JPH0690161B2 - Method and apparatus for measuring concentration of analyte in solution or dispersion - Google Patents
Method and apparatus for measuring concentration of analyte in solution or dispersionInfo
- Publication number
- JPH0690161B2 JPH0690161B2 JP1224235A JP22423589A JPH0690161B2 JP H0690161 B2 JPH0690161 B2 JP H0690161B2 JP 1224235 A JP1224235 A JP 1224235A JP 22423589 A JP22423589 A JP 22423589A JP H0690161 B2 JPH0690161 B2 JP H0690161B2
- Authority
- JP
- Japan
- Prior art keywords
- solution
- temperature
- concentration
- dispersion
- sensor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 239000006185 dispersion Substances 0.000 title claims description 41
- 238000000034 method Methods 0.000 title claims description 26
- 239000012491 analyte Substances 0.000 title claims description 15
- 244000005700 microbiome Species 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 24
- 238000005259 measurement Methods 0.000 claims description 22
- 230000020169 heat generation Effects 0.000 claims description 16
- 230000035945 sensitivity Effects 0.000 claims description 7
- 239000012530 fluid Substances 0.000 description 21
- 210000004027 cell Anatomy 0.000 description 12
- 238000010438 heat treatment Methods 0.000 description 12
- 238000000691 measurement method Methods 0.000 description 7
- 238000012546 transfer Methods 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 230000001276 controlling effect Effects 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000004140 cleaning Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000005611 electricity Effects 0.000 description 2
- 238000002848 electrochemical method Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 229940039696 lactobacillus Drugs 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 239000013307 optical fiber Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- WABPQHHGFIMREM-UHFFFAOYSA-N lead(0) Chemical group [Pb] WABPQHHGFIMREM-UHFFFAOYSA-N 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004848 nephelometry Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000009931 pascalization Methods 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analyzing Materials Using Thermal Means (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は食品化学における微生物もしくはそれらの生成
物の濃度測定に利用される、被検体の濃度測定方法とそ
の装置に関する。Description: TECHNICAL FIELD The present invention relates to a method for measuring the concentration of an analyte, which is used for measuring the concentration of microorganisms or their products in food chemistry, and an apparatus therefor.
各種微生物の培養は、微生物の大量増殖自体を目的とす
るもの、微生物の生成物を得ることを目的とする等の目
的によって行われているが、培養工程中に微生物または
それらの生成物の濃度を知ることは、培養の管理上栄養
供給の管理や溶液中の許容濃度を管理する上で重要な問
題である。Cultivation of various microorganisms is carried out for the purpose of mass growth itself of microorganisms, the purpose of obtaining products of microorganisms, etc., but the concentration of microorganisms or their products during the culture process It is an important problem to control the nutrient supply and the permissible concentration in the solution for controlling the culture.
従来これらの濃度を知るために、槽より一部の溶液を取
り出して種々の方法で濃度測定していたが近年培養の効
率化のため、連続的かつ経時的でさらにインラインでの
無菌的測定が行なえる方法と装置が望まれている。Conventionally, in order to know these concentrations, a part of the solution was taken out of the tank and the concentration was measured by various methods, but in recent years, in order to improve the efficiency of culture, continuous and time-lapse, and in-line aseptic measurement is possible. What is needed is a method and apparatus that can be done.
（従来の技術） 微生物の濃度を測定する方法は、菌体の乾燥重量を求め
る方法、比濁測定法、菌体数測定法等の基本的測定法か
ら、光学的測定法、反応測定法、電気化学的測定法など
が提案されている。(Prior Art) Methods for measuring the concentration of microorganisms include basic measurement methods such as a method for obtaining a dry weight of cells, a nephelometry, a cell number measurement method, an optical measurement method and a reaction measurement method. Electrochemical measurement methods have been proposed.
基本的測定法は測定時間がかかることや、労力を多大に
必要とすること等の問題から、後記したようなインライ
ンでの機器計測的な方法が提案されてきた。Since the basic measurement method has such problems that it takes a long measurement time and requires a lot of labor, an in-line instrument measurement method as described later has been proposed.
投光器と受光器がセンサーに組み込まれた光学的装置
による測定方法。A measuring method using an optical device in which a sender and a receiver are incorporated in a sensor.
実開昭62-16457号のものは、受光量の変化を受光器で電
気量の変化に転換し演算処理して被検体の濃度を測定す
る装置で、被検体の流れを妨害せず小型で、かつ高温高
圧下でも使用可能である。No. 62-16457 is a device that converts the change in the amount of received light into a change in the amount of electricity with a light receiver and performs arithmetic processing to measure the concentration of the sample, which is compact and does not interfere with the flow of the sample. Also, it can be used under high temperature and high pressure.
特開昭51-49787号のものは、培養基中で微生物等が光学
繊維を介して光の透過量を測定することで濃度を測定す
るとともに、紫外線を通して壁面付着する微生物を殺す
ようにしたものである。また、光学繊維を容器内に収納
して開閉蓋を取付け他の光や泡の影響を防止した手段で
ある。In JP-A-51-49787, the concentration of microorganisms in a culture medium is measured by measuring the amount of light transmitted through the optical fiber, and the microorganisms attached to the wall surface are killed by ultraviolet rays. is there. Further, it is a means for storing the optical fiber in a container and attaching an opening / closing lid to prevent the influence of other light and bubbles.
反応測定方法 特開昭62-64934号のものは、電極表面に抗体を固定化し
た水晶振動子バイオセンサーを作成し、微生物の検知及
び濃度の測定を行う手段である。Reaction measuring method The method disclosed in JP-A-62-64934 is a means for producing a quartz oscillator biosensor in which an antibody is immobilized on the electrode surface, and detecting a microorganism and measuring its concentration.
特開昭50-36198号のものは、プローブに微生物又は酵素
を被覆させた感温装置にして、微生物または酵素の基質
である分子の濃度を測定する手段である。Japanese Patent Application Laid-Open No. 50-36198 is a means for measuring the concentration of a molecule which is a substrate of a microorganism or an enzyme by using a temperature-sensitive device in which a probe is coated with the microorganism or the enzyme.
電気化学的測定法 特開昭60-135754号のものは、溶媒が流れる通路におけ
る作用極配置部分と対極配置部分の太さが可変で、両極
間で発生する電気量の変化により被測定物の濃度を測定
する手段である。The electrochemical measurement method disclosed in JP-A-60-135754 is that the thickness of the working electrode arrangement portion and the counter electrode arrangement portion in the passage through which the solvent flows is variable, and the object to be measured is changed by the change in the amount of electricity generated between both electrodes. It is a means for measuring the concentration.
特開昭59-81551号のものは、生細胞が電極に直接接触す
ると電流が発生することから、細胞懸濁液に一対の電極
を設け、周期的電位を印加して生起する電流値を測定
し、その細胞数を測定する手段である。In Japanese Patent Laid-Open No. 59-81551, a current is generated when a living cell comes into direct contact with the electrode, so a pair of electrodes is provided in the cell suspension and a periodic potential is applied to measure the current value generated. Then, it is a means for measuring the number of cells.
特開昭61-48755号のものは、基準濃度培養液中の一対の
電極と培養液流路の一対の電極を含む交流ブリッジ回路
の不平衡出力を位相検波する位相検波回路とを具備した
培養液の電気伝導度を測定することによって、培養液の
濃度を最適に保つ手段である。Japanese Patent Laid-Open No. 61-48755 discloses a culture provided with a phase detection circuit for phase-detecting an unbalanced output of an AC bridge circuit including a pair of electrodes in a culture medium with a reference concentration and a pair of electrodes in a culture fluid channel. It is a means for keeping the concentration of the culture solution optimal by measuring the electrical conductivity of the solution.
（従来技術の問題点） 従来実施されてきた基本的測定法は測定時間や労力に無
駄が多く、かつ被検体の汚染防止上から密閉系培養機器
では利用できるものが少ない。(Problems of the prior art) The basic measurement methods that have been conventionally used are wasteful in measurement time and labor, and few can be used in closed culture devices in order to prevent contamination of the analyte.
滅菌処理される密閉系培養機器ではインラインでの計測
が実施されるが、従来技術に記載した光学的測定機器で
は、センサーへの微生物の付着を完全に防止することが
できず、特に光の発光及び受光器面に付着が進行すると
光量に影響がでて測定能力が低下することを避けられな
いうえ、培地に色がある場合や培養時間の経過とともに
色がつきだす場合なども光の透過に影響を与えるし、ま
た培養槽に設けられた液面窓や照明等による外光が発光
器の光量に影響を与え、測定誤差の原因となることもあ
る。In-line measurement is performed with a closed culture device that is sterilized, but the optical measurement device described in the prior art cannot completely prevent microorganisms from adhering to the sensor. In addition, it is unavoidable that the adhering to the surface of the light receiver will affect the amount of light and reduce the measuring ability.In addition, the light transmission will not be possible even if the medium has color or if color develops over time. In addition, the external light from the liquid level window provided in the culture tank or the illumination may affect the light amount of the light emitter, which may cause a measurement error.
さらに培地の粘度が高い場合などは、受光器・発光器部
分の特異的構造から発光・受光表面に培地がまとわりつ
き、常に変化している培地の状態が正確につかめないこ
とや、培地や菌体の高濃度部分では測定誤差が大きくな
り利用できないという問題がある。In addition, when the viscosity of the medium is high, the medium is clinging to the light-emitting / light-receiving surface due to the specific structure of the light-receiving / light-emitting portion, and the constantly changing state of the medium cannot be accurately grasped. There is a problem that the measurement error becomes large in the high-concentration area of and cannot be used.
反応測定機器では、センサー表面の固定化担体の破壊や
微生物、酵素等の剥離を防止するため、高温高圧処理に
よる滅菌が不可能であるとともに、機器の振動なども避
ける必要がある。また、センサーに固定化した抗体によ
って結合された微生物は生産に利用されず無駄である
し、センサーの再使用には洗浄と抗体固定化などのセン
サー能力復帰操作が必要となる。In a reaction measuring device, in order to prevent destruction of the immobilized carrier on the sensor surface and peeling of microorganisms, enzymes and the like, it is not possible to sterilize by high temperature and high pressure processing, and it is necessary to avoid vibration of the device. In addition, the microorganisms bound by the antibody immobilized on the sensor are not used for production and are wasteful, and the reuse of the sensor requires washing and antibody immobilization operations such as antibody immobilization.
さらに本機器では反応体の反応もしくは反応による触媒
作用を必要とする酵素・微生物に利用がかぎられるとと
もに、菌体が高濃度になると触媒作用に限界があり、測
定範囲も限られてしまうという問題がある。Furthermore, this device can be used only for enzymes and microorganisms that require the reaction of the reactants or the catalytic action of the reaction, and at high concentrations of bacteria, the catalytic action is limited and the measurement range is also limited. There is.
電気化学的測定機器の問題としては、電極へ酵素、微生
物等が付着すると電気障害が生じ測定誤差の原因となる
ことや、電極を洗浄後で洗浄する場合、洗浄液の種類は
腐食等の問題から限定され、装置に装着したままで洗浄
すると洗浄剤が制約されて機器の洗浄不良の原因とな
り、滅菌維持による洗浄をして長時間の培養を行うシス
テムには適しない。Electrochemical measuring equipment has a problem that if an enzyme or microorganism adheres to the electrode, it may cause an electrical error and cause a measurement error. It is limited, and if it is washed while it is attached to the device, the cleaning agent is limited, which causes poor cleaning of the device, and is not suitable for a system in which cleaning is performed by maintaining sterilization and long-term culture is performed.
このように従来技術にはそれぞれ独特の問題があるが、
共通する問題として、培地に気泡が生じることは培養工
程において避けられないことであるが、この気泡がセン
サーに付着することによって検知能力を低下させたり測
定を阻害したりする。特に電極を用いるものは、気泡が
電極に付着することによって電食の原因となることもあ
る。As described above, each of the conventional techniques has its own problems,
A common problem is that air bubbles are unavoidable in the culture process in the culture process, but the air bubbles adhere to the sensor to reduce the detection ability or hinder the measurement. In particular, in the case of using an electrode, bubbles may adhere to the electrode to cause electrolytic corrosion.
本発明は以上の従来技術の問題点を解決するべく金属細
線加熱法を用いた、微生物などの濃度を測定する方法と
装置を提供するものである。The present invention provides a method and an apparatus for measuring the concentration of microorganisms and the like using a metal wire heating method in order to solve the above problems of the prior art.
（課題を解決するための手段） しかして、本発明は以下のごとく構成するものである。(Means for Solving the Problem) The present invention is configured as follows.
溶液もしくは分散液中に発熱可能な素子を内蔵する発熱
センサーを設置して、該センサーを発熱させたときの発
熱センサーの温度、もくはそのときの発熱センサーの温
度と溶液もしくは分散液の温度との差を測定することに
より、溶液もしくは分散液中の被検体の濃度を測定する
方法であって、このとき発熱センサーは一定電流もしく
は一定の発熱量になるよう、電流を制御することで発熱
させ、測定中における発熱している該センサーの温度も
しくは該センサーの温度と溶液もしくは分散液の温度と
の差の変化に追従した溶液もしくは分散液中の被検体の
濃度を測定する方法である。The temperature of the heat generating sensor when the heat generating sensor having a heat generating element is installed in the solution or the dispersion, and the temperature of the heat generating sensor when the sensor is heated, or the temperature of the heat generating sensor at that time and the temperature of the solution or the dispersion liquid It is a method to measure the concentration of the analyte in the solution or dispersion by measuring the difference between the heat generation sensor and the heat generation sensor. Then, the concentration of the analyte in the solution or dispersion is measured in accordance with a change in the temperature of the sensor that is generating heat during the measurement or a difference between the temperature of the sensor and the temperature of the solution or the dispersion.
さらに、培養槽内における流体の乱流などによる測定障
害を避けるため、溶液もしくは分散液の循環ラインを構
成し、該循環ライン内に加熱センサーを配置することを
特徴とする方法である。Furthermore, in order to avoid a measurement obstacle due to a turbulent flow of a fluid in the culture tank, a solution or dispersion liquid circulation line is configured, and a heating sensor is arranged in the circulation line.
なお、循環ラインの液流速は一定流速であることが望ま
しく、流速を測定して測定値の補正をしないかぎりは、
0.01〜1.0m/sの範囲で行なうことによって広域における
濃度測定が可能であるが、感度を高く要求されるときは
低流速で、ノイズを小さく要求される時は高流速でとい
うように、流速を多段階に変化させて測定する方法を用
いてもよい。In addition, it is desirable that the liquid flow velocity in the circulation line be a constant flow velocity, and unless the flow velocity is measured and the measured value is corrected,
The concentration can be measured in a wide range by performing the measurement in the range of 0.01 to 1.0 m / s, but when the sensitivity is required to be high, the flow velocity is low, and when the noise is required to be low, the flow velocity is high. May be measured in various stages.
