JP2725745B2 - Methods for measuring nitrogen oxides and neurotransmitters - Google Patents
Methods for measuring nitrogen oxides and neurotransmittersInfo
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- JP2725745B2 JP2725745B2 JP7288438A JP28843895A JP2725745B2 JP 2725745 B2 JP2725745 B2 JP 2725745B2 JP 7288438 A JP7288438 A JP 7288438A JP 28843895 A JP28843895 A JP 28843895A JP 2725745 B2 JP2725745 B2 JP 2725745B2
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Description
【０００１】[0001]
【産業上の利用分野】この発明は、細径の炭素を作用電
極とする微小炭素センサー電極を用いた一酸化窒素及び
二酸化窒素イオンとドーパミン等の神経伝達物質の検出
方法に関するものである。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method of detecting neurotransmitters such as nitric oxide and nitrogen dioxide ions and dopamine using a fine carbon sensor electrode using fine carbon as a working electrode.
【０００２】[0002]
【従来の技術】近年、生体中の循環器系、神経系、泌尿
器系などあらゆる器官で生体反応を制御する物質として
微量の窒素酸化物とその派生物質及び神経伝達物質が注
目されている。2. Description of the Related Art In recent years, trace amounts of nitrogen oxides and their derivatives and neurotransmitters have been attracting attention as substances that control biological reactions in all organs of the living body, such as the circulatory system, nervous system, and urinary system.
【０００３】即ち、生体中における微量の一酸化窒素(N
O), 二酸化窒素イオン(NO2 -)等の窒素酸化物がドーパミ
ン等の神経伝達物質の放出を制御することが知られてい
る。That is, a trace amount of nitric oxide (N
O), 2 nitrogen dioxide ion (NO - nitrogen oxides) and the like are known to control the release of the neurotransmitter dopamine, and the like.
【０００４】したがって、生体中におけるこれら微量の
窒素酸化物及びドーパミン等の神経伝達物質を感度良
く、迅速に検出可能なセンサー電極を用いて生体系など
での局所で、生きたまま(in vivo) 、その場(in situ)
での情報を得ることが極めて重要である。Therefore, these trace amounts of neurotransmitters such as nitrogen oxides and dopamine in a living body are locally and alive (in vivo) in a biological system or the like by using a sensor electrode capable of detecting a sensitive and quick neurosensor. , In situ
It is extremely important to get information at
【０００５】このような目的に使用する電極としては、
電極を生体の目的とする生体神経組織に刺入したり、神
経細胞の近傍に配置して物理的、化学的、電気的に刺激
を与えた時に神経組織や神経細胞からの化学伝達物質等
の放出量変化の時間的経過を測定できることが要求さ
れ、また細胞に対する毒性がなく極めて細い（μm 程
度）電極が必要とされている。The electrodes used for such purposes include:
When the electrode is inserted into the target biological nerve tissue of a living body or placed near a nerve cell to physically, chemically, or electrically stimulate the nerve, it may cause a chemical messenger from the nerve tissue or nerve cell. It is required to be able to measure the time course of the change in the amount of release, and there is a need for an extremely fine (about μm) electrode having no toxicity to cells.
【０００６】一方、従来計測用電極としては、水素イオ
ンを計測するｐＨ計、ナトリウムイオンなどの無機イオ
ンを検出するイオンセンサー、炭素繊維（ＣＦ）やガラ
ス状炭素（ＧＣ）などの表面に錯体、有機物などを化学
修飾したセンサー、カーボンペースト（ＣＰ）電極のペ
ーストにグルコースオキシダーゼなどの酵素を混合し、
イオン透過膜で覆ったグルコースセンサーなどの各種の
酵素センサー、免疫代謝物を保持したバイオセンサー等
が検討され、実用化されているが、上述の目的に適うセ
ンサーはない。On the other hand, conventional electrodes for measurement include a pH meter for measuring hydrogen ions, an ion sensor for detecting inorganic ions such as sodium ions, and a complex on the surface of carbon fiber (CF) or glassy carbon (GC). Mixing enzymes such as glucose oxidase into the paste of the sensor, carbon paste (CP) electrode, which chemically modified organic substances,
Various enzyme sensors such as a glucose sensor covered with an ion-permeable membrane, a biosensor holding an immune metabolite, and the like have been studied and put to practical use, but there is no sensor suitable for the above purpose.
