200804803 九、發明說明： 【發明所屬之技彳衧領域】 特別有關於一種具有 c酵素感測器。 本每明有關於一種酵素感測器， 延伸式感測場效電晶體結構之維生素 【先前技術】 、准生素c因可用於治療及預 血酸(_blcacid),近料广：：血病，故又稱為抗壞 體中錐4打錢旦 床研究結果發現人 碎 I ./里關係著許多疾病之發病率，例如痒 症〜臟病、糖尿病、風濕性關二 症候群，因此測量攝取食物中維生；；：=性疲勞 性，又因其與強化免疫力Γ堅:增強白血球之活 關，並且能阻止亞侧與胺二:=:自由基有 生素C含量之測量方法有如-般進行維 的氧化還原反應以進行維生素c之^^子及维生素c 破酸鉀溶液（κιο3) v找,⑴^析，此方法係以 化鉀㈤先反應 維峰音Γ、隹—长儿碘刀子可與待測溶液中的 、准生素C進耗化還原反應，當溶液 完畢後，過量的诚分子"、齒〜、六山 京匚凡王反應 中硬離子(Γ)生成破離子 (3)亚和預先加入的澱粉指示劑產生 酸之化與疋劑之滴定體積及其與抗壞血 -夂之化子㈣係即可計算出待 〇619-A21621TWF(N2)；cIaire 200804803 然而’以此方法進行維生素c的定量分析並不能快速得知 分析結果，而還需要更方便及快速的測量方法。 此外’亦有一些研究係針對維生素c之感測器，例如 Erol Akyilmaz荨人在國際期刊gel (Talanta，第50卷，第 87-93頁）揭示一種黃瓜中之抗壞血酸氧化酶與分離式氧探 針所結合之生物感測器，並利用其測量果汁及維生素C片 内所含有之維生素C濃度。此感測器係以戊二搭膠體及預 先處理完成之鐵氟龍（Telflon)膜固定抗壞血酸氧化酶做為 酵素電極。另外，該文獻亦將測量果汁及維生素C片内之 維生素 C 濃度的結果與遇弱氧化劑（2,6 dichloroplienolindophenol)方法加以比較。200804803 IX. Description of the invention: [Technical field to which the invention pertains] Particularly relates to a sensor having a c-enzyme. Each of these has an enzyme sensor, an extended sensing field-effect transistor structure of the vitamin [previous technology], the quasi-suppressant c can be used for treatment and pre-ascorbic acid (_blcacid), near the material:: blood disease Therefore, it is also known as the anti-bad body cone 4 hit money bed research results found that human broken I. / Li is related to the incidence of many diseases, such as itching ~ visceral disease, diabetes, rheumatic sex syndrome, therefore measuring food intake Zhongweisheng;;:=sexual fatigue, and because of its strong immunity to strengthening the immune system: to enhance the activity of white blood cells, and to prevent the sub-side and amine II: =: free radicals, the content of C is measured - The dimensional redox reaction is carried out to carry out vitamin C and vitamin C potassium bromate solution (κιο3) v, (1) analysis, this method is based on potassium (5) first response Weifeng sound Γ, 隹 - long children The iodine knife can be reduced and reacted with the quasi-biotic C in the solution to be tested. When the solution is completed, the excessive amount of the hard molecule (Γ) is broken in the reaction of the "Cheng Molecular", the tooth~, and the Liushan Jingyu Fanwang reaction. Ionic (3) sub-and pre-added starch indicator to produce acidification and bismuth The fixed volume and its anti-scurvy-sputum scorpion (4) system can be calculated as 619-A21621TWF (N2); cIaire 200804803 However, quantitative analysis of vitamin C by this method does not quickly know the analysis results. There is also a need for more convenient and fast measurement methods. In addition, there are also some researches on vitamin C sensors, such as Erol Akyilmaz. In the international journal gel (Talanta, Vol. 50, pp. 87-93), an ascorbate oxidase and a separate oxygen probe are disclosed in a cucumber. A biosensor combined with a needle and used to measure the concentration of vitamin C contained in juice and vitamin C tablets. The sensor is a ferrite electrode with a glutarylene colloid and a pretreated Teflon membrane immobilized ascorbate oxidase. In addition, the literature also compares the results of measuring the concentration of vitamin C in juice and vitamin C tablets with the method of 2,6 dichloroplienolindophenol.
Philip J· O’Connell 等人在國際期刊 SCI (Analytica Chemica Acta，第431卷，第239-247頁）揭示一種使用低 電壓之感測器，其用以測量尿酸、檸檬酸及抗壞血酸（維生 素C)之濃度。該感測器使用電極為多苯胺聚合於玻璃碳電 極及網版印刷電極。該文獻亦探討工作電壓之選擇以及電 聚合時間對響應電壓之影響。Philip J. O'Connell et al., International Journal SCI (Analytica Chemica Acta, Vol. 431, pp. 239-247) discloses a low voltage sensor for measuring uric acid, citric acid and ascorbic acid (vitamin C). The concentration of ). The sensor uses an electrode to polymerize polyaniline to a glassy carbon electrode and a screen printing electrode. The literature also explores the choice of operating voltage and the effect of electropolymerization time on the response voltage.
