CN2674449Y - Real-time on-line tester for live bacterial concentration in fermentation liquid - Google Patents
Real-time on-line tester for live bacterial concentration in fermentation liquid Download PDFInfo
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- CN2674449Y CN2674449Y CN 200420005114 CN200420005114U CN2674449Y CN 2674449 Y CN2674449 Y CN 2674449Y CN 200420005114 CN200420005114 CN 200420005114 CN 200420005114 U CN200420005114 U CN 200420005114U CN 2674449 Y CN2674449 Y CN 2674449Y
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Abstract
The utility model relates to a real-time on-line tester for live bacterial concentration in fermentation liquid, comprising an electrode capacitive sensor, a lock-in amplifier, an A/D converter and a computer. The capacitive sensor (2) comprises a coaxial column-shaped inner electrode (8), an outer electrode (9), the cover of polytetrafluoroethylene (12) and an electrode lead (11). The electrode is arranged on the cover of the polytetrafluoroethylene (12), and the electrode is penetrated through the cover of the connected with the polytetrafluoroethylene (12) so as to connect with the electrode lead (11), and a small hole is drilled on the outer electrode (9). A circular casing cap (10) is fixed on the lower part of the column-shaped inner electrode (8). The real-time on-line tester for live bacterial concentration in fermentation liquid has cheap cost and convenient and simple testing method. The testing result can reflect the growth state of the microbe in the fermentation production process exactly. The real-time on-line tester for live bacterial concentration in fermentation liquid is convenient for the popularization and the enforcement of the industry.
Description
One, technical field
The utility model relates to the detection of microorganism, is specially the device that biological fermentation process fermentation liquor viable bacteria concentration real-time online detects.
Two, background technology
Fermentation process is extremely complicated biochemical reaction process, and the factor that influences sweat is a lot, as the kind of the composition of fermentation liquor and concentration, temperature, pH value, stirring rate, viable bacteria and concentration or the like.In above-mentioned factor, viable bacteria concentration is most important technological parameter, because under certain condition, the yield of fermentation and viable bacteria concentration are closely related.Therefore, the real-time online of sweat viable bacteria concentration detect to grasp and the control biological fermentation process, hydrolysis and fermentation information, improve fermented quality, realize that the robotization of biofermentation is significant.
At present, the off-line measurement method of viable bacteria concentration has: dry weight method, optical densitometric method, cell counting, ultrasonic Method for Measuring.The dry weight method is based on the method for terrestrial gravitation, at first needs filtering fermentation liquor is used distilled water flushing, and it is dry to put into micro-wave oven and exsiccator then, and several times are done in weighing on electronic balance more continuously, average as measurement result.The dry weight method has many shortcomings, at first, generally contains multiple granular organism and inorganics in the biological fermentation process, and they mix with microorganism, is difficult to they and microorganism are separated.Secondly thalline of living and dead thalline are all influential to measurement result, are the thalline numbers of living and we wish to obtain.Though the dry weight method has these shortcomings owing to can not find other better reference method, so at present people still the dry weight method as a kind of reference method.Optical densitometric method (being OD) is another kind of reference method, when with the irradiate light fermentation liquor of certain wavelength, thalline and graininess suspension have reflection and scattering process to light, between reflected light or scattered light and the somatic cells quantity certain corresponding relation is arranged, can be made into the biomass detector according to this principle.But the reading of this quasi-instrument is subject to the influence of fermentation liquor color, turbidity and graininess suspension, can not get rid of the influence of dead thalline, and this quasi-instrument is difficult to be used for measuring mud shape fermentate.For number and the cell activity that detects viable bacteria, adopt cell counter usually or use methylene blue staining, count with microscopic examination then.But this method will be done repeatedly and dilute, observes counting, wastes time and energy very much.Ultrasonic Method for Measuring is that its amplitude can decay when utilizing ultrasound wave to pass through fermentation liquor, and velocity of propagation can change to some extent, and principle can be made into biomass and measures instrument in view of the above.But this quasi-instrument is subject to the influence of the variation of fermentation liquor composition, bubble, temperature variation and viscosity.
Said method is an off-line measurement, all will be from fermentation tank draw samples, off-line measurement brings two problems: bring assorted bacterium in the sampling process easily on the one hand and infect fermentation liquor in the fermentation tank; Off-line measurement has very big hysteresis quality on the other hand, brings difficulty for the automatic control of sweat.