【０００７】そこで、本願発明者等は、先に、シャープ
ペンシル芯として使用されている複合炭素材料ＧＲＣ(G
raphite Reinforcement Carbon) が、電気化学的計測用
電極として優れた特性を示すことを発見し、芯を電極と
して使用する試みに成功し、使用し易いグラファイト複
合炭素電極（ＧＲＣ）を提案した（特開平1-250854
号）。Therefore, the inventors of the present application have previously reported that the composite carbon material GRC (G
raphite Reinforcement Carbon) has been found to exhibit excellent characteristics as an electrode for electrochemical measurement, and succeeded in using the core as an electrode, and proposed an easy-to-use graphite composite carbon electrode (GRC) (Japanese Patent Laid-Open No. 1-250854
issue).
【０００８】また、本願発明者等は、特願平1-250772号
で生体系の微小な組織の局所領域に対して、電流、電
圧、機械的刺激のいずれかを加えることが可能な微小電
極を作成することができる、計測系を被毒しないこ
と、即ち生体内に残存しても安全であること、生体及
び食品などに刺して、極微小（微量）部分の電気化学的
検出を可能とする機械的強度を具備すること、センサ
ー特性にばらつきが少なく、データの再現性があり、信
頼性ある計測が可能であること、特殊な前処理を必要
とせず、電気化学的前処理程度で安定に電極反応を測定
可能であること、安価で使い捨てができる、などの条
件を損なわずに特定物質の検出を可能とする炭素センサ
ー電極を提案した。Further, the present inventors have disclosed in Japanese Patent Application No. 1-250772 a microelectrode capable of applying any one of current, voltage and mechanical stimulation to a local region of a minute tissue of a biological system. Not to poison the measurement system, that is, it is safe to remain in the living body, and it is possible to electrochemically detect very small (trace) parts by piercing the living body and food. High mechanical strength, small fluctuations in sensor characteristics, reproducible data, reliable measurement, no special pre-treatment, stable at electrochemical pre-treatment level We proposed a carbon sensor electrode that can detect a specific substance without impairing the conditions such as being able to measure the electrode reaction and being inexpensive and disposable.
【０００９】更に、本願発明者等は、特開平4-74958 号
で上記〜の他に、炭素繊維電極（ＣＦ）や細いＧ
ＲＣ電極に比べて、電気抵抗が低く（導電性が良好
で）、刺入強度（腰）があり、且つ支柱部分は遥かに丈
夫な（折れにくい）電極として、有機物材料自体、若し
くは有機物材料を粘結剤として結晶性炭素微粉末を高度
に分散複合させた組成物の何れかを所望する線状に押し
出し成形した後、不活性雰囲気中又は非酸化性雰囲気中
で高温度まで焼成することにより、有機物材料自体又は
該組成物中に含まれる有機物粘結剤を炭素化して得られ
る純粋な炭素線を正極として、電解液中で先端部を電解
酸化しつつ徐々に引き上げ先端部を円錐状に尖らせて極
細径とした後、太い方の先端部をリード線に導通させ、
極細先端部の所望部分を除いた全表面を絶縁物で被覆し
てなる先細炭素微小電極を提案した。Further, the present inventors have disclosed in Japanese Patent Application Laid-Open No. 4-74958 a carbon fiber electrode (CF) and a thin G
Compared with the RC electrode, the electrode material has a lower electric resistance (good conductivity), has a piercing strength (waist), and the pole portion is a much more durable (hard to break) electrode. After extruding any of the compositions obtained by highly dispersing and compounding the crystalline carbon fine powder as a binder into a desired linear form, by firing to a high temperature in an inert atmosphere or a non-oxidizing atmosphere, Using a pure carbon wire obtained by carbonizing the organic material itself or the organic binder contained in the composition as a positive electrode, gradually raising the tip while electrolytically oxidizing the tip in an electrolytic solution, and forming the tip into a conical shape. After sharpening to an ultra-fine diameter, connect the thicker end to the lead wire,
We have proposed a tapered carbon microelectrode in which the entire surface except the desired portion of the ultrafine tip is covered with an insulator.
