CN1367382A - Non-enzymic disposable detection electrode bar, its production method and its application - Google Patents
Non-enzymic disposable detection electrode bar, its production method and its application Download PDFInfo
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- CN1367382A CN1367382A CN 01116298 CN01116298A CN1367382A CN 1367382 A CN1367382 A CN 1367382A CN 01116298 CN01116298 CN 01116298 CN 01116298 A CN01116298 A CN 01116298A CN 1367382 A CN1367382 A CN 1367382A
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Abstract
The present invention relates to a non-enzymetic disposable electrode strip which can be used for directly detecting biological molecule being in liquid sample, for example uric acid and concentration of hemoglobin. When it is used for detecting biomolecule concentration being in human body, it can prevent interference, of signal initiated by other component being in blood. Said ivnention also provides the production method of said electrode strip and its detection device containing said electrode strip.
Description
The present invention relates to be used for the disposable electrode bar of detection of biological molecule, be specially the electrode strip that is used to detect uric acid and haemoglobin.
The detection of biomolecule can be carried out with two class electrodes: enzyme electrode and non-enzyme electrode.At clinical biochemistry analytically, biomolecule detects by its minimizing or the method for enzyme.Enzyme electrode is because strict protective condition, and complicated manufacture method and controlled condition cause and involves great expense and can not be produced in a large number.Enzyme electrode is not disposable and is unsuitable for family expenses.On the contrary, non-enzyme electrode can be made by disposable material, and produce easily, preserve and use; Therefore, they are useful biological analysers for detection of biological molecule such as uric acid and haemoglobin.
Up to the present, also there is not effectively to detect the electrode strip appearance of uric acid and haemoglobin.Ai-min Yu, Hai-li Zhang and Hong-yuan Chen (Analyst, 1997; 122:839 ~ 841) the electric current behavior that is described to uric acid on the electrode of polyglycine chemical modification in improves and its determination of trace.This electrode only can detect the uric acid sample in the 0.1M phosphate buffer, and can't measure whole blood sample.Such electrode has following defective: 1) range of linearity has exceeded common diagnostic region; 2) be not suitable for detecting blood sample; 3) complicate fabrication process of electrode and consuming timely be not suitable for a large amount of productions; 4) cost height is unfavorable for making disposable articles for use.
Jyh-Myng Zen and Jen-Sen Tang (Anal.Chem.1995; 67:1892 ~ 1895) use Ru
2-xPb
xO
7-x(massicot ruthenium) modified glassy carbon electrode and detected urea by Osteryoung square wave current device.Electrode is that detection in 1 the aciduria acid solution is fit at pH.This electrode is unsuitable for detecting blood sample.In addition, also there is following defective in kind electrode: 1) cost an arm and a leg; 2) surveyed area does not coincide with diagnostic region; 3) complicate fabrication process of electrode is unsuitable for a large amount of productions; And 4) can not be used to make disposable electrode strip, therefore be unsuitable for using at home.
For the detection of haemoglobin, the method for cyanohemoglobin is well known in the art.Described method is meant the solution dissolving haemoglobin with Drabkin, just uses K
3Fe (CN)
6With the Fe in the haemoglobin
2+Be oxidized to Fe
3+Form inclined to one side haemoglobin (MHb).Further, described MHb combines with KCN and forms stable cyanohemoglobin.The concentration of haemoglobin can obtain by survey its uptake at the 540nm place with colourimetry.At K
3Fe (CN)
6To Fe in the haemoglobin
2+The basis of oxidation on, haemoglobin can detect by suitable electrode and detectable oxidizing potential.Yet there is the defective that causes high toxicity KCN pollution in said method.
Based on above narration, need a kind of advanced person's disposable, easy to use and economic electrode to be used for detection of biological molecule, particularly uric acid and haemoglobin.
An object of the present invention is to provide a kind of electrode band of disposable detection uric acid of non-enzyme, its by electrically insulating substrate at the bottom of, the conduction rete, electrical isolation rete and reaction rete are formed.
An object of the present invention is to provide a kind of electrode band of disposable detection haemoglobin, its by electrically insulating substrate at the bottom of, the conduction rete, electrical isolation rete and reaction rete are formed.
Another object of the present invention provides a kind of method disposable non-enzyme, that detect the electrode band of uric acid and haemoglobin of producing.
Another object of the present invention provides a kind of device that detects uric acid and haemoglobin.
Another object of the present invention provides a kind of method that detects the method for uric acid concentration in fluid sample and detect haemoglobin concentration in fluid sample.
The vertical view of Fig. 1 a. electrode band of the present invention.
The front view of Fig. 1 b. electrode band of the present invention.
Fig. 2 a. will conduct rete 2 and be applied at the bottom of the electrically insulating substrate step synoptic diagram on 1, and this coating comprises an anode and a negative electrode at least.
Fig. 2 a illustrates following step: will conduct rete 2 bags and be formed at least one anode and a negative electrode at the bottom of the electrically insulating substrate.
Fig. 2 b. covers the synoptic diagram of electric adiabatic rete step on conduction rete top.The conduction rete that is not capped comprises anode tie point 6, cathode connection 7, working electrode 8 and contrast electrode 9.
Fig. 2 c. will react rete 4 and be applied to conversion zone, the step synoptic diagram that working electrode and reaction electrode are capped.
Fig. 2 d. is applied to protective film the step synoptic diagram that reacts on the rete.
Haemocylolysis synoptic diagram on the reaction rete of the electrode band of Fig. 3 a. detection haemoglobin.
