CN100480695C - Electrochemical screen printing electrode sensing test piece and making process thereof - Google Patents
Electrochemical screen printing electrode sensing test piece and making process thereof Download PDFInfo
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- CN100480695C CN100480695C CNB2004100084179A CN200410008417A CN100480695C CN 100480695 C CN100480695 C CN 100480695C CN B2004100084179 A CNB2004100084179 A CN B2004100084179A CN 200410008417 A CN200410008417 A CN 200410008417A CN 100480695 C CN100480695 C CN 100480695C
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Abstract
The invention relates to an electrochemical screen printing electric pole sensing test piece and its process method.The test piece comprises an insulating bottom board with such two or three electric poles as working electrode, auxiliary electrode and reference electrode. The electrode region surface directly absorbs a reaction layer comprising reactant, reaction catalyzer, electron transmitter substance, wetting agent and interfacial active agent, wherein, on the top of electrode region is a physical diversion region comprising an interlayer slice with a U-shaped channel on top of electron region and an upper cover plate on the electron region directing line, the head of which has closed convex space.
Description
Technical field
The present invention is about a kind of galvanochemistry or test piece of screen painting electrode senses and method for making thereof.This test piece utilizes the principle of capillary siphon, a minute quantity corpse or other object for laboratory examination and chemical testing can be sucked the electrode reaction district rapidly, utilizes and decides potential method or decide the quantitative test that current method carries out corpse or other object for laboratory examination and chemical testing composition.
Background technology
In recent years the electric pole type inspection test piece since its make easily, the popularizing of the with low cost and inexpensive detector that is convenient for carrying, successfully be applied to various products and detected and commercially produce; Various biochemistry detection devices such as blood sugar, uric acid, cholesterol for example.Maximum and to use maximum blood-sugar detecting instruments be example with market, technical control manufacturer comprises companies such as Luo Shi (Roche), Ya Pei (Abbott), Bayer and Therasense on the market, all is with the test piece of electric chemical formula mode production blood sugar test.Blood volume demand height of first generation electric chemical formula blood sugar test sheet (more than the 5-10 μ l) and time are also grown (30-60 does not wait second), though it is, still unsatisfactory in the use having surpassed optics development process classic methods such as (colorimetric) aspect blood volume and detection time.Along with development of technology, present latest generation product, as long as blood volume is 0.3 μ l (Freestyle of Therasense company product) or 1 μ l (the oneTouch Ultra of Lifescan company product), required time has also foreshortened to 5-10 second, so blood sampling volume is few and detection speed is fast, become the direction of this series products and technical development, and the research and development of various electrode structures develop towards this direction all also.People such as Diebold have disclosed a capillary guiding area and upward opening are made in a kind of utilization to the hand held electrode design and between two electrodes circular air guide port in No. 5,437,999, the United States Patent (USP) of nineteen ninety-five, a corpse or other object for laboratory examination and chemical testing can be accurately in a small amount the reaction zone effect that enters.This invention can be adopted mask and screen painting processing procedure, no matter but its composition of which kind of method for making all is to contain single stranded electrode base version subtend clipping by two to form, complex steps not only, and cost is also very high.Shieh was in United States Patent (USP) 5 in 1998,779, No. 867, also disclosed a kind of electric pole type blood glucose test design, this invention utilizes two corresponding electrodes up and down, and intermediate course is reaction zone and leaves a circular hole and splash into mouth as a corpse or other object for laboratory examination and chemical testing that the interlayer district includes a blood cell separating film to filter red blood cell, this method is controlled the interference that blood volume and phase remove blood cell with separating film with the folded film of two test pieces up and down, but can't reach optimum condition on blood volume and speed because of the obstruct of filtering membrane.Ya Pei company (United States Patent (USP) 6,129,823) utilizes the single or multiple lift stratum reticulare to cover working electrode surface, and the superiors are the lid layer loam cake again, this loam cake directly over working electrode or contiguous working electrode place open a hole, the position of injecting as a corpse or other object for laboratory examination and chemical testing.This design also reaches the purpose that only needs a 2.0-2.5 μ l small size corpse or other object for laboratory examination and chemical testing.But this invention is as the method that reduces a blood volume and a distribution corpse or other object for laboratory examination and chemical testing with stratum reticulare.Therasense company is in United States Patent (USP) 6,299, and 757 and 6, disclose same utilization to the hand held electrode design in 338,790 liang of patents, between two electrodes, seen through meticulous structure dress, put into a slice and have splendid absorptive thin slice, a corpse or other object for laboratory examination and chemical testing is drawn between two electrodes reacts.Utilizing the control of tight suction film volume, the required minimum volume of a corpse or other object for laboratory examination and chemical testing can be reduced to 0.3 μ l, is the minimum design of whole world blood volume at present, but this invention then at canned program than the high weakness of cost is arranged under the complicated situation.People such as Mr. Shen Yanshi are also in Taiwan patent 208, disclose the method for making and the application thereof of screen painting electrode in No. 095, wherein emphasize to finish conducting film and electric insulation layer with screen painting earlier, finish metal level with electro-plating method again, and with insulating gel in work and the reference electrode place form a circular depression, and in this depressed part, adsorb so-called biostrome, and directly a corpse or other object for laboratory examination and chemical testing is splashed into herein during detection, need a corpse or other object for laboratory examination and chemical testing 10 μ l approximately.The method loaded down with trivial details step such as need electroplate and the blood volume demand also higher. other five ancient cooking vessel companies are in Taiwan patent 124, then be disclosed in the electrode district top in No. 332 and make a water conservancy diversion district, the top also covers the nethike embrane that a slice contains surfactant, utilizing capillary and siphon principle that a corpse or other object for laboratory examination and chemical testing is drawn on the electrode reacts, this method with the design as water conservancy diversion of the nethike embrane that covers, also is subjected to the quantitative limitation of minimum corpse or other object for laboratory examination and chemical testing demand as the method for Ya Pei except that increasing cost.