The existence of microbial cell can influence the electrical feature of fermentation, there is the scholar to attempt to survey living bacterial cells concentration by impedance measurement, but the research of this method does not reach realistic scale, even if because under laboratory condition, the complicated variation also can appear in the impedance of fermentation liquor.When bigger variations took place for the composition of fermentation liquor, particularly during the bigger variation of fermentation liquor intermediate ion concentration generation, the accuracy of impedance measurement was very poor.
The scholar is also arranged according to the permittivity of fermentation liquor dependence for viable bacteria concentration and survey frequency, adopt the permittivity mensuration that the yeast viable cell concentrations is carried out real-time online and detect (the research of yeast concentration real-time on-line detecting method, Food Science, 1999 12 phases), detection system is made up of fermentor, platinum electrode, Hewlett-Packard's network analyzer, A/D converter, computing machine etc.Wherein network analyzer and platinum electrode cost height, be not easy to popularize.
Three, summary of the invention
The purpose of this utility model be to provide a kind of with low cost, technology simple, be convenient to the fermentation liquor viable bacteria concentration real time on-line detection device of industrial implementation.
The utility model provides a kind of design concept of fermentation liquor viable bacteria concentration real time on-line detection device as follows: because the existence of microorganism can influence the electrical characteristics of fermentation liquor, the electrical characteristics of fermentation liquor in radio-frequency band can characterize with permittivity (specific inductive capacity).The permittivity increment of fermentation liquor in radio-frequency band is the function of its survey frequency and viable bacteria concentration.This means and to measure viable bacteria concentration by the permittivity of measuring fermentation liquor.But in general, the permittivity of fermentation liquor can not directly record, and we adopt the method for measuring electric capacity to determine the permittivity of two electrode indirect fermentation liquid.If in fermentation liquor, place a pair of coaxle cylinder shape electrode, to establish the internal and external electrode radius and be respectively r, R, electrode length is l, then the electric permittivity epsilon of fermentation liquor
f, electrode capacitance C
fBetween following relation arranged:
Therefore, can determine the changes delta ε of fermentation liquor permittivity by the variation of potential electrode electric capacity
fIn the fermentation process, when living bacterial cells concentration changes, according to above-mentioned detection principle, the electric permittivity epsilon of fermentation liquor
fCan change, thereby the electric capacity on the capacitive transducer changes.Work as capacitor C
fWhen subtle change takes place, the applied signal voltage V of lock-in amplifier
iAmplitude and phase place generation respective change.Because input signal
(having ignored the input impedance that lock is put), wherein V
sBe the high frequency sinusoidal signal.Being locked on the reference signal frequency when the frequency input signal of lock-in amplifier is ω=ω
R, regulate V
iWith V
RBetween phase differential when equalling zero, lock-in amplifier output voltage amplitude V
o=KV
iWherein K is an enlargement factor, and after the lock-in amplifier parameter was selected, it was a constant.Therefore, final available output voltage changes the variation that reflects viable bacteria concentration.
The bio-fermented liquid viable bacteria concentration real time on-line detection device that the utility model provides, it is made up of electrode capacitance sensor, intermediate conversion device, A/D converter, computing machine, display and printer.
The effect of capacitive transducer is that non-electric charge quantity signalling (viable bacteria concentration) is converted to electric signal (electric capacity), since the detected output signal of capacitive transducer a little less than, so accurate to measure the small capacitance variable quantity that is caused by the viable bacteria concentration change delicately be the main task that the intermediate conversion device will solve under the capacitive.
The intermediate conversion device adopts lock-in amplifier to realize that lock-in amplifier is based on coherent detection technology, utilizes reference signal frequency relevant with frequency input signal, and is uncorrelated with noise frequency, thereby extracts useful signal from noise background.Sort circuit has extremely strong anti-interference, high sensitivity, very wide dynamic range, can be used as Detection of weak.The voltage signal of lock-in amplifier output is given computing machine and is carried out the data filtering processing after the A/D conversion, obtain required data.Data processing adopts software to finish, and used filtering method is an intermediate value average filter method.
Above-mentioned capacitive transducer is made of electrode, external electrode and contact conductor in teflon cover, the coaxial cylindrical; Electrode is installed in teflon and puts, and passes teflon cover and be connected with contact conductor; The coaxial cylindrical internal and external electrode is made by nontoxic, the harmless resistant to elevated temperatures molding powder insulating material of electrostatic spraying on the stainless-steel tube, below interior electrode, fix a circular plug, on external electrode, be drilled with aperture, make living bacterial cells pass through easily on the one hand, make the internal and external electrode area equate on the other hand as far as possible; Internal and external electrode is done respectively and is installed after insulation is handled again.Described intermediate conversion device is a lock-in amplifier, and lock-in amplifier is connected with capacitive transducer and function signal generator.
