1276799 九、發明說明： 【發明所屬之技術領域】 本發明係關於一種電極試片及包含其之生物感測器，其 經由併入修飾過之奈米碳管層，以增加氧化還原電流訊號。 【先前技術】 ·)BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an electrode test piece and a biosensor comprising the same, which incorporates a modified carbon nanotube layer to increase a redox current signal. [Prior Art] ·)
由於生物感測器與電極試片可提供具有高準確度的快速 檢測，因此其可在研究及臨床領域上，用以處理大批的檢 體。美國專利第5，120,420號揭示一種生化感測電極，其包 括電極部份、絕緣層、反應層，以及檢體承載空間（其係在 遠反應層上放置樹脂板及親水性蓋板而成）。該生化感測電 極的特徵在於.，需要絕緣上蓋以完成檢體承載空間之縮小 化。美國專利第6,413,394號揭示一種生化感測電極，其包 括電極部份、絕緣層、反應層，以及保護網。該生化感測 電極的特徵在於，以網版或其他相似方式一次塗佈含有親 水性高分子載體之反應層，其雖可克服加工繁瑣性，但唯 有精密網印條件的控制，才能適當縮小其尺寸。美國專利 申請公開案第2002/0,100,685號揭示一種生化感測電極 條，其包括電極部份、絕緣層、反應層、墊高層、保護網。 該生化感測電極的特徵在於，以網目取代絕緣上蓋以完成 檢體承載空間之縮小化，且可多方位採血。美國專利申請 公開案第2004/0,081，752號揭示一種生化感測電極，其包括 電極部份、絕緣層、反應層、墊高層、保護網。該生化感 測電極的特徵在於，以揮發性有機水溶液，溶解非生物活 性物質，以利分散於反應電極區。上述生化感測電極，於 103662.doc 1276799 實際應用時，均面臨為因應使用者對於採樣體積縮小化之 期待而縮小檢體反應區面積，從而使得電流訊號/雜訊比 (S/Ν)降低之問題。 因此，在微小化電化學氧化還原檢測元件之過程中，需 要發展一種具有強化氧化還原電流訊號之生物感測電極試 片，其於縮小化採樣體積之同時，仍可保留夠大、甚至強 化之電訊號（如^ μΑ)，而提升偵測之準確度。 美國專利第6,491，803號與中國大陸專利申請公開案第 1462880Α號及第1462881Α號即為奈米尺寸材質於生化感 測電極之相關應用。然而，該等專利仍不脫先前技術，其 仍需要繁複之製備過程。美國專利第6,49號需先行混 合所有反應物質（包含奈米金屬粒子），從而隨後的網印條件 舄相當嚴苛才能維持網印的均一性。中國大陸專利申請公 開案第1462880A號及第1462881A號則需依序塗佈並乾燥 羧甲基纖維素等水溶性高分子載體、修飾過之奈米碳管及 酵素反應層（包含酵素、電子媒介物、穩定劑、緩衝液等） 等至少二層物質之加工，電極試片製作過程甚為繁複。 因此，對此業界而言，仍存有不需如先前技藝般繁瑣之 加工步驟且同時仍能保留足夠大之電流訊號之電極試片之 需求。 【發明内容】 本發明之一目的在於提供一種併入修飾過之奈米碳管 層以増加氧化還原電流訊號之用以檢測檢體中生化成分 的電極試片，其包含： 103662.doc 1276799 —電絕緣基板； y導電膜之電極系統，其塗覆於該電絕緣基板之—面， 以形成分離且不連接之一工作電極與一參考電極； :配置於該基板上之電絕緣層，其部分覆蓋該電極系 統’電極系統之裸露部分分別形成包含工作電極及參考 電極之接線端及電化學反應端； 多 -修飾過之奈米碳管層，其至少部分覆蓋該電化 端之工作電極；以及 一塗佈於該電化學反應端及奈米碳管層上之反應層，其 至少包含一電子媒介物。 本發明之另一目的在於提供一種電極試片，其除了併入 乜飾過 < 奈米碳管層外，另外包含可減低檢體需求量之辅 助元件。 本發明之又再一目的在提供一種生物感測器，其包含本 發明的電極試片及檢測裝置。 【實施方式】 本發明係關於一種電極試片及包含其之生物感測器，其 特徵為具有併入修飾過之奈米碳管層，以增加反應層和電 極系統間的氧化還原電流訊號。不同以往先前技藝的繁瑣 加工步驟，本發明電極試片之電化學反應區僅需二層物質 之加工程序，因而，可降低製作成本，且同時仍能保留足 夠大之電流訊號。 本發明之一目的係提供一種電極試片，其包含： 一電絕緣基板； 103662.doc 1276799 -導電膜之電極系統’其塗覆於該電絕緣基板之 以形成分離且不連接之一工作電極 ， Θ 參考電極； 一配置於該基板上之電絕緣層，雪 豕層％極系統被該電絕緣爲 邵分覆蓋，而其裸露部分分別形成 曰 工作電極及泉去 電極之接線端及電化學反應端； ，可 —修飾過之奈米碳管層，其至少 / 1刀覆盍孩電化學及虛Because biosensors and electrode test strips provide fast detection with high accuracy, they can be used to process large numbers of specimens in research and clinical fields. U.S. Patent No. 5,120,420 discloses a biochemical sensing electrode comprising an electrode portion, an insulating layer, a reaction layer, and a sample carrying space (which is formed by placing a resin plate and a hydrophilic cover plate on the far reaction layer) . The biochemical sensing electrode is characterized in that an insulating cover is required to complete the reduction of the specimen carrying space. U.S. Patent No. 6,413,394 discloses a biochemical sensing electrode comprising an electrode portion, an insulating layer, a reactive layer, and a protective mesh. The biochemical sensing electrode is characterized in that a reaction layer containing a hydrophilic polymer carrier is coated at a time in a screen or other similar manner, which can overcome the cumbersome processing, but can only be appropriately reduced by the control of precision screen printing conditions. Its size. A biochemical sensing electrode strip comprising an electrode portion, an insulating layer, a reaction layer, a pad high layer, and a protective mesh is disclosed in U.S. Patent Application Publication No. 2002/0,100,685. The biochemical sensing electrode is characterized in that the insulating cover is replaced by a mesh to complete the reduction of the sample carrying space, and the blood can be collected in multiple directions. U.S. Patent Application Publication No. 2004/0,081,752 discloses a biochemical sensing electrode comprising an electrode portion, an insulating layer, a reaction layer, a high layer of a pad, and a protective mesh. The biochemical sensing electrode is characterized in that a non-bioactive substance is dissolved in a volatile organic aqueous solution to facilitate dispersion in the reaction electrode region. The above biochemical sensing electrode, in the practical application of 103662.doc 1276799, faces a reduction in the area of the sample reaction area in response to the user's expectation of shrinking the sampling volume, thereby reducing the current signal/noise ratio (S/Ν). The problem. Therefore, in the process of miniaturizing the electrochemical redox detection element, it is necessary to develop a biosensing electrode test piece having an enhanced redox current signal, which can still retain a large enough or even strengthen the volume while reducing the sampling volume. The electrical signal (such as ^ μΑ) improves the accuracy of detection. U.S. Patent No. 6,491,803 and the Chinese Patent Application Publication No. 1462880 and No. 1461881 are the applications of nano-sized materials for biochemical sensing electrodes. However, these patents still do not deviate from the prior art, which still requires a complicated preparation process. U.S. Patent No. 6,49 requires the mixing of all reactive materials (including nano-metal particles) so that the subsequent screen printing conditions are quite severe to maintain the uniformity of screen printing. In the Chinese Patent Application Publication No. 1462880A and No. 1462881A, it is necessary to sequentially coat and dry a water-soluble polymer carrier such as carboxymethyl cellulose, a modified nanocarbon tube and an enzyme reaction layer (including an enzyme, an electronic medium). Processing of at least two layers of materials, stabilizers, buffers, etc., the electrode test piece production process is very complicated. Therefore, there is still a need in the industry for an electrode test piece that does not require the cumbersome processing steps as prior art while still retaining a sufficiently large current signal. SUMMARY OF THE INVENTION An object of the present invention is to provide an electrode test piece for injecting a modified carbon nanotube layer to detect a biochemical component in a sample by adding a redox current signal, which comprises: 103662.doc 1276799 — An