さらにこの方法を実現するには、溶液もしくは分散液槽
から分岐して一定流速に制御されるポンプを介して前記
槽に戻る溶液もしくは分散液の循環ラインを構成し、前
記槽内もしくは前記循環ライン内において溶液もしくは
分散液の温度を測定するセンサーと前記循環ライン内に
設けた発熱センサーを発熱させたときの該センサーの温
度を測定することにより、溶液もしくは分散液中の被検
体の濃度を測定する装置であって、特には溶液もしくは
分散液中の被検体の濃度の変化とともに変化するセンサ
ーの感度やノイズの状況にあわせて、感度を高く要求さ
れるときは低流速で、ノイズを小さく要求されるときは
高流速でというように、流速を多段階に制御する溶液も
しくは分散液中の被検体の濃度を測定する装置である。In order to realize this method, a solution or dispersion liquid circulation line that branches from the solution or dispersion liquid tank and returns to the tank via a pump controlled at a constant flow rate is formed in the tank or the circulation line. A sensor for measuring the temperature of the solution or the dispersion liquid in the chamber and the temperature of the sensor when the heat generation sensor provided in the circulation line is heated to measure the concentration of the analyte in the solution or the dispersion liquid. In particular, the device requires a low flow rate and low noise when high sensitivity is required in accordance with the sensor sensitivity and noise conditions that change with changes in the concentration of the analyte in the solution or dispersion. It is an apparatus for measuring the concentration of an analyte in a solution or dispersion liquid in which the flow velocity is controlled in multiple stages, such as when the flow velocity is high.
（作用） 本発明は流体のみかけの粘性変化を測定する金属細線加
熱法を用いて、流体内の微生物や微生物の生産物の濃度
を測定しようとするものであって、本出願人が先に出願
した特開昭62-185146号「流体の状態の計測方法」によ
れば、流体の状態変化は、流体のみかけの粘性変化によ
り知りえるが、培養における生産物や菌体の濃度は濃度
の上昇とともに、みかけの粘性が変化することにより同
じく知ることができる。(Operation) The present invention is intended to measure the concentration of microorganisms or products of microorganisms in a fluid by using a metal thin wire heating method for measuring the apparent viscosity change of the fluid. According to the Japanese Patent Application Laid-Open No. 62-185146, “Method of measuring the state of fluid”, the change in the state of the fluid can be known by the apparent viscosity change of the fluid. This can also be seen by the apparent viscosity changing with rising.
例えば熱の伝達状況を表す熱伝達率αは下式で与えられ
る。For example, the heat transfer coefficient α representing the heat transfer state is given by the following equation.
α＝Q/S（θs−θ∞） Q:発熱量 S:センサー表面積 θs：センサー表面温度 θ∞：流体温度 すなわち、結局は発熱体と流体の温度差と菌体の温度と
は特定の関係がある。α = Q / S (θ s −θ ∞) Q: Calorific value S: Sensor surface area θ s : Sensor surface temperature θ ∞: Fluid temperature In other words, the temperature difference between the heating element and the fluid and the temperature of the fungus body are ultimately specified. Have a relationship.
従って、結果的に流体の温度変化が小さく、発熱量が一
定とみなせる時は発熱体の温度もしくは発熱体と流体の
温度差を経時的に計測し、その変化を計測することで流
体の濃度と関連させ、濃度を測定することが可能とな
る。また、流体の温度が大きく変化する時で、発熱セン
サーが電流で制御されるときは、 Ｑ＝Ｒ・i2 から、抵抗Ｒが温度変化により変化して発熱量が変化
し、熱伝達率が流体の温度変化のみで変化してしまうた
め、電流ｉを発熱量が一定となるよう制御して発熱体の
温度をモニターすればよい。Therefore, when the temperature change of the fluid is small as a result and the calorific value can be regarded as constant, the temperature of the heating element or the temperature difference between the heating element and the fluid is measured over time, and the change is measured to determine the concentration of the fluid. It is possible to correlate and measure the concentration. Also, when the temperature of the fluid changes significantly and the heat generation sensor is controlled by the current, from Q = R · i 2 , the resistance R changes due to the temperature change, the heat generation amount changes, and the heat transfer coefficient changes. Since it changes only by the temperature change of the fluid, the temperature of the heating element may be monitored by controlling the current i so that the calorific value is constant.
ちなみに、上記センサーの表面温度（θs）の測定方法
は本出願人が先に出願した特願昭62-51520号（特開昭63
-217261号公報）を利用し、発熱センサー温度（θw）か
ら容易に算出することができる。こうして知りえた熱伝
達率と溶液もしくは分散液中の被検体の濃度とが相関す
る関係において、熱伝達率の変化から濃度変化を数値的
に測定することができ、上式よりセンサー表面温度と流
体温度の差の変化を計測することとなるが、センサー表
面温度は発熱体の温度から算出されることから、発熱体
の温度変化もしくは発熱体の温度と流体の温度との差の
変化と、濃度の相関を見ればよいこととなる。By the way, the method for measuring the surface temperature (θ s ) of the sensor is described in Japanese Patent Application No. 62-51520 filed by the applicant of the present application (Japanese Patent Application Laid-Open No. 63-51520).