Compared with the prior art, the utlity model has following characteristics and effect: at first, the utility model intermediate conversion device adopts lock-in amplifier, nontoxic, the harmless resistant to elevated temperatures molding powder insulating material of electrostatic spraying is made on the electrode capacitance sensor employing stainless-steel tube, greatly reduce the cost of pick-up unit, detection method is simple and easy to do, is convenient to promotion and implementation.Secondly, detection method that the utility model provides and device can real-time online detection of biological sweat living bacterial cells concentration, testing result is not influenced by dead somatic cells, be not subjected to the influence that the fermentation liquor composition changes in the sweat yet, can accurately reflect microbial growth state in the fermentation production process.
Four, description of drawings
Fig. 1 is the utility model pick-up unit synoptic diagram
Fig. 2 is the utility model capacitor sensor structure synoptic diagram
Fig. 3 is the utility model viable bacteria concentration testing circuit figure
Fig. 4 is that the utility model detects the voltage curve of output: ordinate is represented voltage (volt), horizontal ordinate represent fermentation time (my god).
Fig. 5 is the different growth phase family curves of microbial cell: ordinate is represented the logarithm of viable count (solid line) and total bacteria count (dotted line), horizontal ordinate represent fermentation time (hour).
Five, embodiment
The utility model viable bacteria concentration detection apparatus as shown in Figure 1, it is made up of fermentation tank 1, capacitive transducer 2, intermediate conversion device 3, A/D converter 4, computing machine 5, display 6, printer 7.
The structure of capacitive transducer 2 as shown in Figure 2, it is made of electrode in the coaxial cylindrical 8, external electrode 9, teflon cover 12 and contact conductor 11; Electrode is installed on the teflon cover 12, and passes teflon cover 12 and contact conductor 11 connections mutually; Electrode 8, external electrode 9 are made by nontoxic, the harmless resistant to elevated temperatures molding powder insulating material of electrostatic spraying on the stainless-steel tube in the coaxial cylindrical, below interior electrode 8, fix a circular plug 10, on external electrode 9, be drilled with aperture, make living bacterial cells pass through easily on the one hand, make the internal and external electrode area equate on the other hand as far as possible.
Described intermediate conversion device 3 is a lock-in amplifier, and lock-in amplifier is connected with capacitive transducer 2 and function signal generator 13.The diameter of external electrode is 16mm, and inner electrode diameter is 6mm, and internal and external electrode thickness is 1mm, and electrode length is 135mm.
The connection of lock-in amplifier as shown in Figure 3, the output terminal of function generator 13 links to each other with a lead-in wire of capacitive transducer 2 by the first end of Q9, the other end links to each other with the reference-input signal end port2 of lock-in amplifier 3, another output lead of capacitive transducer 2 links to each other with the signal input part port1 of lock-in amplifier 3, and the ground end port4 of function generator 13 links to each other with the ground end port5 of lock-in amplifier 3; The output of lock-in amplifier 3 is connected with the input end of A/D converter 4, and the output of A/D converter 4 is inserted on the interface slot of computing machine 5 by 25 core cables.A/D converter 4 is selected 9014 mould incoming interface cards for use, and it comprises 12 A/D converters of a slice, multicircuit switch, sampling holder, bus interface controller etc., has address switch on the plate I/O address can be set arbitrarily.Conversion from the analog quantity to the digital quantity can followingly be tried to achieve:
V＝N×5000/2048-5000
Wherein V represents digital quantity, and N represents the corresponding simulating magnitude of voltage.
Described lock-in amplifier is selected the accurate lock-in amplifier of ND-201 type two-phase for use, and function generator 13 is selected 1163W type function generator for use.
Adjustment function generator 13, output frequency are that 50KHZ, amplitude are the sinusoidal signal of 1V; Selecting the sensitivity of lock-in amplifier 3 is 1200 * 120, and the cutoff frequency of bandpass filter is 25KHZ～62.5KHZ, and time constant is 300ms; Resistance R=25K Ω.