electrically insulating substrate; an electrode system of the y conductive film, coated on the surface of the electrically insulating substrate to form a separated and not connected one of the working electrode and a reference electrode; an electrical insulating layer disposed on the substrate, a portion of the electrode system partially covering the electrode system forms a terminal including a working electrode and a reference electrode, and an electrochemical reaction terminal; a multi-modified carbon nanotube layer covering at least partially the working electrode of the electrochemical terminal; And a reaction layer coated on the electrochemical reaction end and the carbon nanotube layer, which comprises at least one electron vehicle. Another object of the present invention is to provide an electrode test piece which, in addition to being incorporated into a <nanocarbon tube layer, additionally comprises an auxiliary element which reduces the amount of sample required. Still another object of the present invention is to provide a biosensor comprising the electrode test piece and the detecting device of the present invention. [Embodiment] The present invention relates to an electrode test piece and a biosensor comprising the same, characterized in that it has a modified carbon nanotube layer incorporated to increase the redox current signal between the reaction layer and the electrode system. Different from the cumbersome processing steps of the prior art, the electrochemical reaction zone of the electrode test piece of the present invention requires only a processing procedure of two layers of materials, thereby reducing the manufacturing cost while still retaining a sufficiently large current signal. An object of the present invention is to provide an electrode test piece comprising: an electrically insulating substrate; 103662.doc 1276799 - an electrode system of a conductive film applied to the electrically insulating substrate to form a separate and not connected one of the working electrodes , Θ reference electrode; an electrical insulating layer disposed on the substrate, the skim layer % pole system is covered by the electrical insulation, and the exposed portions thereof form the terminal of the working electrode and the spring electrode and the electrochemical Reaction end; , can be - modified nano carbon tube layer, at least / 1 knife cover 电化学 child electrochemical and virtual
端之工作電極；以及 久I 一塗佈於該電化學反應端及奈米碳管層上之反應層4 至少包含一電子媒介物。 /、 根據本發明，該電絕緣基板具有 ^ 特性以及可耐40°C〜200。(：力、w # φ 、 私絕緣的 、” 觀加溫處理的耐熱能力，以便於加 :處理/合作為本發明之絲切料包括，但不限於加 =(PVC)、破璃纖維、聚㈣、㈣樹脂板、聚對苯 -:乙二^ (PE)、聚醯胺（PA)、嘹贫7此/ΤΛ。、 )永冬乙埽（PS)、破璃或陶瓷。 根據本發明，該電猙 、 、、、，表基板 < 一面塗覆導電膜以形成分 離且不連接之工作雷 °、、乡考電極。於根據本發明之一較 1芏丹植貫施悲樣中， 至 、、 乍电極炙面積為參考電極之面積的2 主3倍。孩導電膜之雨4τ <、 腾夕、^ ^ 电極系統較佳係以網板印刷或黏附金屬 腰又万式，塗覆於該雷 、通、，彖基板 < 一面。適合之金屬膜包 括，但不限於，金、銀七 m , t 銷或免，而週合網板印刷之印刷 墨包括，但不限於， 人弘 人W、至墨、銀墨、碳墨及銀墨的混 口物、揮發性石墨、. 聖、 ]、工或以上的組合（如先印銀墨再印碳 、置、)0 103662.doc 1276799 根據本發明，該電絕緣層厚度一般約為0.01至0.6毫米。 適合作為該電絕緣層之材料包括，但不限於，PP、PVC、 PET、PC 或 PE。The working electrode of the end; and the reaction layer 4 coated on the electrochemical reaction end and the carbon nanotube layer at least contains an electron mediator. According to the present invention, the electrically insulating substrate has a characteristic and is resistant to 40 ° C to 200. (: force, w # φ, privately insulated, "heat-resistant ability to heat treatment, in order to facilitate the addition: processing / cooperation for the wire cutting of the invention includes, but not limited to, plus = (PVC), glass fiber, Poly (4), (4) resin board, poly-p-benzene-: Ethylene (PE), polydecylamine (PA), 嘹 7 7 this / ΤΛ., ) Yongdong 埽 (PS), broken glass or ceramic. According to the invention, the electric circuit, the surface substrate < the surface of the surface of the substrate is coated with a conductive film to form a separated and unconnected working mine, and a test electrode. In the middle, the area of the 炙 electrode is 3 times the area of the reference electrode. The rain of the conductive film 4τ <, Tengxi, ^ ^ electrode system is better to screen printing or sticking the metal waist a type of coating, applied to the surface of the ray, the 、, and the 彖 substrate. Suitable metal films include, but are not limited to, gold, silver, seven m, t pin or exempt, and the printing ink of the stencil screen includes However, it is not limited to a combination of people, people, ink, silver ink, carbon ink and silver ink, volatile graphite, saint, ], work or above (such as silver first) Ink reprinted carbon, placed,) 0 103662.doc 1276799 According to the invention, the thickness of the electrically insulating layer is generally about 0.01 to 0.6 mm. Suitable materials for the electrically insulating layer include, but are not limited to, PP, PVC, PET, PC or PE.
根據本發明，該至少部分覆蓋電化學反應端之工作電極 之奈米碳管層（如圖1(A)中之6a、6b或6c所示）係由奈米尺寸 且具有-OH或=00修飾之奈米碳管及視需要之載體所組 成，其可用於增加生物感測電極之氧化還原電流。該經修 飾之奈米碳管可為單層壁奈米碳管（Single Wall Carbon Nanometer Tube ; SWCNT)或為多層壁奈米碳管（Multiple Walls Carbon Nanometer Tube ; SWCNT)，或其混合。該視 需要之載體則包含水溶性高分子聚合物並選擇性含有界面 活性劑。適合之高分子載體包括，但不限於，聚乙烯醇 (PVA)、聚乙晞吡咯酮（PVP)、聚乙二醇（PEG)、白明膠 (gelatin)、叛甲基纖維素（CMC; carboxymethyl cellulose)、 甲基纖維素（methyl cellulose)、或其混合物。適合之界面活 性劑包括，但不限於，三通X-l〇〇(Triton X-100)、三通 C-405、三通X-114、十二烷基硫酸鈉、聚氧乙晞山梨糖醇 肝單月桂酸酉旨（例如，Tween 20、Tween 40、Tween 60及 Tween 80)、或其它水溶性界面活性劑。 根據本發明，可以滴加或網板印刷方式將一包含修飾過 之奈米碳管之漿體施加於電化學反應端上，以形成至少覆 蓋一部分裸露之工作電極之該修飾過之奈米碳管層。