【００１０】そして、これらの炭素材の電極上で、検出
電位をシフトするだけで生体系に存在するドーパミン等
の神経伝達物質を容易に検出可能なことが明らかにされ
た(Anal.Sci 1990,6,439参照）。[0010] It has been clarified that a neurotransmitter such as dopamine existing in a biological system can be easily detected on these carbon material electrodes only by shifting the detection potential (Anal. Sci 1990, 6,439).
【００１１】[0011]
【発明が解決しようとする課題】しかし、これ迄ドーパ
ミン等の神経伝達物質の制御物質である一酸化窒素(N
O), 二酸化窒素イオン(NO2 -)等の窒素酸化物とドーパミ
ン等の神経伝達物質とを同時に計測することは行われて
いない。そこで、この発明では炭素電極によるこれら窒
素酸化物と神経伝達物質の同時計測を目的とするもので
あり、更に詳しくは生体組織などより具体的な対象に有
効な構成の炭素電極を使用して上記化合物の同時測定を
目的とするものである。However, to date, nitric oxide (N), which is a regulator of neurotransmitters such as dopamine, has been used.
O), nitrogen dioxide ion (NO 2 -) simultaneously measuring the neurotransmitter nitrogen oxide and dopamine such as has not been performed. Therefore, the present invention aims to simultaneously measure these nitrogen oxides and neurotransmitters using a carbon electrode. More specifically, the present invention uses a carbon electrode having a configuration effective for a more specific object such as a living tissue. It is intended for simultaneous measurement of compounds.
【００１２】具体的対象に使用する有効な電極構成とし
ては、電極の中心である作用電極、電流を受け取る対
極、電位を決める参照電極のいずれの極も、電極特性を
落とすことなく、出来るだけ生体組織を損傷しないよう
に細くしてしかも丈夫であることが望まれる。As an effective electrode configuration used for a specific object, any of the working electrode at the center of the electrode, the counter electrode for receiving current, and the reference electrode for determining the potential can be used as much as possible without reducing the electrode characteristics. It is desirable to be thin and strong so as not to damage the tissue.
【００１３】また、測定対象である生体系には、タンパ
ク質、脂質などの吸着性の物質が多量に共存している上
に、炭素面に一度吸着したこれらの有機物は酸化除去し
ない限り取り除きにくいので、センサー電極は使い捨て
か、既に使った電極部分は折り取ることができるものが
望ましく、更に臨床検査などの面からも使い捨ての特性
を持つ構成のセンサー及びセンサー電極であることが望
まれる。In addition, a large amount of adsorptive substances such as proteins and lipids coexist in the biological system to be measured, and these organic substances once adsorbed on the carbon surface are difficult to remove unless they are oxidized and removed. It is desirable that the sensor electrode is disposable or that the electrode portion already used can be cut off. Further, it is desirable that the sensor and the sensor electrode have a disposable characteristic in view of clinical examination and the like.
【００１４】その要求される条件を列挙すると、(1) 生
体系に優しい炭素を電極とする、(2) その炭素材は目的
物質の検出電極としての活性が高いこと、(3) 一度使用
済みの電極部分を折り取るか或は切り取れること、(4)
計測部分に作用電極（３電極）が容易に配置できるこ
と、(5) 使い捨てられる程度に安価であること。[0014] The required conditions are listed as follows: (1) using carbon which is kind to a living body as an electrode; (2) the carbon material has high activity as a detection electrode for a target substance; and (3) once used. (4) Folding or cutting off the electrode part of
The working electrodes (three electrodes) can be easily arranged on the measurement part, and (5) Inexpensive enough to be disposable.
【００１５】使い捨て或は使用済み部分を除去する構成
のセンサー電極は、少なくとも上記５項目の条件を満た
すものである必要があり、このセンサー電極を手軽に使
用するには、これらの要求のうち、特に(2)(4)の機能を
満たした炭素センサー電極の構成が重要である。A sensor electrode configured to remove disposable or used parts must satisfy at least the above five conditions. In order to easily use this sensor electrode, it is necessary to satisfy the following requirements. In particular, the configuration of the carbon sensor electrode satisfying the functions (2) and (4) is important.
