CN101303357A

Movatterモバイル変換

Info

Publication number: CN101303357A
Application number: CNA2007100404999A
Authority: CN
Inventors: 胡军
Original assignee: Individual
Current assignee: Individual
Priority date: 2007-05-10
Filing date: 2007-05-10
Publication date: 2008-11-12

Abstract

The invention discloses a matching test paper for a glucose tester and a preparation method thereof, which pertain to the technical field of bio-sensor. The substrate of the test paper is printed with a working electrode, a reference electrode and an electrical circuit; one end of the substrate is printed with a conducting wire connected with a read-out instrument; the working electrode and the reference electrode are carbon coatings which are printed on the substrate at once and printed with biological enzyme, a medium and an adhesive. The method adopts bio-sensor and electrochemical principle, produces the test paper by screen printing which is not restricted to a conductive circuit as well as prints relatively sensitive bio-components directly on an electrode holder; screen printing area is fully controlled by a screen printing machine. The encapsulation of the test paper adopts siphon encapsulation which limits reaction space. Once the test paper of the invention contacts human blood, a limited quantity of blood is sucked into a reaction tank and accurate results can be obtained conveniently and visually from a corresponding matching testing instrument. The glucose test paper has the advantages of simple production method, high accuracy and low cost and is convenient for later process and encapsulation and good for scale production.

Description

The facture of rapid blood sugar test paper

Technical field

What the present invention relates to is a kind of method for making of rapid blood sugar test paper, and the particularly a kind of and supporting test paper method for making of blood glucose monitoring system belongs to the biosensor technique field.

Background technology

China's diabetic surpasses 4,500 ten thousand according to statistics, and also increase with 3000 patients' speed of development every year.One of effective means of control of diabetes development at present is to implement blood sugar test every day, thereby the stable of patient's glycemic index kept in strict control invalid diet and medication.Its supporting test paper of some blood glucose meter that occur on the home market adopts complicated technology production, this manufacture craft is after the basic electrode silk screen printing is finished, also need to implement the point sample processing of biology enzyme material, yield poorly, production cost and rejection rate height, can only implement small-scale production, be difficult to carry out industrialization and produce continuously.Find by literature search, U.S. Patent Publication No.: 2004/0146958, patent name is: Glucose strip sensor and glucose measurement usingthe glucose strip sensor (utilizing glucose strip sensor determination glucose), introduced a kind of method for making of three-electrode system glucose strip sensor in this patent.Include a working electrode in the test macro, a reference electrode also has a verification electrode, its objective is to make system's working sample glucose more accurately.Thereby also having introduced employing control reaction zone space, system make the reaction mixture volume in the reaction chamber obtain the methods such as accuracy that strict control raising is measured.China Patent No.: ZL03116172.3, the glucose of having introduced a kind of two electrode systems is tested the production method of strip.Its conductive coating adopts the double-deck print process of conductive silver and carbon, after the conduction base electrode is printed and finished, needs to adopt spraying equipment that biology enzyme is coated on the base electrode surface, finishes the making of blood sugar test paper.These methods all adopt the silk-screen technology of multilayer conductive coating, also need to use biological elements point sample equipment with the surface of biology enzyme solution coat to electrode, cause complex manufacturing technologyization, but also will use conductive silver printing ink, the expensive industrial-scale production that is unfavorable for product of material price.

Summary of the invention

The objective of the invention is to overcome deficiency of the prior art, the method for making of a kind of blood sugar test paper and simplification serigraphy thereof is provided, the base electrode silk-screen one-step print of test paper is finished, the biochemical reaction enzyme reagent of test paper also adopts the printing of silk-screen method, simplified manufacture craft, reduce the production equipment input, improved the utilization factor of screen printing device.Method for making is simple and easy to control, more is adapted to large-scale industrial production.The quick test paper of the blood sugar of making adopts the siphonage encapsulation, in case behind the contact blood of human body, can just read test result intuitively above corresponding blood sugar test instrument.

The present invention is achieved by the following technical solutions, and the quick test paper of blood sugar of the present invention comprises: a substrate, as the PP synthesized paper sheet; Disposablely on the scraps of paper print a working electrode simultaneously; A reference electrode; Two electrodes constitute the blood sugar test loop with the sample circuit of blood sugar test instrument.Working electrode and reference electrode all are made of the conductive carbon coating, stamp one deck on the working electrode again and contain the biology enzyme that the catalysis blood-glucose reacts, as glucose oxidase, glucose dehydrogenase or glucokinase, and a kind of water-soluble conducting medium such as the potassium ferricyanide or benzoquinones that can between enzymic catalytic reaction center and electrode, carry out electron transport; Bonding agent such as carboxymethyl cellulose, hydroxyethyl cellulose, sodium alginate or edible gelatin so constitute a test paper electricity loop.According to the height of concentration of glucose in the blood, produce the redox current signal of corresponding size during reaction, the rapid tested instrument of this signal detects to be read.This test paper has been simplified the typography of the multilayer conductive coating of base electrode, has omitted biological elements point sample equipment, and technology is simpler.Tested material is a blood-glucose, and the enzyme of use can be glucose oxidase, glucose dehydrogenase or glucokinase.

Working electrode is the single carbon coating conductive film of a kind of serigraphy on holder, comprises the enzyme of catalysis biochemical reaction and the conducting medium and the bonding agent that can carry out electron transport between enzymic catalytic reaction is with electrode republishing one deck on the conductive film.This working electrode can transmit the oxidation current that produces behind the enzymic catalytic reaction.Working electrode and contrast electrode are close, and two electrodes constitute an effecting reaction zone, after connecting with tester, form effective blood sugar test system.

The present invention adopts bio-sensing and electrochemical principle, make above-mentioned test paper by pure silk reticulated printing technology, the conducting wire is not only printed in serigraphy, also will compare responsive biological elements such as enzyme, conducting medium and bonding agent etc. and be imprinted on the electrode surface by the silk-screen method.

Below the inventive method is further described, concrete steps are as follows:

(1) silk-screen conductive carbon ink, standby with the low-speed mixer mixing;

(2) the nylon polyester material silk screen of 300 order numbers is fixed on square aluminum metal frame and adds, in the darkroom, coat photoresists, drying for standby;

(3) working electrode and the reference electrode film sheet with computer drawing places on the photoresists, removes the unhardened photoresists that covered by the electrode negative film with giant flushing after exposure under the uviol lamp, and dry back forms the silk-screen template;

(4) the silk-screen template that will have working electrode and a reference electrode figure is installed on the automatic screen printer standby;

(5) be that the glucose oxidase of 250 international units or glucose dehydrogenase or activity that activity is 350 international units are the glucokinase of 120 international units with activity, allocate in the phosphate buffer of every liter of 90 mM, add every liter of 20-30 gram of the benzoquinones or the potassium ferricyanide and bonding agent simultaneously, it is standby fully to shelve 4 ℃ of refrigerators behind the mixing.With conductive carbon ink with working electrode and the disposable serigraphy of reference electrode on substrate; Print enzyme and conducting medium potpourri then.The ink mixture that is used for second layer printing comprises: bonding agent, conducting medium and enzyme.The material of substrate can be a kind of good toughness, the polymeric material that insulativity is high, and such as Polyvinylchloride, polyester, polycarbonate or PP synthetic paper.

The present invention has substantive distinguishing features and marked improvement, and the inventive method has been simplified the multilayer of conductive coating and printed repeatedly, and screen printing technique has been expanded to the printing of biological elements enzyme.Optimized production process, reduced production technology and made requirement; Reduced production equipment; Improved the industrialization level of product.Wire printing material and screen printing device needn't specially be processed, and can obtain by commercially available channel, and cost is very low; Relatively more responsive biological elements such as enzyme, conducting medium and bonding agent etc. are imprinted on the electrode base sheet with screen printing mode, the precision of final products and repeatability are fully ensured.Test paper of the present invention adopts the encapsulation of hydrocone type method, in case behind the contact blood of human body, can conveniently read test result intuitively on corresponding supporting testing tool; method for making is simple, the precision height, and cost is low; make things convenient for post-production and encapsulation, help large-scale industrial production.

Description of drawings

Fig. 1 test paper structural representation of the present invention

Embodiment

As shown in Figure 1, test paper of the present invention comprises: a substrate 1, be printed on a working electrode 2 on the substrate 1, with a reference electrode 3, end at substrate 1 is printed on the lead 4 that connects with the readout meter epiphase, working electrode 2 and reference electrode 3 are respectively by being imprinted on the substrate 1, republish the biology enzyme that contains the reaction of catalysis blood-glucose on working electrode 2 and the reference electrode 3, as glucose oxidase, glucose dehydrogenase or glucokinase 6, and the medium and the bonding agent that can between enzymic catalytic reaction center and electrode, carry out electron transport, so constitute a circuit loop.According to the height of concentration of glucose in the blood, produce the current signal of corresponding size during reaction, and the detection of tested instrument is read.Tested material is a blood-glucose.