當以 滴加方式進行時，係將包含修飾過之奈米碳管及溶劑（例 如，但不限於，甲醇、乙醇、乙醚、丙酮或其混合物）之漿 103662.doc -10-According to the present invention, the carbon nanotube layer (shown as 6a, 6b or 6c in Fig. 1(A)) at least partially covering the working electrode of the electrochemical reaction end is modified by a nanometer size and has -OH or =00 The carbon nanotube and the optional carrier can be used to increase the redox current of the biosensing electrode. The modified carbon nanotube may be a Single Wall Carbon Nanometer Tube (SWCNT) or a Multiple Walls Carbon Nanometer Tube (SWCNT), or a mixture thereof. The optional carrier comprises a water soluble high molecular polymer and optionally contains an interfacial active agent. Suitable polymeric carriers include, but are not limited to, polyvinyl alcohol (PVA), polypyridone (PVP), polyethylene glycol (PEG), gelatin, methyl cellulose (CMC; carboxymethyl) Cellulose), methyl cellulose, or a mixture thereof. Suitable surfactants include, but are not limited to, Triton X-100, Tee C-405, Tee X-114, sodium lauryl sulfate, polyoxyethylene sorbitol liver Monolaurate (eg, Tween 20, Tween 40, Tween 60, and Tween 80), or other water soluble surfactants. According to the present invention, a slurry comprising a modified carbon nanotube can be applied to the electrochemical reaction end by drop or screen printing to form the modified nanocarbon covering at least a portion of the exposed working electrode. Pipe layer. When carried out in a dropwise manner, it will contain a slurry of modified carbon nanotubes and a solvent (for example, but not limited to, methanol, ethanol, diethyl ether, acetone or a mixture thereof) 103662.doc -10-
1276799 於該電化學反應端之工作電極上，然後使溶劑揮 形成該修飾過之奈米碳管層。於根據本發明之另一 ^體實施態樣中’亦可以網板特方式形錢修飾過之夺 ，管層’此時係使用由修部過之奈米碳管、載體及落劑 組成之漿體，而以網板印刷方式塗覆於電化學反應端之 工作電極。 " 、根據本發明’该塗佈於該電化學反應端及奈米碳管層上 之反，少包含一電子媒介物’其組成對熟習此項技術 者而Τ，係顯而易知的，端視電極試片之最終用途而定。 :般而言，若電極試片係供酵素型電流式生物感測器用之 電極試片，該&應層典型含有針對特定待測物之氧化還原 2素（例如，但不限於，葡萄糖氧化酵素）、氧化還原電子媒 介物、载體及共輛酸驗混合組成之緩衝劑。若電極試片係 供非酵素型電流式生物感測器用之電極試片，該反應層則 典型含有氧化還原電子媒介物與載體。載體一般包含水溶 性高分子聚合物並選擇性含有界面活性劑。適合之高分子 載體包括’但不限^，聚乙料（PVA)、聚乙^比嗔明 (PVP)、聚乙二醇（PEG)、白明膠（gelatin)、羧甲基纖維素 (CMC; Carb〇xymethyl cellul〇se)、甲基纖維素（邮邮 cellulose)或其混合物。適合之界面活性劑包括，但不限 三通X-114、 於，三通 X_100(Trit〇n x_1〇〇)、三通 C4〇5、 十二烷基硫酸鈉、聚氧乙烯山梨糖醇酐單月桂酸酯（例如，1276799 is applied to the working electrode of the electrochemical reaction end, and then the solvent is evaporated to form the modified carbon nanotube layer. In another embodiment of the present invention, the stencil can be modified by a special method, and the tube layer is composed of a carbon nanotube, a carrier and a falling agent. The slurry is applied to the working electrode of the electrochemical reaction end by screen printing. " According to the invention, the coating on the electrochemical reaction end and the carbon nanotube layer contains less than an electron mediator', and its composition is obvious to those skilled in the art. The end-use electrode test piece depends on the end use. In general, if the electrode test piece is used for an electrode test piece for an enzyme type current type biosensor, the & layer should typically contain a redox element for a specific test substance (for example, but not limited to, glucose oxidation). A buffer composed of an enzyme), a redox electron carrier, a carrier, and a total acid mixture. If the electrode test piece is an electrode test piece for a non-enzyme type current type biosensor, the reaction layer typically contains a redox electron carrier and a carrier. The carrier generally comprises a water soluble high molecular polymer and optionally a surfactant. Suitable polymer carriers include 'but not limited to', poly (PVA), polyethyl bromide (PVP), polyethylene glycol (PEG), gelatin, carboxymethyl cellulose (CMC) Carb〇xymethyl cellul〇se), methylcellulose (postal) or a mixture thereof. Suitable surfactants include, but are not limited to, three-way X-114, three-way X_100 (Trit〇n x_1〇〇), three-way C4〇5, sodium lauryl sulfate, polyoxyethylene sorbitan Monolaurate (for example,
Tween 20、Tween 40、Tween 60及Tween 80)、或其它水溶 性界面活性劑。氧化還原電子媒介物一般係使用鐵氰化鉀。 103662.doc 1276799 根據本發明> — 較佳具體實施態樣，電極試片可另外包 含用於減低檢體需电旦、 ^ , 而求里疋辅助元件。該等輔助元件可形成 具有毛細作用士、ΤΓ W、 、 <取樣空間，從而檢體得以迅速而全面地 被吸往反應居， 發π 、 曰不僅传以確保有效的檢測結果，而且可以 減少檢體的滴姑县 > I°以糖尿病患者為例，其在使用血糖試 4寺知血夏夕，尤其是較嚴重的患者，一天内常要進行多 W樣以進仃監控。若能在血液需求量上減少，便能使 車乂戴、、田的採血針，使患者的傷口及疼痛減到S低。再者， =樣^電極試片上時以因為血量少而塗抹試片，進而 L成里測上的誤差。本發明之電極試片可降低血糖電極試 片在量測時的血液需求量，以及改良採血樣的方式。 圖1(A) 1(B)及1(c)顯示根據本發明之具有減低檢體需求 Υ <辅助το件之電極試片之一具體實施態樣，其包含·· 一電絕緣基板（1); 一導電膜之電極系統（2)，其塗覆於該電絕緣基板（丨）之一Tween 20, Tween 40, Tween 60 and Tween 80), or other water soluble surfactants. Redox electron mediators typically use potassium ferricyanide. 103662.doc 1276799 According to the present invention, in a preferred embodiment, the electrode test piece may additionally comprise an auxiliary element for reducing the need for the sample to be charged, ^. The auxiliary elements can form a sampling space, a ΤΓ W, and a sampling space, so that the sample can be quickly and comprehensively sucked into the reaction, and the π and 曰 are transmitted not only to ensure effective detection results but also to reduce In the case of Diabetes in the sample, I° takes diabetes patients as an example. It uses blood sugar test 4 to know the blood and summer, especially for the more serious patients. It is often necessary to carry out multiple monitoring in one day. If the blood demand can be reduced, the lancet can be worn by the rut, and the blood and needle of the patient can be reduced to a low S. Furthermore, when the test piece is on the test piece, the test piece is applied because of the small amount of blood, and the error in the measurement is L. The electrode test piece of the present invention can reduce the blood requirement of the blood glucose electrode sample at the time of measurement, and improve the manner of blood sampling. 1(A) 1(B) and 1(c) show a specific embodiment of an electrode test piece having a reduced sample requirement Υ < auxiliary το according to the present invention, which comprises an electrically insulating substrate ( 1); an electrode system (2) of a conductive film coated on one of the electrically insulating substrates (丨)
面’以形成分離且不連接之一工作電極（2a)與一參考電極 (2b)； 一配置於該基板上之電絕緣層（3)，其部分覆蓋該電極系 統（2)，電極系統之裸露部分分別形成包含工作電極（2&) 及參考電極（2b)之接線端（4)及電化學反應端（5); 一修飾過之奈米碳管層（6a、6b或6c)，其至少部分覆蓋該 電化學反應端（5)之工作電極； 一塗佈於該電化學反應端（5)及奈米碳管層（6a、6b或6c) 上之反應層（7)，其至少包含一電子媒介物； 103662.doc -12- 1276799 一配置於該反應層（7)上之絕緣墊高層（8)，該絕緣墊高層 (8)形成一用以容納檢體至該反應層（7)之具有二個開口 >而（9)及（9a)之缺口；及 一配置於該絕緣墊高層（8)之上且覆蓋該缺口之無孔絕 _ 緣上蓋（10a)，其中該無孔絕緣上蓋（10a)與反應層（7)及絕 . 緣墊高層（8)共同構成一檢體容納槽道（1 la)。 其中，該檢體容納槽道（11a)係具有毛細作用之取樣空 馨） 間，從而可使檢體由該檢體容納槽道（11a)的開口端（9)或（9a) 到達反應層（7)。 一根據本發明，該絕緣墊高層（8)之厚度一般約為〇ι至〇·5 毫米，而該無孔絕緣上蓋（10a)之厚度一般約為〇〇5至〇5毫 米。適合作為孩絕緣墊高層（8)及無孔絕緣上蓋（l〇a)之材 、斗L括仁不限於，pVc、玻璃纖維、聚酯楓、盼酸樹脂 板、PET、PC、PP、PE、pA、ps、玻璃或陶瓷。 圖2顯示根據本發明之具有減低檢體需求量之輔助元件 _ <電極試片之另一具體實施態樣，其包含·· • 一電絕緣基板（1); ' —I電膜之電極系統⑺，其塗覆於該電絕緣基板⑴之一 _ ㊆，以形成分離且不連接之-工作電極(2a)與一參考電極 (2b); _ —配置於該基板上之電絕緣層（3)，其部分覆蓋該電極系 _ 、统⑺’電㈣統之裸露部分分別形成包含工作電極㈣ 及參考電極（2b)之接線端（4)及電化學反應端（5); 一修飾過之奈米碳管層（6&、61)或6〇，其至少部分覆蓋該 103662.doc -13· 1276799 電化學反應端（5)之工作電極； 一塗佈於該電化學反應端（5)及奈米碳管層（6a、6b或6c) 上之反應層（7)，其至少包含一電子媒介物； 一配置於該反應層（7)上之絕緣墊高層（8a)，該絕緣墊高 層（8a)形成一用以容納檢體至該反應層之缺口，其具 有一個開口端（9);及a surface 'to separate and not connect one of the working electrode (2a) and one reference electrode (2b); an electrically insulating layer (3) disposed on the substrate, partially covering the electrode system (2), the electrode system The exposed portions respectively form a terminal (4) including a working electrode (2&) and a reference electrode (2b) and an electrochemical reaction end (5); a modified carbon nanotube layer (6a, 6b or 6c), a working electrode at least partially covering the electrochemical reaction end (5); a reaction layer (7) coated on the electrochemical reaction end (5) and the carbon nanotube layer (6a, 6b or 6c), at least Including an electron mediator; 103662.doc -12- 1276799 A high-rise (8) insulating pad disposed on the reaction layer (7), the upper layer (8) of the insulating pad is formed to accommodate the sample to the reaction layer ( 7) having two openings > and a notch of (9) and (9a); and a non-porous rim upper cover (10a) disposed on the upper layer (8) of the insulating pad and covering the notch, wherein The non-porous insulating upper cover (10a) and the reaction layer (7) and the rim pad (8) together constitute a sample receiving channel (1 la). Wherein, the sample receiving channel (11a) has a capillary action between the sampling spaces, so that the sample reaches the reaction layer from the open end (9) or (9a) of the sample receiving channel (11a). (7). According to the present invention, the thickness of the upper layer (8) of the insulating mat is generally from about 1 to about 5 mm, and the thickness of the non-porous insulating upper cover (10a) is generally from about 5 to about 5 mm. Suitable for children's insulation pad high-rise (8) and non-porous insulation cover (l〇a) material, bucket L is not limited to, pVc, fiberglass, polyester maple, acid resin board, PET, PC, PP, PE , pA, ps, glass or ceramic. Fig. 2 shows another embodiment of an auxiliary component having an improved sample requirement according to the present invention, which comprises an electrically insulating substrate (1); an electrode of the -I film a system (7) applied to one of the electrically insulating substrates (1) to form a separate and unconnected-working electrode (2a) and a reference electrode (2b); _ an electrically insulating layer disposed on the substrate ( 3), partially covering the exposed portions of the electrode system _, system (7) 'electric (four) system respectively forming a working electrode (four) and reference electrode (2b) terminal (4) and electrochemical reaction end (5); a carbon nanotube layer (6&, 61) or 6〇, which at least partially covers the working electrode of the electrochemical reaction end (5) of 103662.doc -13· 1276799; one coated on the electrochemical reaction end (5 And a reaction layer (7) on the carbon nanotube layer (6a, 6b or 6c) comprising at least one electron medium; a high-rise insulating layer (8a) disposed on the reaction layer (7), the insulation The upper layer (8a) of the mat forms a gap for accommodating the sample to the reaction layer, and has an open end (9);
一配置於該絕緣墊高層（8a)之上且覆蓋該缺口之網狀覆 蓋層（l〇b)，其中該網狀覆蓋層（1〇b)與反應層（7)及絕緣墊 高層（8a)共同構成一檢體容納槽道（丨lb)。 根據本發明，該網狀覆蓋層（10b)可選自親水性網狀材料 或疏水性網狀材料或金屬網，較佳係選自親水性網狀材 料’其網目數目包括’但不限於，6 〇至3 0 0目。適合作為網 狀覆蓋層之材料係為此技術領域具有通常知識者所熟知， 例如，親水性網狀材料可為但不限於經界面活性劑處理過 之聚醚颯（polyethersulfone ; PES)網；疏水性網狀材料可為 但不限於聚醚砜（PES)網；金屬網可為但不限於不銹鋼網。 禹该網狀覆盍層（1 〇b)係由親水性網狀材料所組成時，對親 水性的檢體而言，檢體可由開口端（9)導入或由該網狀覆蓋 層（10b)上方導人。當該網狀覆蓋層（1〇b)係由疏水性網狀材 料所組成時，對於親水性的血液檢體而言，由於其親和性 較弱，因此檢體主要是由開口端（9)藉毛細現象導入。該網 狀覆蓋層（1Gb)亦可為經過界面活性劑處理之疏水性網狀材 料，從而使得檢體亦可由該網狀覆蓋層（1〇b)上方導入，其 中該界面活性劑係選自，但不限於，三通χ_ι〇〇(τ岀⑽ 103662.doc -14·a mesh covering layer (10b) disposed on the upper layer (8a) of the insulating pad and covering the notch, wherein the mesh covering layer (1〇b) and the reaction layer (7) and the insulating layer upper layer (8a) Together, they constitute a sample holding channel (丨lb). According to the invention, the reticulated cover layer (10b) may be selected from a hydrophilic reticulated material or a hydrophobic reticulated material or a metal mesh, preferably selected from the group consisting of hydrophilic reticulated materials, the number of which includes, but is not limited to, 6 〇 to 300 mesh. Materials suitable as a network cover are well known to those of ordinary skill in the art. For example, the hydrophilic mesh material can be, but is not limited to, a polyethersulfone (PES) network treated with a surfactant; hydrophobic The mesh material can be, but is not limited to, a polyethersulfone (PES) mesh; the metal mesh can be, but is not limited to, a stainless steel mesh.禹 When the mesh-like covering layer (1 〇b) is composed of a hydrophilic mesh material, for a hydrophilic sample, the sample may be introduced from the open end (9) or by the mesh covering layer (10b) ) The top guide. When the network cover layer (1〇b) is composed of a hydrophobic network material, for a hydrophilic blood sample, since the affinity is weak, the sample is mainly composed of an open end (9). Imported by capillary phenomenon. The mesh covering layer (1Gb) may also be a surfactant-treated hydrophobic mesh material, so that the sample may also be introduced from above the mesh covering layer (1〇b), wherein the surfactant is selected from the group consisting of , but not limited to, three-way χ _ι〇〇 (τ岀 (10) 103662.doc -14·
