail gas into 16 way word signals with 16 A/D conversions; The maximal value of 1 gas sensor voltage responsive in this 60 seconds data acquisition times and minimum value are the olfactory analog instrument carries out qualitative and quantitative analysis to biological fermentation process foundations finally by record and be kept in the data file of computing machine.
The cascade model that becomes the neural network composition when change neural network and modularization are non-during with modularization carries out qualitative and quantitative analysis to biological fermentation process; One 16 dimension sample is gas sensor array peak response to fermentation tail gas in the gas sampling one-period, the i.e. digitized representations of biological fermentation process current state; The structure that becomes neural network when change neural network and modularization are non-during modularization all is the single layers that conceal of single output; Become neural network module in the time of one and represent a gas sensor, be responsible for next response constantly of this gas sensor of prediction; A plurality of neural network modules that become when non-are discerned the fermentation tail gas key component with the gas sensor array current time to the response of fermentation tail gas and are predicted its concentration, prediction concentration of substrate, cell concentration, production concentration, DO value, pH value.
The olfactory analog instrument is following to the step that fermentation tail gas carries out online detection and analysis:
(1) start preheating; Two-position two- way solenoid valve 105 and 107 is connected; Two-position three way magnetic valve 103 breaks off with two-position two-way solenoid valve 111; Under the swabbing action of minipump, surrounding air flows through gas sensor array annular working chamber through two-position three way magnetic valve 103 with the flow of 5000～6500 ml/min; Be 30 minutes preheating time;
(2) accurately demarcate; Two-position two-way solenoid valve 111 is connected; Two-position three way magnetic valve 103 breaks off with two-position two- way solenoid valve 105 and 107; Pure air oppositely flows into gas sensor array annular working chamber through throttling valve 110, two-position two-way solenoid valve 111 from venthole with 600 ml/min and above flow; Be discharged to the surrounding air through two-position three way magnetic valve 103 from air admission hole then, last 40 seconds; Thus, gas sensor array is able to accurate demarcation, makes the repeatedly measurement of fermentation tail gas is carried out on same benchmark;
(3) balance; Two-position three way magnetic valve 103 all breaks off with two-position two-way solenoid valve 105,107 and 111, and the indoor gas of gas sensor array annular working remains static, the balance period that gas sensor array got into for 5 seconds;
(4) tail gas sampling; Two-position three way magnetic valve 103 is connected with two-position two-way solenoid valve 105; Two-position two- way solenoid valve 107 and 111 breaks off; Minipump sucks gas sensor array annular working chamber with 600 ml/min and above flow via two-position three way magnetic valve 103 with fermentation tail gas; Make it to skim over gas sensor sensitive membrane surface, be discharged into the surrounding air from waste gas outlet through two-position two-way solenoid valve 105, throttling valve 106 and flowmeter then; When fermentation tail gas accumulated samples amount reaches 300 milliliters and when above, the tail gas sampling process finishes, and lasts for 30 seconds;
(5) the preliminary recovery; After the tail gas sampling finishes; Two-position two- way solenoid valve 105 and 107 is connected; Two-position three way magnetic valve 103 breaks off with two-position two-way solenoid valve 111; Minipump sucks gas sensor array annular working chamber via two-position three way magnetic valve 103 from air admission hole with surrounding air with the flow of 5000～6500 ml/min, lasts 225 seconds; Gas sensor array is tentatively recovered under the effect of surrounding air;
(6) data acquisition; In fermentation tail gas was flowed through the process of gas sensor array annular working chamber, 16 gas sensors produced the responsive response voltages of simulation, and these responsive responses are sent to computing machine through cable; Computing machine obtains 16 dimension word response vectors through 16 tunnel and 16 data collecting card; Become the cascade model analyzing and processing that neural network is formed when change neural network and modularization are non-when modularization; Provide fermentation tail gas bulk strength, key component classification and concentration estimated value within 10 seconds after data acquisition finishes; Provide the estimated value of concentration of substrate, cell concentration, production concentration, DO value, pH value, and show through display;
(7) remote data transmission; When a data gatherer process finished, computing machine passed to the maximal value and the minimum value of gas sensor array response voltage on the Internet, realized the remote monitoring and the analysis of biological fermentation process;
(8) repeat (2)～(7), repeatedly sample;
Under the control of computing machine, online detection, prediction and the optimization of the whole process of biofermentation are accomplished by the olfactory analog instrument automatically, and a complete gas sampling cycle is 300 seconds, and the data acquisition time length in this cycle is 60 seconds;
Olfactory analog instrument of the present invention has that volume is little, constant temperature precision and automaticity is high, long term repeatability and good, the easy to use advantage of long-time stability; Become during modularization of the present invention neural network become when non-with modularization cascade model that neural network forms have analysis speed soon, advantage that the on-line prediction performance is good.