Viatcheslav Volotovsky 等人在國際期刊 SCI (Analytica Chemica Acta，第43 1卷，第239-247頁）揭示一種以離子 感測場效電晶體（ion-sensitive field effect transisitor，ISFET) 為感測基底之感測器，其並結合酵素固定化技術與酵素反 應之單一性而形成。文獻中並以該感測器測量果汁與市售 飲料内之維生素C濃度。 然而，上述維生素C感測器仍有其限制，例如，離子 0619-A21621TWF(N2)；claire 6 200804803 感測場效電晶體易受待測溶液中訊號之干擾，因此此種酵 素感測器仍亟待改善之道。並且未有利用延伸式離子感測 場效電晶體（Extended Gate Field Effect Transistor, EGFET) 架構製備離子選擇電極，使用維生素C酵素薄膜固定化於 一基板上之氧化層上而延伸出來做為感測膜者。 【發明内容】 有鑑於此，本發明的目的在於提供一種可量測維生素 C ;辰度之感測器，其可改善上述離子感測場效電晶體之缺 點5將維生素C酵素感測薄膜固定於延伸式離子感測場效 電晶體之感測元件部分。本發明又一目的在於提供_種測 量一溶液中維生素C濃度的系統，可利用上述之維生素c 酵素感測器進行維生素c濃度之量測。 根據上述目的，本發明提供一種維生素c酵素感測 器’其係一延伸式閘極場效電晶體結構，包括··一金氧半 場效電晶體，位於一半導體基底上；一感測元件，其包括 一導電層，一感測薄膜位於該導電層上，以及一維生素c 酵素薄膜固定於該氧化層上；以及一導線，連接該金氧半 ¥政電晶體與該感測元件。 根據上述目的，本發明提供一種測量一溶液中維生素 C /辰度的系統，其包括：一如申請專利範圍第1項之維生 素c酵素感測器；一參考電極以提供穩定電壓；以及—半 導體特性量測儀，其分別與該維生素C酵素感測器與該參 考電極連接。 06l9'-A2l621TWF(N2)；claire 200804803 【實施方式】 為使本發明之上述和其他目的、特徵、和優點能更明 顯易懂，請參考第1圖做詳細說明如下： 本發明實施例之維生素c酵素感測器，其係一延伸式 感測場效電晶體（Extended Gate Field Effect Transistor, EGFET)結構l〇，包括一金氧半場效電晶體(M〇SFET)21、 一感測元件20、及一導線π。由於感測元件2Q係自延伸 式感測％效電晶體1 〇之閘極延伸出來，亦即，只有感測元 件與待測溶液（未顯示）接觸，而金氧半場效電晶體21元件 口 P刀則14彳寺測洛液隔離，如此可避免半導體元件之不穩定 性與受到溶液中之訊號干擾。 本發明實施例所使用之金氧半場效電晶體21，如一般 結構之N型或p型場效電晶體，位於一半導體基底上，具 有一對源極/汲極區位於閘極兩侧之半導體基底中，閘極可 包括一氧化層。閘極經由導線i 7與本發明所使用之感測元 件20包相連，以獲得來自感測元件2〇之電訊號。可利用 -般商品化之金氧半場效電晶體或場效電晶體CD4〇〇7做 為上述之金氧半場效電晶體。 15上。可利用市售之商品化二氧化錫/氧化銦錫/玻璃作為 感測元件之基底12、導電層18及感測薄膜15 ’並且藉由 本發明實施例之維生素C酵素感測器 件20,包括-基底12，例如麵；—導電層18二= 化铜錫（mdiurn tin oxide ’ IT0) ·，一感測薄膜i 5，例如二I 化錫；以及-維生素C酵素薄膜19固定化於該感測薄^ 〇619-A21621TWF(N2)；ciaire 8 200804803 導線17與金氧半場效電晶體21之閘極端電性連接。上述 感測元件20外並以一絕緣層16包覆，僅露出部分感測薄 膜15與待測溶液（未顯示）接觸。在一實施例中，絕緣層16 為環氧樹脂（epoxy)，導線17為銘導線。由於感測元件需 浸泡於酸鹼溶液中，故封裝於感測元件外部之絕緣層16 以具備良好之絕緣性及抗酸鹼腐蝕能力為較佳，此外，其 亦具備烘乾後體積小、加工性佳的特性為更佳。舉例而言， 以環氧樹指為封裝材料，由於其固化前具流動性且固化後 硬度佳，因此具有良好之加工性。 感測薄膜15未被絕緣層16包覆的部分係感測窗口 14 。於感測窗口 14 内以丙基三曱氧基矽 (3-glycidixypropyltrimethoxysilane，GPTS)固定法進行維生 素C酵素薄膜之固定化，該方法包括：先取丙基三曱氧基 矽與曱苯之混合液滴於該感測窗口内，之後對感測元件實 施烘烤步驟以將上述混合液烘乾，待烘乾後之感測元件冷 卻後，可使用磷酸鹽溶液清洗以將未鍵結之丙基三曱氧基 矽與曱苯之混合液去除。接著，調製一維生素C酵素溶液 並滴於感測窗口 14内，並靜置一段時間以完成酵素固定化 步驟，其中維生素C酵素溶液之調製方法可將維生素C氧 化酶溶於一溶劑中，例如碟酸鹽溶液。於一實施例中，上 述維生素c氧化酶係粉末狀。 請參照第2圖，其係繪示以本發明實施例製作完成的 維生素C酵素感測器之感測元件20部分。感測元件20之 外部以絕緣層16所包覆，僅露出感測窗口 14用以接觸待 0619-A21621TWF(N2);claire 9 200804803 ，♦液。導線η分職接至感測元件2Q及錢半場效+ 晶體(未顯示)，且部分之導線17外可另包覆一毛 广 可在進行測量時達到防水及絕緣之功用。 本發明實施例之維生素c酵素感測器的量測方法 如第4圖之電流·電塵(!_ v)量測系統，將本發明實施例之維 生素c酵素感測器之感測元件2〇浸入待測溶液μ中 測溶液24係置於-容器（未標示）中。該感測器20係經導 Ή ’例如紹導線，將其與金氧半場效電晶體幻相連接， 而該金氧半場效電晶體之汲極與源極分別與半導體特性 (電流-電壓）量測儀Μ，例如Keithley 236相連，以進—I 將電晶體所測得之電訊號做數據處理。 / η另外，在待測溶液24中可同時浸入一參考電極22以 提供疋電流，其可為一銀/氯化銀(Ag/AgC1)參考電極， 且亦經由一導線30與上述半導體特性（電流_電壓）量測儀 31相連；而在容器之外部有一組加熱器25，連接於一溫度 控制器（溫度控制中樞)2 6，而待測溶液2 4之溫度高低則由 連接於該溫度控制器26之熱耦合器27做感應。當埶耦人 器^偵'測到待測溶液24之溫度上升或下降時，該溫度才; 制器（溫度控制中樞)26負責控制該加熱器25進行停止加熱 或加熱之動作。前述之待測溶液24、與該待測溶液24有 接觸之各元件以及加熱器25等皆置放於一光隔絕容器（如 曰曰箱）23中，以隔絕光對量測數值之影響。 上述系統量測維生素C酵素感測器之感測度的方法係 如下所述··首先，將上述維生素c酵素感測器之維生素c 〇619-A21621TWF(N2)；claire 200804803 酵素薄膜與一含有維生素c之溶液接觸。接著，在一 :度下為室溫25。〇改變該維生素W液之酸驗值， 例如_2、邱4、阳6、阳8、阳1〇。此時，上 卿於該維生素C酵素感測器之場效電晶體閑 =纖由1伏糊至6伏特⑺，並對於該維生素C 雕失之場效電晶體源/沒端固t電壓。再以該半導 里測儀3!測量，並記錄該維生素c酵素感測器之 源極琶流對閘極電壓的曲線，如第5 _示 :極電流(w對閘極電壓(VGS)之曲線，取—固定電流以;; 出在该固定溫度下該維生素cg#素感測器之感測度，如第 圖所7τ 第5圖’其係|示本發明實施例之 維生素C酵素感測器於pH 2、pH 4、pH 6、pH 8、阳1〇 以本發明貫施例之量測系統測量維生素c酵素感測器 的最佳量測條件方法如下所述：首先，將上述維生素C酵 素感測器之維生素C酵素薄膜與—含有維生素〇之填酸鹽 綾衝液(phosphate buffer solution，PBS)接觸，接著，在一 固定鱗酸鹽缓衝液濃度下，例如5福，改變該磷酸鹽緩衝 液=酸鹼值，例如 ρΗ 6、ρΗ 6.5、pH 7、pH 7 5、pH 8， 亚量測胃輸出電壓與響應時間之關係，其結果如第7圖所 不’測量結果顯示當5mM之構酸鹽緩衝液酸驗值為pH 7 5 0619-A21621TWF(N2)；claire C溶液’以閘極電壓節制源極電流之機制所得到 t源極電流（lDS)對閘極電壓（Vgs)曲、線。經由第6圖顯示之 置測結果得知本發明實施例之維生素c感測器於p h 2至 PH 10之間的溶液中，感測度約為59.8 mV/pH。 200804803 時，維生素C感測器於量測時較快達到飽和電壓即具有最 快之響應時間。 為使本發明之上述目的、特徵和優點能更明顯易懂， 下文特舉一較佳實施例，並配合所附圖示，作詳細說明如 下： 實施例 本實施例係以依照本發明方法所製備之維生素C酵素 感測器為例，實際測量含有維生素C之待測液，並詳細說 明如何實施本發明。於後述測量條件之實驗參數值乃至於 測量裝置均僅用以舉例說明，但並非用以限定本發明之範 圍。 1.