1276799 tiOO)，三通C-405，三通Χ-Π4，十二烷基硫酸鈉、聚氧 乙缔山梨糖醇奸單月桂酸酯（例如，Tween 20、Tween 40、 Tween 60、Tween 80)，和其它水溶性界面活性劑。 圖3顯示根據本發明之具有減低檢體需求量之輔助元件 ‘ '^電極試片之另一具體實施態樣，其包含： 、 一電絕緣基板（1); 導電膜之電極系統（2)，其塗覆於該電絕緣基板（丨）之一 面，以形成分離且不連接之一工作電極（2a)與一參考電極 (2b); 一配置於該基板上之電絕緣層（3)，其部分覆蓋該電極系 統（2)，電極系統之裸露部分分別形成包含工作電極（2a) 及參考電極（2b)之接線端（4)及電化學反應端（5); 一修飾過之奈米碳管層（6a、6b或6c)，其至少部分覆蓋該 電化學反應端（5)之工作電極； 一塗佈於該電化學反應端（5)及奈米碳管層（6a、讣或以) ^ 上之反應層（7)，其至少包含一電子媒介物； 配置於忒反應層（7)上之絕緣塾高層（8a)，該絕緣墊高 、 層（8a)形成一用以容納檢體至該反應層（7)之缺口，其具 . 有一個開口端（9);及 一配置於該絕緣墊高層（8a)之上且覆蓋該缺口之有孔絕 _ 緣上蓋（10c)’其中該有孔絕緣上蓋（l〇c)與反應層（7)及絕 緣墊高層（8a)共同構成一檢體容納槽道（llc)。 於此一根據本發明之實施態樣中，檢體可以藉由有孔絕 緣上蓋（l〇c)與反應層（7)間所產生之毛細吸引力而由該檢 103662.doc -15· 1276799 體容納槽道（Ue)的開口端（9)到達反應層⑺。 根據本發明，該絕緣塾高層（8a)之厚度-般約為〇·!至〇.5 而該有孔絕緣上蓋⑽)為—具有通氣孔之絕緣上 盖’其厚度-般約為〇.〇5至〇.5毫米。適合作為該絕緣塾高 層（8a)及有孔絕緣上蓋（1()e)之材料包括，但不限於，pvc、 玻璃，截維、聚酿颯、酚醛樹脂板、PET、pc、pp、、PA、 ps、玻璃或陶瓷。1276799 tiOO), tee C-405, three-way Χ-Π4, sodium lauryl sulfate, polyoxyethylene sorbitan monolaurate (eg, Tween 20, Tween 40, Tween 60, Tween 80) , and other water soluble surfactants. 3 shows another embodiment of an auxiliary component of an auxiliary component having a reduced sample requirement according to the present invention, comprising: an electrically insulating substrate (1); an electrode system of a conductive film (2) Applying to one side of the electrically insulating substrate (丨) to form a separate and not connected one working electrode (2a) and one reference electrode (2b); an electrically insulating layer (3) disposed on the substrate, Partially covering the electrode system (2), the exposed portions of the electrode system respectively form a terminal (4) including a working electrode (2a) and a reference electrode (2b) and an electrochemical reaction end (5); a modified nanometer a carbon tube layer (6a, 6b or 6c) at least partially covering the working electrode of the electrochemical reaction end (5); a coating on the electrochemical reaction end (5) and a carbon nanotube layer (6a, 讣 or a reaction layer (7) comprising at least an electron medium; an insulating high layer (8a) disposed on the ruthenium reaction layer (7), the insulating mat height and the layer (8a) forming a accommodating a gap from the sample to the reaction layer (7), having an open end (9); and a height disposed on the insulating mat The upper cover (10c) above the (8a) and covering the notch, wherein the perforated insulating upper cover (10c) and the reaction layer (7) and the insulating pad upper layer (8a) together constitute a sample housing Channel (llc). According to the embodiment of the present invention, the sample can be detected by the capillary attraction between the perforated insulating cover (10c) and the reaction layer (7). The test 103662.doc -15· 1276799 The open end (9) of the body receiving channel (Ue) reaches the reaction layer (7). According to the present invention, the thickness of the high-rise insulating layer (8a) is generally about !·! to 〇.5 and the perforated insulating upper cover (10) is an insulating cover having a venting opening, the thickness of which is about 〇. 〇5 to 〇.5 mm. Suitable materials for the insulating high-rise (8a) and perforated insulating upper cover (1()e) include, but are not limited to, pvc, glass, truncation, polystyrene, phenolic resin sheet, PET, pc, pp, PA, ps, glass or ceramic.
圖4顯7F根據本發明之具有減低檢體需求量之輔助元件 之奈米碳管層之電極試片之一具體實施態樣，其包含： 一電絕緣基板（1); 一導電膜之電極系統（2)，其塗覆於該電絕緣基板（1)之一 面，以开> 成分離且不連接之一工作電極（2a)與一參考電極 (2b)； 一配置於該基板上之電絕緣層，其部分覆蓋該電極系 統（2)，電極系統之裸露部分分別形成包含工作電極（2a) 及參考電極（2b)之接線端（4)及電化學反應端（5); 一修飾過之奈米碳管層（6a、6b或6c)，其至少部分覆蓋該 電化學反應端（5)之工作電極； 一塗佈於該電化學反應端（5)及奈米碳管層（6a、6b或6c) 上之反應層（7)，其至少包含一電子媒介物； 一配置於該反應層（7)上之絕緣墊高層（8a)，該絕緣墊高 層（8a)形成一用以容納檢體至該反應層（7)之缺口，其具 有一個開口端（9); 一配置於該絕緣墊高層（8a)之上且覆蓋該缺口之無孔絕 103662.doc 16 1276799 I (1〇d)’其中該無孔絕緣上蓋（l〇d)與反應層（7)及絕 ♦ 緣墊高層（8a)共同構成一檢體容納槽道（lid);及 仏於孩電絕緣基板（1)上之通氣孔（12)，其與該檢體容 納槽道（lid)相連。 . 、此一根據本發明之實施態樣中，該檢體容納槽遒（lid) • '、八有毛細作用之取樣空間，可使檢體由該檢體容納槽 道（Ud)的開口端（9)到達反應層（7)。 ·) 一根據本發明，該絕緣墊高層（8a)之厚度一般約為0.1至0.5 毫米而忒供孔絕緣上蓋（10d)之厚度一般約為〇〇5至〇·5毫 米。適合作為該絕緣墊高層（8a)及無孔絕緣上蓋（10d)之材 、斗已括仁不限於，PVC、玻璃纖維、聚酯楓、酴酸樹脂 板 PET、PC、pp、PE、PA、PS ' 玻璃或陶瓷。 本發明之再一目的為提供一生物感測器，其包含本文所 述的電極試片及檢測裝置。較佳的是，該檢測裝置為由一 黾i輸出裝置’ 一訊號接收裝置’以及一顯示裝置所組成 ϋ 之電流感測器。該電壓輸出裝置能輸出400 mV以下之電壓 至根據本發明之電極試片之電化學反應區，促使反應層與 、 檢體中特定待測物反應後而使充滿於電化學反應區之電子 媒介物由還原狀態氧化成為氧化狀態。該訊號接收裝置能 將此狀態改變時的電流、電壓或電阻值接收，並傳回顯示 - 裝置’藉此顯示檢體中特定待測物的含量。 本發明之電極試片並不需要如先前技藝般繁瑣之加工步 驟’同時仍能保留足夠大之電流訊號。此外，本發明之電 極式片具有可減低檢體需求量之設計，且可具有多個取樣 103662.doc 1276799 位置（例如：將電極試片靠近檢體或將檢體滴在電極試片 上因此’本發明之電極試片不僅製備較為簡便，可更方 便而有效的自待分析物取樣，以使患者的不便可減至最 低’且同時仍能滿足保留足夠大之電流訊號的需求。 • 以下實施例將對本發明作進一步之說明，唯非用以限制 本發明之範圍’任何於此項技術領域中具有通常知識者可 輕易達成之修飾及改變，均涵蓋於本發明之保護範圍内。 ·) 實例 實例1 將導電碳漿，以網版印在一 PVC板基材（1)之扁平表面 上，以形成由各自分離之一工作電極（2a)與一參考電極（2b) 所組成之電極系統（2)，然後在40〜150°C之下烘乾；隨即在 印有該電極系統（2)之同一侧，塗佈一電絕緣漆（3)，且保留 部份裸露的工作電極與參考電極以形成接線端（4)及電化學 反應端（5)，然後在40〜150°C之下烘乾；再將含有0.1 %修飾 過之單層壁奈米碳管（Single Wall Carbon Nanometer • Tube; SWCNT)之丙酮懸浮液，滴加於該電化學反應端（5)， . 以形成一修飾過之奈米碳管層（如6a)，然後使丙酮乾燥而完 成併入修飾過之奈米碳管之生物感測電極試片。 以相同方式，惟，不使用修飾過之奈米碳管，完成一作 - 為控制組之用之未併入修飾過之奈米碳管之電極試片。 分別以（Α)0.1Μ ρΗ7·0之磷酸緩衝液（PBS)與 （Β)20〇 mg/dL葡萄糖水作為檢體，對上述根據本發明之併入修飾過 之奈米碳管之電極試片（b組）與未併入修飾過之奈米碳管之 103662.doc -18- 1276799 控制組電極試片（a組），進行循 (cyclic voltammetry) 分析，結果顯示於圖5。其顯 根據本發明之併入修飾過 之奈米碳管電極試片的氧化還 遂原，皮峰電流明顯被強化（如 圖5-B)，f景電流的增加則不明顯（如圖μ卜 續以如下組成之反應層溶液滴加於上述根據本發明之電 極試片之修飾過之奈米碳管層上·· 0.3% 3.0% 0.3% 96.4% 葡萄糖氧化酵素 鐵氰化鉀 三通X-100(第三辛基苯氧基聚乙醇) 擰檬酸鹽緩衝液（PH6.5) 然後於贼下烘乾以形成一反應層，而完成根據本發明 之併入修飾過之奈米碳管之葡萄糖感測電極試片。 以如上相同方式，於上述未併入修飾過之奈米碳管之控 制組電極試片上形成一反應層，而完成作為控制組用之未 併入修飾過之奈米碳管之葡萄糖感測電極試片。