The current measurement uric acid of Fig. 3 b. by adjusting pick-up unit and the synoptic diagram of the concentration of haemoglobin.
Fig. 4. the method that combines with the EPAC biological analyser with electrode band of the present invention and uricase detects the synoptic diagram of the concentration of uric acid in blood respectively.
Fig. 5. detect the synoptic diagram of the concentration of haemoglobin in blood respectively with the method for electrode band of the present invention and cyanohemoglobin.
Fig. 6. with electrode band of the present invention and and centrifugation method detect hematocrit in the whole blood respectively.
Characteristics of the present invention be with contain the redox electronics water-soluble medium and surfactant modified after the disposable detection electrode band. Electrode band comes the concentration of biomolecule in the tracer liquid sample by catching the signal that is produced by biomolecule such as the oxidation current that contains the redox electronic media and uric acid and hemoglobins.
The electrode band that we are surprised to find after the low concentration cationic surfactant is modified has improved uric acid detection reaction speed and levels of precision. From another point of view, the electrode band after the high concentration neutral surface active agent modifies has improved speed and the accuracy of hemoglobin detection reaction. Electrode band
An object of the present invention is to provide electrode band a kind of non-enzyme, disposable detection uric acid, it is comprised of following part:
1) at the bottom of a kind of electrically insulating substrate;
2) a kind of conduction rete, this conduction rete is applied to a side of above-mentioned megohmite insulant, forms independently, disjunct anode and negative electrode;
3) be applied to a part of a kind of electrical isolation rete that conducts on the rete, wherein form a contrast electrode and another terminal anode connector that forms at least, negative electrode one terminal at least one working electrode and another the terminal cathode connector that forms of forming that be not coated with conducting stratum at anode one end that is not coated with conducting stratum;
4) a reaction rete, this reaction rete comprises that carrier, conductive medium and concentration are lower than 0.05% cationic surfactant, the zone that it was coated with comprises described working electrode and described contrast electrode at least, so that connect working electrode and contrast electrode separately, wherein said reaction rete does not comprise any enzyme.
The electrode band of another object of the present invention provides detection haemoglobin a kind of non-enzyme, disposable, it is by forming with the lower part:
1) at the bottom of a kind of electrically insulating substrate;
2) a kind ofly conduct the side that rete is applied to above-mentioned megohmite insulant, form independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a working electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) the reaction rete is higher than 1% neutral surface active agent by carrier, conductive medium and concentration and forms, the zone that it was coated with comprises described working electrode and described reference electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, how reaction rete described here does not comprise enzyme.
In the present invention, a flat surface and have the characteristic of insulation is arranged at the bottom of the electrically insulating substrate.The heat hardiness of this material is higher than 40 ℃, and preferred 40-80 ℃, its conduction can increase by thermal treatment.Any material with above-mentioned characteristic is suitable for as at the bottom of the electrically insulating substrate of the present invention.Preferably, these materials can be selected from following material: Polyvinylchloride (PVC), optical fiber glass (FR-4), polyester suphone, insulated electro plank, polyethylene terephthalate (PET) plate, polycarbonate (PC) plate, glass plate and ceramic wafer (CEM-1).
As described herein, the conduction rete is applied to a side of described megohmite insulant, forms independently disjunct anode and negative electrode.Preferably with marking ink or the coating of adherent metal mould sieve seal.More preferably, metallic diaphragm is selected from following metal: gold, silver, platinum and padium.Marking ink is selected from burnt black ink, golden ink, silver-colored ink or carbon element and silver-colored Heisui River potpourri, unstable graphite, copper ink or the potpourri of above-mentioned various inks (for example, first-selected using printed silver-colored ink, then with printing burnt black ink).Described negative electrode is covered by the electrical insulating film layer segment, and its end that is not capped forms contrast electrode and cathode connector respectively.Described anode is covered by the electrical insulating film layer segment, and its end that is not capped forms contrast electrode and anode connector respectively.The contrast electrode of negative electrode is reacted that rete covers and is found out the influence of electro-induction jointly with the working electrode of anode.Anode often links to each other with current sensor with cathode connector.
As described herein, the electrical isolation rete is applied at the bottom of the described electrically insulating substrate on the face, but does not cover above-mentioned cathode connector, anode connector, working electrode and contrast electrode.Do not comprised working electrode and contrast electrode, form conversion zone, coat the reaction rete then in this zone and be used for test sample by electrical isolation rete overlay area.Preferably the thickness of electrical isolation rete is greater than 0.6mm.
As described herein, the reaction rete is applied on the conversion zone by any suitable method, preferably by moving the method for liquid or sieve seal.The reaction rete reduces dielectric, carrier and surfactant by water soluble oxidized and forms.The reaction rete generation electrochemical reaction that contacts with sample.And the reaction rete can cover by protected rete.
As described herein, conductive medium comprises the electrolytic solution that redox ability is lower than detected sample.Become reducing condition from the state of oxidation behind conductive medium and the example reaction.Conductive medium may revert back to the state of oxidation again after being applied in certain coercive voltage.The change of the electromotive force that electrochemical reaction causes, resistance and electric current can be moved by the conduction rete, promptly from the reaction working electrode that links to each other of rete with to the connector of contrast electrode to anode and negative electrode.The preferred potassium ferricyanide of described conductive medium.Its consumption accounts for the 0.3%-5% of the weight ratio of reaction rete.
As described herein, because of sample is hydrophilic, difficult and hydrophobic conduction rete is approaching, thereby needs to absorb the carrier of sample.Carrier can increase the uptake of sample, thereby strengthens the signal that is migrated on the conduction rete.The reaction rete contacts with sample and electrochemical reaction takes place with it.Therefore, thus carrier can be spread all over entire reaction rete district can make signal all shift.Preferably, carrier is selected from crystallite shape cellulose, carboxymethyl cellulose or methylcellulose.More preferably less than the cellulose of 100um size.The weight ratio of cellulose comprises reaction rete approximately is 0.01%-0.1%.
As described herein, carrier can randomly comprise a polymkeric substance.This polymkeric substance helps reacting rete and has certain viscosity.Thereby the reaction rete can be distributed fully.Described polymkeric substance is selected from: polyvinyl alcohol (PVA) (PVA), polyvinylpyrrolidone (PVP), polyoxyethylene (PEG), gel and composition thereof.More preferably polymkeric substance accounts for the 0-15% of reaction rete weight.
As described herein, in the detection of uric acid, used surfactant is an anionic surface active agent in the reaction rete, and its concentration accounts for weight ratio will be less than 0.05%.Anionic surface active agent well known in the art all is suitable for the present invention.Preferably, the surfactant of described anionic comprises the surfactant that contains ammonium.More preferably, described surfactant is selected from N-acetyl-N, N, N-trimethyl-ammonium bromide and alkyl dimethyl ammonium halogenide.The preferred consumption of its anionic surfactant amount is 0.01%-0.03%.
From another point of view, as described herein, in the detection of haemoglobin, used surfactant is the neutral surface active agent in the reaction rete, and its concentration accounts for weight ratio and is greater than 1%.Arbitrary suitable neutral-surface activity well known in the art all can be used in the reaction rete of hemoglobin detecting electrode of the present invention.Preferably, described neutral surface active agent comprises: polyethylene oxide, oxirane, aliphatic alcohol, tertiary amine oxide and triphosphoric acid hydrogen oxide.More preferably, described surfactant is selected from trinitro-toluene and polyoxyethylene alkyl ether composition.Most preferably, described neutral surface active agent's concentration range is 1%-5%.
As described herein, the electrode band of the disposable detection haemoglobin of non-enzyme can be used for detecting the concentration of haemoglobin, and the ratio that well known to a person skilled in the art hematocrit/haemoglobin is 2.9-3.1.Therefore, hematocrit can be calculated by the concentration that the electrode band that utilizes detection haemoglobin of the present invention is surveyed haemoglobin.
The preparation of electrode band
Another object of the present invention provides a kind of method of electrode band of the detection uric acid (or haemoglobin) of producing disposable, non-enzyme, is made up of following steps:
A) conduct the side that rete is applied to above-mentioned megohmite insulant with a kind of, form independently, disjunct anode and negative electrode;
B) a kind of electric insulation coating layer is applied on the part conduction rete, the end that wherein said conducting stratum does not cover anode is that at least one contrast electrode and another end are an anode connector.The unlapped cathode end of described conducting stratum is that at least one working electrode and another end are a cathode connector.
C) comprising the regional top sieve seal or the dropwise reaction rete of described at least working electrode and contrast electrode, so that described working electrode is linked to each other separately with contrast electrode.
As described herein, at first, the conduction rete is applied to a side of flat megohmite insulant, thereby forms at least one anode separated from one another and negative electrode, after the coating, oven dry conduction rete under 35 ℃ of-80 ℃ of conditions.
As described herein, then, the electrical insulating film layer segment that thickness is equal to or greater than 0.6mm is printed on the conduction rete.The unmasked portion of described conduction rete forms an anode connector, a cathode connector, a working electrode and a contrast electrode.The zone that annular working electrode and contrast electrode surround or other arbitrary suitable zones that is shaped as the reaction rete.
As described herein, in the 3rd step, will react rete sieve and print or be added drop-wise on the conversion zone, thereby single working electrode is linked to each other with contrast electrode.Described zone can be annular or other arbitrary suitable shape.Multiple common sieve seal technology all can be used for the present invention.And a kind of new sieve seal technology has been open in 85,109,554 the R.O.C patent at application number by an inventor of the present invention, also this method can be used for preparation method of the present invention.
As described herein, in the 4th step, will react rete 40 ℃-80 ℃ further oven dry down.Protective film can randomly reacted on the rete or its dressing.The electrode band of the detection uric acid of disposable so non-enzyme is produced.
Pick-up unit
Another object of the present invention provides a kind of device that detects uric acid, and this device comprises the current sensor of uric acid in the electrode band of detection uric acid of a disposable non-enzyme and the direct analyzing liquid sample.
Wherein said electrode band comprises:
1) at the bottom of the electrically insulating substrate;
2) a conduction rete, this rete is applied to a side of above-mentioned megohmite insulant, forms independently, disjunct anode and negative electrode;
3) electric insulation coating layer, this electric insulation coating layer is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a working electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) a reaction rete, this rete comprises that a carrier, a conductive medium and concentration are lower than 0.05% cationic surfactant composition, the zone that it was coated with comprises described working electrode and described contrast electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, reaction rete described here does not comprise any enzyme.The cathode connector of the electrode band of described current sensor joint detection uric acid and anode connector, comprise a voltage output device, a signal receiver and a display device, wherein voltage output device can provide the voltage that is lower than 400mv to the reaction rete of uric acid detecting electrode bar, thereby behind reaction rete on this uric acid detecting electrode band and uric acid example reaction conductive medium is being oxidized to the state of oxidation from reducing condition; Described signal receiver is received in electric current, voltage or the changes in resistance that produces in the redox reaction process, and this is sent on the display device concentration of uric acid in the show sample.
Another object of the present invention provides a kind of device that detects haemoglobin, this device: the current sensor of haemoglobin in the electrode band of the detection uric acid of disposable non-enzyme and the direct analyzing liquid sample.
Described electrode band comprises
1) at the bottom of a kind of electrically insulating substrate;
2) a kind ofly conduct the side that rete is applied to above-mentioned megohmite insulant, form independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a working electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) the reaction rete is made up of greater than 0.05% neutral surface active agent carrier, conductive medium and concentration, the zone that it was coated with comprises described working electrode and described reference electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, reaction rete described here does not comprise any enzyme.The cathode connector of the electrode band of described current sensor joint detection haemoglobin and anode connector, it is made up of voltage output device, signal receiver and display device, and wherein voltage output device provides a voltage that is lower than 400mv.Behind reaction rete on the electrode band of this detection haemoglobin and the uric acid example reaction conductive medium is oxidized to the state of oxidation from reducing condition, signal receiver is received in electric current, voltage or the changes in resistance that produces in the redox reaction process, and this is sent on the display device concentration of haemoglobin in the show sample.
As described herein, testing conditions is that operating voltage is lower than 400mv and pH value between 5.0-10.0, can avoid the interference of other oxidisability composition.Detection method
Another object of the present invention provides the method for uric acid concentration in a kind of tracer liquid, this method comprises: with fluid drips on the disposable uric acid detecting electrode band of non-enzyme of the present invention, and the Control current sensor below 400mv low operating voltage and the pH value for operating in the scope of 7.0-10.0.
Another object of the present invention provides the method for hemoglobin concentration in a kind of tracer liquid, this method comprises: on the disposable uric acid detecting electrode band of non-enzyme of the present invention, and the low operating voltage below 400mv and pH value are current sensor of scope inner control of 5.0-8.0 with fluid drips.
Of the present invention, adopt electrochemical detection device can carry out the detection method of non-enzyme electrode band easily.Drip one respectively and drip off whole whole blood sample on the conversion zone of the electrode band of the disposable detection uric acid of non-enzyme of the present invention or haemoglobin, uric acid pick-up unit or haemoglobin pick-up unit can detect the redox reaction electric current of uric acid or haemoglobin respectively.This electrochemical reaction technology is generally used for detecting the galvanochemistry blood-sugar detecting instrument of blood sugar.It is 7.0-10.0 that the present invention adopts the redox electronic media to transmit the redox reaction information of uric acid and reaction conditions is controlled at the pH value, and the following low operating voltage of 400mv, to avoid the glucose in the blood and the interference of ascorbic acid vitamin c, this method that uric acid in the whole blood is directly detected is brand-new, is disclosed among the present invention and have first.Equally, the method that directly detects haemoglobin in the whole blood also is brand-new, and it is open first in the present invention, this method transmits the redox reaction information of haemoglobin and controls reaction pH value between 5.0-8.0 with the redox electronic media, the low operating voltage that 400mv is following is to avoid the glucose in the blood and the interference of ascorbic acid vitamin c.
Use
The present invention can be used for detecting biomolecule such as arbitrary fluid sample uric acid or haemoglobin.When adopting whole blood to detect uric acid or haemoglobin as sample, other composition in the blood particularly ascorbic acid vitamin c will produce interference.Pick-up unit of the present invention and disposable detection electrode band can directly detect uric acid or the haemoglobin in the conventional whole blood, suitable family expenses.
A significant advantage of the present invention is to have got rid of to use for example bioactivator of enzyme and so on.Therefore, the present invention has simplified production technology, has reduced production cost, has prolonged the holding time, has relaxed the preservation condition requirement of detecting electrode band.
Fig. 1 a and Fig. 1 b are respectively the vertical view and the front views of electrode band of the present invention.This electrode band by electrically insulating substrate at the bottom of 1, the conduction rete 2 on being coated at the bottom of the electrically insulating substrate is coated in the electrical isolation rete 3 on the conduction rete 2 and forms with the reaction rete 4 of example reaction.
At the bottom of the electrically insulating substrate of the present invention 1 with the conduction rete 2 link to each other.The negative electrode of conduction rete 2 is covered by electrical isolation rete 3 parts, and two end points that are not capped of negative electrode are respectively working electrode 8 and cathode connector 6.Reaction rete 4 has covered negative electrode working electrode 8, and working electrode 8 is used to detect the electrical effect of inducing of when electrochemical reaction of uric acid sample.Cathode connector 6 is used for being connected with current sensor.Anode is also covered by the electrical insulating film layer segment, and two unlapped end points of anode are respectively contrast electrode 9 and anode connector 7.The contrast electrode 9 of anode is covered by reaction rete 4, and induces electrical effect with the working electrode synergy detection of negative electrode.
The preparation of electrode band such as Fig. 2 a, 2b is shown in 2c and the 2d.As shown in Figure 2, at first adopt method such as sieve seal will conduct on the face that rete 2 is coated on plane material 1, form at least one anode and a negative electrode, their intervals independently of one another.
Shown in Fig. 2 b, the unlapped part of conduction rete forms cathode connector 6, anode connector 7, working electrode 8 and contrast electrode 9.The working electrode 8 and the contrast electrode 9 of annular have surrounded a zone.
Shown in Fig. 2 c and 2d, conductive medium sieve is imprinted on the border circular areas of above-mentioned reaction rete 4, and on described reaction rete border circular areas or randomly wrap by a protective film on every side.
Embodiment given below is used for describing and unrestricted the present invention.