The experiment of viable bacteria concentration is as follows in employing the utility model pick-up unit real-time online detection fermentation liquor:
Used bacterial classification is a bread microzyme, the nutrient culture media yellow seriflux, experiment is carried out with one liter fermentor, 700 milliliters of nutrient solutions of interior Sheng, for making the yeast cells in the fermentor even, used magnetic stirring apparatus continuous stirring fermentation liquor, stirring rate is 140 rev/mins, the fermentation temperature basic controlling is at 28 ℃, and fermentating liquid PH value is controlled at about 5.0.
According to the detected experimental data of bread microzyme sweat real-time online, simulated fermentation different phase detection system output voltage curve (as shown in Figure 4), this magnitude of voltage has reflected the variation of viable bacteria concentration.This curve reflects several different phases of sweat living bacterial cells growth experience, and online test experience begins to carry out after inoculation, and this moment, detected output voltage values was less and remain unchanged substantially, illustrate that cell is grown to be in the lag phase; Fermentation is through after 1 day, and output voltage begins to increase, and illustrates that cell grows into increased logarithmic phase; Fermentation is through after 4 days, and output voltage reaches maximum and one period maximum plateau is arranged, and illustrates that cell grows into stationary phase; After entering the 5th day, detect output voltage and descend rapidly gradually, cell has grown into decline phase.Its different growth phase family curve with the sweat microbial cell (as shown in Figure 5) conforms to substantially, and the feasibility and the validity of this detection method has been described.
Owing to contain a small amount of lactic acid bacteria in the used nutrient culture media yellow seriflux in this fermenting experiment, and this detection method detected be the concentration of all living bacterial cells, this brings error to testing result, makes the viable yeast bacteria concentration testing result and the off-line cell counting measurement result of cell growth decline phase that some deviations be arranged.It is single culture that but the bacterial classification of general biofermentation requires, and does not allow other assorted bacterium and exists, and so just can eliminate this error.
Claims (3)
1, a kind of fermentation liquor viable bacteria concentration real time on-line detection device, it is made up of capacitive transducer (2), intermediate conversion device (3), A/D converter (4), computing machine (5), it is characterized in that described intermediate conversion device (3) is a lock-in amplifier, and adopt the reference-input signal of the output signal of function generator (13) as lock-in amplifier.
2,, it is characterized in that described capacitive transducer (2) is made of coaxial cylindrical internal and external electrode (8), cylindrical outer electrode (9), teflon cover (12) and contact conductor (11) according to the described a kind of fermentation liquor viable bacteria concentration real time on-line detection device of claim 1; Electrode is installed on the teflon cover (12), and passes teflon cover (12) and be connected with contact conductor (11); And on external electrode (9), being drilled with aperture, a circular plug (10) is fixed in electrode in column type (8) bottom.
3,, it is characterized in that electrode (8) in the described coaxial cylindrical, external electrode (9) made by spraying insulating material on the stainless-steel tube according to the described a kind of fermentation liquor viable bacteria concentration real time on-line detection device of claim 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200420005114 CN2674449Y (en) | 2004-02-11 | 2004-02-11 | Real-time on-line tester for live bacterial concentration in fermentation liquid |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200420005114 CN2674449Y (en) | 2004-02-11 | 2004-02-11 | Real-time on-line tester for live bacterial concentration in fermentation liquid |
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| Publication Number | Publication Date |
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| CN2674449Y true CN2674449Y (en) | 2005-01-26 |
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| CN 200420005114 Expired - Fee Related CN2674449Y (en) | 2004-02-11 | 2004-02-11 | Real-time on-line tester for live bacterial concentration in fermentation liquid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009140796A1 (en) * | 2008-05-23 | 2009-11-26 | 西门子公司 | A particle concentration measuring device and method |
| US20100276501A1 (en) * | 2008-01-10 | 2010-11-04 | Akita University | Temperature measuring method using temperature-sensitive magnetic substance and temperature controlling method |
-
2004
- 2004-02-11 CN CN 200420005114 patent/CN2674449Y/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100276501A1 (en) * | 2008-01-10 | 2010-11-04 | Akita University | Temperature measuring method using temperature-sensitive magnetic substance and temperature controlling method |
| US8801280B2 (en) * | 2008-01-10 | 2014-08-12 | Akita University | Temperature measuring method using temperature-sensitive magnetic substance and temperature controlling method |
| WO2009140796A1 (en) * | 2008-05-23 | 2009-11-26 | 西门子公司 | A particle concentration measuring device and method |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |