TWI244550B

TWI244550B - Electrochemistry test unit, biological sensor, the manufacturing method, and the detector

Info

Publication number: TWI244550B
Application number: TW90115075A
Authority: TW
Inventors: Tien-Tsai Hsu
Original assignee: Hmd Biomedical Inc
Priority date: 2001-06-21
Filing date: 2001-06-21
Publication date: 2005-12-01

Abstract

An electrochemistry test unit made by a photo-lithography process and a biological sensor using the test unit are disclosed. The sensor comprises an insulated substrate and a positive electrode unit on the substrate. The two ends of the positive electrode unit separately form a work electrode and a positive electrode contact. There is a negative electrode unit on the substrate. The two ends of the negative electrode unit separately form a reference electrode and a negative electrode contact. The work electrode and the reference electrode form a defined area. There is a restrict circle out of the defined area. There is an active biological layer on the surface of the defined area. The active biological layer is for contacting a specimen to introduce an electrochemistry reaction. On the substrate, there is a hydrophobic insulation layer having a gap and a window of the gap for introducing a specimen into the reactive layer. On the active layer, there is a hydrophilic cover layer at least covering the gap. There is also a detector using a new algorithm disclosed in this invention.

Description

1244550 ---—-- 五、發明說明（1) 發明領域本發明是有關於新穎之電化學及包括该試片之生物感測器。特別地及其製備方法，以微影蝕刻製程製備片狀電化學電流’本發明是有關於以塗（dispensing)的方式製備之，片，以及以分散喷品中之生化成份含量或濃度；本發明^剛器，用以檢測樣測器之反應結果之檢測裝置以及包括罝測該生物感發明背景 7、、、°果之記憶方式。一般而言，生化分析可分為渴式红三種。傳統的濕式生化分析儀是將^式及生物感測器色劑）混合反應，再利用光學測：與試劑（例如’呈< :光光度計等）判讀其特定波長的‘色化，以作為計量的橡準。濕式生化分化里或顏色的k 設備昂貴’其特點為速度快、分析的】f的功能齊全’ 'f 較適合於檢驗中心或大型醫療診所，但：J f :樣化’= 家檢驗之使用。乾式生化分析儀則是；：用者可省去試劑添加步驟並具有少^ ° 口進仃反應，使 =與專業因f，因此’並不適合於居家檢驗之單樣檢驗 2物=器2生物元件、薄膜元件及傳感器所組成· 杪痗π栌耸且士* 1生物、細胞、組織、酵素、抗原、抗體荨具有專一性辨識能力的生物性材料元件一般係高分子材料’用以固定上述生物元件並隔離干 0713-6437TWF;Frank.ptd 第4頁 1244550 五、發明說明（2) 擾物質；傳感器則包括電極、離子選擇性場效晶體、熱敏電阻器、壓電裝置、光纖、光電管及聲波計數器等。^中電流式電化學電極是目前應用最廣的生物感測器用傳^器之一。在現有的生物感測器中，若是使用可拋棄式電極者，其電極及生物元件層，均為利用網版印刷技術而製備（例如，參見中華民國專利第374 1 1 6號；美國專利第 5,951,836、5, 770,028 及 5,711,868 號等），網版印刷製程雖可大量製造，但其精確度約僅能達到〇· 1毫米的範圍，且若網版放置的位置稍有偏差，則會明顯地影響塗佈介層的厚度均勻性，進而影響後續檢測的準確性。此外，若要達到相當之精密度，則所使用之網版印刷機械的成本亦會相對提高。如此將無法滿足居家檢驗用的要求，既不經濟又失去準確性。採取侵入式（invasive )採樣的生物感測器，由於需要穿刺使用者皮膚以抽出樣品（例如，血液或組織間液） ’通常會造成使用者的恐懼，進而排斥使用，特別是對於需要較高頻率使用的糖尿病患者而言。美國專利第5，6 2 8， 8 9 0號揭示一種片狀檢驗電極，其製作程序不僅繁複，更重要的是’其所需血液樣品量高達1 0微升（// L )以上；< 即便如中華民國專利第416005號，其所需血量亦要5微升。究其原因都是為了要降低網印製程精確度低所造成之誤差’所以需要較多量之樣品以改進其信號雜訊比。由此可知’樣品需要量小又準確，是重要且必須克服的技術。1244550 ------ 5. Description of the invention (1) Field of the invention The present invention relates to novel electrochemistry and a biosensor including the test strip. In particular, and a preparation method thereof, a sheet-shaped electrochemical current is prepared by a lithographic etching process. The present invention relates to a sheet prepared by dispensing, and a content or concentration of a biochemical component in a dispersed spray product; The invention is a rigid device, a detection device for detecting a response result of a sample detector, and a memory method including detecting the background of the biosensor invention. Generally speaking, biochemical analysis can be divided into three types of thirsty red. The traditional wet-type biochemical analyzer is a mixed reaction of ^ -type and bio-sensor colorants, and then uses optical measurement: and reagents (such as' present <: photometer, etc.) to determine the coloration of its specific wavelength, Used as a measure of rubber standard. Wet biochemical differentiation or color k equipment is expensive 'Its characteristics are fast and analytical] f's full-featured' 'f is more suitable for inspection centers or large medical clinics, but: J f: likeness' = home inspection use. The dry-type biochemical analyzer is: The user can omit the reagent addition step and have less ^ ° mouth reaction, so = and professional factors f, so 'single sample test 2 is not suitable for home inspection 2 device = biological 2 Components, thin-film components, and sensors · 杪痗 π 栌起士 * 1 biological, cells, tissues, enzymes, antigens, antibodies, biological materials with specific identification capabilities, generally high-molecular materials used to fix the above Biological components and isolation stem 0713-6437TWF; Frank.ptd Page 4 1244550 5. Description of the invention (2) Disturbing substances; sensors include electrodes, ion-selective field-effect crystals, thermistors, piezoelectric devices, optical fibers, and photoelectric tubes And sonic counters. The current type electrochemical electrode is one of the most widely used sensors for biosensors. In the existing biosensors, if disposable electrodes are used, the electrodes and biological element layers are prepared using screen printing technology (for example, see the Republic of China Patent No. 374 1 16; US Patent No. 5,951,836, 5, 770,028, 5,711,868, etc.) Although the screen printing process can be manufactured in large quantities, its accuracy can only reach the range of 0.1 mm, and if the position of the screen is slightly deviated, it will obviously be Affects the thickness uniformity of the coating interlayer, which in turn affects the accuracy of subsequent detection. In addition, if considerable precision is to be achieved, the cost of the screen printing machinery used will also be relatively high. This will not meet the requirements for home inspection, which is neither economical nor accurate. Biosensors that use invasive sampling require puncturing the user's skin to extract a sample (eg, blood or interstitial fluid). 'Usually it will cause the user's fear and then reject it, especially for those that require high Frequently used by diabetics. U.S. Patent No. 5,6 28.8,900 discloses a sheet-shaped test electrode. The manufacturing procedure is not only complicated, but more importantly, 'the amount of blood sample required is as high as 10 microliters (// L) or more; < Even if the patent of the Republic of China No. 416005, the blood volume required is 5 microliters. The reason is to reduce the error caused by the low accuracy of the screen printing process, so a larger number of samples are needed to improve the signal-to-noise ratio. It can be known from this that the small amount of sample required and accurate is an important and must be overcome technology.

0713-6437TWF;Frank.ptd 第5頁 1244550 ------------------- 五、發明說明（3) ^ 此外，一般居家檢驗用的儀器設備之使用者，大都是十=性病患者’檢驗的結果與測量時間有密切的關聯，因此檢驗儀器在紀錄檢驗結果同時，最好是將檢驗的時間一併記錄起來。為了要記錄時間，儀器必然需要設定時間。習知的時間設定方法有：（1 )在儀器上設計按鍵直接設定；或（2)由另外的設備輸入設定。此外，因世界各地的時區不同，也造成無法在儀為出廠前就先行設定，因此也會造成生產時的困擾。上述缺點均為居家檢驗展一種精確度高、需要樣品感测器，以及可以有效紀錄備’仍有其急迫的需要。發明概述儀器設備所共通者，因此，發量少及價格低廉的居家用生物籲檢驗結果與檢驗時間的儀器設有鑑於此，感測器中之電極到相當之精確度於3微升的樣品' 外’本發明也使時間參數需要設本發明的第 ’包括提供一複導電層；以及在作電極、陽極接中該工作電極及 …衣低，用以裂造生物 70件層’可大幅降低生產成本並達0713-6437TWF; Frank.ptd Page 5 1244550 ------------------- 5. Description of the invention (3) ^ In addition, users of general home inspection equipment Most of the test results are closely related to the measurement time. Therefore, it is best to record the test time together with the test equipment while recording the test results. To record time, the instrument must set the time. The conventional time setting methods are: (1) designing keys directly on the instrument for setting; or (2) inputting settings by another device. In addition, due to the different time zones around the world, it is also impossible to set the instrument before it leaves the factory, which will cause trouble during production. The above disadvantages are all in-home inspections. There is still an urgent need for high accuracy, the need for a sample sensor, and the ability to record effectively. SUMMARY OF THE INVENTION The instruments and equipment are common to all. Therefore, the instruments with low output and low cost of biological testing results and testing time are provided in view of this. The electrodes in the sensor are equivalent to a sample with a precision of 3 microliters. The 'outside' of the present invention also requires that the time parameter of the present invention include the provision of a complex conductive layer; and the working electrode in the electrode and anode connection and ... Reduce production costs and achieve

Vm本丨發明之生物感測器僅需使用少叮有效減少使用者的痛苦與恐懼感。此用新穎的訊號處踩古、上定的不便。處方法’以解決傳統使用一形悲疋提供一種電彳卜合基板，“少片之製備方法1 該第-導電層二：邑緣層以及-第-頭、參考電極二刻法製作出一工參考電極形成一 ΓΓίΓ;限流圈’其義區域，該限流圈形成The biosensor invented by Vm only requires less bite to effectively reduce the user's pain and fear. This is an inconvenience to follow the ancient times with a novel signal. "Processing method" to solve the traditional use of a form of sadness to provide an electrical coupling substrate, "the preparation method of the first piece of the first-conductive layer two: Yi margin layer and-first-head, reference electrode two-cut method to produce a The reference electrode forms a ΓΓίΓ; its limiting area, the current limiting circle forms

12445501244550

五、發明說明（4) 於該定義區域外部在一較佳具體式將一第二導電層鍍在該第一導電層本發明的第二緣基板；一位於該工作電極及一陽極兩端分別形成一參及參考電極形成一外部；一位於該定接觸以產生化學反其具有一用以容納開口端；以及一位覆盖該缺口。本發明的第三，包括提供一複合導電層；在該第一、陽極接頭、參考作電極及參考電極義區域外部；以及 (dispensing )的一生物活性層，其會溢流。丄以形成所需之電化學試片以電錢的方或石墨）電，包括一絕分別形成一極部份，其該工作電極該定義區域用於與樣品< 性絕緣層，及一缺口的層，其至少之製備方法以及一第_ 一工作電極 ’其中該工形成於該定噴塗彳面，以形成限流圈而不實施例中，μ、+、士^ 上述方法更包括 C例如，白八 b 入曰金、把、金、銀上。形怨疋提供_種生物感測器基板上之陽極部份，其兩端接頭；一位於該基板上之陰考電極及一陰極接頭，其中定義區域；一限流圈，位於義表面上之生物活性層，係應；一位於該基板上之疏水該樣品至該反應層之缺口以於該反應層上之親水性覆蓋形態是提供一種生物感測器基板，其至少包括一絕緣層導電層上以微影蝕刻法製成電極、陰極接頭以及限流圈形成一定義區域，該限流圈將一生物活性混合物以分散方式，喷塗在該定義區域表中該生物活性混合物會因該本發明的第四形態是提供一種偵測生化物只 < 才双測裝V. Description of the invention (4) A second specific conductive substrate is plated on the first conductive layer of the second edge substrate of the present invention in a preferred embodiment outside the defined area; one is located at each end of the working electrode and one anode Forming a reference and reference electrode forms an exterior; one is located at the fixed contact to generate a chemical reaction and has an open end to accommodate the opening; and one bit covers the gap. The third aspect of the present invention includes providing a composite conductive layer; outside the first, anode terminal, reference electrode, and reference electrode definition region; and a biologically active layer that will overflow.丄 to form the required electrochemical test strips (e.g., square or graphite), including a pole to form a pole part, a defined area of the working electrode for the sample < insulating layer, and a gap Layer, its at least preparation method, and a first _ working electrode, where the process is formed on the fixed spraying surface to form a current-limiting ring. In the embodiment, μ, +, and ^ The above method further includes C, for example , Bai Ba b into gold, silver, gold, silver. Xingyu provides _ a kind of anode part on the substrate of the biosensor, its two ends of the connector; a negative electrode and a cathode connector on the substrate, which define the area; a current-limiting ring, located on the surface of the sense A biologically active layer should be provided; a gap between the sample hydrophobic and the reaction layer on the substrate to form a hydrophilic covering on the reaction layer is to provide a biosensor substrate including at least an insulating layer and a conductive layer The electrode, cathode connector, and current limiting ring are made by lithographic etching to form a defined area. The current limiting ring uses a biologically active mixture in a dispersed manner. The biologically active mixture is sprayed on the table of the defined area. A fourth aspect of the invention is to provide a dual detection device for detecting biochemicals.

1244550 、五、發明說明（5) 置，包括：（1)如上所述之生物感測器，其可將生化物質轉換成電子訊號；以及（2 ) —訊號處理裝置，其包括（i ) 一邏輯運算單元，用以將該等電子訊號轉換成對應之生化物質之濃度；以及（i i ) 一時間運算單元。當測量第η次樣品之後，該時間運算單元即記錄時間tn，並將時間自動歸零，於測量第n+ 1次樣品時，該時間運算單元即記錄時間 tn+1，將時間自動歸零，使得所測量的時間參數，均以△ t 的形式儲存在該訊號處理裝置中。在一較佳具體實施例中，上述偵測生化物質之檢測裝置更包括一資料輸出埠，以將所計算之生化物質濃度及時φ 間參數傳輸至一資料處理裝置（例如，個人電腦、高速電腦或伺服器）。為了讓本發明之上述和其他目的、特徵及優點能更明顯易懂，下文特舉較佳實施例並配合圖示，做詳細說明如下：圖示之簡單說明第1圖是本發明之生物感測器之示意圖。第2圖是第1圖沿著a-a’之剖面圖。第3圖是利用本發明之生物感測器及偵測生化物質之裝置所測得葡萄糖之結果。翁參考標號說明 1〜生物感測器； 1 0〜工作電極； 1 2〜陽極接頭； 1 4〜參考電極； 1 6〜陰極接頭； 1 8〜限流圈；1244550, V. Description of the invention (5) device, including: (1) the biosensor as described above, which can convert biochemical substances into electronic signals; and (2)-a signal processing device, which includes (i) a A logic operation unit for converting the electronic signals into corresponding concentrations of biochemical substances; and (ii) a time operation unit. After measuring the n-th sample, the time calculation unit records the time tn and automatically resets the time to zero. When measuring the n + 1th sample, the time calculation unit records the time tn + 1 and automatically resets the time to zero. The measured time parameters are stored in the signal processing device in the form of Δt. In a preferred embodiment, the above-mentioned detection device for detecting biochemical substances further includes a data output port to transmit the calculated biochemical substance concentration and time parameter to a data processing device (for example, a personal computer, a high-speed computer). Or server). In order to make the above and other objects, features, and advantages of the present invention more comprehensible, the preferred embodiments are illustrated below with detailed descriptions as follows: Brief description of the illustrations Figure 1 is the biological sense of the present invention Schematic diagram of the tester. Fig. 2 is a sectional view taken along a-a 'in Fig. 1. Fig. 3 is a result of measuring glucose using the biosensor and the device for detecting biochemical substances of the present invention. Description of reference numerals 1 ~ biosensor; 10 ~ working electrode; 12 ~ anode connector; 14 ~ reference electrode; 16 ~ cathode connector; 18 ~ current limiting coil;

0713-6437TWF;Frank.ptd 第8頁 1244550 五發明說明（6) 1 9〜缺口； 2卜缺口之開口端；“〜：：J板； 26〜疏水性絕緣層；28〜親水性^層；較佳具體實施例之詳細說明、χ丨生復蓋層。本發明將藉由以下的較佳且詳細說明’但這些具體實施例僅是；；：；；作更進一步地以限定本發明之範嘴。疋作為舉例說明，而非用請參見第1圖，根據本發測器1 ’係可用於偵測樣品中 ^體實施例之生物感度，包括：一絕緣基板2〇 ; 質的含量及/或浪其兩端分別形成-工作電極10及=;上之陽極部份基板上之陰極部份，Α Λ 騎極接頭1 2 ; —位於該極接頭16，其分別形成-參考電極14及一陰區域；一限流圈i 8，位於該 ^電和形成一疋義表面上之生物活性層24 (參見第;圖或外部一位於該定義觸以產生化學反應；一位 θ ，係用於與樣品接 ^ ^ ^ , 4基板上之疏水性絕緣層26，的開！端f該樣品至該反應層之缺口19以及-缺口的開口 1 ，以及一位於其至少覆蓋該缺口。反應層上之親水性覆蓋層28，有別於傳統網版印刷的製程，本發明首弁利用瞧#料碰影蝕刻製程，於一遴人I ^ ^%月百先利用曝先微餐、陽極接頭、的導電層上製作出一工作電極微影蝕刻掣r'丌Γ極、陰極接頭以及限流圈；藉由導入化學試片，以用於後續生物感測器的；；有= 0713-6437TWF;Frank.ptd 第9頁 1244550 五、發明說明（8) 藉此可大幅降低生產成本， ± 0.1%微升（//L) 並維持應有之準確性。絕緣基板2 0具有平軟電絕緣的特性，適合刻製程）以及 PVC、PET、PC、PE、P =包括’但並不限峨-4、pp、。 S '玻璃、陶兗或其他絕緣塑膠材質陽極邛伤包括工作電極丨〇及陽極接頭1 2 品在化學或生化反應時所乂傳導樣如下所述）。陰極部份包括又二效應至檢測裝置（上…= 的電化學效應進行檢測。- ’生物活性混合物係由酵素、載體 < 界面活性劑所組成。所選 /子媒"物及質而決定，在本發明之較佳具體實施例中，例如的^匕^ 驗血糖之目的，所選用的醅妾0 # 馬了才双 )。適合的載體包括不具活性之蛋白質（例如，牛血清白蛋白（BSA))、超微粒纖維素、甲基纖維素（Mc)、羧基甲基纖維素（CMC)、殿粉、乙婦醇、聚乙稀醇（pvA)、聚乙稀㈣酮（PVP)、聚乙二醇（PEG)或動物明勝等。適合的電子媒介物例如赤血鹽（p〇tassium ferricyanide )。適合的界面活性劑包括卜丨t〇n?系列（例如，t〇n ^ X-100、Triton x_405、Trit0n χ_114 等）、Tween?系列 (例如，Tween 20、Tween 40、Tween 60、Tween 80 等）、十二烷硫酸鈉或其他親水性清潔劑。為了減少測试者感覺恐懼之目的（亦即儘可能減少所 0713-6437TWF;Frank.ptd 第11頁 1244550 五、發明說明（9) 需之樣品量），本發明設計如第1圖及第2圖所示之妙構。請參考第1圖，可容納樣品至反應層24 (生物活性層'的缺口 1 9以及缺口的開口端2 1，藉由使電極靠近缺口 9端，使得所需的樣品量可盡量的減少。再請參考第2圖，疏水性絕緣層26 (材質同絕緣基板，包括FR —4、pp、pvc PC、PE、PS、玻璃、陶莞或其他絕緣塑膠材質）作為間隔區（spacer )，於頂端覆蓋一層親水性覆蓋層“，所產生的開口 19配合親水性覆蓋層28，可形成吏得進入的液體產生毛細作用，有助=== 帶至生物活性層24進行反應。藉由上述的設計，本發明生物感測器僅需使用少於3微升的樣品’可有效減少 ^ 者的疼痛與恐懼。為了解本發明生產之生物感測器的特性，將本發明 ==測器’以分別含有50、2。。及4〇〇毫克"0。毫升之私準U萄糖溶液模擬全血中的血糖進圖的^果顯示’所得到的定量線之迴歸隸㈣），生物戊、則写且：準偏差（SD)僅有0.38%，顯示本發明之生物感測器可具有準確之測量結果。所ΐ ? 1月更提供一種檢測生化物質之裝置，包括⑴如以及⑻-物ΐ測器，其可將生化物質轉換成電子訊號；· ，用以將所處理裝置，#包括：⑴—邏輯運算單元、二將所產生的該等電子訊號轉換成對應之生化物質之辰又 11 日守間運异單元’當測量第η次摄口之德 # 時間運算單元即々鉾ρ “ 列里弟η久樣σ口之後，該 Ρ Ζ錄時間tn，並將時間自動歸零，於測量 0713-6437TWF;Frank.ptd 第12頁 12445500713-6437TWF; Frank.ptd Page 8 1244550 Fifth invention description (6) 1 9 ~ notch; 2 open end of the notch; "~ :: J plate; 26 ~ hydrophobic insulation layer; 28 ~ hydrophilic ^ layer; The detailed description of the preferred embodiments, the cover layer. The present invention will be described by the following better and detailed 'but these specific embodiments are only ;;;; further to limit the invention Fan mouth. 疋 As an example, please refer to Figure 1 instead. According to the tester 1 'can be used to detect the biological sensitivity of the embodiment of the sample, including: an insulating substrate 20; And / or the two ends of the wave are formed respectively-the working electrode 10 and the cathode portion on the substrate, Α Λ riding pole connector 1 2;-located at the pole connector 16, which respectively form-the reference electrode 14 And a negative region; a current-limiting circle i 8, which is located on the surface and forms a biologically active layer 24 on the sense surface (see FIG .; or the exterior is located on the definition touch to generate a chemical reaction; a bit θ, is used The opening ^ of the hydrophobic insulating layer 26 on the substrate ^ ^ ^, 4 The gap 19 from the sample to the reaction layer and the opening 1 of the gap, and a gap covering at least the gap. The hydrophilic covering layer 28 on the reaction layer is different from the traditional screen printing process. # Material touch shadow etching process, a working electrode lithography etcher r '丌 Γ pole, cathode connector, and limiter were fabricated on a conductive layer of the first micro-meal, anode connector, and conductive layer on a first person. Flow coil; by introducing chemical test strips for subsequent biosensors; yes = 0713-6437TWF; Frank.ptd Page 9 1244550 V. Description of the invention (8) This can greatly reduce production costs, ± 0.1% microliter (// L) and maintain the required accuracy. Insulating substrate 20 has the characteristics of flat soft electrical insulation, suitable for engraving process) and PVC, PET, PC, PE, P = including 'but not limited E-4, pp, .S 'glass, ceramics or other insulating plastic material anode damage including working electrode and anode connector 12 in the chemical or biochemical reaction of the sample is described below). Cathode part Including the electrochemical effect of the second effect to the detection device (up ... = The test is performed.-'The biologically active mixture is composed of an enzyme, a carrier < surfactant. The choice / sub-media " is determined by the nature and nature of the material. In a preferred embodiment of the present invention, for example, ^^ For the purpose of blood glucose testing, 醅妾 0 # horses are only used.) Suitable carriers include inactive proteins (for example, bovine serum albumin (BSA)), ultrafine cellulose, methyl cellulose (Mc) , Carboxymethyl cellulose (CMC), Dianfen, ethyl alcohol, polyvinyl alcohol (pvA), polyethylene phenone (PVP), polyethylene glycol (PEG), or animal Mingsheng. A suitable electronic vehicle is, for example, potassium ferricyanide. Suitable surfactants include the Ton ™ series (for example, ton ^ X-100, Triton x_405, Triton 0x 114, etc.), the Tween® series (for example, Tween 20, Tween 40, Tween 60, Tween 80, etc.) ), Sodium lauryl sulfate or other hydrophilic cleaning agents. In order to reduce the fear of the tester (that is, to reduce as much as possible 0713-6437TWF; Frank.ptd page 11 1244550 V. Description of the invention (9) required sample size), the present invention is designed as shown in Figure 1 and Figure 2 The wonderful structure shown in the figure. Please refer to Fig. 1. The sample can be accommodated in the reaction layer 24 (notch 19 of the bioactive layer 'and the open end 21 of the notch. By bringing the electrode close to the notch 9 end, the required sample amount can be reduced as much as possible. Please refer to FIG. 2 again. The hydrophobic insulating layer 26 (same material as the insulating substrate, including FR-4, pp, pvc PC, PE, PS, glass, ceramic or other insulating plastic materials) is used as a spacer. The top is covered with a hydrophilic covering layer. The opening 19 produced in conjunction with the hydrophilic covering layer 28 can form a capillary fluid that enters and produces capillary action, which helps === bring it to the biologically active layer 24 for reaction. Design, the biosensor of the present invention only needs to use less than 3 microliters of sample 'can effectively reduce the pain and fear of people. In order to understand the characteristics of the biosensor produced by the present invention, the present invention == tester' The results of the quantitative lines obtained by simulating the blood glucose in the whole blood with 50, 2, and 400 milligrams of private quasi-glucose solution are shown in the figure below. E. Write and: the quasi deviation (SD) is only 0.38% It shows that the biosensor of the present invention can have accurate measurement results. Therefore, in January, a device for detecting biochemical substances was provided, including a biochemical detector and a biochemical detector, which can convert biochemical substances into electronic signals. ;, Used to process the device, # includes: ⑴-logical operation unit, two will convert the electronic signals generated into the corresponding biochemical substances and another 11 days between the different units when the measurement η口口之德 # The time operation unit is 々鉾 ρ “After the Riley η long sample σ mouth, this ρ z records the time tn, and automatically resets the time to zero, measured 0713-6437TWF; Frank.ptd Page 12 1244550