No. 217261), it can be easily calculated from the heat generation sensor temperature (θ w ). In the relationship where the heat transfer coefficient thus found and the concentration of the analyte in the solution or dispersion correlate, it is possible to numerically measure the change in the concentration from the change in the heat transfer coefficient. The change in temperature difference is measured, but since the sensor surface temperature is calculated from the temperature of the heating element, the temperature change of the heating element or the change in the difference between the temperature of the heating element and the temperature of the fluid, and the concentration It is only necessary to look at the correlation of.
（実施例） 以下本発明の実施例を説明する。(Examples) Examples of the present invention will be described below.
第１図はこの発明の方法を実施する濃度測定装置の構成
説明図である。図に於いて、微生物等の分散液（１）を
攪拌羽根（３）を装着した容器（２）の中に入れ、配管
（４）を通じてポンプ（５）で一定流速で循環させる。
循環ラインで、流体の温度を計測するセンサー（６）と
発熱センサー（７）を配管（４）中に配置し、発熱セン
サー（７）の温度と流体温度との差を計測する。FIG. 1 is a structural explanatory view of a concentration measuring apparatus for carrying out the method of the present invention. In the figure, a dispersion liquid (1) of microorganisms or the like is placed in a container (2) equipped with a stirring blade (3) and circulated at a constant flow rate by a pump (5) through a pipe (4).
A sensor (6) for measuring the temperature of the fluid and a heat generation sensor (7) are arranged in the pipe (4) in the circulation line, and the difference between the temperature of the heat generation sensor (7) and the fluid temperature is measured.
（８）は電流源、（９）は電圧計、（10）は制御装置で
あり、これらはGPIB〔ゼネラルインターフェースバス〕
（11）で連絡されている。（12）は各センサー（６）
（７）と電流源（８）及び電圧計（９）を結ぶリード線
である。(8) is a current source, (9) is a voltmeter, (10) is a controller, and these are GPIB (General Interface Bus).
Contacted at (11). (12) is each sensor (6)
It is a lead wire connecting (7) with the current source (8) and the voltmeter (9).
循環ラインにおいて一定流速で循環させるのは、コンピ
ューター等でポンプ（５）の出力を制御することにより
容易に行われる。Circulation at a constant flow rate in the circulation line is easily performed by controlling the output of the pump (5) with a computer or the like.
溶液もしくは分散液の流速は一般に0.01〜1.0m/sが測定
上、広範囲の濃度流体に対処できる巾であるが、流体濃
度が非常に低い時や逆に高い時は、流速を前記以外の巾
においても多段階に制御することで測定は可能である。The flow velocity of the solution or dispersion is generally 0.01 to 1.0 m / s, which is wide enough to handle a wide range of concentration fluids, but when the fluid concentration is extremely low or conversely high, the flow velocity should be within the range other than the above. In the case of, measurement is possible by controlling in multiple stages.
ここにおいて流速は段階的に変更するのであって、無段
階的ではない。これは、加熱センサー周囲流速が測定中
は一定であることが望ましいからである。Here, the flow velocity is changed stepwise and is not stepless. This is because it is desirable that the flow rate around the heating sensor be constant during the measurement.
配管（４）内において発熱センサー（７）の設置形態は
垂直でもよいし水平でもよく、また精度よく温度測定す
るためには各センサー（６）（７）は白金抵抗体で構成
されたものを用いるのが好ましいが、精度よく温度が測
定されればその手段は問わない。The heat generating sensor (7) may be installed vertically or horizontally in the pipe (4), and each sensor (6) (7) should be composed of a platinum resistor in order to measure the temperature accurately. It is preferably used, but any means can be used as long as the temperature can be measured accurately.
そして各センサー（６）（７）に印加した電圧Ｖと電流
ｉを計測して、その抵抗値Ｒを調べ、センサーの温度θ
wを関係式 θ＝（R/Ro−１）／α Ro:0℃におけるセンサーの抵抗値 α：電気抵抗の温度係数 を用いて求める。Then, the voltage V and the current i applied to each sensor (6) (7) are measured, the resistance value R thereof is checked, and the temperature θ of the sensor is measured.
w is calculated using the relational expression θ = (R / R o −1) / α R o : 0 sensor resistance value α: temperature coefficient of electrical resistance.
各センサー（６）（７）には安定した電流を通電するよ
うにし、流体温度を測るセンサー（６）には、発熱を生
じぬよう微少電流を供給する。A stable current is applied to each of the sensors (6) and (7), and a minute current is supplied to the sensor (6) for measuring the fluid temperature so as not to generate heat.
以上のようにして行った実験例を以下に示す。An example of the experiment conducted as described above is shown below.