維生素C酵素感測器之製備 本實施例係採用維生素C酵素薄膜做為偵測維生素C 濃度之感測器，其元件之剖面如第1圖所示，首先取商品 化之二氧化錫/氧化銦錫/玻璃（Sn02/IT0 glass)作為感測元 件之感測薄膜15、導電層18及基底12使用，其中該二氧 化錫之片電阻約為18.34 Ω/□，大小約為1公分χθ.4公分， 將該二氧化錫/氧化銦錫/玻璃置於燒杯中並倒入適量之丙 酮以超音波震盪器震盪5分鐘，再以去離子水(D.I water) 將該二氧化錫/氧化銦錫/玻璃清洗乾淨。接著，倒入適量之 乙醇於燒杯中並以超音波震盪器震盪5分鐘，再以去離子 水清洗該將二氧化錫/氧化銦錫/玻璃，之後以氮氣將該二氧 化錫/氧化姻錫/玻璃吹乾。利用銀膠將導線17之一端固定 於該二氧化錫/氧化銦錫/玻璃上，並置於130°C之烤箱約10 0619-A21621TWF(N2)；claire 200804803 刀名里。待導線17固定完成後，將導 ★ 示)中，並利用環氧樹脂16(EP=Viatcheslav Volotovsky et al., International Journal of SCI (Analytica Chemica Acta, Vol. 43 1 , pp. 239-247) discloses an ion-sensitive field effect transisitor (ISFET) for sensing the sense of the substrate. The detector is formed in combination with the singularity of the enzyme immobilization technique and the enzyme reaction. The sensor measures the concentration of vitamin C in juices and commercially available beverages in the literature. However, the above vitamin C sensor still has its limitations, for example, ion 0619-A21621TWF (N2); claire 6 200804803 sensing field effect transistor is susceptible to interference in the signal to be tested, so this enzyme sensor is still There is a need for improvement. And the ion selective electrode is not prepared by using an Extended Gate Field Effect Transistor (EGFET) structure, and is fixed on the oxide layer on a substrate by using a vitamin C enzyme film to be sensed. Membrane. SUMMARY OF THE INVENTION In view of the above, an object of the present invention is to provide a sensor capable of measuring vitamin C, which can improve the shortcomings of the above-mentioned ion sensing field effect transistor, and fix the vitamin C enzyme sensing film. The portion of the sensing element of the extended ion sensing field effect transistor. Still another object of the present invention is to provide a system for measuring the concentration of vitamin C in a solution, which can be measured by the above-mentioned vitamin c enzyme sensor. According to the above object, the present invention provides a vitamin c enzyme sensor, which is an extended gate field effect transistor structure, comprising a gold oxide half field effect transistor, located on a semiconductor substrate; a sensing element, The utility model comprises a conductive layer, a sensing film is disposed on the conductive layer, and a vitamin C enzyme film is fixed on the oxide layer; and a wire connecting the MOS transistor and the sensing element. In accordance with the above objects, the present invention provides a system for measuring vitamin C / Chen in a solution comprising: a vitamin c enzyme sensor as in claim 1; a reference electrode to provide a stable voltage; and - a semiconductor A characteristic measuring instrument is respectively connected to the vitamin C enzyme sensor and the reference electrode. The above and other objects, features, and advantages of the present invention will become more apparent and understood. The enzyme sensor is an extended gate field effect transistor (EGFET) structure, including a metal oxide half field effect transistor (M〇SFET) 21, and a sensing element 20 And a wire π. Since the sensing element 2Q extends from the gate of the extended sensing % effect transistor 1 , that is, only the sensing element is in contact with the solution to be tested (not shown), and the gold oxide half field effect transistor 21 element port The P-knife is isolated from the 14-inch temple, which avoids the instability of the semiconductor components and the signal interference in the solution. The gold-oxygen half field effect transistor 21 used in the embodiment of the present invention, such as a general structure N-type or p-type field effect transistor, is located on a semiconductor substrate and has a pair of source/drain regions on both sides of the gate. In a semiconductor substrate, the gate may include an oxide layer. The gate is connected via a wire i 7 to the sensing element 20 used in the present invention to obtain an electrical signal from the sensing element 2 . A commercially available gold-oxygen half-field effect transistor or field effect transistor CD4〇〇7 can be used as the above-mentioned gold-oxygen half-field effect transistor. 