People such as Winarta are in calendar year 2001 United States Patent (USP) 6,258, disclosing a kind of deserted pick-up unit in No. 229, only need to declare a liquid corpse or other object for laboratory examination and chemical testing less than 1 μ l. its method for making is that a end on a strip sheet gold/polystyrene (gold/polyester) or tin oxide/gold/polystyrene (tin oxide/gold/polyester) top layer is with CO2 Lasers (CO
2Laser), this strip sheet of etching is to form three blocks that do not conduct mutually mutually as three electric pole types; On as working electrode, the middle layer of containing U type opening along the front end stickup last of test piece, this moment, working electrode was positioned at U type groove, on the middle layer, paste a loam cake that contains a square ventilation holes again. therefore formed one in the working electrode upper end by the fluid passage of liquid inlet to the air hole, this fluid passage has limited the liquid volume that can bear less than 1 μ l.This design just like people such as Nankai early than disclosed in the United States Patent (USP) 5,120,420 in 1992, electrode method for making difference in both conductive layers just.The disclosed electrode test piece method for making of people such as Nankai is the conductive layer with method brush system bipolar electrode form on an insulation back of the body version of screen painting, one of mode that its fluid passage is formed, be horizontally in rear and front end, working electrode place to stick two walls, stick the loam cake that a slice does not contain hole thereon again, so formed a horizontal open channel by working electrode.Use amount with this method corpse or other object for laboratory examination and chemical testing admission passage is then uncontrollable, and can pollute from the vent port outflow.Another kind of composition mode, as mentioned above, a middle layer of containing U type opening is promptly sticked in people's such as Winarta design in working electrode upper end, stick a slice centre more thereon and contain the loam cake of a ventilation holes, so formed one by the fluid passage of liquid inlet to the blow vent centre.This kind structure, too small when blow vent, the misgivings that still have a corpse or other object for laboratory examination and chemical testing to flow out by blow vent; Moderate when blow vent, a corpse or other object for laboratory examination and chemical testing then can by chance rest on the blow vent edge, especially in today of stressing the test piece downsizing, when getting test piece with hand, is easy to cause corpse or other object for laboratory examination and chemical testing outflow to pollute because of touching blow vent because of carelessness.
Summary of the invention
As mentioned above, want to reduce the volume and the express-analysis simultaneously of a corpse or other object for laboratory examination and chemical testing, must avoid causing corpse or other object for laboratory examination and chemical testing contamination of heavy, need utilization capillary and siphon principle, and cooperate the design of electrode just can reach above-mentioned purpose.The present invention promptly is the design according to above-mentioned electrode district, obtain the electrode reaction district of a quick water conservancy diversion, so the invention provides not only fast but also accurate electrochemica biological detecting device, easy production simple in structure and need not cover nethike embrane fully, and can avoid a corpse or other object for laboratory examination and chemical testing to leak outside polluting.
The object of the present invention is to provide a kind of electric chemical formula biological sensing test sheet, it is in conjunction with the design of capillary siphon principle, can reach to use the minute quantity corpse or other object for laboratory examination and chemical testing not only can be fast but also accurately carry out the purpose that electrochemica biological detects.
The present invention also aims to provide a kind of method for making of electric chemical formula biological sensing test sheet, the test piece that obtains is carried out biological detection not only fast but also accurate, and is simple in structure and can avoid a corpse or other object for laboratory examination and chemical testing to leak outside polluting.
Electric chemical formula biological sensing test sheet provided by the invention is made of following assembly:
An insulating base;
One conductor layer, it is to have bipolar electrode formula or three electric pole type leads with what the screen painting mode system of brushing on insulating base formed;
One electrode layer, it is to be covered on the conductor layer in the screen painting mode, contain the bipolar electrode formula of a working electrode and a reference electrode or contain three electric pole types of a working electrode, a reference electrode and an auxiliary electrode with formation, one end of this electrode layer is an electrode district, the other end is the contact that links to each other with detector, wherein except that the reference electrode district, conductor layer is covered by electrode layer fully;
The electrical isolation middle layer, it is directly to paste with the insulating gel system of directly brushing or with insulating material to be covered on the electrode layer, makes and exposes described electrode district and contact portions, this electrical isolation middle layer has a U type or T type duct in the part with respect to described electrode district;
The reactivity layer, it contains materials such as buffer solution, filling material, catalytic reaction thing, electron mediator and surfactant, and is coated on electrode layer surface in aforementioned U type or T type duct;
Loam cake, it has a confined space that raises up with respect to the electrical isolation middle layer, this loam cake is covered on the electrical isolation middle layer, make and form a capillary water conservancy diversion district between loam cake and aforementioned U type or the T type duct, at the bottom of the medial extremity in the corresponding U type of the described confined space that raises up duct or the co-ordination intersection point place in T type duct.
The present invention also provides a kind of method for making of described electric chemical formula biological sensing test sheet, and step is as follows:
A, with screen printing technology on the one side of insulating base, print out a conductor layer, then under 40 ℃ to 120 ℃ the oven dry; Wherein said conductor layer is bipolar electrode formula or three electric pole types;
B, on aforesaid conductor layer, brush electrode layer with screen printing technology, again in 40 ℃ to 120 ℃ following oven dry; Wherein said electrode layer is three electric pole types that contain the bipolar electrode formula of a working electrode and a reference electrode or contain a working electrode, a reference electrode and an auxiliary electrode, and an end of this electrode layer is an electrode district, the other end is the contact that links to each other with detector, wherein except that the reference electrode district, conductor layer is covered by electrode layer fully;
C, utilize the insulation glue-line that the one electrical isolation middle layer of containing U type or T type duct is set on the aforementioned insulating base that is printed on electrode layer, make the electrode district of electrode layer one end be positioned at U type or T type duct, and the contact of the electrode layer other end is exposed;
D, in described U type or T type duct, apply a reactivity layer;
E, on the electrical isolation middle layer, paste a loam cake, this loam cake has a confined space that raises up with respect to the electrical isolation middle layer, and this confined space that raises up is positioned at the bottom of the medial extremity in described U type duct or the co-ordination infall in T type duct, and makes and form a capillary water conservancy diversion district between this loam cake and U type or the T type duct.
The present invention is according to the design of capillary and siphon combination principle and electrode district, obtain the electrode reaction district of a quick water conservancy diversion, it only needs a few corpse or other object for laboratory examination and chemical testing (about 0.5-0.8 μ l) just can allow the electrode operate as normal, all be about 5-10 second detection time, this test piece be one not only fast but also accurate electrochemica biological detecting device, simple in structure, need not cover nethike embrane fully, and can avoid a corpse or other object for laboratory examination and chemical testing to leak outside polluting.
Above-mentioned insulating base is the arbitrary material in polycarbonate, polyester, polyethers, polyamide, polyurethane, poly-imines, Polyvinylchloride, glass, glass mat, pottery, polypropylene and the tygon; Conductor layer (conductive layer) is any material in silver, silver chloride or the gold; Electrode layer is any material in carbon, silver, gold or the platinum; The catalytic reaction thing is biological catalytic materials such as ferment or other inanimate catalytic materials.
Above-mentioned electrical isolation intermediate layer thickness is between the 20-400 micron, and most preferred thickness is the 50-200 micron; The breach orifice throat length in this electrical isolation middle layer is between the 2-8 millimeter, and width is between the 0.5-5 millimeter; The confined space volume that raises up with respect to the electrical isolation middle layer is between the 0.5-4 microlitre.
The method for making in above-mentioned middle layer can directly be pasted insulating intermediate layer or in the screen painting mode; The breach duct in this middle layer can be U type or T type, and the horizontal cut of this T type is the exhausr port to the test piece both sides.
This test piece has the characteristic that a fast Absorption corpse or other object for laboratory examination and chemical testing, effective control detection corpse or other object for laboratory examination and chemical testing volume and a corpse or other object for laboratory examination and chemical testing are filled the location.The structure examination of this test piece has two or three electrodes on an insulating base, be respectively working electrode, auxiliary electrode and reference electrode (three-pole); Directly adsorb one deck responding layer on the electrode district surface; Responding layer is made up of buffer solution, catalytic reaction thing, electron transport substance, wetting agent and surfactant etc.; Then there is a corpse or other object for laboratory examination and chemical testing water conservancy diversion district electrode district top, respectively since the middle layer thin slice that electrode district top has a U type passage with and on upper cover plate combine. formed hollow channels has capillary and siphonage, can be rapidly the corpse or other object for laboratory examination and chemical testing of minute quantity be sucked into the electrode reaction district, utilize then and electrochemically decide potential method or decide current method, measure the reaction that the catalytic reaction thing and the electron transport substance of a corpse or other object for laboratory examination and chemical testing and reaction zone are produced, and then can analyze the target composition in the quantitative corpse or other object for laboratory examination and chemical testing.Face the lead front end in electrode district on the upper cover plate and have an airtight convex space, have the function that an effective control corpse or other object for laboratory examination and chemical testing volume and a corpse or other object for laboratory examination and chemical testing are filled the location.
Description of drawings
Fig. 1: screen painting electrode test piece tool U type pore space structure synoptic diagram of the present invention;
Fig. 2: screen painting electrode test piece tool T type pore space structure synoptic diagram of the present invention;
Fig. 3: screen painting electrode test piece profile synoptic diagram of the present invention;
Fig. 4: internal of the present invention is to upper process closed space superstructure synoptic diagram;
Fig. 5: internal of the present invention is to upper process closed space superstructure profile synoptic diagram;
Fig. 6: internal of the present invention is to the upper process closed space half tone test piece profile structural representation that prints electrode;
Fig. 7: when using electrode test piece of the present invention, the influence of whole blood corpse or other object for laboratory examination and chemical testing volume to detecting.
Embodiment
Describe the present invention in detail below in conjunction with specific embodiment, but do not limit practical range of the present invention.
Sensing test piece
Galvanochemistry three-pole screen painting biological sensing test sheet structure of the present invention as shown in Figure 1, at insulativity base plate 1, as Polyvinylchloride (PVC), polyester (PE), polycarbonate (PC) etc., above with conductive material, colloidal material such as silver, gold for example, brush in the screen painting mode and to make lead 2, republish with the gluey material of electric conductivity such as carbon, gold or platinum again and be covered on the lead 2, form the rectangular of electrode, one end such as working electrode 3, reference electrode 4, auxiliary electrode 5 (then not having reference electrode during the two plate design); The other end then is contact mutually with detector as 3 ', 4 ' with the 5 ' end points in comparative electrode, and 6 is the automatic identification line of test piece of detecting instrument.Paste or brush a non-conductive middle layer or an insulation glue-line 7 that contains U type hole in the screen painting mode being printed on the base plate of electrode, it has the interval function concurrently for insulation dielectric layer.Groove 7a is a corpse or other object for laboratory examination and chemical testing diversion trench, loam cake 8 form in relative diversion trench bottom one airtight and to the space of upper process 8a, the about 2 μ l of its internal volume.Electrode reaction district coated agent prescription has comprised buffer solution, catalytic reaction thing (for example ferment), electronic conduction material (for example dimethyl ferrocene, tetrathiofulvalene etc.), wetting agent (cellulose cellulose, hydroxyethyl cellulose hydroxyethyl cellulose, polyvinyl alcohol (PVA) polyvinyl alcohol, ethene polymers polyvinyl, pyrrolidone pyrrolidone, gelatin gelatin etc.) and surfactant (tween20, triton X-100, surfynol, mega8 etc.).Wall 7 forms capillary flow-guiding channel 7a on every side with last cap rock 8 applying backs in the electrode workspace, after allowing the leading edge contact of a corpse or other object for laboratory examination and chemical testing such as blood by 7a, be filled up to the electrode reaction district via capillarity rapidly, this moment by a corpse or other object for laboratory examination and chemical testing and reactant via catalysis of catalytic reaction thing and electron transport substance effect, again via the signal of electrode and detector detecting generation electric current.Above-mentioned water conservancy diversion groove can and need in (less than 1 second) very fast to allow electrode carry out normal work of detection and examination under the situation of blood (less than 1 microlitre) seldom.