先ず乳酸桿菌菌体の濃度を測定結果を示す。容器（２）
内で乳酸桿菌を所定菌体濃度になるように水に分散し、
分散液（１）の液温を35℃とした。攪拌羽根（３）を用
い回転数250rpmで菌体を均一に分散しつつ、ポンプ
（５）で流速0.3m/sになるよう分散液（４）をラインに
循環させた。この時、発熱センサー（７）には0.3Aの直
流電流を供給し、発熱センサー（７）内部の白金線温度
（θw）と液体温度（θ∞）の温度差（θw−θ∞）を計
測した。この温度差と菌体濃度との関係を第２図に示
す。この図より、乾燥重量で０〜25g/lの領域でその温
度差から容易に菌体濃度が予測されることがわかる。First, the measurement results of the concentration of Lactobacillus cells are shown. Container (2)
Disperse the lactobacillus in water so that the concentration of cells will be the specified concentration.
The liquid temperature of the dispersion liquid (1) was set to 35 ° C. The dispersion liquid (4) was circulated through the line at a flow rate of 0.3 m / s by a pump (5) while uniformly dispersing the bacterial cells at a rotation speed of 250 rpm using a stirring blade (3). At this time, a DC current of 0.3 A is supplied to the heat generation sensor (7), and the temperature difference (θ w −θ ∞) between the platinum wire temperature (θ w ) inside the heat generation sensor (7) and the liquid temperature (θ ∞). Was measured. The relationship between this temperature difference and the bacterial cell concentration is shown in FIG. From this figure, it is understood that the cell concentration can be easily predicted from the temperature difference in the dry weight range of 0 to 25 g / l.
なお、センサー表面温度（θs）と流体の温度（θ∞）
の差（θs−θ∞）でも同じ結果を得られるが、この時
の（θs）は作用で記載したように、本出願人の先の出
願特願昭62-51520号によって算出し、（θs−θ∞）と
濃度との相関を取って利用すれば良い。The sensor surface temperature (θ s ) and fluid temperature (θ ∞)
Although the same result can be obtained with the difference of (θ s −θ ∞), (θ s ) at this time is calculated by the applicant's earlier application Japanese Patent Application No. 62-51520 as described in the action, The correlation between (θ s −θ ∞) and the concentration may be used.
実施例では、発熱体温度（θw）と流体温度（θ∞）の
差（θw−θ∞）と濃度との相関を示すものである。In the example, the correlation between the difference (θ w −θ ∞) between the heating element temperature (θ w ) and the fluid temperature (θ ∞) and the concentration is shown.
次に酵母菌体の濃度の測定結果を示す。Next, the measurement results of the yeast cell concentration are shown.
酵母を所定濃度になるように水に分散し、分散液（１）
を25℃とした。攪拌条件を250rpmとし、均一分散させつ
つポンプ（５）で0.5m/sになるよう分散液（１）を循環
させた。発熱サンサー（７）には0.5Aの直流電流を流し
発熱した。この時の発熱サンサー（７）内部の白金線温
度（θw）と液体温度（θ∞）との温度差（θw−θ∞）
を計測した。この温度と酵母菌体濃度との関係を第３図
に示す。この図より酵母菌体濃度は、乾燥重量160g/lも
の高濃度領域でもこの温度差から容易に精度よく測定で
きることが判明した。Disperse the yeast in water to a specified concentration to obtain a dispersion (1)
Was set to 25 ° C. The stirring condition was 250 rpm, and the dispersion liquid (1) was circulated by the pump (5) at 0.5 m / s while uniformly dispersing. A direct current of 0.5 A was applied to the heat generation sensor (7) to generate heat. At this time, the temperature difference (θ w −θ ∞) between the temperature of the platinum wire (θ w ) inside the heat generation sensor (7) and the liquid temperature (θ ∞)
Was measured. The relationship between this temperature and the yeast cell concentration is shown in FIG. From this figure, it was found that the yeast cell concentration can be easily and accurately measured from this temperature difference even in the high concentration range of 160 g / l dry weight.
以上のように、微生物の濃度もしくは微生物の生産物の
濃度を測定する時の電流制御範囲は、0.3〜0.9Aが本実
験では適当であった。しかし、センサーの設計によりこ
の値は変化し一定のものではない。また実験例では温度
差と菌体濃度の例をあげたが、溶液もしくは分散液の温
度が一定である時は、発熱センサー（７）の温度のみを
計測してもよいし、また発熱センサー（７）の温度と分
散液（１）の温度から熱伝達率αを計算し、この熱伝達
率の変化を計測して濃度と関連ずけてもよいこととな
る。As described above, the current control range for measuring the concentration of the microorganism or the concentration of the product of the microorganism was 0.3 to 0.9 A, which was appropriate in this experiment. However, this value changes due to the design of the sensor and is not constant. In the experimental example, the temperature difference and the bacterial cell concentration are given as examples. However, when the temperature of the solution or dispersion is constant, only the temperature of the heat generation sensor (7) may be measured, or the heat generation sensor ( The heat transfer coefficient α may be calculated from the temperature of 7) and the temperature of the dispersion liquid (1), and the change in this heat transfer coefficient may be measured and correlated with the concentration.