Figure 4 shows a specific embodiment of an electrode test piece of a carbon nanotube layer having an auxiliary component for reducing the amount of sample required according to the present invention, comprising: an electrically insulating substrate (1); an electrode of a conductive film a system (2) coated on one side of the electrically insulating substrate (1) to be separated and not connected to one of the working electrode (2a) and one reference electrode (2b); one disposed on the substrate An electrically insulating layer partially covering the electrode system (2), wherein the exposed portions of the electrode system respectively form a terminal (4) including a working electrode (2a) and a reference electrode (2b) and an electrochemical reaction end (5); a carbon nanotube layer (6a, 6b or 6c) which at least partially covers the working electrode of the electrochemical reaction end (5); a coating on the electrochemical reaction end (5) and a carbon nanotube layer ( a reaction layer (7) on 6a, 6b or 6c), comprising at least one electron medium; a high-rise (8a) insulating spacer disposed on the reaction layer (7), the upper layer (8a) of the insulating pad is formed To accommodate the gap of the sample to the reaction layer (7), which has an open end (9); a high layer disposed on the insulating mat 8a) above and covering the gap without holes 103662.doc 16 1276799 I (1〇d)' where the non-porous insulating upper cover (l〇d) and the reaction layer (7) and the rim edge of the upper layer (8a) Together, a sample receiving channel (lid) is formed; and a vent hole (12) on the child's electrically insulating substrate (1) is connected to the sample receiving channel (lid). According to the embodiment of the present invention, the sample receiving slot (lid) • ', eight capillary processing space, the specimen can be accommodated by the specimen receiving channel (Ud) open end (9) Arrive at the reaction layer (7). According to the present invention, the upper layer of the insulating mat (8a) is generally about 0.1 to 0.5 mm thick and the thickness of the insulating perforated upper cover (10d) is generally about 〇〇5 to 〇·5 mm. Suitable for the high-rise (8a) and non-porous insulating cover (10d) of the insulating mat, the bucket is not limited to PVC, glass fiber, polyester maple, tantalum resin sheet PET, PC, pp, PE, PA, PS 'glass or ceramic. It is still another object of the present invention to provide a biosensor comprising the electrode test strip and detection device described herein. Preferably, the detecting means is a current sensor comprising a 输出i output device 'a signal receiving device' and a display device. The voltage output device can output a voltage of 400 mV or less to the electrochemical reaction zone of the electrode test piece according to the present invention, and cause the reaction layer to react with a specific analyte in the sample to make the electronic medium filled in the electrochemical reaction zone The substance is oxidized from a reduced state to an oxidized state. The signal receiving device can receive the current, voltage or resistance value when the state is changed, and return it to the display device to thereby display the content of the specific object to be tested in the sample. The electrode test strip of the present invention does not require the cumbersome processing steps of the prior art while still retaining a sufficiently large current signal. In addition, the electrode type sheet of the present invention has a design capable of reducing the amount of sample required, and can have a plurality of sampling positions 103662.doc 1276799 (for example, bringing the electrode test piece close to the sample or dropping the sample onto the electrode test piece) The electrode test piece of the invention is not only simple to prepare, but also more convenient and effective for sampling the analyte to be used, so that the patient's inability can be minimized while still meeting the requirement of retaining a sufficiently large current signal. The invention is further described, but is not intended to limit the scope of the invention. Any modifications and variations that may be readily made by those skilled in the art are intended to be included within the scope of the invention. EXAMPLES Example 1 A conductive carbon paste was screen-printed on a flat surface of a PVC board substrate (1) to form an electrode system composed of a working electrode (2a) and a reference electrode (2b) separated from each other. (2), then drying at 40~150 ° C; then on the same side printed with the electrode system (2), coating an electrically insulating varnish (3), and retaining part of the bare working electrode and the reference The electrode is formed to form a terminal (4) and an electrochemical reaction end (5), and then dried at 40 to 150 ° C; and then a 0.1% modified single-walled carbon nanotube (Single Wall Carbon Nanometer • An acetone suspension of Tube; SWCNT) is added dropwise to the electrochemical reaction end (5) to form a modified carbon nanotube layer (such as 6a), and then the acetone is dried to complete the incorporation of the modified neat Biosensing electrode test piece of rice carbon tube. In the same manner, except that the modified carbon nanotubes were not used, one was performed - an electrode test piece for the control group that was not incorporated into the modified carbon nanotubes. An electrode test of the above-described modified carbon nanotubes according to the present invention was carried out by using (Α) 0.1Μ ρΗ7·0 phosphate buffer (PBS) and (Β) 20 〇mg/dL glucose water as the samples, respectively. The tablets (group b) and the 103662.doc -18-1276799 control group electrode test piece (group a) which was not incorporated into the modified carbon nanotubes were subjected to cyclic voltammetry analysis, and the results are shown in Fig. 5. According to the present invention, the oxidation of the modified carbon nanotube electrode test piece is further reduced, and the peak current is obviously enhanced (Fig. 5-B), and the increase of the f-state current is not obvious (Fig. μ Further, a reaction layer solution having the following composition is dropwise added to the modified carbon nanotube layer of the above-mentioned electrode test piece according to the present invention · 0.3% 3.0% 0.3% 96.4% Glucose oxidase potassium ferricyanide tee X -100 (trioctylphenoxypolyethanol) citrate buffer (pH 6.5) and then dried under thieves to form a reaction layer, and the modified nano carbon according to the present invention is completed. Glucose sensing electrode test piece of tube. In the same manner as above, a reaction layer was formed on the control group electrode test piece which was not incorporated into the modified carbon nanotube, and was not incorporated into the control group. Glucose sensing electrode test piece of carbon nanotubes.
另進衧不同葡萄糖濃度檢體（0, 25, 50, 1〇〇5 4()()5 mo mg/dL)之時序-安培（chronoamperometry)分析（如圖 6A)。其 結果顯示（如圖6B)，於特定供電時間下，根據本發明之併 入修飾過之奈米碳管之葡萄糖感測電極試片（6B_b組），其還 原態電子媒介物之氧化電流於扣除背景電流後之電流值 (△ current)，相較於控制組（6B-a組）明顯被強化。 實例2 將導電板聚’以網版印在一 PVC板基材（1)之扁平表面 上’以形成由各自分離之一工作電極（2 a)與一參考電極（2b) 103662.doc -19-A sequence-chronoamperometry analysis of different glucose concentration samples (0, 25, 50, 1〇〇5 4()() 5 mo mg/dL) was also performed (Fig. 6A). The results show (as shown in FIG. 6B) that, under a specific power supply time, the glucose sensing electrode test piece (Group 6B_b) incorporating the modified carbon nanotube according to the present invention has an oxidation current of the reduced electron carrier. The current value (Δ current) after subtracting the background current was significantly enhanced compared to the control group (Group 6B-a). Example 2 The conductive sheets were collectively printed on a flat surface of a PVC sheet substrate (1) to form a working electrode (2 a) separated from each other and a reference electrode (2b) 103662.doc -19 -
1276799 所組成之電極系統（2)，然後在4〇〜15(TC之下烘乾；隨即在 印有該電極系統（2)之同一側，塗佈一電絕緣漆（3)，且保留 部份裸露的工作電極與參考電極以形成接線端（4)及電化學 反應端（5)，然後在4〇〜i5(rc之下烘乾；再將含有以下組合 之漿體： 修飾過之多層壁奈米碳管 0.1〜0.4% (Multiple Walls Carbon Nanometer Tube ； MWCNT) 羧曱基纖維素（CMC) 21.5% 二通X-100(第三辛基苯氧基聚乙醇） 0.5% 7jC 77·9 〜77.6% 以網印方式塗佈於該電化學反應端（5)之一部份工作電 極（2a)上，以形成一修飾過之奈米碳管與高分子載體複合層 (如6c) ’然後使該複合層乾燥。依序操作下列製備步驟，以 形成如圖2所示之可使檢體微小化之感測電極試片： 以如實例1之反應層溶液，滴加於已乾燥之修飾過之奈米 碳管與高分子載體複合層之上，然後於5〇〇c下烘乾以形 成一反應層（7); 毛v亥反應層（7)上形成一絕緣塾局層（ga)(pET、厚户〇 h 釐米），該絕緣墊高層（8a)形成一用以容納檢體至該反應 層（7)之缺口以及一缺口的開口端（9);及 於該絕緣墊高層（8a)之上形成一覆蓋該缺口之親水性網 狀覆蓋層（l〇b)(經界面活性劑處理之聚酸石風 (polyethersulfone ; PES)網，從而該親水性網狀覆苗層 (1 〇b)與反應層（7)及絕緣墊高層（8a)共同構成一檢體^二 103662.doc -20- 1276799 槽道（lib) ’藉此，可以藉由該網狀覆蓋層（1〇b)之親水性 由該網狀覆蓋層（l〇b)上方加入檢體，或藉由網狀覆蓋層 (10b)與反應層（7)間所產生之毛細吸引力而使檢體由該 檢體春納槽道（1 lb)的開口端（9)到達反應層（？）。 實例3 將實例2之製備步驟改以： 以如實例1之反應層溶液，滴加於已乾燥之修飾過之奈米 碳管與局分子載體複合層之上，然後於5下烘乾以形 成一反應層（7); 於該反應層（7)上形成一絕緣墊高層（8a)( pET、厚度〇·35 釐米），該絕緣墊高層（8a)形成一用以容納檢體至該反應 層（7)之缺口以及一缺口的開口端（9);及 於戒絕緣墊咼層（8a)之上形成一覆蓋該缺口之有孔絕緣 上蓋（PET、厚度0.1釐米）（10c)，從而該有孔絕緣上蓋（1〇c) 與反應層（7)及絕緣墊高層（8a)共同構成一檢體容納槽道 (1 lc)，藉此，可以藉由該有孔絕緣上蓋（1〇c)與反應層（7) 間所產生之毛細吸引力而使檢體由該檢體容納槽道（丨i c) 的開口端（9)到達反應層（7); 以形成如圖3所示之可使檢體微小化之感測電極試片。 實例4 將實例2之製備步驟改以： 以如貫例1之反應層溶液，滴加於已乾燥之修飾過之奈米 石反f與尚分子載體複合層之上，然後於5〇π下烘乾以形 成一反應層（7); 103662.doc -21 -The electrode system (2) consisting of 1276799 is then dried at 4 〇 to 15 (TC); then an electrically insulating varnish (3) is applied on the same side of the electrode system (2), and the retaining portion is retained. a bare working electrode and a reference electrode to form a terminal (4) and an electrochemical reaction end (5), and then dried at 4 〇 to i5 (rc; and then a slurry containing the following combination: modified multilayer 0.1% to 0.4% (Multiple Walls Carbon Nanometer Tube; MWCNT) Carboxymethyl Cellulose (CMC) 21.5% Two-way X-100 (Third Octylphenoxy Polyethanol) 0.5% 7jC 77·9 ~77.6% is applied by screen printing on a part of the working electrode (2a) of the electrochemical reaction end (5) to form a modified nanocarbon tube and polymer carrier composite layer (such as 6c). Then, the composite layer is dried. The following preparation steps are sequentially operated to form a sensing electrode test piece which can make the sample miniaturized as shown in FIG. 2: the reaction layer solution as in Example 1 is added dropwise to the dried The modified nano carbon tube and the polymer carrier composite layer are then dried at 5 ° C to form a reaction layer (7); An insulating layer (ga) (pET, thick 〇h cm) is formed on the v-reaction layer (7), and the upper layer (8a) of the insulating pad forms a gap for accommodating the sample to the reaction layer (7) And a notched open end (9); and a hydrophilic mesh covering layer (10) covering the notch (8a) of the insulating mat (the surfactant-treated polycide wind ( Polyethersulfone; PES) network, such that the hydrophilic network layer (1 〇b) and the reaction layer (7) and the insulating layer (8a) together form a sample ^ 2, 032, 662. doc -20 - 1276799 channel ( Lib) ' Thereby, the sample can be added from above the network cover layer (1〇b) by the hydrophilicity of the network cover layer (1〇b), or by the network cover layer (10b) and the reaction The capillary attraction generated between the layers (7) causes the specimen to reach the reaction layer (?) from the open end (9) of the specimen Chunna channel (1 lb). Example 3 The preparation procedure of Example 2 was changed : The reaction layer solution as in Example 1 was dropped on the dried modified carbon nanotube and the local molecular carrier composite layer, and then dried at 5 to form a reaction layer (7). An insulating mat high layer (8a) (pET, thickness 〇·35 cm) is formed on the reaction layer (7), and the insulating mat upper layer (8a) is formed to accommodate the sample to the reaction layer (7). a notch and a notched open end (9); and a perforated insulating cover (PET, 0.1 cm thick) (10c) covering the notch on the insulating pad layer (8a), so that the perforated insulating cover (1〇c) together with the reaction layer (7) and the upper layer of the insulating mat (8a) constitute a sample receiving channel (1 lc), whereby the porous insulating cover (1〇c) and the reaction layer can be used (7) The capillary attraction generated between the specimens reaches the reaction layer (7) from the open end (9) of the sample receiving channel (丨ic); to form a sample as shown in FIG. Miniaturized sensing electrode test piece. Example 4 The preparation procedure of Example 2 was changed to: using a reaction layer solution as in Example 1, dropwise addition to the dried modified nanostone anti-f and the molecular carrier composite layer, and then at 5 〇π Drying to form a reaction layer (7); 103662.doc -21 -