15 on. Commercially available commercial tin dioxide/indium tin oxide/glass can be utilized as the substrate 12 of the sensing element, the conductive layer 18 and the sensing film 15' and by the vitamin C enzyme sensing device 20 of the embodiment of the invention, including - Substrate 12, such as a face; - conductive layer 18 = mdiurn tin oxide 'IT0', a sensing film i 5, such as tin-tin; and - vitamin C enzyme film 19 is immobilized on the sensing Thin ^ 〇 619-A21621TWF (N2); ciaire 8 200804803 The wire 17 is electrically connected to the gate of the MOS half-effect transistor 21. The sensing element 20 is externally covered with an insulating layer 16, and only a portion of the sensing film 15 is exposed to contact with the solution to be tested (not shown). In one embodiment, the insulating layer 16 is epoxy and the wire 17 is an inscribed wire. Since the sensing element needs to be immersed in the acid-base solution, the insulating layer 16 encapsulated on the outside of the sensing element is preferably provided with good insulating property and acid and alkali corrosion resistance, and further has a small volume after drying. Better processing properties are better. For example, epoxy resin is used as the encapsulating material, and has good processability because of its fluidity before curing and good hardness after curing. The portion of the sensing film 15 that is not covered by the insulating layer 16 is a sensing window 14 . Immobilization of the vitamin C enzyme film by a 3-glycidixypropyltrimethoxysilane (GPTS) immobilization method in the sensing window 14, the method comprising: first taking a mixture of propyltrimethoxy hydrazine and hydrazine Drip into the sensing window, and then perform a baking step on the sensing element to dry the mixture, and after the sensing element after drying is cooled, it can be washed with a phosphate solution to remove the unbonded propyl group. The mixture of trimethoxy hydrazine and hydrazine is removed. Next, a vitamin C enzyme solution is prepared and dropped in the sensing window 14 and allowed to stand for a period of time to complete the enzyme immobilization step, wherein the vitamin C enzyme solution is prepared by dissolving the vitamin C oxidase in a solvent, for example, Dish acid salt solution. In one embodiment, the vitamin c oxidase is in the form of a powder. Referring to Figure 2, there is shown a portion of the sensing element 20 of the vitamin C enzyme sensor fabricated in accordance with an embodiment of the present invention. The outer portion of the sensing element 20 is covered with an insulating layer 16, and only the sensing window 14 is exposed for contact with 0619-A21621TWF(N2); claire 9 200804803, ♦ liquid. The wire η is connected to the sensing element 2Q and the half-field effect + crystal (not shown), and part of the wire 17 can be covered with a smear to achieve waterproof and insulating functions when measuring. The measuring method of the vitamin c enzyme sensor of the embodiment of the present invention is the current/electric dust (!_ v) measuring system of FIG. 4, and the sensing element 2 of the vitamin c enzyme sensor of the embodiment of the present invention is used. The immersion test solution 24 in the solution to be tested is placed in a container (not labeled). The sensor 20 is connected to a metal oxide half field effect transistor by means of a conductor, for example, a wire, and the drain and source of the metal oxide half field effect transistor are respectively related to semiconductor characteristics (current-voltage). A measuring instrument, such as Keithley 236, is connected to the data signal processed by the transistor. / η In addition, a reference electrode 22 may be simultaneously immersed in the solution to be tested 24 to provide a krypton current, which may be a silver/silver chloride (Ag/AgC1) reference electrode, and also via a wire 30 and the above semiconductor characteristics ( The current_voltage measuring device 31 is connected; and a heater 25 is connected outside the container to a temperature controller (temperature control center) 2 6, and the temperature of the solution to be tested 24 is connected to the temperature. The thermal coupler 27 of the controller 26 senses. When the temperature of the solution to be tested 24 rises or falls, the temperature is changed; the controller (temperature control center) 26 is responsible for controlling the heater 25 to stop heating or heating. The aforementioned solution to be tested 24, the components in contact with the solution 24 to be tested, and the heater 25 are placed in a light-insulating container (e.g., a box) 23 to isolate the influence of light on the measured value. The method for measuring the sensitivity of the vitamin C enzyme sensor by the above system is as follows: First, the vitamin c 〇619-A21621TWF (N2); claire 200804803 enzyme film and a vitamin containing vitamin C enzyme sensor The solution of c is in contact. Next, at room temperature, it was room temperature 25. 〇 Change the acid value of the vitamin W solution, such as _2, Qiu 4, Yang 6, Yang 8, Yang 1 〇. At this time, Shang Qing in the vitamin C enzyme sensor of the field effect transistor = fiber from 1 volt paste to 6 volts (7), and for the vitamin C carved field effect transistor source / no fixed t voltage. Then measure with the semi-inductance meter 3! and record the source turbulence vs. gate voltage curve of the vitamin C enzyme sensor, as shown in the fifth _: pole current (w versus gate voltage (VGS) The curve, taking a fixed current to;; the sensitivity of the vitamin cg# sensor at the fixed temperature, as shown in the figure 7τ, FIG. 5, which shows the vitamin C enzyme sensation of the embodiment of the present invention. The method for measuring the optimal measurement condition of the vitamin C enzyme sensor by using the measuring system of the present invention at pH 2, pH 4, pH 6, pH 8, and pH 1 is as follows: First, the above The vitamin C enzyme film of the vitamin C enzyme sensor is contacted with a phosphate buffer solution (PBS), and then changed at a fixed sulphate buffer concentration, for example, 5 福Phosphate buffer = pH value, such as ρ Η 6, ρ Η 6.5, pH 7, pH 7 5, pH 8, sub-measurement of gastric output voltage and response time, the results as shown in Figure 7 'measurement results show When the 5 mM acid buffer buffer acid value is pH 7 5 0619-A21621TWF (N2); claire C solution 'gate The voltage source is controlled by the source current (tDS) to the gate voltage (Vgs) curve and line. The measurement result shown in Fig. 6 shows that the vitamin c sensor of the embodiment of the present invention is in ph In the solution between 2 and PH 10, the sensitivity is about 59.8 mV/pH. At 200804803, the vitamin C sensor reaches the saturation voltage faster when it is measured, which has the fastest response time. The objects, features, and advantages will be more apparent and understood. The following detailed description of the preferred embodiments, together with the accompanying drawings, are illustrated as follows: EXAMPLES This example is a vitamin C enzyme sensation prepared according to the method of the present invention. The measuring device is taken as an example, and the test liquid containing vitamin C is actually measured, and the present invention is described in detail. The experimental parameter values of the measurement conditions described later and even the measuring device are only for exemplification, but are not intended to limit the present invention. 1. Preparation of Vitamin C Enzyme Sensor In this example, a vitamin C enzyme film is used as a sensor for detecting the concentration of vitamin C, and the cross section of the element is as shown in Fig. 1, first commercialized. Tin oxide/indium tin oxide/glass (Sn02/IT0 glass) is used as the sensing film 15, the conductive layer 18 and the substrate 12 of the sensing element, wherein the sheet resistance of the tin dioxide is about 18.34 Ω/□, and the size is about 1 cm χθ.4 cm, place the tin dioxide/indium tin oxide/glass in a beaker and pour an appropriate amount of acetone into the ultrasonic oscillator for 5 minutes, then dip the oxidation with DI water. Tin/indium tin oxide/glass is cleaned. Then, pour the appropriate amount of ethanol into the beaker and shake it with an ultrasonic oscillator for 5 minutes, then rinse the tin dioxide/indium tin oxide/glass with deionized water, then The tin dioxide/oxidized sulphur tin/glass was blown dry with nitrogen. One end of the wire 17 was fixed to the tin dioxide/indium tin oxide/glass using silver glue and placed in an oven at 130 ° C about 10 0619-A21621TWF (N2); claire 200804803. After the wire 17 is fixed, it will be guided and used with epoxy resin 16 (EP=
Epoxy)將感測元件整個封裝，僅露出部分之二氧^g 為感測窗口 14,感測窗口 14約為2 乍 著置於1物料鐘以烤乾二I::後: 、，權 j压王則冉以&乳樹脂將孔洞填平， =、、.買以13(TC之烤箱烘烤，使感;収件於水溶液中進行 夏測時有良好之防水及絕緣效果，封 測元件如第2、3圖所示。 封&錢之延伸式感 =測元件進行封裝後，將維生素〇酵素薄膜固定化 高t測^ 14内之二氧化錫15上，其步驟如下：以微量 滴官吸取體積比約^ 1 ·· 9 t丙基三甲氧基矽 (3-glycid〇Xypropyltrimeth〇xysilane, GpTs)與甲苯ς 合 :…乂將該溶液滴於感測窗口 14内，用以連結二氧：： (η〇2)溥膜及維生素c氧化酶，接著將感測元件置於肋^ 之烤箱中烘烤約24Q分鐘。待烘烤後之❹』元件冷卻後， 將其置於雜鹽溶液巾2次，且每次各約1()分鐘左右，如 此則可將未鍵結之丙基三甲氧基碎與甲笨之混合液移除。 =微量滴管吸取約G.()2ml維生素c酵素溶液滴於感測窗口 内，其中維生素c酵素溶液之調配方法係取2 5mg之維生 素^氧化酶粉末溶於〇.8ml之鱗酸鹽緩衝溶液，該鱗酸鹽 綾衝溶液濃度約5mM，酸鹼值約pH7 5。將維生素c酵素 岭液滴於感測窗口後，將感測元件置於冰箱内約I]小時^ 即完成維生素C酵薄膜固定化步驟。此時維生素c酵素感 0619-A21621TWF(N2)；claire 13 200804803 測元件20藉由導線17與金氧半場效電晶體21之閘極連 接，而形成一延伸式感測場效電晶體結構10。 2.以維生素C濃度量測系統進行量測 使用如實施例1所得之維生素C酵素感測器，量測方 法係採用如弟4圖之電流-電堡（I-V)置測糸統’將感測兀件 20與銀/氯化銀(Ag/AgCl)參考電極22 —同置入待測溶液 24中，利用半導體特性量測儀21，可量測感測元件於各待 測液中所響應之電流-電壓（I-V)曲線。且利用溫度控制器將 溫度控制於室溫25°C。 取5mM pH 7.5之構酸鹽緩衝溶液80毫升加入14克 之維生素C，調配出1M之維生素待測溶液。再以半稀釋 法，將磷酸鹽缓衝溶液與維生素C溶液以1 : 1比例調和， 持續9次，即可調出2mM之維生素C待測液。 磷酸缓衝溶液可利用下列方法備製不同之酸鹼值以供 量測：取分子量為136.09之磷酸氫鉀(KH2P〇4)約680.45mg 加入水以調配濃度5mM且pH 8.5之磷酸鹽緩衝溶液。取 分子量為174.18之磷酸氫鉀約870.9mg加入水以調配濃度 5mM且pH 4.0之鱗酸鹽緩衝溶液。混合上述之pH 8.5、pH 4.0之磷酸鹽緩衝液，再配合酸鹼儀（pH meter)則可調配 ρΗ6·0、ρΗ6·5、ρΗ7·0、ρΗ7·5、ρΗ8·0 之 5mM 石粦酸鹽緩衝 溶液。 第5圖係繪示本發明實施例之維生素C酵素感測器於 不同酸鹼值之pH標準液(pH 2、pH 4、pH 6、pH 8、pH 10)， 以閘極電壓節制源極電流之機制所得到之源極電流（IDS)對 0619-A21621TWF(N2)；claire 14 200804803 間極電觸曲線。經由第6圖可計 感測器之感測度為編摩，線性度為〇、、= 弟7圖係：示於不同酸驗值之碟酸鹽 Η =Γ干::5 t ΡΗ 8)之輪出電壓與響應時間之關係。 :由=現5_之侧緩衝液酸鹼值為pH Μ時， 、准生素C感測盗於量測時較快達 之響應時間。 屋，即具有最快 以本發明實施例之維生素叫^ 准生素^相溶液及量測市售果汁之響應電黯響應 :二於2:中A係為1 M之維生素C待測溶液的量測值： β k為2mM之維生辛c往、、目丨卜、六、、右g t f CH夜的置測值’c係為市售果 /里d值由弟8圖顯不，當元件於1M之維生 測溶液中其響應電㈣1幻· 21 m V，於2 mM之维生素吝 測溶液中其響應電壓為65.61mV，且於市售果汁卜塑應 電塵為WtoV。於1M之響應時間為％秒，於城為一 幻秒，於市售果汁中為120秒。由此得知維生素^測; 之#性為隨著濃度之升高響應電壓增加、響應時間則減少。 、由^述實施例結果顯示，本發明實施例之維生素c酵 素感測器具有感測度佳、讀取之量測值正確、及快速之塑 應時間等優點’且元件長期連_試亦表現良好之重現ς 與穩定性。此外本發明之間極延伸式感測器亦具有成本低 廉之優點，可作為可拋棄式藥物感測器。 —雖然本發明已以較佳實施例揭露如上，然其並非用以 限定本發明，任何熟習此技藝者，在不脫離本發明之精神 0619-Α21621TWF(N2)；clain 15 200804803 和範圍内，當可作各種之更動與潤飾，故本發明之保護範 圍當視後附之申請專利範圍所界定者為準。 0619-A21621TWF(N2)；claire 16 200804803 【圖式簡單說明】 第1圖係顯示依照本發明之實施例，延伸式感測場效 電晶體的剖面圖； 第2圖係顯示依照本發明之實施例，延伸式感測場效 電晶體之感測元件及導線部分； 第3圖係顯示依照本發明之實施例，封裝完成之感測 元件； 第4圖係顯示依照本發明之實施例，量測感測元件感 測度之電流-電壓（I_v)量測系統； 一第5®制示依照本發明之實施例，維生素c酵素感 測益於不同酸驗值待測溶液之輸出電壓曲線圖； 第6圖絲示依照本發明之實_，維生素C酵素感 第7圖係顯示依照本發明之實施例，維 酵素感 測器之響應時間圖； ” 第8圖係繪示以本發明之每 ^ . x月之男、施例之維生素C酵素感測 待測溶液及量測市售果汁之響應電歷及響 【主要元件符號說明】 1〇〜延伸式感測場效電晶體； 12〜基底； $ 13〜毛細管； 14〜感測窗口； 〇619-A21621TWF(N2)；claire 200804803 15〜感測薄膜； 16〜絕緣層； 17〜導線； 18〜導電層； 19〜維生素C酵素薄膜； 20〜感測元件； 21〜金氧半場效電晶體， 22〜參考電極； 2 3〜光隔絕容器； 24〜待測溶液； 25〜加熱器； 26〜溫度控制器； 27〜熱耦合器； 3 0〜導線， 31〜半導體特性（電流-電壓）夏測儀。 0619-A21621TWF(N2)；claire 18Epoxy) will sense the entire package, only part of the dioxin is exposed as the sensing window 14, the sensing window 14 is about 2 置于 placed in the 1 material clock to dry the two I:: after:,, right j Press the king to fill the hole with & latex resin, =,,. Buy 13 (TC oven to bake, make sense; receive a good waterproof and insulation effect when taking the summer test in aqueous solution, sealing and testing The components are shown in Figures 2 and 3. The extension of the seal & money = after the test component is packaged, the vitamin 〇 enzyme film is fixed on the tin dioxide 15 in the high t test 14, the steps are as follows: The micropipette is taken up in a volume ratio of about 1 to 9 g of 3-glycid(R) Xypropyltrimeth〇xysilane (GpTs) and toluene: (乂) the solution is dropped into the sensing window 14 for Connect the dioxo:: (η〇2) decidua and vitamin c oxidase, then place the sensing element in the oven of the ribs for about 24Q minutes. After the component is cooled, place it. The mixed salt solution towel is used twice, and each time is about 1 () minutes, so that the unbonded mixture of propyltrimethoxy sulphate and scorpion can be removed. A micropipette draws about 2 g of vitamin C enzyme solution into the sensing window, wherein the vitamin c enzyme solution is prepared by taking 25 mg of vitamin oxidase powder dissolved in 8.8 ml of sulphate buffer solution. The concentration of the sulphate buffer solution is about 5 mM, and the pH value is about pH 7. 5. After the vitamin C enzyme ridge is dropped into the sensing window, the sensing element is placed in the refrigerator for about 1 hour. Film immobilization step. At this time, the vitamin C enzyme sense 0619-A21621TWF (N2); claire 13 200804803 The measuring element 20 is connected to the gate of the gold-oxygen half field effect transistor 21 by the wire 17 to form an extended sensing field effect. The crystal structure is 10. 2. The vitamin C concentration measuring system is used for measurement. The vitamin C enzyme sensor obtained in the first embodiment is used, and the measuring method is based on the electric current-electric (IV) measurement of the figure 4 The sensation element 20 is placed in the solution 24 to be tested together with the silver/silver chloride (Ag/AgCl) reference electrode 22, and the sensing element can be measured by the semiconductor characteristic measuring instrument 21 The current-voltage (IV) curve responsive to the measurement, and using a temperature controller Control the temperature at room temperature 25 ° C. Take 5 mM pH 7.5 acid buffer solution 80 ml to add 14 grams of vitamin C, formulate 1M vitamin test solution. Then use half-dilution method, phosphate buffer solution and vitamins The C solution is adjusted in a ratio of 1:1 for 9 times, and 2 mM of the vitamin C test solution can be transferred. The phosphate buffer solution can be prepared by the following methods for measurement: the molecular weight is 136.09. Potassium hydrogen phosphate (KH2P〇4) about 680.45 mg was added to water to prepare a phosphate buffer solution having a concentration of 5 mM and a pH of 8.5. Approximately 870.9 mg of potassium hydrogen phosphate having a molecular weight of 174.18 was added to water to prepare a sulphate buffer solution having a concentration of 5 mM and a pH of 4.0. Mix the above phosphate buffer with pH 8.5 and pH 4.0, and mix with pH meter to adjust 5 mM sulphuric acid of ρΗ6·0, ρΗ6·5, ρΗ7·0, ρΗ7·5, ρΗ8·0. Salt buffer solution. Figure 5 is a diagram showing the vitamin C enzyme sensor of the embodiment of the present invention in pH standard solutions (pH 2, pH 4, pH 6, pH 8, pH 10) with different pH values, and the gate voltage is used to control the source. The source current (IDS) obtained by the current mechanism is 0619-A21621TWF(N2); claire 14 200804803. The sensitivity of the sensor can be edited by the graph of Fig. 6, the linearity is 〇, , = = brother 7: the disc is shown in different acid values Γ = Γ dry:: 5 t ΡΗ 8) The relationship between the voltage of the turn-off and the response time. : When the pH value of the side buffer of the current 5_ is pH Μ, the quasi-gene C senses the response time of the stolen measurement. House, that is, having the fastest response to the vitamins in the embodiment of the present invention and measuring the response of the commercially available juice: 2: 2 in the A system is 1 M of the vitamin C test solution Measured value: β k is 2 mM of the vitamin C, 、, 丨, 、, 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 The component responded to electricity (4) 1 phanta 21 m V in a 1 M vitamin solution, and the response voltage was 65.61 mV in a 2 mM vitamin oxime solution, and the commercially available juice blister was WtoV. The response time at 1M is % seconds, which is a magic second in the city, and 120 seconds in commercially available juice. It is known that the vitamin is measured; the response voltage increases as the concentration increases, and the response time decreases. The result of the embodiment shows that the vitamin c enzyme sensor of the embodiment of the invention has the advantages of good sensitivity, correct reading value, and rapid plasticizing time, and the long-term connection of the component is also performed. Good reproduction and stability. In addition, the pole-extending sensor of the present invention has the advantage of being low in cost and can be used as a disposable drug sensor. The present invention has been disclosed in the above preferred embodiments, but it is not intended to limit the invention, and any person skilled in the art, without departing from the spirit of the invention, 0619-Α21621TWF(N2); clain 15 200804803 and Various modifications and refinements may be made, and the scope of the invention is defined by the scope of the appended claims. 0619-A21621TWF(N2); claire 16 200804803 [Simplified Schematic] FIG. 1 is a cross-sectional view showing an extended sensing field effect transistor according to an embodiment of the present invention; and FIG. 2 is a view showing an implementation according to the present invention. For example, the sensing element and the wire portion of the extended field-effect transistor; FIG. 3 shows the packaged sensing element in accordance with an embodiment of the present invention; and FIG. 4 shows the amount according to an embodiment of the present invention. A current-voltage (I_v) measurement system for sensing the sensing element; a fifth product is shown in accordance with an embodiment of the present invention, and the vitamin C enzyme senses the output voltage curve of the solution to be tested for different acid values; Figure 6 is a diagram showing the response time of a vitamin enzyme sensor according to an embodiment of the present invention, in accordance with the present invention. Fig. 8 is a diagram showing the present invention. ^ . x month of the male, the case of vitamin C enzyme sensing test solution and measuring the response of the commercially available juice electronic calendar and ring [main component symbol description] 1〇 ~ extended sensing field effect transistor; 12 ~ Substrate; $13~capillary; 14~ sensing window 〇619-A21621TWF(N2); claire 200804803 15~sensing film; 16~insulation layer; 17~ wire; 18~conductive layer; 19~vitamin C enzyme film; 20~ sensing element; 21~gold oxygen half field Effect transistor, 22~ reference electrode; 2 3~ light isolation container; 24~ test solution; 25~ heater; 26~ temperature controller; 27~ thermal coupler; 3 0~ wire, 31~ semiconductor characteristic (current -voltage) summer tester 0619-A21621TWF(N2);claire 18