【００１６】本願発明者等はこれらを考慮して作製した
細径の炭素を作用電極とする微小炭素センサー電極を使
用して一酸化窒素、二酸化窒素イオン及びドーパミン等
の神経伝達物質の同時検出を試みたところ、これが可能
であることを見出した。The inventors of the present application have performed simultaneous detection of neurotransmitters such as nitric oxide, nitrogen dioxide ion and dopamine using a fine carbon sensor electrode using a small diameter carbon prepared as a working electrode in consideration of the above. I have tried and found that this is possible.
【００１７】そこで、この発明では、細径の炭素から成
る作用電極、参照電極及び対極電極から構成される微小
炭素複合センサー電極を用いて、前記作用電極をスライ
ドして計測部分の望みの位置に設定し、生体内の神経系
を制御する一酸化窒素、二酸化窒素イオン等の窒素酸化
物及びドーパミン、セロトニン等の神経伝達系の化学種
即ち複数の神経伝達物質を同時にしかも経時的に計測す
ることを特徴とする神経伝達物質及び窒素酸化物の計測
方法を提案するものである。 Therefore, in the present invention, carbon is formed from small diameter carbon.
Micro electrode consisting of working electrode, reference electrode and counter electrode
Using a carbon composite sensor electrode, slide the working electrode
To the desired position of the measurement part,
Control of nitrogen monoxide, nitrogen dioxide ion, etc. nitrogen oxidation
Substances and neurotransmitter species such as dopamine and serotonin
That is, multiple neurotransmitters are measured simultaneously and over time.
Measurement of neurotransmitters and nitrogen oxides
It proposes a method.
【００１８】この発明で作用電極として使用する細径の
炭素は炭素繊維で構成することができ、また特開平4-74
958 号で開示されているアモルファス炭素と結晶性炭素
の棒状焼成品で構成することもできる。The fine carbon used as a working electrode in the present invention can be composed of carbon fibers.
No. 958 discloses a rod-shaped fired product of amorphous carbon and crystalline carbon.
【００１９】なお、作用電極の電極特性は、使用する炭
素材の電気化学的特性に負おうところが大きいので、使
用する炭素細棒或は炭素繊維の電気化学的選択を行わな
ければならない（TANSO 1991、303参照）。Since the electrode characteristics of the working electrode largely depend on the electrochemical characteristics of the carbon material used, the electrochemical selection of the carbon rod or carbon fiber to be used must be performed (TANSO 1991). , 303).
【００２０】細径の炭素表面の微細孔にシリコーン油、
ワセリン、レジン等を含浸処理しても良く、また細径の
炭素棒表面を化学的、電気化学的に処理或は修飾しても
良い。Silicon oil is introduced into the fine pores on the surface of the small-diameter carbon,
Vaseline, resin or the like may be impregnated, or the surface of the small-diameter carbon rod may be chemically or electrochemically treated or modified.
【００２１】また、この発明に使用する微小炭素センサ
ー電極は、細径の炭素を作用電極に対極及び／或は参照
電極を複合した２電極或は３電極一体型の構造の複合セ
ンサー電極とした方が実用上使い易い。The fine carbon sensor electrode used in the present invention is a composite sensor electrode having a two- or three-electrode integrated structure in which fine carbon is combined with a working electrode and a counter electrode and / or a reference electrode. It is easier to use practically.
【００２２】このような構造の微小炭素複合センサー電
極としては、計測部の望みの位置に電極を配置するため
に、図１に示すように作用電極の位置を対極や参照極の
位置からずらして配置する機構を持つ３電極一体型電極
とすることが好ましい。In the microcarbon composite sensor electrode having such a structure, the position of the working electrode is shifted from the position of the counter electrode or the reference electrode as shown in FIG. 1 in order to arrange the electrode at a desired position of the measuring section. It is preferable to use a three-electrode integrated electrode having a mechanism for disposing.