Working electrode 2 and reference electrode 3 be a kind of serigraphy at on-chip carbon coating conductive film, comprise the enzyme of catalysis biochemical reaction and the medium 6 that can between enzymic catalytic reaction is with electrode, carry out electron transport in serigraphy on the conductive film.Working electrode 2 is close with contrast electrode 3, and two electrodes constitute an effecting reaction zone, after connecting with tester, forms effective body fluid components test macro.

Content in conjunction with the inventive method provides following examples, and the present invention is further illustrated.

Embodiment 1: polyphenyl (PP) synthetic paper, it is standby to be cut into 300X300 centimetre substrate in advance.

(1) 1 gram silk-screen conductive carbon ink (ACHESON Acheson423SS) mechanical raking 30 minutes, mixing is standby.

(2) metal of 300 order numbers or nylon polyester material silk screen are fixed on square light-weight metal frame and add, in the darkroom, coat Kodak's photoresists, drying for standby.

(3) working electrode and the reference electrode negative film with computer drawing places on the photoresists, and exposure is 15 minutes under 20 watts of uviol lamps.Remove the unhardened photoresists that covered by the electrode negative film with giant flushing.Dry back forms the silk-screen template.

(4) the silk-screen template that will have working electrode and a reference electrode figure is installed on the automatic screen printer standby.

(5) be that the glucose oxidase of 250 international units is allocated in the phosphate buffer of every liter of 50 mM with activity, add every liter 12 gram of benzoquinones simultaneously, every liter 20 gram of bonding agent.It is standby fully to shelve 4 ℃ of refrigerators behind the mixing.

On above-mentioned automatic screen printing machine, add conductive carbon ink, conduct electricity the printing of base electrode, make working electrode and reference electrode, the electrode of having printed need be held in 80 ℃ of baking ovens and solidify 10 minutes, on each sheet PP substrate, form 250 pairs of work and reference electrode, take out standby then.When making the enzyme test peper of test glucose, take out Manufactured element task electrode and reference electrode, with the mixed slurry serigraphy in above-mentioned (5) in the setting of working electrode of making and reference electrode position all.This position is positioned at the interval of the about 3 * 3mm of working reaction end of working electrode and reference electrode.

With Manufactured test-strips, place tester and apply the 300mV operating voltage for two sections at working electrode and reference electrode, measure the glucose of variable concentrations, the result shows that reaction took place after 10 seconds, shows to make good test repeatability and the correlativity of electrode.Through relatively with Biochemical Analyzer, get 50 parts of glucose sugar control liquids and 50 parts of patient's blood sugar contrast tests respectively, the relative standard deviation that its correlativity γ reaches 0.9567 test-strips is CV≤6.5%.

Embodiment 2: polyester (PET) sheet material, it is standby to be cut into 300 * 300 centimeters substrates in advance.

(1) silk-screen conductive carbon ink mechanical raking mixing is standby.

(3) working electrode and the reference electrode negative film with computer drawing places on the photoresists, and exposure is 25 minutes under 20 watts of uviol lamps.Remove the unhardened photoresists that covered by the electrode negative film with giant flushing.Dry back forms the silk-screen template.

(5) be that the glucose dehydrogenase of 350 international units is allocated in the phosphate buffer of every liter of 50 mM with activity, add every liter 18 gram of the potassium ferricyanide simultaneously, bonding agent 30 grams.It is standby fully to shelve 4 ℃ of refrigerators behind the mixing.

Method for makings such as silk-screen are with example 1, with the test-strips of making, place tester and apply the 300mV operating voltage for two sections at working electrode and reference electrode, measure the blood-glucose of variable concentrations respectively, the result shows that reaction took place after 10 seconds, shows to make good test repeatability and the correlativity of electrode.Through relatively with Biochemical Analyzer, get 50 parts of glucose control liquids and 50 parts of patient's blood sample blood sugar contrast tests, its correlativity γ reaches 0.967, and the relative standard deviation of test-strips is CV≤5.7%.

Embodiment 3: polyphenol ester (PP) sheet material, it is standby to be cut into 300 * 300 centimeters substrates in advance.