また濃度を測定するとき、本発明の測定方法では循環ラ
インの溶液もしくは分散液の流速を低くすると感度があ
がるがノイズが多くなり、逆に流速を高くするとノイズ
は少ないが感度が鈍るという傾向があるため、感度を高
く要求されるときは流速を高く、ノイズを少なくしたい
ときは流速を低くするというように、流速を多段階に制
御して測定することによって、巾の広い濃度領域におけ
る測定を可能とすることができる。Further, when measuring the concentration, in the measuring method of the present invention, when the flow rate of the solution or dispersion in the circulation line is lowered, the sensitivity is increased but the noise is increased. Conversely, when the flow rate is increased, the noise is decreased but the sensitivity is decreased. Therefore, by controlling the flow rate in multiple stages, such as increasing the flow rate when high sensitivity is required and decreasing the flow rate when reducing noise, measurement in a wide concentration range can be performed. It can be possible.
またこの制御を付加させることによって、多種類の溶液
もしくは分散液の測定に応用することができ、従来のご
とく色々の測定機器を測定目的にあわせて選択するとい
うわずらわしさを解消することができる。Further, by adding this control, it can be applied to the measurement of many kinds of solutions or dispersions, and the troublesomeness of conventionally selecting various measuring instruments according to the measuring purpose can be eliminated.
（発明の効果） 本発明によれば、以下の効果を奏することができる。(Effects of the Invention) According to the present invention, the following effects can be achieved.
・ライン洗浄が可能で無菌維持による洗浄や長時間の培
養などが可能である。・ Line cleaning is possible, and cleaning by maintaining sterility and long-term culture are possible.
・手洗い洗浄等をしてもセンサーを損傷することがな
い。-The sensor will not be damaged even if it is washed by hand.
・高温高圧での滅菌処理が可能である。すなわち培養ラ
インの無菌化を容易に実現できる。・ Sterilization at high temperature and high pressure is possible. That is, sterilization of the culture line can be easily realized.
・溶液もしくは分散液の色や濃度に影響されずに測定が
可能で、長時間にわたって菌体濃度の管理が可能にな
る。従って培養システムの管理が容易になる。・ Measurement is possible without being affected by the color or concentration of the solution or dispersion, and it is possible to control the cell concentration over a long period of time. Therefore, the management of the culture system becomes easy.
・センサー表面の仕上げ加工が容易で、微生物の付着を
極力小さくできる。また、溶媒もしくは分散液の泡発生
による影響も小さくなる。-The sensor surface is easy to finish, and the adhesion of microorganisms can be minimized. Further, the influence of bubbles generated in the solvent or the dispersion liquid is also reduced.
・菌体の高濃度領域においても計測が可能。・ Measurement is possible even in the high concentration area of bacterial cells.
・メンテナンスが容易である。・ Easy maintenance.
第１図は本発明の説明図 第２、３図は温度差と濃度の関係を示すグラフである。 FIG. 1 is an explanatory diagram of the present invention. FIGS. 2 and 3 are graphs showing the relationship between temperature difference and concentration.
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭59−84145（ＪＰ，Ａ) ─────────────────────────────────────────────────── ─── Continuation of front page (56) References JP-A-59-84145 (JP, A)
Claims (5)
内蔵するセンサーを設け、該センサーを一定電流、もし
くは一定の発熱量となるよう電流を制御して発熱させた
ときの発熱センサーの温度、もしくはそのときの発熱セ
ンサーの温度と溶液もしくは分散液の温度との差を測定
することにより、溶液もしくは分散液中の被検体の濃度
を測定する方法。1. A temperature of a heat-generating sensor when a sensor having a heat-generating element is provided in a solution or a dispersion and the sensor is heated at a constant current or by controlling the current so that a constant amount of heat is generated. Alternatively, a method of measuring the concentration of the analyte in the solution or dispersion by measuring the difference between the temperature of the exothermic sensor and the temperature of the solution or dispersion at that time.
しくは微生物の生産物であることを特徴とする、請求項
（１）に記載の溶液もしくは分散液中の被検体の濃度を
測定する方法。2. The method for measuring the concentration of an analyte in a solution or dispersion according to claim 1, wherein the analyte in the solution or dispersion is a microorganism or a product of the microorganism. .
し、該循環ライン内に発熱センサーを配置すると共に、
循環ラインの流速が0.01〜1.0m/sであることを特徴とす
る請求項（１）〜（２）いずれかに記載の溶液もしくは
分散液中の被検体の濃度を測定する方法。3. A solution or dispersion liquid circulation line is formed, and a heat generation sensor is arranged in the circulation line, and
The method for measuring the concentration of an analyte in a solution or dispersion according to any one of claims (1) and (2), characterized in that the flow rate of the circulation line is 0.01 to 1.0 m / s.