【００２３】具体的には金属管、炭素管、導電性プラス
チック管等の対極管１内に、ガラス管２を挿入し、この
ガラス管２内をガラス壁３で縦方向に２分割して室４、
５を形成し、室４、５の先端部をガラスレジン等の封止
材６で塞ぎ、室４内には炭素材で構成される作用電極７
を挿入し、その先端部を室４より突出させ、一方室５に
は塩化銀で飽和した飽和塩化カリウム溶液８を満たして
内部に参照電極９を挿入し、更にガラス管２は固定され
た対極管１に対して上下に可変出来るように対極管１と
ガラス管２の間にはワセリンなどの潤滑剤を充填して構
成する。Specifically, a glass tube 2 is inserted into a counter electrode 1 such as a metal tube, a carbon tube, or a conductive plastic tube, and the inside of the glass tube 2 is vertically divided into two parts by a glass wall 3. 4,
5 is formed, and the tips of the chambers 4 and 5 are closed with a sealing material 6 such as a glass resin.
, The tip of which protrudes from the chamber 4, while the chamber 5 is filled with a saturated potassium chloride solution 8 saturated with silver chloride, a reference electrode 9 is inserted therein, and the glass tube 2 is further fixed with a fixed counter electrode. The space between the counter electrode tube 1 and the glass tube 2 is filled with a lubricant such as petrolatum so that the tube 1 can be moved up and down.
【００２４】これら３電極一体型電極は少なくとも３電
極まとめてその電極の先端部の直径が1 〜0.5mm φ以下
であることが望ましい。In these three-electrode-integrated electrodes, it is desirable that the diameter of the tip of the at least three electrodes is 1 to 0.5 mmφ or less.
【００２５】対極管１としては、そこそこのサイズ(0.5
〜0.2mm φ）の炭素棒、被毒の心配のない系では白金な
どの貴金属がそのまま使用可能である。The counter electrode 1 has a reasonable size (0.5
Precious metals such as platinum can be used as they are in carbon rods of up to 0.2 mm in diameter and in systems that do not pose a risk of poisoning.
【００２６】ガラス管２としては、その先端部を細く引
いて縮径したものが使用される。As the glass tube 2, a tube whose diameter is reduced by pulling its tip portion thin is used.
【００２７】なお、生体系から放出される化合物を検出
するためには、作用電極７としては炭素棒先端の円盤部
分を使用するのみでは感度不足であり、側面も電極とし
て使用するのが有効である。In order to detect a compound released from a biological system, it is not sufficient to use only the disk portion at the tip of the carbon rod as the working electrode 7, and it is effective to use the side surface as an electrode. is there.
【００２８】そこで、作用電極７は図示するように、実
用計測には主に、計測対象に合わせて1 〜2mm の一定長
の炭素線を作用極とすることが好ましい。Therefore, as shown in the figure, it is preferable that the working electrode be a carbon electrode of a fixed length of 1 to 2 mm mainly for practical measurement, as shown in FIG.
【００２９】参照電極９としては、出来るだけ細径のも
のが好ましいが、従来実用化されている最も細径のもの
でもその先端部が1mm φあり、この発明の目的に沿うも
のでない。The reference electrode 9 preferably has a diameter as small as possible. However, even the thinnest electrode which has been practically used in the past has a tip of 1 mmφ, which does not meet the purpose of the present invention.
【００３０】そこで、この実施例では参照電極として0.
1 〜0.3mm φの銀線を使用し、この銀線の先端部約5mm
長の部分を電気化学的に、0.1 〜0.3Vで塩化銀化し、こ
の部分を塩化銀で飽和した飽和塩化カリウム溶液を満た
した室５内に挿入すると共に、室５の先端部をガラスレ
ジン等の封止材６で塞いで高抵抗部分を作成した構成の
ものを参照極とする。Therefore, in this embodiment, the reference electrode is set to 0.1.
Use a silver wire with a diameter of 1 to 0.3 mm φ, and the tip of this silver wire is about 5 mm
The long part is electrochemically silver-chlorinated at 0.1 to 0.3 V, and this part is inserted into a chamber 5 filled with a saturated potassium chloride solution saturated with silver chloride, and the tip of the chamber 5 is made of glass resin or the like. The reference electrode has a configuration in which a high-resistance portion is formed by closing with the sealing material 6 of FIG.