Another galvanochemistry three-pole electrode test piece structure of the present invention as shown in Figure 2.Sketch it, on insulating base 1, brush conductive layer 2 as materials such as silver, silver chloride, gold in the screen painting mode, be covered in brush systems such as conductive material such as carbon, gold or platinum again and form working electrode 3, reference electrode 4 and auxiliary electrode 5 on the conductive layer with conductive material.The end 3 ', 4 ' and 5 ' that its each electrode is relative is the contact that joins with detector; And 6 be the automatic identification line of test piece of detecting instrument.Next step is for finishing the wall 7 of pasting or brush in the screen painting mode with insulating gel a T font hole on the base plate of electrode layer; Last cap rock 8 comprises the airtight and space 8a that raises up, the about 2 μ l of the internal volume of this protruding space 8a, be positioned at T word co-ordination intersection point directly over; And the groove 7a that forms between loam cake and wall is a corpse or other object for laboratory examination and chemical testing diversion trench, and 7b and 7c are the exhaust passage towards both sides.A corpse or other object for laboratory examination and chemical testing such as blood etc., after the contact of the leading edge of 7a, being flowed through rapidly by the capillary principle is filled up to the electrode reaction district, and the leading edge of a corpse or other object for laboratory examination and chemical testing is no more than the front end of 8a also as the design of Fig. 1; In addition, this design can be deleted the exhaust passage of 7b or 7c either side, also can obtain identical effect.
The base plate base material
Base plate 1 can be made up of various insulativity materials, macromolecule for example, plastics and stupalith etc., the demand that the kind of the selection of material must cooperate electrode material and use, if the intrusive mood purposes then should select for use flexible material to ease the pain and to avoid injuring tissue, this type of purposes can be used for example polycarbonate of insulativity macromolecular material usually, polyester (for example Mylar series of products of E.I.Du Pont Company), polyethylene terephthalate (polyethylene terephthalate, PET), Polyvinylchloride (PVC), polyethers, polyamide, polyurethane, poly-imines etc.; Rigid material can use not easy fracture or incliner such as stupalith as silicon dioxide or aluminium dioxide etc. on the other hand.With the vitro detection purposes, the dimension width of base plate is general common between the 3-15 millimeter, more accurate can be between the 5-10 millimeter, thickness is about 50-800 micron or more accurate between the 200-400 micron, floor length is influenced by various factors then may be between 1-8 centimetre, more accurate between 2-5 centimetre.
Conductor layer and electrode
As shown in Figure 1, conductive material forms conductor section 2 as materials such as silver, gold, platinum through the screen painting mode on base plate 1, as the purposes that connects each electrode and proving installation, the material of use high conductivity and low resistance characteristic can reduce the impedance of electrode, improves the detecting current signal.Conductive material such as carbon paste are then brushed on lead 2,6 as the automatic identification purposes of the test piece of detecting instrument, except reference electrode 4, the lead 2 of the system of brushing is covered fully, 4 the silver-colored conductor part that exposes utilizes electrochemical principle can silver surface be processed into the plan reference electrode of silver chloride again, perhaps be covered on the silver-colored lead, or, so just do not need to do in addition silver chloride and handle directly with silver/silver chloride printing ink brush system lead with silver/silver chloride printing ink brush system.
Insulation course
Insulating intermediate layer 7 can be formed by the dielectric material brush system of tool insulativity, or is covered in electrode surface with bonding method, except covering the carbon surface that needn't expose, also provides the reaction zone of a fixed-area.
The reaction reagent district
Reaction reagent is coated on the electrode district, has comprised the catalytic reaction thing, buffer solution, filling material, electron transport substance, surfactant of reaction usefulness etc.For example use that the catalytic reaction thing can be glucose oxidase or dehydrogenasa when glucose detection, the filling material composition has comprised macromolecule or wetting agent for example cellulose, poly-ethanol, gelatin etc., surfactant component, for example Tween-80, Triton X-100, Surfynol or Mega 8 or the like, provide a corpse or other object for laboratory examination and chemical testing and reagent to return molten divergent function, and the water wettability and the divergent function in capillary water conservancy diversion district.Therefore, reaction reagent layer can provide reaction and capillary function simultaneously, not only allows a corpse or other object for laboratory examination and chemical testing can be full of electrode district and analytical reactions are carried out, and provides electrode reaction current as the quantitative function of corpse or other object for laboratory examination and chemical testing composition.Better suited electron transport substance should have the oxidation-reduction potential between-100 to+500 millivolts, use different electron transport substances according to different detection demands. can use the dimethyl ferrocene (dimethyl ferrocene) of ferrocene series and the derivant or the misfit thing of tetrathiafulvalene (tetrathiafulvalene) or said two devices when for example detecting glucose, lower current potential can be avoided the interfering material in the corpse or other object for laboratory examination and chemical testing, and higher electron transport efficient then can provide stronger current signal.The buffer solution effect is to keep the pH-value of certain limit, general pH-value is between PH4-9, being fit to pH-value between PH5-8. operable damping fluid composition comprises phosphate, acetate, citrate etc., concentration range is between the 10-1000 millimolar concentration, the 30-1000 millimolar concentration of optimum concentration.
Capillary water conservancy diversion layer
The water conservancy diversion layer is covered in the electrode top by interval thin slice 7 and loam cake 8 and is formed; 7a is a corpse or other object for laboratory examination and chemical testing capillary groove, as have to both sides degassing function (design of T font) then 7b and 7c be exhausr port, but 7b or 7c individualism.Utilize the width of the thickness of thin slice 7 and 7a groove can adjust the size of water conservancy diversion district volume, in general thickness is between the 20-400 micron, preferred thickness 50-200 micron, hollow area length is between the 2-8 millimeter, width is between the 0.5-5 millimeter, and preferable width is then between the 1-2 millimeter, and formed hollow region volume is between the 0.2-160 microlitre, optimal dose is approximately between the 0.5-2 microlitre during operation of an actual corpse or other object for laboratory examination and chemical testing, during corpse or other object for laboratory examination and chemical testing contact water conservancy diversion layer edge to being full of required time less than 1 second.
The closed protruding 8a of last cap rock 8 can be circular, square or other geometric configuration, suitable size is between the 0.5-4 microlitre, the position of breach is positioned at the back upper place of working electrode flow-guiding channel, and this breach halts after making a blood corpse or other object for laboratory examination and chemical testing that enters fill up reaction zone.Distance piece 7 can be made up of transparent or opaque insulativity material with upper cover plate 8; for example plastics or macromolecular material such as PVC Polyvinylchloride or Mylar etc.; the 8a of water conservancy diversion district upper area can be transparence, except whether the user can fill up by a visual corpse or other object for laboratory examination and chemical testing smoothly, also has the function of protection test piece.Last cap rock can form by two steps.First step is that a hole is opened in figure one so-called upper end of going up cap rock, that is 8a, covers a complete thin slice 9 (as Fig. 4 and shown in Figure 5), the profile of shown Fig. 1 of Fig. 3 and Fig. 6 and sheet-containing 9 test pieces again on this sheet.
Fill with sensor
Filling with Design of Sensor is used for detecting three groups of electrodes top and whether is full of a corpse or other object for laboratory examination and chemical testing, in three-pole design if working electrode during in water conservancy diversion district outermost, utilize auxiliary electrode and working electrode to finish and fill with sensing function, the utilization electric current, the monitoring of current potential or resistance value, when a corpse or other object for laboratory examination and chemical testing does not fill up three electrodes as yet, the resistance value of deciding between potential monitoring work and auxiliary electrode should be infinitely great, and when a corpse or other object for laboratory examination and chemical testing fills up electrode, the impedance meeting sharply reduces, through determine that thus a corpse or other object for laboratory examination and chemical testing just starts the analytical parameters of galvanochemistry part when filling up, also can utilize with principle and monitor when two electrode design.Utilize electrode as filling with sensor, must make the distribution of electrode consistent with corpse or other object for laboratory examination and chemical testing flow direction, that is to say that working electrode contacts a corpse or other object for laboratory examination and chemical testing at first, auxiliary electrode must contact a corpse or other object for laboratory examination and chemical testing at last can correctly declare to know whether fill up, and equally also auxiliary electrode can be come foremost contacting position.
Electrochemical analysis
When combination of electrodes is finished via cutting or after impact style cuts, utilize external hand held galvanochemistry instrument can carry out specimen analyzing, analysis mode can be carried out in many ways, for example decide potential method (0-0.6 volt) and measure steady-state current, perhaps to decide the total charge value in the current potential measurement certain hour, charge value is the integration of electric current and time, the direct ratio that all can be used as constituent concentration in the corpse or other object for laboratory examination and chemical testing with steady-state current is examined magnitude relation, the sensing function of filling with that cooperates electrode slice in the detector, read to know when detector and start the electrochemical analysis parameter when filling with signal automatically, so increase the accuracy that detects, especially when the whole detection time had been lower than 10 seconds, the error of a little time tended to cause very big variation.
Following embodiment is to be example with the blood sugar test, and its purpose is not just limited to the scope of the invention to some extent setting forth practicability of the present invention.
Embodiment one, screen painting glucose detection specimen preparation
The Dampler Ace base plate after under 50 ℃ dry 30 minutes, brushes three groups of electrodes (working electrode, reference electrode and auxiliary electrode) with carbon paste with the screen painting conductive silver glue-line of 300 meshes again.In 90 ℃ of bakings 15 minutes, form insulation course with insulating gel brush system back dry through under ultraviolet light then once more, and expose inspection water conservancy diversion reaction zone 7a, 7b and 7c (design of air hole is arranged).Reaction reagent 2-6 microlitre (includes 0.5-3 unit's glucose oxidase, 0.1-1% polyvinyl alcohol (PVA), pH4.0-9.0,100mM kaliumphosphate buffer, 10-100mM potassium chloride, dimethyl ferrocene 0.05-0.5%, tween-20
0.005-0.2%, surfynol0.005-0.2% and carbonic acid methyl fiber 0.1%-1.0%), be coated on aforementioned formed depression water conservancy diversion district 7a, take out after 1 hour again to fit in 45 ℃ of dryings and contain the upper cover plate 8 of central hole, after the stickup again to cut the product of finishing that are again after a transparent upper cover sheet 9 pressing sets.
Embodiment two, standard glucose solution and whole blood test
Configuration standard buffer solution of potassium phosphate (PH7.4) includes concentration of glucose 0-400 milligram/100 milliliters, with the embodiment one prepared screen painting electrode glucose test piece that forms, at electrochemical analyser (CHInstrument Co.650A) under the situation of 100 millivolts of test potential, test duration was 8 seconds, during each the test, supply corpse or other object for laboratory examination and chemical testing volume is 3 microlitres, and the volume that each test piece sucks is all less than 3 microlitres.Its test result is as shown in table 1.
Table 1 standard glucose test result
| Concentration of glucose (mg/dl) | Voltage (microcoulomb) |
| 0 | 0.690 |
| 25 | 1.532 |
| 50 | 2.952 |
| 100 | 5.248 |
| 200 | 7.400 |
| 400 | 9.577 |
A whole blood corpse or other object for laboratory examination and chemical testing also can be measured by this test piece.Fresh adopt the vein whole blood add gained inspection amount with standard glucose, data are as shown in table 2, test potential 100 millivolts, it is 2 microlitres that the test volume of whole blood is provided.
The whole blood test result of the different glucose contents of table 2
| Concentration of glucose (mg/dl) | Electric weight (microcoulomb) |
| 80 | 1.556 |
| 105 | 2.636 |
| 130 | 3.440 |
| 180 | 5.946 |
| 280 | 9.707 |
| 380 | 11.733 |
| 480 | 12.464 |
| 580 | 13.945 |
The blood sugar test of embodiment three, different botal blood volumes
Utilize the made electrode test piece of embodiment one, the demand blood volume of the present invention of the whole blood test of supply different volumes, the vein whole blood is mixed with the concentration of 300mg/dl with standard glucose liquid.
The method of this test makes test piece suck with the principle of siphon, and detects with the testing conditions as embodiment two for the whole blood of different volumes is provided.Show that as Fig. 7 result when sucking corpse or other object for laboratory examination and chemical testing quantity not sufficient (as 0.5 μ l), it is on the low side to detect concentration of glucose; But when the corpse or other object for laboratory examination and chemical testing volume of providing suction reaches 0.8 μ l when above, glucose detects concentration and then levels off to the concentration of being prepared, and a whole blood corpse or other object for laboratory examination and chemical testing that is provided also can't all be inhaled into; The corpse or other object for laboratory examination and chemical testing volume that is provided is higher, remaining volume then the more, this expression corpse or other object for laboratory examination and chemical testing water conservancy diversion reaction zone has reached saturated, can't suck a more corpse or other object for laboratory examination and chemical testing.The leading edge of the corpse or other object for laboratory examination and chemical testing that sucks is neither to surpass 8a and the water conservancy diversion district part of joining, and suffices to show that the function of the restricted corpse or other object for laboratory examination and chemical testing volume of design of the present invention.
More than described the preferred embodiments of the present invention, so it is not in order to limit the present invention.Those skilled in the art can not depart from the improvement and the variation of category of the present invention and spirit to embodiment disclosed herein.
Claims (17)
1, a kind of electric chemical formula biological sensing test sheet is made of following assembly:
An insulating base;
-conductor layer, it is to have bipolar electrode formula or three electric pole type leads with what the screen painting mode system of brushing on insulating base formed;
One electrode layer, it is to be covered on the conductor layer in the screen painting mode, contain the bipolar electrode formula of a working electrode and a reference electrode or contain three electric pole types of a working electrode, a reference electrode and an auxiliary electrode with formation, one end of this electrode layer is an electrode district, the other end is the contact that links to each other with detector, wherein except that the reference electrode district, conductor layer is covered by electrode layer fully;
The electrical isolation middle layer, it is directly to paste with the insulating gel system of directly brushing or with insulating material to be covered on the electrode layer, makes and exposes described electrode district and contact portions, this electrical isolation middle layer has a U type or T type duct in the part with respect to described electrode district;
The reactivity layer, it contains buffer solution, filling material, catalytic reaction thing, electron mediator and surfactant, and is coated on electrode layer surface in aforementioned U type or T type duct;
Loam cake, it has a confined space that raises up with respect to the electrical isolation middle layer, this loam cake is covered on the electrical isolation middle layer, make and form a capillary water conservancy diversion district between loam cake and aforementioned U type or the T type duct, at the bottom of the medial extremity in the corresponding U type of the described confined space that raises up duct or the co-ordination intersection point place in T type duct.
2, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein insulating base is any material in polycarbonate, polyester, polyethers, polyamide, polyurethane, poly-imines, Polyvinylchloride, glass, glass mat, pottery, polypropylene and the tygon.
3, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein conductor layer is any material in silver, silver chloride or the gold.
4, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein electrode layer is a carbon, any material in silver, gold or the platinum.
5, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein the catalytic reaction thing is biological catalytic materials or inanimate catalytic materials.
6, electric chemical formula biological sensing test sheet as claimed in claim 5, wherein biological catalytic materials is a ferment.
7, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein the electrical isolation intermediate layer thickness is between the 20-400 micron.
8, electric chemical formula biological sensing test sheet as claimed in claim 7, wherein the electrical isolation intermediate layer thickness is the 50-200 micron.
9, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein the U type in electrical isolation middle layer or T type orifice throat length are between the 2-8 millimeter, and width is between the 0.5-5 millimeter.
10, electric chemical formula biological sensing test sheet as claimed in claim 1, wherein the confined space volume that raises up with respect to the electrical isolation middle layer of loam cake is between the 0.5-4 microlitre.
11, a kind of method for making of electric chemical formula biological sensing test sheet as claimed in claim 1, finish by following step:
A, with screen printing technology on the one side of insulating base, print out a conductor layer, then under 40 ℃ to 120 ℃ the oven dry; Wherein said conductor layer is bipolar electrode formula or three electric pole types;
B, on aforesaid conductor layer, brush electrode layer with screen printing technology, again in 40 ℃ to 120 ℃ following oven dry; Wherein said electrode layer is three electric pole types that contain the bipolar electrode formula of a working electrode and a reference electrode or contain a working electrode, a reference electrode and an auxiliary electrode, and an end of this electrode layer is an electrode district, the other end is the contact that links to each other with detector, wherein except that the reference electrode district, conductor layer is covered by electrode layer fully;
C, utilize the insulation glue-line that the one electrical isolation middle layer of containing U type or T type duct is set on the aforementioned insulating base that is printed on electrode layer, make the electrode district of electrode layer one end be positioned at U type or T type duct, and the contact of the electrode layer other end is exposed;
D, in described U type or T type duct, apply a reactivity layer;
E, on the electrical isolation middle layer, paste a loam cake, this loam cake has a confined space that raises up with respect to the electrical isolation middle layer, and this confined space that raises up is positioned at the bottom of the medial extremity in described U type duct or the co-ordination infall in T type duct, and makes and form a capillary water conservancy diversion district between this loam cake and U type or the T type duct.
12, method for making as claimed in claim 11, wherein the method for making in electrical isolation middle layer is to be provided with direct stickup insulating intermediate layer or in the screen painting mode.
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| TWI645185B (en) * | 2017-08-24 | 2018-12-21 | 鄭原輝 | Biosensor test strips, method for making same and test strip receiving device |
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| CN101261270B (en) * | 2007-03-09 | 2012-11-07 | 中国人民解放军军事医学科学院微生物流行病研究所 | Immunity-chromatography multiple detection test paper disk and immunity-chromatography multiple detection method |
| CN101430336B (en) * | 2007-11-05 | 2012-12-19 | 勤立生物科技股份有限公司 | Method for hemachrome or hematocrit detection by electro-chemistry method, and detection test piece thereof |
| CN101344501B (en) * | 2008-08-22 | 2011-10-05 | 中南大学 | Silk screen printing electrode, manufacturing technique and use thereof |
| CN102297885A (en) * | 2010-06-25 | 2011-12-28 | 厚美德生物科技股份有限公司 | Biological testing chip structure with micro-channel |
| CN103308675B (en) * | 2013-05-08 | 2015-08-12 | 北京工业大学 | The preparation of the screen printing electrode immunosensor of quick detection Microcystin and detection method |
| CN103412026A (en) * | 2013-08-30 | 2013-11-27 | 江苏鱼跃医疗设备股份有限公司 | Biosensor and manufacturing method thereof |
| CN104345074B (en) * | 2014-11-13 | 2017-01-18 | 东莞市莞信企业管理咨询有限公司 | Blood sugar test strip |
| CN111175357A (en) * | 2020-01-20 | 2020-05-19 | 杭州联晟生物科技有限公司 | Prothrombin time detection test strip and preparation method thereof |
| CN112229883A (en) * | 2020-09-04 | 2021-01-15 | 中芯维康医疗科技(重庆)有限责任公司 | Step protection structure, detection printed electrode, preparation process and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| TWI645185B (en) * | 2017-08-24 | 2018-12-21 | 鄭原輝 | Biosensor test strips, method for making same and test strip receiving device |
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