Description of drawings
Fig. 1 is the present invention-a kind of olfactory analog instrument and on-line analysis method-olfactory analog instrument component units and biofermentation system relevant portion that is connected and pure air bottle synoptic diagram towards biological fermentation process.
Fig. 2 is the present invention-a kind of component units-test box and computer operation principle schematic towards the olfactory analog instrument and the on-line analysis method-olfactory analog instrument of biological fermentation process.
Fig. 3 is the present invention-a kind of olfactory analog instrument and on-line analysis method-gas sensor array annular working chamber enlarged drawing towards biological fermentation process; Gas sensor is installed sectional view; The air admission hole sectional view, the sectional view of air admission hole and venthole and dividing plate relation; Wherein, (a) gas sensor array annular working chamber enlarged drawing, (b) gas sensor is installed sectional view A-A, (c) air admission hole sectional view B-B, (d) the sectional view C-C of air admission hole and venthole and dividing plate relation.
When Fig. 4 is the present invention-a kind of towards the olfactory analog instrument of biological fermentation process and on-line analysis method-tail gas sampling, olfactory analog instrument and fermentation tank, industrial computer concern synoptic diagram.
Fig. 5 is the present invention-a kind of towards the olfactory analog instrument of biological fermentation process and on-line analysis method-in the gas sampling cycle, gas flow and single gas sensor response change situation map; Wherein, (a) in a gas sampling cycle, the interval synoptic diagram of single gas sensor response voltage situation of change and data acquisition; (b) in a gas sampling cycle, gas flow situation of change synoptic diagram.
Fig. 6 becomes the neural network cascade model synoptic diagram that the change neural network is formed when non-with modularization when being the present invention-a kind of towards the olfactory analog instrument of biological fermentation process and on-line analysis method-by modularization; Wherein, become the neural network module synoptic diagram when (a) a plurality of, (b) a plurality of neural network module synoptic diagram that become when non-.
Fig. 7 is the present invention-a kind of, and (polyhydroxyalkanoates is PHA) with the online detection instance of cephalosporin sweat to polyhydroxyalkanoate towards the olfactory analog instrument of biological fermentation process and 2 component units of on-line analysis method-gas sensor array; Wherein, (a) gas sensor TGS813 and TGS832 are to the response curve of PHA sweat, and (b) gas sensor TGS813 and TGS826 are to the response curve of cephalosporin sweat.
Embodiment
Below in conjunction with accompanying drawing the present invention is described in further detail.
The present invention-a kind of olfactory analog instrument component units and the biofermentation system relevant portion that is connected and pure air bottle synoptic diagram are characterized in shown in accompanying drawing 1, comprise test box 1 and computing machine 2 two large divisions; When with this olfactory analog instrument biological fermentation process being carried out online detection and analyzing, user's configuration section comprises: the industrial computer 3 and the modulator-demodular unit 4 that are connected with computing machine 2, and what be connected with test box 1 has pure air bottle 5 and a tail gas bottle 6; Wherein, test box 1 all is connected through cable with modulator-demodular unit 4 with industrial computer 3 with computing machine 2, computing machine 2, and test box 1 all is connected through gas circuit with tail gas bottle 6 with pure air bottle 5.