1276799 於該反應層（7)上形成一絕緣墊高層（8a)( PET、厚度0.35 釐米），該絕緣墊高層（8a)形成一用以容納檢體至該反應 層（7)之缺口以及一缺口的開口端（9);及 於該絕緣塾高層（8a)之上形成一覆蓋該缺口之（pet、厚 度〇· 1釐米）無孔絕緣上蓋（10d)，從而該無孔絕緣上蓋 (1 〇d)與反應層（7)及絕緣墊高層（8a)共同構成一檢體容納 槽道（lid);及 於該PVC板基板（1)上形成一與該檢體容納槽道（lld)相 連之通氣孔（12)，藉此，該檢體容納槽道（lid)成為具有 毛細作用之取樣空間，可使檢體由該檢體容納槽道（lld) 的開口端（9)到達反應層（7); 以形成如圖4所示之可使檢體微小化之感測電極試片。 依據上述實例2〜4之製備步驟，完成根據本發明之併入修 飾過之奈米碳管之葡萄糖感測電極試片之其它實施態樣。 使該等電極試片進行與實例1相似之測試後發現，該等併入 修飾過之奈米碳管之葡萄糖感測感測電極試片，具有與實 例1相似之強化特徵，其約可增加15〜4〇%之 chronoamperometry氧化電流。 經由以上實施方式顯示，併入修飾過之奈米碳管，可使 生物感測電極試片之氧化還原電流訊號增強，不僅可有效 提升訊號/雜訊比（S/N比），更能改善量測之精確度與準確 度。此外’所有實施例所揭示的生物感測電極試片，不僅 製造較為簡便’且具有檢體承載空間縮小化（即，減少檢體 的需求量以降低病患的痛楚）之優點，並且可滿足保留夠大 103662.doc -22- 1276799 而便於偵測之電流訊號（> π(==βΑ)’甚至強化電流訊號的需 求。對於欲录顧電極試片 儀器製造廠商而…與品質之相關電化學 • Η m本發明之併人修飾過奈米碳管之 生物感測電極試片確實可提供方便且有效的解決方案。 本發明的技術内容和锆傅 争徵’已无分描述於上述實例及說 明中’應了解的是，任材 、 仃不脫離本發明精神下所為之修飾 或改變，皆應涵蓋於本發明之保護範圍中。 【圖式簡單說明】 圖1(A)為根據本發明具體實施例的電極試片之分層結構 圖。 圖1(B)係圖1(A)電極試片之立體圖。 圖1(C)係圖1(B)電極試片之a-A線的剖面圖。 圖2為根據本發明具體實施例的電極試片之結構分解固 圖3為根據本發明具體實施例的電極試片之結構分解圖 圖4為根據本發明具體實施例的電極試片之結構分解固 圖5為根據本發明之併入修飾過之奈米碳管之電極1片 與未併入修飾過之奈米碳管之控制組電極試片，進彳_ $ 伏安（cyclic voltammetry)之分析結果。 衰 圖6根據本發明之併入修飾過之奈米碳管之電極$ # 未併入修飾過之奈米碳管之控制組電極試片，進行時序、 培（chronoamperometry)之分析結果。 【主要元件符號說明】 1 2 電絕緣基板 電極系統 103662.doc -23- 1276799 ·) 2a 2b 3 4 5 6a, 6b，6c 7 8，8a 9, 9a 10a，lOd 10b 10c 1 la-1 Id 12 工作電極 參考電極 電絕緣層 接線端 電化學反應端 修飾過之奈米碳管層 反應層 絕緣塾1¾層 絕緣塾高層之開口端 無孔絕緣上蓋 網狀覆蓋層 有孔絕緣上蓋 具毛細作用力之檢體容納槽道 通氣孔1276799, an insulating mat high layer (8a) (PET, thickness 0.35 cm) is formed on the reaction layer (7), and the insulating mat high layer (8a) forms a gap for accommodating the sample to the reaction layer (7) and a a notched open end (9); and a non-porous insulating upper cover (10d) covering the notch (pet, thickness 〇·1 cm) is formed on the insulating raft upper layer (8a), so that the non-porous insulating cover (1) 〇d) together with the reaction layer (7) and the upper layer of the insulating mat (8a) constitute a sample receiving channel (lid); and forming a sample receiving channel (lld) on the PVC board substrate (1) The vent hole (12) is connected, whereby the sample accommodating channel (lid) becomes a sampling space having a capillary action, and the sample is allowed to reach the reaction from the open end (9) of the sample accommodating channel (11d). Layer (7); to form a sensing electrode test piece which can make the sample miniaturized as shown in FIG. According to the preparation steps of the above Examples 2 to 4, other embodiments of the glucose sensing electrode test piece incorporating the modified carbon nanotube according to the present invention were completed. The electrode test pieces were subjected to tests similar to those of Example 1 and found that the glucose sensing sensing electrode test pieces incorporating the modified carbon nanotubes had similar strengthening characteristics as in Example 1, which may be increased. 15~4〇% of the chronoamperometry oxidation current. According to the above embodiment, the incorporation of the modified carbon nanotubes can enhance the redox current signal of the bio-sensing electrode test piece, which can not only effectively improve the signal/noise ratio (S/N ratio), but also improve the performance. The accuracy and accuracy of the measurement. In addition, the biosensing electrode test piece disclosed in all the embodiments is not only simple to manufacture, but also has the advantages of reducing the carrying capacity of the sample (that is, reducing the demand of the sample to reduce the pain of the patient), and can satisfy Retain large enough 103662.doc -22- 1276799 for easy detection of current signals (> π(==βΑ)' and even enhance the need for current signals. For the manufacturer of the electrode test instrument to be consulted... Electrochemistry • Η m The biosensing electrode test piece of the present invention modified by a carbon nanotube does provide a convenient and effective solution. The technical content of the present invention and the zirconium sequel' have been described in the above examples and In the description, it should be understood that any modifications or changes that may be made without departing from the spirit of the invention are intended to be included in the scope of the invention. FIG. 1(A) is in accordance with the present invention. Fig. 1(B) is a perspective view of the electrode test piece of Fig. 1(A). Fig. 1(C) is a cross-sectional view of the aA line of the electrode test piece of Fig. 1(B). Figure 2 is a concrete embodiment in accordance with the present invention 3 is a structural exploded view of an electrode test piece according to an embodiment of the present invention. FIG. 4 is a structural exploded view of an electrode test piece according to an embodiment of the present invention. FIG. One piece of the electrode incorporating the modified carbon nanotube tube and the control group electrode piece not incorporated into the modified carbon nanotube tube, the analysis result of the cyclic voltammetry. The fading pattern 6 according to the present The invention incorporates the modified nanocarbon tube electrode ## The control group electrode test piece which has not been incorporated into the modified nano carbon tube is subjected to the analysis of the timing and chronoamperometry. [Main component symbol description] 1 2 Electrically insulating substrate electrode system 103662.doc -23- 1276799 ·) 2a 2b 3 4 5 6a, 6b, 6c 7 8,8a 9, 9a 10a, lOd 10b 10c 1 la-1 Id 12 Working electrode reference electrode electrical insulation layer Terminal electrochemical reaction end modified nanocarbon tube layer reaction layer insulation 塾13⁄4 layer insulation 塾 high-rise open end non-porous insulation upper cover mesh cover layer perforated insulation cover with capillary force sample receiving channel vent
103662.doc -24-103662.doc -24-
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW94132200ATWI276799B (en) | 2005-09-16 | 2005-09-16 | Electrode trip and biosensor comprising the same |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW94132200ATWI276799B (en) | 2005-09-16 | 2005-09-16 | Electrode trip and biosensor comprising the same |
| Publication Number | Publication Date |
|---|---|
| TWI276799Btrue TWI276799B (en) | 2007-03-21 |
| TW200712485A TW200712485A (en) | 2007-04-01 |
| Application Number | Title | Priority Date | Filing Date |
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| TW94132200ATWI276799B (en) | 2005-09-16 | 2005-09-16 | Electrode trip and biosensor comprising the same |
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| TW (1) | TWI276799B (en) |
| Publication number | Priority date | Publication date | Assignee | Title |
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| US8236154B2 (en) | 2007-07-05 | 2012-08-07 | Apex Biotechnology Corp. | Composite modified electrode strip |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109030595B (en)* | 2017-06-09 | 2023-09-26 | 清华大学 | Biosensor electrode and biosensor |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8236154B2 (en) | 2007-07-05 | 2012-08-07 | Apex Biotechnology Corp. | Composite modified electrode strip |
| Publication number | Publication date |
|---|---|
| TW200712485A (en) | 2007-04-01 |
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