It is 7.0-10.0 and the operating voltage that is lower than 400mv to avoid the interference of glucose and ascorbic acid in the blood that the detection uric acid of embodiment 1 biochemical substances and the detection of haemoglobin are controlled at PH.
The present invention finds that the vitamin c of normal concentration can not influence the detection of uric acid and haemoglobin.Therefore, biochemical substances can accurately be detected.What Fig. 3 a showed is the hemolytic reaction that causes on the hemoglobin detecting electrode band reaction rete.Fig. 3 b shows by the interference of adjusting electric current minimizing uric acid can amplify the signal of haemoglobin reaction.
The detection of embodiment 2 uric acid
On a plane of Polyvinylchloride (PVC) matrix, sieve seal burnt black ink forms conduction rete 2, and is comprising one group of independently disjunct anode and negative electrode, dry down at 40 ℃-80 ℃ then.The electrical isolation rete 3 parts sieve that with thickness is 0.6mm then is imprinted on the conduction rete 2.The part that the guided modes layer is not capped forms cathode connector 6, anode connector 7, working electrode 8 and contrast electrode 9.The border circular areas that working electrode 8 and contrast electrode 9 form is the scope of reaction rete 4.
To drop in the encircled of reaction rete 4 by the slurry like material that following ingredients and ratio are formed with suction pipe.
Methylcellulose 0.01%
CTAB (N-acetyl-N, N, N-trimethyl-amine bromide) 0.01%
H
2O?????????????????????????????????99.68％
The potassium ferricyanide 0.3%
Above-mentioned substance is dripped after reacting on the rete 4,35 ℃ of-80 ℃ of dryings.Protective film 5 is coated in reaction rete 4 top and on every side.Uric acid detecting electrode band by the disposable non-enzyme of above-mentioned steps has just prepared successfully.
The uric acid detecting electrode band of the disposable non-enzyme of the present invention that the employing said method obtains, the uric acid that detects in the whole blood needed for 20 seconds.Test findings and any conventional method detect coming to the same thing of uric acid.Fig. 4 is the comparing result of uric acid concentration during it was starched after uricase combined with the EPAC Biochemical Analyzer.
The result shows that disposable non-enzymuria acid detecting electrode band of the present invention can accurately detect the uric acid concentration in the blood, adopts whole blood, without any need for pre-treatment.
The detection of embodiment 3 uric acid
The step that repeats example 2 will be dropped in the encircled of reaction rete 4 by the slurry like material that following ingredients and ratio are formed with suction pipe.
Methylcellulose 0.05%
PEG polyglycol 15.00%
CTAB (N-acetyl-N, N, N-trimethyl-amine bromide) 0.03%
H
2O?????????????????????????????????83.092％
The potassium ferricyanide 1.00%
The uric acid detecting electrode band of the disposable non-enzyme of the present invention that the employing said method obtains, the uric acid that detects in the whole blood needed for 30 seconds.Test findings detects coming to the same thing of uric acid with the method that adopts the EPAC Biochemical Analyzer.The result shows that disposable non-enzymuria acid detecting electrode band of the present invention can accurately detect the uric acid concentration in the blood, adopts whole blood, without any need for pre-treatment.
The detection of embodiment 4 haemoglobins and hemolytic
The slurry like material that the step of repetition example 2 is made up of following ingredients and ratio drops in the encircled of reaction rete 4 with suction pipe:
Methylcellulose 0.05%
Triton 100 (Triton 10) 4.00%
H
2O????????????????????????????92.98％
The potassium ferricyanide 3.00%
The hemoglobin detecting electrode band of the disposable non-enzyme of the present invention that the employing said method obtains, the haemoglobin that detects in the whole blood needed for 30 seconds.As shown in Figure 5, test findings detects coming to the same thing of haemoglobin with the method that adopts cyaniding-haemoglobin.And the hematocrit of electrode band of the present invention detects and centrifugal testing result identical (as shown in Figure 6).
The result shows that disposable non-enzyme hemoglobin detecting electrode band of the present invention can accurately detect haemoglobin and the hematocrit in the blood, adopts whole blood, without any need for pre-treatment.
The detection of embodiment 5 haemoglobins and hemolytic
Repeat the step of example 3, difference is the composition of slurry like material and ratio are done following change:
Ultra micro optical fiber (the on average about 20um of diameter) 0.05%
PVC (polyvinyl alcohol (PVA)) 10.00%
Gel 3.50%
H
2O??????????????????????????????81.40％
The potassium ferricyanide 5.00%
The hemoglobin detecting electrode band of the disposable non-enzyme of the present invention that the employing said method obtains, the haemoglobin that detects in the whole blood needed for 80 seconds.As shown in Figure 5, test findings detects coming to the same thing of haemoglobin with the method that adopts cyaniding-haemoglobin.The result shows that disposable non-enzyme hemoglobin detecting electrode band of the present invention can accurately detect haemoglobin and the hematocrit in the blood, adopts whole blood, without any need for pre-treatment.
The foregoing description clearlys show that the electrode band of detection haemoglobin of the present invention is without any need for bioactive material, and production run is simple, quick.And the present invention is the hemoglobin concentration that can detect whole blood under the condition of 5.0-8.0 accurately at the low-work voltage that is lower than 400mv and pH value.
The detection of comparative example 6 uric acid
Adopt electrode band of the present invention and applicant at the 1999/04/21 patented claim USSN09/295 that submits to, the electrode band described in 400 detects uric acid respectively.The test findings of the sample of five variable concentrations is as shown in the table:
| The whole blood sample | ????1 | ????2 | ????3 | ????4 | ????5 | ?????6 | |
| The uricase method of standard | Mean value (mg/dl) | ???3.55 | ??5.11 | ??7.35 | ??9.10 | ??10.20 | ???11.85 |
| Electrode band of the present invention | Mean value (mg/dl) | ???3.60 | ??5.20 | ??7.10 | ??8.90 | ??10.40 | ???11.90 |
| ??％CV | ???3.80 | ??4.50 | ??3.40 | ??3.40 | ??5.40 | ???2.00 | |
| USSN09/295, the electrode band in 400 | Mean value (mg/dl) | ???3.50 | ??5.60 | ??6.80 | ??8.90 | ??10.50 | ???12.00 |
| ??％CV | ???12.00 | ??8.50 | ??7.30 | ??6.00 | ??4.00 | ???4.60 | |
As above shown in the table, the electrode band of the present invention and USSN 09/295,400 all has high precision.Yet, shown CV% aspect, electrode band of the present invention is more superior than the electrode strip of USSN 09/295,400.And, the detection time of electrode band shown in the present only be among the USSN 09/295,400 electrode strip 1/10.
Claims (39)
1. a non-enzyme, disposable uric acid detecting electrode band, this electrode band comprises:
1) at the bottom of a kind of electrically insulating substrate;
2) a kind of conduction rete, this rete is applied to a side of above-mentioned megohmite insulant, forms independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer, this coating is applied on the part of described conduction rete, wherein said conducting stratum does not cover terminal at least one contrast electrode and anode connector of another terminal formation of forming of anode, is not capped terminal at least one contrast electrode and cathode connector of another terminal formation of forming of negative electrode at conducting stratum;
4) a reaction rete, this rete is lower than 0.05% anionic surface active agent by carrier, conductive medium and concentration and forms, the zone that it was coated with comprises described working electrode and described reference electrode at least, so that working electrode is connected separately with contrast electrode, wherein said reaction rete does not comprise any enzyme.
2. the electrode band of the described detection uric acid of claim 1 has the flat surface and the characteristic of insulation at the bottom of the wherein said electrically insulating substrate.
3. the electrode band of the described detection uric acid of claim 1, the heat impedance at the bottom of the wherein said electrically insulating substrate is higher than 40 ℃, is preferably 40 ℃-80 ℃.
4. the electrode band of the described detection uric acid of claim 1, it is to make at the bottom of the wherein said electrically insulating substrate: Polyvinylchloride (PVC) by following substances, optical fiber glass (FR-4), polyester suphone, the insulated electro plank, polyethylene terephthalate (PET) plate, polycarbonate (PC) plate, glass plate or ceramic wafer (CEM-1).
5. the electrode band of the described detection uric acid of claim 1, wherein said reaction rete contacts with sample and electrochemical reaction takes place.
6. the electrode band of the described detection uric acid of claim 1, wherein said reaction rete adopt the mode of moving liquid or sieve seal to be applied to conversion zone.
7. the electrode band of the described detection uric acid of claim 1, wherein said reaction rete further covers with protective film.
8. the electrode band of the described detection uric acid of claim 1, wherein said anionic surface active agent is N-acetyl-N, N, N-trimethyl-amine bromide.
9. the electrode band of the described detection uric acid of claim 1, the weight ratio of the concentration of wherein said anionic surface active agent is 0.01%-0.03%.
10. the electrode band of the described detection uric acid of claim 1, wherein said carrier is selected from crystallite shape cellulose, carboxymethyl cellulose and methylcellulose and composition thereof.
11. the electrode band of the described detection uric acid of claim 1, the optional polymkeric substance that is selected from following substances that comprises of wherein said carrier: polyvinyl alcohol (PVA) (PVA), polyvinylpyrrolidone (PVP), polyglycol (PEG), gelatin and composition thereof.
12. the electrode band of the described detection uric acid of claim 1, wherein said carrier are the crystallite shape celluloses of big slight ground 100 μ m
13. the electrode band of the described detection uric acid of claim 1, the consumption of wherein said cellulose carrier account for about 0.01%-about 0.1% of reaction rete weight ratio.
14. the electrode band of the described detection uric acid of claim 11, the consumption of its polymkeric substance account for about 0%-15% of the weight ratio of reaction rete.
15. the electrode band of the described detection uric acid of claim 1, its conductive medium is K
3Fe (CN)
6
16. the electrode band of the described detection uric acid of claim 1, the consumption of its conductive medium account for the 0.3%-5% of the weight ratio of reaction rete.
17. the electrode band of a detection haemoglobin non-enzyme, disposable, it is by forming with the lower part:
1) at the bottom of a kind of electrically insulating substrate;
2) a kind ofly conduct the side that rete is applied to above-mentioned megohmite insulant, form independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a contrast electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) the reaction rete is lower than 0.05% anionic surface active agent by carrier, conductive medium and concentration and forms, the zone that it was coated with comprises described working electrode and described reference electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, reaction rete described here does not comprise any enzyme.
18. the electrode band of the described detection haemoglobin of claim 17 has the flat surface and the characteristic of insulation at the bottom of the wherein said electrically insulating substrate.
19. the electrode band of the described detection haemoglobin of claim 17, the heat impedance at the bottom of the wherein said electrically insulating substrate are higher than 40 ℃, preferred 40 ℃-80 ℃.
20. the electrode band of the described detection uric acid of claim 17, be by Polyvinylchloride (PVC) at the bottom of the wherein said electrically insulating substrate, optical fiber glass (FR-4), polyester suphone, the insulated electro plank, polyethylene terephthalate (PET) plate, polycarbonate (PC) plate, glass plate and ceramic wafer (CEM-1) are formed.
21. the electrode band of the described detection haemoglobin of claim 17, wherein said reaction rete contacts with sample and electrochemical reaction takes place.
22. the electrode band of the described detection haemoglobin of claim 17, wherein said reaction rete adopt the mode of moving liquid or sieve seal to be applied to conversion zone.
23. the electrode band of the described detection haemoglobin of claim 17, wherein said neutral surface active agent is triton (Triton).
24. the electrode band of the described detection haemoglobin of claim 17, the shared weight ratio of wherein said cationic surfactant is 1%-5%.
25. the electrode band of the described detection haemoglobin of claim 17, wherein said carrier are from crystallite shape cellulose, carboxymethyl cellulose and methylcellulose potpourri.
26. the electrode band of the described detection haemoglobin of claim 17, the optional polymkeric substance that is selected from following material that comprises of wherein said carrier: polyvinyl alcohol (PVA) (PVA), polyvinylpyrrolidone (PVP), polyoxyethylene (PEG), gelatin and composition thereof.
27. the electrode band of the described detection haemoglobin of claim 17, wherein carrier is the crystallite shape cellulose of size less than 100um.
28. the electrode band of the described detection haemoglobin of claim 17, the consumption of its cellulose carrier account for about 0.01%-0.1% of the weight ratio of reaction rete.
29. the electrode band of the described detection haemoglobin of claim 17, the consumption of wherein said polymkeric substance account for about 0%-15% of the weight ratio of reaction rete.
30. the electrode band of the described detection haemoglobin of claim 17, wherein said conductive medium is K
3Fe (CN)
6
31. the electrode band of the described detection haemoglobin of claim 17, the consumption of its conductive medium account for the 0.3%-5% of reaction rete weight ratio.
32. the electrode band of the described detection haemoglobin of claim 17 often is used in the detection of hematocrit.
33. the method for a production such as claim 1 or 17 described electrode bands, it is made up of following steps:
A) conduct the side that rete is applied to above-mentioned megohmite insulant with a kind of, form independently, disjunct anode and negative electrode;
B) a kind of electric insulation coating layer is applied on the part of conduction rete, wherein said conducting stratum does not cover terminal at least one contrast electrode and anode connector of another end formation of forming of anode, terminal at least one working electrode and cathode connector of another terminal formation of forming of unlapped formation negative electrode;
C) sieve seal or dropwise reaction rete in the zone that comprises working electrode and ginseng at least than electrode, so that working electrode is connected separately with contrast electrode.
34. a device that detects uric acid is by forming with the lower part: the current sensor of uric acid in the electrode band of the detection uric acid of disposable non-enzyme and the analyzing liquid sample;
Described electrode band is by forming with the lower part
1) at the bottom of a kind of electrically insulating substrate;
2) a kind of conduction rete, this rete is applied to a side of above-mentioned megohmite insulant, forms independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer, this coating is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a working electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) reaction rete, this rete is lower than 0.05% cationic surfactant by carrier, conductive medium and concentration and forms, the zone that it was coated with comprises described working electrode and described contrast electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, reaction rete described here does not comprise any enzyme.
The cathode connector of the electrode band of described current sensor joint detection uric acid and anode connector, it is made up of voltage output device, signal receiver and display device, wherein voltage output device can provide the voltage that is lower than 400mv to the reaction rete of described uric acid detecting electrode bar, thus make with the uric acid example reaction after conductive medium is oxidized to the state of oxidation from reducing condition; Signal receiver is received in electric current, voltage or the changes in resistance that produces in the redox reaction process, and this is sent on the display device concentration of uric acid in the show sample.
A kind of device that detects haemoglobin is by forming with the lower part: the current sensor of haemoglobin in the electrode band of the detection uric acid of disposable non-enzyme and the analyzing liquid sample.
Described electrode band is by forming with the lower part
1) at the bottom of a kind of electrically insulating substrate;
2) a kind ofly conduct the side that rete is applied to above-mentioned megohmite insulant, form independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a working electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) the reaction rete is made up of greater than 0.05% neutral surface active agent carrier, conductive medium and concentration, the zone that it was coated with comprises described working electrode and described reference electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, reaction rete described here does not comprise any enzyme.
The cathode connector of the electrode band of described current sensor joint detection haemoglobin and anode connector, it is made up of voltage output device, signal receiver and display device, and wherein voltage output device provides a voltage that is lower than 400mv.Behind reaction rete on the electrode band of this detection haemoglobin and the uric acid example reaction conductive medium is oxidized to the state of oxidation from reducing condition, signal receiver is received in electric current, voltage or the changes in resistance that produces in the redox reaction process, and this is sent on the display device concentration of haemoglobin in the show sample.
35. one kind detect haemoglobin by forming with the lower part: the current sensor of haemoglobin in the electrode band of the detection haemoglobin of disposable non-enzyme and the analyzing liquid sample;
Described electrode band is by forming with the lower part
1) at the bottom of a kind of electrically insulating substrate;
2) a kind ofly conduct the side that rete is applied to above-mentioned megohmite insulant, form independently, disjunct anode and negative electrode;
3) a kind of electric insulation coating layer is applied on the part conduction rete, at least comprise the end of a contrast electrode and another anode connector in the anode end that is not coated with conducting stratum, comprise the end of a working electrode and another cathode connector in the cathode end that is not coated with conducting stratum at least;
4) the reaction rete is lower than 0.05% cationic surfactant by carrier, conductive medium and concentration and forms, the zone that it was coated with comprises described working electrode and described contrast electrode at least, that is to say that being connected of working electrode and contrast electrode is distinct, reaction rete described here does not comprise any enzyme.
The cathode connector of the electrode band of described current sensor joint detection haemoglobin and anode connector, it is made up of voltage output device, signal receiver and display device, wherein voltage output device can provide the voltage that is lower than 400mv, behind reaction rete on the electrode band of this detection uric acid and the haemoglobin example reaction conductive medium is oxidized to the state of oxidation from reducing condition, signal receiver is received in electric current, voltage or the changes in resistance that produces in the redox reaction process, and this is sent on the display device concentration of haemoglobin in the show sample.
36. as claim 39 or 40 described devices, its detecting electrode band is disposable.
37. as claim 39 or 40 described devices, wherein said test sample is a blood.
38. the method for uric acid concentration in the tracer liquid, comprising following step: the liquid sample is dripped on electrode band as claimed in claim 1, and the Control current sensor is being lower than the low-work voltage of 400mv, and the pH value is operated under the condition of 7.0-10.0.
39. the method for hemoglobin concentration in the tracer liquid, comprising following step: the liquid sample is dripped on electrode band as claimed in claim 17, and the Control current sensor is being lower than the low-work voltage of 400mv, and the pH value is operated under the condition of 5.0-8.0.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01116298 CN1367382A (en) | 2001-01-23 | 2001-01-23 | Non-enzymic disposable detection electrode bar, its production method and its application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01116298 CN1367382A (en) | 2001-01-23 | 2001-01-23 | Non-enzymic disposable detection electrode bar, its production method and its application |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7384659B2 (en) * | 2002-03-29 | 2008-06-10 | Apex Biotechnology Corporation | Method of producing whole blood detecting electrode strip and reaction film formulation and the related products |
| CN1952653B (en) * | 2006-10-01 | 2010-08-11 | 中南大学 | Disposable whole blood uric acid testing electrode indicator bar and manufacturing method thereof |
| CN101191781B (en) * | 2006-11-29 | 2012-03-21 | 合世生医科技股份有限公司 | Non- ferment type uric acid reagent sensing device, sensing test strip and method for making same |
| CN101726520B (en) * | 2008-10-17 | 2014-04-16 | 五鼎生物技术股份有限公司 | Quality control liquid for non-enzyme type electrochemical detection system and usage thereof |
| CN105136885A (en) * | 2015-09-21 | 2015-12-09 | 南京明茂英华生物科技有限公司 | Preparation and application of Prussian blue/N-doped carbon nanometer composite material |
| CN105877694A (en) * | 2014-08-30 | 2016-08-24 | 孙宗正 | Tissue fluid constituent sensor |
| CN106483182A (en) * | 2016-08-22 | 2017-03-08 | 浙江大学 | A kind of dry type determines biology sensor of hemoglobin and preparation method thereof |
| CN107405115A (en) * | 2014-09-08 | 2017-11-28 | 印度科学研究所 | Apparatus and method for detecting hemoglobin and its compound |
| CN113017995A (en) * | 2021-03-03 | 2021-06-25 | 杭州可靠护理用品股份有限公司 | Adult diaper with uric acid prompting function |
-
2001
- 2001-01-23 CN CN 01116298 patent/CN1367382A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7384659B2 (en) * | 2002-03-29 | 2008-06-10 | Apex Biotechnology Corporation | Method of producing whole blood detecting electrode strip and reaction film formulation and the related products |
| US8512532B2 (en) | 2002-03-29 | 2013-08-20 | Apex Biotechnology Corporation | Method of producing whole blood detecting electrode strip and reaction film formulation and the related products |
| CN1952653B (en) * | 2006-10-01 | 2010-08-11 | 中南大学 | Disposable whole blood uric acid testing electrode indicator bar and manufacturing method thereof |
| CN101191781B (en) * | 2006-11-29 | 2012-03-21 | 合世生医科技股份有限公司 | Non- ferment type uric acid reagent sensing device, sensing test strip and method for making same |
| CN101726520B (en) * | 2008-10-17 | 2014-04-16 | 五鼎生物技术股份有限公司 | Quality control liquid for non-enzyme type electrochemical detection system and usage thereof |
| CN105877694A (en) * | 2014-08-30 | 2016-08-24 | 孙宗正 | Tissue fluid constituent sensor |
| CN107405115A (en) * | 2014-09-08 | 2017-11-28 | 印度科学研究所 | Apparatus and method for detecting hemoglobin and its compound |
| CN105136885A (en) * | 2015-09-21 | 2015-12-09 | 南京明茂英华生物科技有限公司 | Preparation and application of Prussian blue/N-doped carbon nanometer composite material |
| CN105136885B (en) * | 2015-09-21 | 2017-12-22 | 南京明茂英华生物科技有限公司 | A kind of preparation and application of Prussian blue/N doping carbon nano-composite materials |
| CN106483182A (en) * | 2016-08-22 | 2017-03-08 | 浙江大学 | A kind of dry type determines biology sensor of hemoglobin and preparation method thereof |
| CN113017995A (en) * | 2021-03-03 | 2021-06-25 | 杭州可靠护理用品股份有限公司 | Adult diaper with uric acid prompting function |
| CN113017995B (en) * | 2021-03-03 | 2022-01-04 | 杭州可靠护理用品股份有限公司 | Adult diaper with uric acid prompting function |
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