【００３１】[0031]
実施例１ 生体系の実組織中でのセンサー電極による計測には、計
測目的と計測位置に従った電極が必要であるが、その前
に先ず、組織を損傷しないため、電気化学計測が遂行で
きる最小限のサイズの電極にする必要があり、図１に示
すように３電極一体型炭素微小電極を作成した。Example 1 An electrode according to a measurement purpose and a measurement position is required for measurement by a sensor electrode in a real tissue of a biological system, but before that, electrochemical measurement can be performed because the tissue is not damaged first. An electrode having a minimum size was required, and a three-electrode-integrated carbon microelectrode was prepared as shown in FIG.
【００３２】なお、電極群はそれぞれの目的に使用可能
なように３電極まとめて先端部が1〜0.5mm φ以下にな
ることが望ましいが、実施例１では、作用電極７は対極
管１、ガラス管２の２重管内に保持されているため、３
電極間が約2mm となるが、この構造では作用電極７を２
重管の中をスライドさせることができるため、作用電極
７を深さ方向の任意の位置に配置することができる。It is desirable that the electrode group has a tip portion of 1 to 0.5 mmφ or less for all the electrodes so that the electrode group can be used for each purpose. Since it is held in the double tube of the glass tube 2, 3
The distance between the electrodes is about 2 mm.
Since the inside of the heavy tube can be slid, the working electrode 7 can be arranged at an arbitrary position in the depth direction.
【００３３】なお、この実施例で使用する複合センサー
電極は図２、図３に示すように対極管１内にガラス管２
を配置し、更にガラス管２内をガラス壁３で縦方向に２
分割して一方の室４に作用電極７を挿入し、他の室５に
参照電極９を挿入して構成する例について説明したが、
複合化する方法はこれに限られない。The composite sensor electrode used in this embodiment has a glass tube 2 inside a counter electrode 1 as shown in FIGS.
And further, the inside of the glass tube 2 is vertically
Although an example has been described in which the working electrode 7 is inserted into one of the chambers 4 and the reference electrode 9 is inserted into the other chamber 5, the structure is divided.
The method of compounding is not limited to this.
【００３４】例えば、図４に示すように対極1aを挿入し
たガラス管2a、作用電極７を挿入したガラス管2b、 参照
電極９を挿入したガラス管2cを夫々接合して構成しても
良い。For example, as shown in FIG. 4, a glass tube 2a into which the counter electrode 1a is inserted, a glass tube 2b into which the working electrode 7 is inserted, and a glass tube 2c into which the reference electrode 9 is inserted may be joined.
【００３５】また、図５に示すように作用電極７を挿入
したガラス管2b、 参照電極９を挿入したガラス管2cを対
極管１内に配置しても良い。As shown in FIG. 5, a glass tube 2b into which the working electrode 7 is inserted and a glass tube 2c into which the reference electrode 9 is inserted may be arranged in the counter electrode 1.
【００３６】更に、図６に示すように作用電極７を挿入
したガラス管2bを参照電極９を挿入したガラス管2cを配
置し、ガラス管2cの外側に対極管１を配置しても良い。Further, as shown in FIG. 6, a glass tube 2b into which the working electrode 7 is inserted, a glass tube 2c into which the reference electrode 9 is inserted may be arranged, and the counter electrode 1 may be arranged outside the glass tube 2c.
【００３７】実施例２ 生体系における実験では、組織を生かしておくために酸
素が共存している場合が多く、生理活性な化合物の多く
が、酸素と相互に係り合い変化する。この変化を追跡す
るのに実施例１で示したセンサー電極を使用した。Example 2 In an experiment in a biological system, oxygen often coexists in order to keep the tissue alive, and many of the physiologically active compounds interact with oxygen and change. The sensor electrode shown in Example 1 was used to track this change.
【００３８】先ず、リンゲル液中に500 μM 以下のNO,N
O2 -,200 μM 以下のドパーミン(DA)，セロトニン(5-HT)
を添加してその検出特性と定量性について検討した。First, NO, N of 500 μM or less in Ringer's solution was used.
O 2 -, 200 μM following dopamine (DA), serotonin (5-HT)
Was added and its detection characteristics and quantitative properties were examined.