第η Η次樣品時，該時間運算ηπ _ 間自動歸零，使得所測量的1早7^即記錄時間tn+1，亦將時存在該訊號處理裝置中。、3參數，均以△ t的形式儲本發明利用裝置中的訊 )，其内含一時間運算單元紀錄測量結果同時紀錄之，將時間運算單元之資料歸零確實的時間，或是出國差旅再輸入為當地的時間。號處理裝置（例如，微處理器 ’ k供一計時之資料，在每次在元成紀錄之指令後，即自動 ’如此，使用者就不需要輸入時’不必因為時差的改變而需本發明之檢測裝置也包括—邏輯運算單元，其至少括-電化學電位差電路，以提供電化學反應所需之能量，以及一電流放大及量測電路，以量測電化學反應所產生之電流。整體而言’該邏輯運算單元可將生物感測器所產生的電子訊號，轉換成對應之生化物質之濃度。當需要時，儲存在訊號處理裝置中之時間差距-生化物質濃度（Δΐ-cone.)資料庫，可藉由一資料輸出埠 (例如’ R S - 2 3 2埠），而傳輸至一資料處理裝置（例如，個人電腦、高速電腦或伺服器）；藉由該資料處理裝置的軟體，而將依序儲存之資料庫還原為真實時間-生化物質濃度的數據圖表，藉此可得到該使用者這一段時間中之病籲史資訊，可供醫師進行研判。此外，藉由時間差距（△ t )的設定，可在本發明的裝置中控制一定時段的△ΐ (例如，4小時、8小時、24小時等等），當超過預定的時間差距而沒有進行下一次測量^，則裝置可發出警告訊息，提At the ηth sample, the time calculation ηπ _ is automatically reset to zero, so that the measured time is as early as 7 ^, that is, the recording time tn + 1, and is also stored in the signal processing device. And 3 parameters are stored in the form of Δt in the utilization device of the present invention.) It contains a time operation unit to record the measurement results and record it at the same time. The data of the time operation unit is reset to zero for the actual time, or the difference in travel time. Enter the travel time as the local time. No. processing device (for example, the microprocessor 'k provides a timed piece of data, and it will automatically' before each time a command is recorded in Yuancheng, so that the user does not need to enter it 'and does not need the invention because of the time difference change. The detection device also includes a logic operation unit, which at least includes an electrochemical potential difference circuit to provide the energy required for the electrochemical reaction, and a current amplification and measurement circuit to measure the current generated by the electrochemical reaction. In terms of 'the logical operation unit can convert the electronic signal generated by the biosensor into the concentration of the corresponding biochemical substance. When needed, the time difference stored in the signal processing device-the concentration of biochemical substance (Δΐ-cone. ) Database, which can be transmitted to a data processing device (for example, a personal computer, a high-speed computer, or a server) through a data output port (for example, 'RS-2 3 2 port); the software of the data processing device , And the sequentially stored database is restored to a real time-biochemical concentration data chart, so that the user ’s disease history can be obtained It can be used by physicians for research and judgment. In addition, by setting the time gap (Δt), the device of the present invention can be controlled for a certain period of time Δΐ (for example, 4 hours, 8 hours, 24 hours, etc.). A predetermined time gap without taking the next measurement ^, the device may issue a warning message to mention