速に制御されるポンプを介して前記槽に戻る溶液もしく
は分散液の循環ラインを構成し、前記槽内もしくは前記
循環ライン内において溶液もしくは分散液の温度を測定
するセンサーと、前記循環ライン内に設けた発熱センサ
ーを発熱させたときの該センサーの温度を測定すること
により溶液もしくは分散液中の被検体の濃度を測定する
装置。4. A solution or dispersion liquid circulation line is branched from the solution or dispersion liquid tank and returned to the tank via a pump controlled at a constant flow rate, and the solution or dispersion liquid is circulated in the tank or in the circulation line. A device for measuring the temperature of the dispersion liquid, and a device for measuring the concentration of the analyte in the solution or dispersion liquid by measuring the temperature of the sensor when the heat generation sensor provided in the circulation line is heated.
度を高く要求されるときは低流速で、ノイズを少なく要
求される時は高流速でというように、溶液もしくは分散
液の流速を多段階に制御することを特徴とした請求項
（４）に記載の溶液もしくは分散液中の被検体の濃度を
測定する装置。5. A multi-step flow rate of a solution or dispersion, such as a low flow rate when high measurement sensitivity is required when circulating a solution or dispersion, and a high flow rate when low noise is required. The device for measuring the concentration of an analyte in a solution or dispersion according to claim 4, which is controlled to
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1224235A JPH0690161B2 (en) | 1989-08-30 | 1989-08-30 | Method and apparatus for measuring concentration of analyte in solution or dispersion |
| DE4026751A DE4026751C2 (en) | 1989-08-30 | 1990-08-24 | Method and device for measuring the concentration of a solution or suspension |
| US07/797,166 US5225334A (en) | 1989-08-30 | 1991-11-26 | Method and apparatus for measuring concentration of solution or suspension using electrically heating method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1224235A JPH0690161B2 (en) | 1989-08-30 | 1989-08-30 | Method and apparatus for measuring concentration of analyte in solution or dispersion |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0385433A JPH0385433A (en) | 1991-04-10 |
| JPH0690161B2 true JPH0690161B2 (en) | 1994-11-14 |
Family
ID=16810616
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1224235A Expired - Fee Related JPH0690161B2 (en) | 1989-08-30 | 1989-08-30 | Method and apparatus for measuring concentration of analyte in solution or dispersion |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPH0690161B2 (en) |
| DE (1) | DE4026751C2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0812142B2 (en) * | 1991-09-03 | 1996-02-07 | 雪印乳業株式会社 | Method for determining apparent thermal conductivity of sensor sheath and method for measuring kinematic viscosity of fluid |
| JP4188287B2 (en) | 2004-07-15 | 2008-11-26 | 三井金属鉱業株式会社 | Thermal sensor and measuring apparatus using the same |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3878049A (en) * | 1973-04-03 | 1975-04-15 | Massachusetts Inst Technology | Biochemical temperature-sensitive probe and method for measuring reactant concentrations thereof |
| SE7410610L (en) * | 1974-08-21 | 1976-02-23 | Lkb Produkter Ab | OPTICAL SYSTEM IN A CULTIVATION TANK. |
| JPS5984145A (en) * | 1982-10-01 | 1984-05-15 | サラソタ・オ−トメイシヨン・リミテツド | Method and device for measuring reynolds number of fluid |
| JPS5981551A (en) * | 1982-11-02 | 1984-05-11 | Tadashi Matsunaga | Electrochemical activity measuring method of cell |
| JPS60135754A (en) * | 1983-12-23 | 1985-07-19 | Matsushita Electric Works Ltd | Concentration measuring device |
| JPS6148755A (en) * | 1984-08-16 | 1986-03-10 | Sigma Denshi Kogyo:Kk | Measuring device for electric conductivity of culture solution |
| JPS6216457A (en) * | 1985-06-12 | 1987-01-24 | イ−・アイ・デユポン・デ・ニモアス・アンド・カンパニ− | Herbicidal sulfonamides |
| JPS6256849A (en) * | 1985-09-06 | 1987-03-12 | Snow Brand Milk Prod Co Ltd | Sensor used for electric heating method |
| JPS6264934A (en) * | 1985-09-17 | 1987-03-24 | Seiko Instr & Electronics Ltd | Quarts oscillator biosensor |
| JPS62185146A (en) * | 1986-02-12 | 1987-08-13 | Snow Brand Milk Prod Co Ltd | Measurement of fluid condition |
| JPH0623700B2 (en) * | 1987-03-06 | 1994-03-30 | 雪印乳業株式会社 | Method for measuring surface temperature of sensor used in electric heating method |
| JPH0774790B2 (en) * | 1987-08-12 | 1995-08-09 | 雪印乳業株式会社 | Sensor used for electric heating method |
-
1989
- 1989-08-30 JP JP1224235A patent/JPH0690161B2/en not_active Expired - Fee Related
-
1990
- 1990-08-24 DE DE4026751A patent/DE4026751C2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0385433A (en) | 1991-04-10 |
| DE4026751C2 (en) | 1995-07-27 |
| DE4026751A1 (en) | 1991-03-14 |
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| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |