CN109085218A - A kind of reagent and electrochemical sensor - Google Patents

A kind of reagent and electrochemical sensor Download PDF

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Publication number
CN109085218A
CN109085218A CN201810063224.5A CN201810063224A CN109085218A CN 109085218 A CN109085218 A CN 109085218A CN 201810063224 A CN201810063224 A CN 201810063224A CN 109085218 A CN109085218 A CN 109085218A
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China
Prior art keywords
urine
reagent
glucose
electrode
buffer
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CN201810063224.5A
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Chinese (zh)
Inventor
王文冲
徐晨龙
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Shanghai Desert Island Technology Co Ltd
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Shanghai Desert Island Technology Co Ltd
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Priority to CN201810063224.5A priority Critical patent/CN109085218A/en
Publication of CN109085218A publication Critical patent/CN109085218A/en
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28Electrolytic cell components
    • G01N27/30Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • G01N27/3272Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N9/00Investigating density or specific gravity of materials; Analysing materials by determining density or specific gravity
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N9/00Investigating density or specific gravity of materials; Analysing materials by determining density or specific gravity
    • G01N9/36Analysing materials by measuring the density or specific gravity, e.g. determining quantity of moisture

Abstract

The present invention provides a kind of reagent and electrochemical sensors, specifically, providing a kind of reagent for detecting urine specific gravity of urine comprising: the electron mediator of 2~10wt%;The stabilizer of 1~5wt%;The anti-agglomerating agent of 0.5~2wt%;The surfactant of 0.1~0.5wt%;Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1;A kind of reagent for detecting urine glucose is additionally provided, the glucose oxidase or glucose dehydrogenase of 0.5~5wt% are increased on the basis of the reagent of above-mentioned detection urine specific gravity of urine.The application passes through the content selected the component of special ratios and control each component, and reagent is made to be respectively used to detection specific gravity of urine or urine glucose accuracy with higher.