See also accompanying drawing 2; The test box 1 of olfactory analog instrument of the present invention comprises gas sensor array 101, gas sensor array annular working chamber 102, two-position three way magnetic valve 103, dividing plate 104, two-position two-way solenoid valve 105, throttling valve 106, two-position two-way solenoid valve 107, flowmeter 108, minipump 109, throttling valve 110, two-position two-way solenoid valve 111, four road D.C. regulated power supplies 112, driving and control circuit 113, thermofin 114, fan 115, resistance heating wire 116, temperature sensor 117; Computing machine 2 comprises main frame 201, is inserted in data collecting card 202, display 203 on the computer motherboard.
Wherein, gas sensor array 101 is made up of TGS-8 series and 16 gas sensors of TGS-26 series, and the sensitive membrane material is a metal-oxide semiconductor (MOS); The concrete model of gas sensor comprises TGS813,816,821,822,823,825,826,830,831,832,842,2600,2602,2610,2611,2620; Present embodiment selects the reason of these two serial gas sensors that Japanese Figaro company produces to be; Their heating voltages are identical; Operating voltage also can be identical, and can use the sensor socket of same type, thereby make gas sensor array annular working cell structure simple.The principle of work of single gas sensor is to regard the gas sensor sensitive membrane as a resistance, with fixing pull-up resistor R
LSeries connection forms the loop, and fixing operating voltage is added on the two ends, loop; When contacting with tested gas, the resistance at gas sensor sensitive membrane two ends changes; Because operating voltage is fixed pull-up resistor R
LTherefore respective change also takes place in the voltage at two ends, and the sensitivity response of Here it is gas sensor is directly exported with voltage signal.
Seeing also accompanying drawing 3, wherein, (a) be gas sensor array annular working chamber 102 enlarged drawings, (b) for gas sensor sectional view A-A is installed, (c) be air admission hole sectional view B-B, (d) is the sectional view C-C of air admission hole and venthole and dividing plate relation.A-A cross sectional illustration, gas sensor array annular working chamber 102 comprise sender unit cap 118, sensor holder 119, O RunddichtringO 120,7 core ceramic sockets 121, tasteless adhesive 122, screw 123, form the annular cavity of sealing; Sensor holder 119 is uniformly-spaced arranged 18 holes along the diameter of 160 millimeters of D=φ 120～φ, comprises 16 holes that are used to install 7 core ceramic sockets, 1 air admission hole and 1 venthole; 16 the 7 tasteless adhesives 122 of core ceramic socket 121 usefulness are directly glued in 16 holes of sensor holder 119; The B-B cross sectional illustration, air admission hole or venthole can be connected with solenoid valve through pipe adapter; The C-C cross sectional illustration, air admission hole and venthole are adjacent, and the centre is separated by dividing plate 104; Dividing plate 104 sizes-wide * height * thick are 20 millimeters * 16 millimeters * 8 millimeters, with the annular inside groove of tasteless adhesive 122 gluings at sender unit cap 118, thereby force gas to flow around gas sensor array 101.
See also accompanying drawing 2 and accompanying drawing 3, " two-position three way magnetic valve 103 is connected " expression air admission hole and fermentation tail gas are connected, and " two-position three way magnetic valve 103 breaks off " expression air admission hole and fermentation tail gas break off but connect with surrounding air.
Fig. 4 is position and the principle of work synoptic diagram of olfactory analog instrument of the present invention in the biofermentation system, and promptly olfactory analog instrument and fermentation tank 7 and industrial computer 3 concerns synoptic diagram.For for simplicity, not drawing among the figure is used to put aside the tail gas bottle 6 and pure air bottle 5 of fermentation tail gas; The fermentation tail gas of direct perception fermentation tank 7 head spaces of olfactory analog instrument; And the measurement result of temperature sensor T, pH value sensor, DO sensor, fermentation liquor weight sensor W, fermentating liquid volume sensor V is passed to industrial computer 3 in jar; Realize the On-line Control and the optimization of biological fermentation process, through the Internet net remote monitoring of modulator-demodular unit 4 realization biological fermentation process.