【００３９】図７はこれら４種の化学種の同時検出例
（酸化波を計測したもの）を示すものであり、各物質の
検出、定量に関しては、各々の物質の酸化反応電位での
電流応答から、図８〜図１０に示すように予め濃度既知
の各化学種を用いて作成した校正曲線と対比して定量を
行った。FIG. 7 shows an example of simultaneous detection of these four species (measurement of oxidation waves). Regarding the detection and quantification of each substance, the current response at the oxidation reaction potential of each substance is shown. Therefore, as shown in FIGS. 8 to 10, quantification was performed in comparison with a calibration curve prepared using each chemical species whose concentration is known in advance.
【００４０】[0040]
【発明の効果】以上で明らかなように、この発明に係る
センサー電極を使用することにより、リンゲル液中に共
存するNO,NO2 - 等の窒素酸化物及びドパーミン(DA)，セ
ロトニン(5-HT)などの神経伝達系の化学種を直接に、望
みの微小領域で同時計測が可能となり、特にこれらの化
学種の経時変化をin vivo で計測することができる。As is evident from the above, according to the present invention, by using a sensor electrode according to the present invention, NO coexisting in Ringer's solution, NO 2 - nitrogen oxides and dopamine (DA), such as, serotonin (5-HT ) Can be simultaneously measured directly in the desired micro-regions of the neurotransmitter chemical species, and in particular, the temporal changes of these chemical species can be measured in vivo.
【図１】 この発明に使用する微小炭素複合センサー電
極の一例を示す斜視図FIG. 1 is a perspective view showing an example of a microcarbon composite sensor electrode used in the present invention.
【図２】 同上の要部を拡大した縦断側面図FIG. 2 is a longitudinal sectional side view in which main parts of the above are enlarged.
【図３】 図１の水平断面での、（ａ）は上面図、
（ｂ）は下面図FIG. 3A is a top view in a horizontal section of FIG.
(B) is a bottom view
【図４】 この発明に使用する微小炭素複合センサー電
極の他の一例を示す水平断面での上面図FIG. 4 is a top view in a horizontal section showing another example of the micro carbon composite sensor electrode used in the present invention.
【図５】 この発明に使用する微小炭素複合センサー電
極の更に他の一例を示す水平断面での上面図FIG. 5 is a top view in horizontal section showing still another example of the micro carbon composite sensor electrode used in the present invention.
【図６】 この発明に使用する微小炭素複合センサー電
極の更に他の一例を示す水平断面での上面図FIG. 6 is a top view in horizontal section showing still another example of the micro carbon composite sensor electrode used in the present invention.
【図７】 NO,NO2 -,ドーパミン(DA), セロトニン(5-HT)
を含むリンゲル液中においてこの発明に係る炭素センサ
ー電極を使用して測定した酸化波であり、各物質の酸化
反応電位と電流応答値( μＡ）との関係を示す図[7] NO, NO 2 -, dopamine (DA), serotonin (5-HT)
FIG. 4 is an oxidation wave measured using a carbon sensor electrode according to the present invention in a Ringer solution containing, and showing a relationship between an oxidation reaction potential of each substance and a current response value (μA).