(1) silk-screen conductive carbon ink machinery mixing is standby;

(2) metal of 300 order numbers or nylon polyester material silk screen are fixed on square light-weight metal frame and add, in the darkroom, coat Kodak's photoresists, drying for standby;

(3) working electrode and the reference electrode negative film with computer drawing places on the photoresists, and exposure is 30 minutes under 40 watts of uviol lamps, removes the unhardened photoresists that covered by the electrode negative film with giant flushing, and dry back forms the silk-screen template;

(5) be that the glucokinase of 120 international units is allocated in the phosphate buffer of every liter of 120 mM with activity, add every liter 16 gram of the potassium ferricyanide simultaneously, bonding agent 30 grams.It is standby fully to shelve 4 ℃ of refrigerators behind the mixing.

Method for makings such as silk-screen are with embodiment 1, with the test-strips of making, place tester and apply the 350mV operating voltage for two sections at working electrode and reference electrode, measure the glucose of variable concentrations respectively, the result shows that reaction took place after 10 seconds, shows to make good test repeatability and the correlativity of electrode.Through relatively with hospital's Biochemical Analyzer, get 50 parts of glucose control liquids and 50 parts of diabetes patient's blood sample glucose contrast tests respectively, its correlativity γ reaches 0.975, and the relative standard deviation of test-strips is CV≤7.1%.

Claims

1, a kind of rapid blood sugar test paper, mainly comprise: a substrate (1), be printed on a working electrode (2) on the substrate (1), be printed on a reference electrode (3) on the substrate (1), substrate is printed on electric loop on (1), end at substrate (1) is printed on the lead (4) that connects with the readout meter epiphase, it is characterized in that, working electrode (2) and reference electrode (3) are that the disposable substrate (1) that is imprinted on is gone up the conductive carbon coating that forms, be printed on the biology enzyme of catalysis blood-glucose reaction and the medium and the adhesive coating (6) that can between enzymic catalytic reaction center and electrode, carry out electron transport on the reaction zone of working electrode (2) and reference electrode (3), so constitute a circuit loop, during reaction according to the height of concentration of glucose in the blood, produce the current signal of corresponding size, and the detection of tested instrument is read.

2, rapid blood sugar test paper according to claim 1 is characterized in that, tested material is a glucose, and the enzyme of use is a glucose oxidase, glucose dehydrogenase, glucokinase.

3, rapid blood sugar test paper according to claim 1, it is characterized in that, working electrode (2) and reference electrode (3) are the monolayer carbon conductive film of a kind of serigraphy on holder, comprise the enzyme of catalysis biochemical reaction and the medium (6) that can carry out electron transport between enzymic catalytic reaction is with electrode at silk-screen on the conductive film.

4, rapid blood sugar test paper according to claim 1 is characterized in that, working electrode (2) is close with contrast electrode (3), and two electrodes constitute an effecting reaction zone, after connecting with tester, forms effective blood constituent test macro.

5, rapid blood sugar test paper according to claim 1 is characterized in that, the polymeric material that the material of substrate (1) is a kind of good toughness, insulativity is high, as: Polyvinylchloride, polyester or polycarbonate.

6, a kind of method for making of rapid blood sugar test paper, it is characterized in that: adopt bio-sensing and electrochemical principle, make above-mentioned test paper by the method for serigraphy, serigraphy not only is confined to print the conducting wire, also the biology enzyme element directly is printed on the electrode support substrate, the silk-screen area is controlled by screen printer fully.

7, the method for making of rapid blood sugar test paper according to claim 6 is characterized in that, concrete steps are as follows:

(1) the silk-screen conductive carbon ink is standby through stirring and evenly mixing;

(3) working electrode and the reference electrode negative film with computer drawing places on the photoresists, and under 20-40 watt of uviol lamp exposure 15-30 minute, remove the unhardened photoresists that covered by the electrode negative film with the giant flushing, dry back forms the silk-screen template;

(5) be that the glucose oxidase of 250 international units or glucose dehydrogenase or activity that activity is 350 international units are the glucokinase of 120 international units with activity, allocate in the phosphate buffer of every liter of 50-120 mM, add every liter of 12-20 gram of the benzoquinones or the potassium ferricyanide simultaneously, bonding agent 20-30 gram, it is standby fully to shelve 4 ℃ of refrigerators behind the mixing.

According to the method for making of claim 6 or 7 described rapid blood sugar test papers, it is characterized in that 8, working electrode adopts screen printing technique that conductive carbon material is imprinted on the substrate earlier; Print enzyme and electron transit mediator then, as the benzoquinones or the potassium ferricyanide, bonding agent.

According to the method for making of claim 6 and 7 described rapid blood sugar test papers, it is characterized in that 9, the autogamy printing ink that is used for serigraphy comprises: adhesive coating, conducting objects and enzyme.