0713-6437BfF;Frank.ptd0713-6437BfF; Frank.ptd

第13頁 1244550Page 13 1244550

五、發明說明（11) 醒使用者定期間的測量；藉此可提供長時々期/不定期^的測量，對於患有慢性疾病（例如，回’尸疒疋的患者而言，有助於其定時追蹤/監測體内生铷：：、曲度，確保其生命安全及健康。 &生化物質的浪V. Description of the invention (11) Periodic measurement of awake users; this can provide long-term / periodical measurement, which is helpful for patients with chronic diseases (eg It regularly tracks / monitors the growth of the body ::, curvature, to ensure the safety and health of its life.

由上述說明可知，本發明不僅有效生罐'"式片的品f，以及改進生產流程血：K 本發明新穎之檢测生化物質的裝£，不省設定鍵之成本，亦可減少使老命 j即名汉置时間府 $ 用者而要輸入時間/轉換時間 ' 、，且，更可定時間提醒使用者測量，這此停點是From the above description, it can be known that the present invention not only effectively produces the product f of the canned film, and improves the production process blood: K The novel detection of biochemical substances in the present invention does not save the cost of setting keys, and can also reduce the cost of Lao Ming j is the name of the Chinese home time user. You need to enter the time / conversion time ', and you can also set the time to remind the user to measure. This stop point is

的先前技藝所沒有教示的，因此，本發。第-ί -人k出這樣的構想並付諸實現。 ’、I 限定發:月已"交佳實施例揭露如上，然其並非用以 " 壬何熟悉此技藝者，在不脫離本發明之於袖 3圍外：當可作各種之更動與潤飾。因*，本：：= 濩粑圍，當視後附之申請專利範圍而所界定者為準。’、The previous art institute did not teach it, so this post. The first-person k came up with such a concept and put it into practice. ', I limited hair: The month has been "explained as above", but it is not intended to be used by those who are familiar with the art, without departing from the scope of the invention outside the sleeve: when various changes and changes can be made Retouch. Because of *, this :: = 濩粑 Wai, which shall be determined by the scope of the patent application attached. ’,

Claims

1244550 六、申請專利範圍 1一種電化學試片提供一複合基板，電層；以及在該第一導電層上極接頭、參考電極、陰極及參考電極形成一定域外部，以形成所需之 2·如申請專利範圍一導電層包括銅。 3 ·如申請專利範圍緣層係 F R - 4、P p、p v ◦、其他絕緣塑膠材質。 ^ 4·如申請專利範圍電鍍的方式將一第二導二、、5 ·如申請專利範圍一導電層包括白金、鈀 6 · —種生物感測器一絕緣基板；位於該基板上之 /形成一工作電極及一、、ι 仇於該基板上之法形# 成—參考電極及_ 、極形成一定義區域；限流圈，位於該於該定義表面之製猶方法’包括：其至少包括一絕緣層以及一第一導以微影蝕刻法定義一工作電極、陽極接頭以及限流圈，其中該工作電義區域，該限流圈形成於該定義區電化學試片。第1項所述之製備方法，其中該第第2項所述之製備方法，其_該嗜 ‘PET、PC、PE、PS、坡‘、陶7或第3項所遂之製備方法’更包括以電層電鍍在該第一導電層上。第4項所述之製備方法，其中該第、金、或銀。，包括：〒極部份’其兩端分別以微影蝕刻陽極接頭；陰極部份，其兩端分別从微影蝕刻陰極接頭’其中該工作電極及參考丈義區域外部，土與樣品接 0713. 6437颇 •Ptc 第15頁 1244550 _案號90115075_年月曰修正_ 六、申請專利範圍觸以產生化學反應；一位於該基板上之疏水性絕緣層，其具有一用以容納該樣品至該反應層之缺口以及一缺口的開口端；以及一位於該反應層上之親水性覆蓋層，其至少覆蓋該缺 7. 如申請專利範圍第6項所述之生物感測器，其中該反應層係由酵素、載體、電子媒介物及界面活性劑所組成。1244550 6. Scope of patent application 1. An electrochemical test strip provides a composite substrate and an electrical layer; and on the first conductive layer, an electrode joint, a reference electrode, a cathode, and a reference electrode form a certain area outside to form a desired 2 · For example, a conductive layer includes copper. 3 · If the scope of the patent application is F R-4, P p, p v ◦, other insulating plastic materials. ^ 4. If the scope of the patent application is electroplating, a second lead will be used. 5 · If the scope of the patent application is a conductive layer including platinum and palladium 6. A kind of biosensor-an insulating substrate; located on / formed on the substrate A working electrode and a method formed on the substrate are formed by the reference electrode and the electrode forming a defined area; the current-limiting ring, which is located on the defined surface, includes: it includes at least An insulating layer and a first lead define a working electrode, an anode joint, and a current-limiting ring by a lithographic etching method, wherein the working electrode region and the current-limiting ring are formed in the electrochemical test strip in the defined area. The preparation method described in item 1, wherein the preparation method described in item 2, which is _ the preparation method of 'PET, PC, PE, PS, Po', Tao 7 or item 3 ' The method includes electroplating an electrical layer on the first conductive layer. The preparation method according to item 4, wherein the first, gold, or silver. Including: 〒 pole part 'the two ends are lithographically etched to the anode connector respectively; cathode part, the two ends are etched to the cathode connector from lithography respectively', where the working electrode and the reference area are outside, soil and sample are connected 0713 . 6437 Po • Ptc Page 15 1244550 _ Case No. 90115075_ year month amendment _ six, the scope of the patent application to produce a chemical reaction; a hydrophobic insulating layer on the substrate, which has a A gap of the reaction layer and an open end of the gap; and a hydrophilic covering layer on the reaction layer, which covers at least the gap 7. The biosensor according to item 6 of the scope of patent application, wherein the reaction The layer system consists of enzymes, carriers, electronic mediators and surfactants.

8. 如申請專利範圍第7項所述之生物感測器，其中該酵素包括葡萄糖氧化酵素。 9. 如申請專利範圍第7項所述之生物感測器，其中該載體包括不具活性之蛋白質、超微粒纖維素、甲基纖維素、羧基曱基纖維素、澱粉、乙烯醇、聚乙烯醇、聚乙烯吡咯酮、聚乙二醇或動物明膠。 1 0.如申請專利範圍第7項所述之生物感測器，其中該電子媒介物包括赤血鹽。8. The biosensor as described in item 7 of the patent application scope, wherein the enzyme comprises a glucose oxidase. 9. The biosensor as described in item 7 of the patent application scope, wherein the carrier comprises inactive protein, ultrafine cellulose, methyl cellulose, carboxymethyl cellulose, starch, vinyl alcohol, polyvinyl alcohol , Polyvinylpyrrolidone, polyethylene glycol or animal gelatin. 10. The biosensor as described in item 7 of the patent application scope, wherein the electronic medium includes red blood salt.

11.如申請專利範圍第7項所述之生物感測器，其中該界面活性劑包括T r i t ο nTM系列、T w e e nTM系列、十二烧硫酸鈉或其他親水性清潔劑。 1 2, —種生物感測器之製備方法，包括：提供一複合基板，其至少包括一絕緣層以及一第一導電層；在該第一導電層上以微影蝕刻法定義一工作電極、陽極接頭、參考電極、陰極接頭以及限流圈，其中該工作電極及參考電極形成一定義區域，該限流圈形成於該定義區11. The biosensor as described in item 7 of the scope of the patent application, wherein the surfactant includes a T r i t nTM series, a Tw e nTM series, sodium dodecyl sulfate or other hydrophilic cleaning agents. 1 2, A method for preparing a biosensor, comprising: providing a composite substrate including at least an insulating layer and a first conductive layer; defining a working electrode on the first conductive layer by lithography etching, Anode connector, reference electrode, cathode connector and current limiting ring, wherein the working electrode and reference electrode form a defined area, and the current limiting ring is formed in the defined area

0713-6437TWFl;jyliu.ptc 第16頁 1244550 修正案號 90115075 六、申請專利範圍域外部；以及將一生物活性混合物以分散喷塗（d i s p e n s i n g )的方式，喷塗在該定義區域表面，以形成一生物活性層，其中該生物活性混合物會因該限流圈而不會溢流。 1 3. —種偵測生化物質之檢測裝置，包括： (1 )如申請專利範圍第6項至第1 1項所述之生物感測器，其可將生化物質轉換成電子訊號；以及 (2 ) —訊號處理裝置，其包括·· (i) 一邏輯運算單元，用以將該等電子訊號轉換成對應之生化物質之濃度； (i i) 一時間運算單元，當測量第η次樣品之後，該時間運算單元即記錄時間tn，並將時間自動歸零，於測量第η + 1次樣品時，該時間運算單元即記錄時間tn+1，將時間自動'歸零，使得所測量的時間參數，均以△ t的形式儲存在該訊號處理裝置中。 1 4.如申請專利範圍第1 3項所述之檢測裝置，更包括一資料輸出璋，以將所計算之生化物質濃度及時間參數傳輸至一資料處理裝置。 1 5.如申請專利範圍第1 4項所述之檢測裝置，其中該資料處理裝置包括個人電腦、高速電腦或伺服器。0713-6437TWFl; jyliu.ptc Page 16 1244550 Amendment No. 90115075 6. Outside the scope of the patent application; and spraying a biologically active mixture on the surface of the defined area in a dispersing manner to form a A biologically active layer, wherein the biologically active mixture will not overflow due to the restriction circle. 1 3. A detection device for detecting biochemical substances, including: (1) the biosensor as described in item 6 to item 11 of the scope of patent application, which can convert biochemical substances into electronic signals; and ( 2) — signal processing device, which includes ... (i) a logical operation unit for converting these electronic signals into the corresponding concentration of biochemical substance; (ii) a time operation unit, after measuring the n-th sample The time calculation unit records the time tn and automatically resets the time to zero. When measuring the η + 1st sample, the time calculation unit records the time tn + 1 and automatically 'zeros' the time to make the measured time The parameters are stored in the signal processing device in the form of Δt. 14. The detection device as described in item 13 of the scope of patent application, further includes a data output unit to transmit the calculated biochemical substance concentration and time parameters to a data processing device. 15. The detection device according to item 14 of the scope of patent application, wherein the data processing device includes a personal computer, a high-speed computer, or a server.

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