【図８】 リンゲル液中でのドーパミンの校正曲線FIG. 8: Calibration curve of dopamine in Ringer's solution
【図９】 セロトニン10μM,NO及びNO2 -を含むリンゲル
液中でのドーパミンの校正曲線[9] Serotonin 10 [mu] M, NO and NO 2 - Calibration curve for dopamine in Ringer's solution containing
【図１０】 リンゲル液中でのNO2 -の校正曲線[10] NO in Ringer's solution 2 - Calibration curve for
１は対極管 1aは対極 ２,2a,2b,2c はガラス管 ３はガラス壁 ４、５は室 ６は封止材 ７は作用電極 ８は飽和塩化カリウム溶液 ９は参照電極 1 is a counter electrode 1a is a counter electrode 2, 2a, 2b, 2c is a glass tube 3 is a glass wall 4, 5 is a chamber 6 is a sealing material 7 is a working electrode 8 is a saturated potassium chloride solution 9 is a reference electrode
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 平１−250854（ＪＰ，Ａ) 特開 平５−26840（ＪＰ，Ａ) 特開 平３−188367（ＪＰ，Ａ) 特開 平４−74958（ＪＰ，Ａ) 実開 平１−148855（ＪＰ，Ｕ) ＲＥＶ．ＰＯＬＡＲＯＧＲ 40〜３ ／６！（1990）Ｐ．64 ＡＮＡＬ．ＳＣＩ．，６〜３！ （1990）Ｐ．439−442 ────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-1-250854 (JP, A) JP-A-5-26840 (JP, A) JP-A-3-188367 (JP, A) JP-A-4- 74958 (JP, A) Hikaru 1-148855 (JP, U) REV. POLAROGR 40-3 / 6! (1990) P.A. 64 ANAL. SCI. , 6-3! (1990) P.A. 439-442
Claims (7)
び対極電極から構成される微小炭素複合センサー電極を
用いて、前記作用電極をスライドして計測部分の望みの
位置に設定し、生体内の神経系を制御する窒素酸化物及
び複数種の神経伝達物質を同時にしかも経時的に計測す
ることを特徴とする神経伝達物質及び窒素酸化物計測方
法。1. A working electrode, a reference electrode and a thin carbon electrode.
A small carbon composite sensor electrode consisting of
Slide the working electrode to the desired
Nitrogen oxides that control the nervous system in vivo
And multiple neurotransmitters simultaneously and over time
A method for measuring neurotransmitters and nitrogen oxides, comprising :
載の方法。2. The method of claim 1, wherein the small diameter carbon comprises carbon fibers.
素の棒状焼成品である請求項１記載の方法。3. The method according to claim 1, wherein the small-diameter carbon is a rod-shaped fired product of amorphous carbon and crystalline carbon.
はレジンを含浸処理した細径の炭素を使用する請求項１
記載の方法。4. Silicon oil, petrolatum or silicone oil
2. Use of small diameter carbon impregnated with resin.
The described method.
飾した細径の炭素を使用する請求項１記載の方法。5. The method according to claim 1, wherein a small-diameter carbon whose surface is chemically or electrochemically treated or modified is used.
び対極電極から一体に構成された微小炭素複合センサー
電極を用いる請求項１、２、３、４又は５記載の方法。6. A working electrode, a reference electrode and a thin carbon electrode.
Carbon composite sensor integrally composed of electrode and counter electrode
The method according to claim 1, 2, 3, 4, or 5 , wherein an electrode is used .
た微小炭素複合センサー電極を用いる請求項１、２、
３、４又は５記載の方法。7. The working electrode is movable with respect to a counter electrode.
1, 2 or 3 using a micro carbon composite sensor electrode.
The method according to 3, 4 or 5 .
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| JP7288438A JP2725745B2 (en) | 1995-11-07 | 1995-11-07 | Methods for measuring nitrogen oxides and neurotransmitters |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7288438A JP2725745B2 (en) | 1995-11-07 | 1995-11-07 | Methods for measuring nitrogen oxides and neurotransmitters |
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| Publication Number | Publication Date |
|---|---|
| JPH09127056A JPH09127056A (en) | 1997-05-16 |
| JP2725745B2 true JP2725745B2 (en) | 1998-03-11 |
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ID=17730220
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| JP7288438A Expired - Lifetime JP2725745B2 (en) | 1995-11-07 | 1995-11-07 | Methods for measuring nitrogen oxides and neurotransmitters |
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| JP4669145B2 (en) * | 2001-03-26 | 2011-04-13 | 日本曹達株式会社 | Improved voltage sensing device |
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| JPH01148855U (en) * | 1988-04-01 | 1989-10-16 | ||
| JP2740587B2 (en) * | 1991-07-18 | 1998-04-15 | 工業技術院長 | Micro composite electrode and method of manufacturing the same |
-
1995
- 1995-11-07 JP JP7288438A patent/JP2725745B2/en not_active Expired - Lifetime
Non-Patent Citations (2)
| Title |
|---|
| ＡＮＡＬ．ＳＣＩ．，６〜３！（１９９０）Ｐ．４３９−４４２ |
| ＲＥＶ．ＰＯＬＡＲＯＧＲ ４０〜３／６！（１９９０）Ｐ．６４ |
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