Fig. 5 is the present invention-a kind of olfactory analog instrument and on-line analysis method towards biological fermentation process, in a gas sampling cycle, and the situation of change of gas flow and the response of single gas sensor.Fig. 5 (b) illustrates that gas flow experiences following 4 changes phases:
(1) the preliminary recovery-under the effect of minipump 109, surrounding air flows through gas sensor array 101 with the flow of 5000～6500 ml/min, lasts 225 seconds, and therefore gas sensor array 101 is able to preliminary recovery;
(2) accurately demarcation-flow is that 600 ml/min and above pure air last 40 seconds, makes gas sensor array 101 accurately return to normal condition; Here, normal condition is meant the responsive responsive state of 101 pairs of pure airs of gas sensor array.
(3) gas in balance-gas sensor array annular working chamber 102 does not flow, and lasts for 5 seconds;
(4) tail gas sampling-fermentation tail gas is surperficial with the sensitive membrane that 600 ml/min and above flow skim over gas sensor array 101, and therefore gas sensor array 101 produces responsive response, lasts for 30 seconds;
In the gas flow process, comprise balance 5 seconds, tail gas sampling 30 seconds and surrounding air be preliminary to recover initial 25 seconds in time of totally 60 seconds, and the response voltage value of gas sensor array can be noted by computing machine 2; Fig. 5 (a) illustrates that a gas sampling cycle is 300 seconds, and a data collection is that the data recording time span is 60 seconds; And in these 60 seconds; The maximal value of gas sensor array response voltage and minimum value are delivered to industrial computer 3; And it is online to pass to Internet through modulator-demodular unit 4, and when modularization, becomes and show neural network becomes the cascade model processing that neural network forms when non-with modularization after.
Become the cascade model synoptic diagram that neural network is formed when change neural network and modularization were non-when Fig. 6 had provided by modularization; Become neural network during m=16 modularization through preceding t-1, t-2 ..., t-k is k response x constantly altogether
j(t-1), x
j(t-2) ..., x
j(t-i) ..., x
j(t-k) next response of t is constantly predicted to 16 gas sensors; Here, subscript j representes j gas sensor; Because gas sampling cycle is 300 seconds and data recording time span is 60 seconds, as variable, time delay value k can be taken as 30, i.e. k=30 with the maximal value of the responsive response in a gas sampling cycle of each gas sensor in the gas sensor array; In other words, the olfactory analog instrument is with preceding 2.5 hours its next state of status predication of biological fermentation process.In order to eliminate the influence that ambient temperature and humidity changes, the olfactory analog instrument can be used in the difference of maximal value and minimum value of each gas sensor response in the gas sampling cycle as variable.Become neural network when a plurality of modularizations are non-and predict production concentration, cell concentration, concentration of substrate, DO value, pH value, respiratory quotient, OUR value, tail gas O successively with the response of gas sensor array current time t
2And CO
2Concentration etc.h
1And h
2Be respectively the latent node number of neural network.
Fig. 7 has provided olfactory analog instrument of the present invention and on-line analysis method to two kinds of online detections of biological fermentation process and analysis example.
Wherein, Fig. 7 (a) and Fig. 7 (b) have provided gas sensor array respectively to the response curve of PHA and cephalosporin sweat.For the purpose of clear, this instance only provides the wherein response curve of 2 gas sensors.
Can find out that by Fig. 7 (a) when the PHA fermentation proceeded to 40～50 hours, tangible, violent response had taken place for gas sensor TGS813 and TGS832, this means that undesired variation possibly appear in sweat; Hold a conference or consultation through the fermentation expert, this variation has meaned the sweat microbiological contamination.As a result, when proceeding to about 69 hours, the PHA sweat is had to because of the microbiological contamination premature termination.The significant information that Fig. 7 (a) transmits is that before about 45 hours, the response of TGS813 is bigger than TGS832 always; After about 45 hours, the response of TGS832 is but big than TGS813; These change olfactory analog instrument and on-line analysis method just carry out status predication, process control and optimization to biological fermentation process foundation.
Fig. 7 (b) tell us; Cephalosporin fermentation beginning about 10 hours; Acute variation has taken place twice in the response of gas sensor TGS813; Warp is compared with the mass spectrometer test result; These two peaks are corresponding to the sudden change of dissolved oxygen DO, and the response of gas sensor TGS826 changes not quite in this section period.Take a broad view of whole fermentation process, the response of these 2 gas sensors is always in undesired increase; When proceeding to about 57 hours, the cephalosporin sweat is had to because of the microbiological contamination premature termination.
The result of comparison diagram 7 (a) and Fig. 7 (b), we see that same gas sensor (like TGS813) is inequality to the response of different material (like PHA and cephalosporin) sweat; Certainly, different gas sensors (like TGS832 and TGS826) are inequality especially to the response of different material sweat; Like this; Olfactory analog instrument of the present invention and on-line analysis method just can change online detection and the analysis that realizes biological fermentation process according to these of gas sensor array response; Comprise status predication and Internet remote monitoring, and realize biological fermentation process control and optimize through industrial computer 3.
Claims (12)
1. an olfactory analog instrument comprises test box and computing machine, it is characterized in that:
Establish constant temperature oven, minipump, two-position three way magnetic valve 103, two-position two-way solenoid valve 105 in the described test box, two-position two-way solenoid valve 107, two-position two-way solenoid valve 105, flowmeter, D.C. regulated power supply, driving and control circuit, throttling valve 106, throttling valve 110; Described constant temperature oven comprises gas sensor array, gas sensor array annular working chamber, resistance heating wire, temperature sensor, fan, thermofin; Described gas sensor array comprises 16 gas sensors that performance overlaps each other;
Described computing machine comprises main frame, display and is inserted in the data collecting card in the main frame; Described test box is connected through cable with computing machine; Described test box is connected with the pure air bottle with fermentation tank respectively through gas circuit, and the tail gas bottle that a capacity is set is 1500～2000 milliliters in the middle of the gas circuit of test box and fermentation tank is with the savings fermentation tail gas; Pure air makes gas sensor array before fermentation tail gas is measured, be in normal condition;
In tail gas when sampling,, under the swabbing action of minipump, the fermentation tail gas of savings in the tail gas bottle is with 600 ml/min and the above flow gas sensor array of flowing through, and gas sensor array produces i.e. 16 road analog voltage signals of responsive response thus; Data collecting card changes into 16 way word signals with this 16 road analog voltage signal, and the olfactory analog instrument carries out on-line analysis, prediction to biological fermentation process in view of the above and optimizes.
2. olfactory analog instrument according to claim 1 is characterized in that, gas sensor array annular working chamber through screw retention, and forms the annular cavity of sealing with 2 O RunddichtringOs by sensor holder and sender unit cap; Sensor holder upper edge diameter uniformly-spaced is provided with 18 holes, comprises 16 7 core ceramic socket mounting holes, 1 air admission hole and 1 venthole; Air admission hole and venthole are adjacent, and the centre is separated by a dividing plate; 16 7 core ceramic sockets are glued one by one in 16 mounting holes of sensor holder with tasteless bonding agent, and are connected with control circuit with driving with D.C. regulated power supply through lead; 16 gas sensors directly are inserted on 16 7 core ceramic sockets, form gas sensor array; Fermentation tail gas gets into gas sensor array annular working chamber from air admission hole, skims over 16 gas sensors successively, discharges from venthole then.
3. olfactory analog instrument according to claim 1; It is characterized in that; The venthole of the air admission hole of gas sensor array annular working chamber and two-position three way magnetic valve 103, gas sensor array annular working chamber and two-position two-way solenoid valve 105 and 111, two-position two-way solenoid valve 105 and throttling valve 106 and two-position two-way solenoid valve 107, throttling valve 106 and flowmeter, minipump and flowmeter and two-position two-way solenoid valve 107, two-position two-way solenoid valve 111 and throttling valve 110, throttling valve 110 and pure air bottle all are connected through gas circuit.
4. olfactory analog instrument according to claim 1 is characterized in that constant temperature oven is heated by the resistance heating wire, and fan makes the temperature inside the box even, and thermofin prevents that heat from distributing, and the temperature sensor detected temperatures changes; Temperature regulating range is 50～60 ℃, and the constant temperature precision is ± 0.1 ℃, and thermostatic process only heats, and does not freeze.
5. olfactory analog instrument according to claim 1 is characterized in that D.C. regulated power supply comprises 4 road voltages, is respectively 5V, 10V, 12V and 36V; Wherein, 5V is the gas sensor array heating voltage, and 10V is the gas sensor array operating voltage, and 12V is 103,3 two-position two-way solenoid valves of two-position three way magnetic valve, minipump, fan work voltage, and 36V is resistance heating wire's heating voltage; 5V and 10V are linear power supply, and 12V and 36V are Switching Power Supply.
6. olfactory analog instrument according to claim 1 is characterized in that minipump is worked in a continuous manner; Computing machine is controlled the energising and the outage of two-position three way magnetic valve 103, two-position two-way solenoid valve 105,107,111 through data collecting card and cable; To control the through and off of gas sensor array annular working chamber and fermentation tail gas, pure air and surrounding air respectively; And on this basis, through the size of throttling valve 106 and 110 pilot-gas flows; Complete gas sampling cycle comprises preliminary recovery 225 seconds successively, accurately demarcates 40 seconds, balance 5 seconds and tail gas sampling 30 seconds totally 4 stages.
7. olfactory analog instrument according to claim 1; It is characterized in that; During the tail gas sampling; Fermentation tail gas is aspirated by minipump; With 600 ml/min and above flow flow through successively two-position three way magnetic valve 103, air admission hole, gas sensor array annular working chamber, venthole, two-position two-way solenoid valve 105, throttling valve 106, flowmeter, be discharged into the surrounding air from waste gas outlet by minipump then; The tail gas sampling duration is 30 seconds.
8. olfactory analog instrument according to claim 1; It is characterized in that; Accurate timing signal; Pure air oppositely flows into gas sensor array annular working chamber through throttling valve 110 and two-position two-way solenoid valve 111 from venthole with 600 ml/min and above flow; And finally be discharged in the surrounding air, thereby make gas sensor array when repeatedly tail gas is sampled, be in same normal condition through air admission hole and two-position three way magnetic valve 103; Accurately demarcating the duration is 40 seconds; During balance, all solenoid valves all are in off-state, and the indoor gas of gas sensor array annular working remains static, and continue 5 seconds.
9. olfactory analog instrument according to claim 1; It is characterized in that; During preliminary the recovery; Under minipump suction, surrounding air with the flow of 5000～6500 ml/min flow through successively two-position three way magnetic valve 103, air admission hole, gas sensor array annular working chamber, venthole, two-position two-way solenoid valve 105, two-position two-way solenoid valve 107, be discharged into the surrounding air from waste gas outlet by minipump then; The effect of surrounding air, the one, make gas sensor array tentatively return to normal condition fast; The 2nd, the heat that accumulation produces when taking away 16 gas sensors and working simultaneously; The 3rd, wash away the scent molecule that is adsorbed on the gas sensor array annular working chamber interior walls; The preliminary recovery duration is 225 seconds.
10. olfactory analog instrument according to claim 1 is characterized in that, in a gas sampling cycle, in 60 seconds of balance zero hour, the gas sensor array response data begins by record, and promptly data acquisition time length is 60 seconds; Data collecting card converts gas sensor array 16 road analog voltage signals that perception obtains to fermentation tail gas into 16 way word signals with 16 A/D conversions; The maximal value of each gas sensor voltage responsive in this 60 seconds data acquisition times and minimum value are the olfactory analog instrument carries out qualitative and quantitative analysis to biological fermentation process foundations finally by record and be kept in the data file of computing machine.
11. olfactory analog instrument according to claim 1 is characterized in that, the cascade model that becomes the neural network composition when change neural network and modularization are non-during with modularization carries out qualitative and quantitative analysis to biological fermentation process; One 16 dimension sample is gas sensor array peak response to fermentation tail gas in a gas sampling cycle, the i.e. digitized representations of biological fermentation process current state; The structure that becomes neural network when change neural network and modularization are non-during modularization all is the single layers that conceal of single output; Become neural network module in the time of one and represent a gas sensor, be responsible for next response constantly of this gas sensor of prediction; A plurality of neural network modules that become when non-are discerned the fermentation tail gas key component with the gas sensor array current time to the response of fermentation tail gas and are predicted its concentration, prediction concentration of substrate, cell concentration, production concentration, DO value, pH value.
12. adopt the olfactory analog instrument of claim 1 that fermentation tail gas is carried out online detection and analysis, it is characterized in that, may further comprise the steps:
(1) start preheating; Two-position two-way solenoid valve 105 and 107 is connected; Two-position three way magnetic valve 103 breaks off with two-position two-way solenoid valve 111; Under the swabbing action of minipump, surrounding air flows through gas sensor array annular working chamber through two-position three way magnetic valve 103 with the flow of 5000～6500 ml/min; Be 30 minutes preheating time.
(2) accurately demarcate; Two-position two-way solenoid valve 111 is connected; Two-position three way magnetic valve 103 breaks off with two-position two-way solenoid valve 105 and 107; Pure air oppositely flows into gas sensor array annular working chamber through throttling valve 110, two-position two-way solenoid valve 111 from venthole with 600 ml/min and above flow; Be discharged to the surrounding air through two-position three way magnetic valve 103 from air admission hole then, last 40 seconds; Thus, gas sensor array is able to accurate demarcation, makes the repeatedly measurement of fermentation tail gas is carried out on same benchmark;
(3) balance; Two-position three way magnetic valve 103 all breaks off with two-position two-way solenoid valve 105,107 and 111, and the indoor gas of gas sensor array annular working remains static, the balance period that gas sensor array got into for 5 seconds;
(4) tail gas sampling; Two-position three way magnetic valve 103 is connected with two-position two-way solenoid valve 105; Two-position two-way solenoid valve 107 and 111 breaks off; Minipump sucks gas sensor array annular working chamber with 600 ml/min and above flow via two-position three way magnetic valve 103 with fermentation tail gas; Make it to skim over gas sensor sensitive membrane surface, be discharged into the surrounding air from waste gas outlet through two-position two-way solenoid valve 105, throttling valve 106 and flowmeter then; When fermentation tail gas accumulated samples amount reached 300 milliliters, the tail gas sampling process finished, and lasts for 30 seconds;
(5) the preliminary recovery; After the tail gas sampling finishes; Two-position two-way solenoid valve 105 and 107 is connected; Two-position three way magnetic valve 103 breaks off with two-position two-way solenoid valve 111; Minipump sucks gas sensor array annular working chamber via two-position three way magnetic valve 103 from air admission hole with surrounding air with the flow of 5000～6500 ml/min, lasts 225 seconds; Gas sensor array is tentatively recovered under the effect of surrounding air;
(6) data acquisition; In fermentation tail gas was flowed through the process of gas sensor array annular working chamber, 16 gas sensors produced the responsive response of simulation, and these responsive responses are sent to computing machine through cable; Computing machine obtains 16 dimension word response vectors through 16 tunnel and 16 data collecting card; Become the cascade model analyzing and processing that neural network is formed when change neural network and modularization are non-when modularization; Provide fermentation tail gas bulk strength, key component classification and concentration estimated value within 10 seconds after data acquisition finishes; Provide the estimated value of concentration of substrate, cell concentration, production concentration, DO value, pH value, and show through display;
(7) remote data transmission; When a data gatherer process finished, computing machine passed to the maximal value and the minimum value of gas sensor array response on the Internet, realized the remote monitoring and the analysis of biological fermentation process;
(8) repeat (2)～(7), repeatedly sample;
Under the control of computing machine, online detection, prediction and the optimization of the whole process of biofermentation are accomplished by the olfactory analog instrument automatically; A complete gas sampling cycle is 300 seconds, and data acquisition time length wherein is 60 seconds.
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