Description

A kind of reagent and electrochemical sensor
Technical field
The present invention relates to urine detection technical field more particularly to a kind of reagents and electrochemical sensor.
Background technique
Routine urinalysis is a basic conventional project of disease detection.Specific gravity of urine and the detections such as glucose in urine in routine urinalysis are normal The inspection item of rule.Example is detected as below with glucose in urine to be illustrated.
Glucose in urine (urine glucose) inspection is the early diagnosis simplest method of diabetes, and blood sugar concentration is more than after renal glucose threshold It will appear glucose in urine, blood sugar concentration is higher, and urine sugar concentrations are higher.Glucose in urine detection is as a kind of simple and noninvasive detection method, to sugar Urinating sick patient is a kind of very favorable method.
Clinical Urinary sugar detection is mostly used urine test paper at present, and urine test paper is by glucose in urine and enzyme reaction on test paper and makes to contaminate Expect reagent discoloration, the variation of color reflects concentration of glucose, but its precision is low, belongs to sxemiquantitative test.In addition, there are also use electrification Test strips are learned to test the scheme of urine sugar concentrations, but actually rare on the market using electrochemical test strip test glucose in urine, it is main former Because being that substance is complicated in urine, to a large amount of electrolyte in glucose in urine test serious interference, such as ascorbic acid, high specific gravity of urine urine Ion, metal ion can test glucose in urine to Electrochemical enzyme biosensor and generate bigger interference, cause test result inaccurate. When being tested using electrochemical detection test strip, measurement accuracy can be preferably such as improved using suitable reagent.
Summary of the invention
Present invention solves the technical problem that being the provision of a kind of reagent for detecting urine specific gravity of urine and glucose, the application The reagent detection urine specific gravity of urine of offer or glucose accuracy are higher.
In view of this, this application provides a kind of reagents for detecting urine specific gravity of urine, comprising:
Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1.
Present invention also provides a kind of reagents for detecting urine glucose, comprising:
Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1.
Preferably, the electron mediator is the iron cyanide, ferrocyanide, ferrocene or derivatives thereof;The stabilization Agent is selected from one or both of trehalose and chitosan;The anti-agglomerating agent in hydrophilic silicon dioxide and xanthan gum one Kind or two kinds;The pH buffer is selected from one of phosphate buffer, MES buffer and citrate buffer solution or a variety of; The surfactant is selected from Tween-20, Tween-80 and Qula logical one of -100 or a variety of.
Preferably, the content of the electron mediator is 4~7wt%.
Preferably, the content of the glucose oxidase or glucose dehydrogenase is 0.8~3.2wt%.
Present invention also provides a kind of electrochemical sensor for urine detection, including substrate, electrode and reagent layer, institutes Stating the corresponding reagent of reagent layer is reagent described in above scheme.
Preferably, the electrochemical sensor further includes insulating film, hydrophilic film, adhesive layer, leads sample slot and gas outlet;
Multiple electrodes are provided in substrate;
At least one electrode surface is provided with insulating film, and the insulating film offers opening, and reagent layer is set to described open In mouthful;
Adhesive layer and hydrophilic film are being disposed with along far from the insulating film surface on the substrate direction;
The adhesive layer, which offers, leads sample slot, and the hydrophilic film offers gas outlet, described to lead some or all of sample slot Reagent layer described in face leads sample slot described in the gas outlet face.
Preferably, the electrode includes working electrode with to electrode, and the working electrode is L-type, and described is L to electrode Type;The end L of the working electrode and the same end that substrate is all set in the end L of electrode.
Preferably, the opening is multiple, and the reagent layer is set to proper pair of L of an opening of the working electrode In.
Present invention also provides a kind of kits for urine detection, including being used for urine detection described in above scheme Reagent or the reagent layer that is formed of the reagent as described in above scheme.
This application provides it is a kind of detect urine specific gravity of urine reagent, due to it include electron mediator, surfactant, Stabilizer, anti-agglomerating agent, buffer and water, and by the content of control mentioned component, so that it is applied to test urine specific gravity of urine With preferable accuracy;It further include Portugal in reagent meanwhile present invention also provides a kind of reagent for detecting urine glucose Grape carbohydrate oxidase or glucose dehydrogenase, by with said components electron mediator, surfactant, stabilizer, anti-agglomeration Agent, buffer and water are used cooperatively, and so that it is applied to test urine glucose has preferable accuracy.
Detailed description of the invention
Fig. 1 is the fractionation structural representation of electrochemical sensor in one embodiment of the invention;
Fig. 2 is the structural schematic diagram of electrochemical sensor in one embodiment of the invention;
Fig. 3 is that the urine specific gravity of urine that embodiment 2 provides and electrochemical sensor test electric current and be fitted Line Chart;
Fig. 4 is that the urine specific gravity of urine that embodiment 3 provides and electrochemical sensor test electric current and be fitted Line Chart;
Fig. 5 is glucose in urine testing background electric current and no enzyme electrode current relationship curve;
Fig. 6 is glucose in urine measurement sensitivity and no enzyme electrode current relationship;
Fig. 7 is that embodiment 4 detects urine sugar concentrations and compound concentration contrast curve chart;
Fig. 8 is that embodiment 5 detects urine sugar concentrations and compound concentration contrast curve chart;
Fig. 9 is the fractionation structural representation for the electrochemical sensor that the embodiment of the present invention 6 uses;
Figure 10 is that the embodiment of the present invention 6 detects urine sugar concentrations and compound concentration contrast curve chart.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described below with reference to embodiment, still It should be appreciated that these descriptions are only further explanation the features and advantages of the present invention, rather than to the claims in the present invention Limitation.
For electrochemical sensor in addition to that can be used to test in urine other than glucose, present inventor passes through experiment hair It is existing: to be the small molecules such as urea and sodium, potassium plasma, electrochemical sensor due to influencing the main substance of specific gravity of urine in urine It can also be used to measurement specific gravity of urine.Usual electrochemical sensor needs to use reagent, and to this, the embodiment of the invention provides one The reagent and a kind of reagent for detecting urine glucose of kind detection urine specific gravity of urine.It is pointed out that those skilled in the art Sometimes also electrochemical sensor can be known as electrochemical detection test strip.
The embodiment of the invention provides the reagents of a kind of reagent for detecting urine specific gravity of urine and detection urine glucose, utilize The reagent can realize the detection of urine specific gravity of urine and urine glucose respectively, and accuracy is higher.Wherein, the embodiment of the present invention A kind of reagent for detecting urine specific gravity of urine is disclosed first, comprising:
Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1.
In another embodiment, the application provides a kind of reagent for detecting urine glucose simultaneously, comprising:
Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1.
In above-described embodiment, for the identical component of detection urine specific gravity of urine and detection urine glucose, including electronics matchmaker Mediator, stabilizer, anti-agglomerating agent, surfactant, pH buffer and water, specifically, the electron mediator be the iron cyanide, Ferrocyanide, ferrocene or derivatives thereof, in a particular embodiment, the electron mediator are the iron cyanide;The electronics The content of mediator is 2~10wt%, and in a particular embodiment, the content of the electron mediator is 4~7wt%;It is directly Influence reaction process electron-transport, detection range, response sensitivity and size of current.
The stabilizer is one or both of trehalose and chitosan, and in a particular embodiment, the stabilizer is Trehalose;Its content is 1~5wt%, and in a particular embodiment, the content of the stabilizer is 1.5~3.5wt%.
The anti-agglomerating agent is selected from one or both of hydrophilic silicon dioxide and xanthan gum, in a particular embodiment, institute Stating anti-agglomerating agent is hydrophilic silicon dioxide;Its content is 0.5~2wt%, in a particular embodiment, the content of the anti-agglomerating agent For 0.8~1.8wt%.The anti-agglomerating agent is used to prevent the agglomeration of reagent layer.
The surfactant is selected from Tween-20, Tween-80 and Qula logical one of -100 or a variety of, specific real It applies in example, the surfactant is Tween-20;Its content is 0.1~0.5wt%.The surfactant make aqueous solution with Hydrophobic electrode can contact well.
The pH buffer is selected from one of phosphate buffer, MES buffer and citrate buffer solution or a variety of, In specific embodiment, the pH buffer is phosphate buffer;The pH buffer changes for buffer reagent pH.It is detecting In the reagent of urine specific gravity of urine, surplus is pH buffer and water, and the mass ratio of the pH buffer and water is 1:1.
It further include glucose oxidase or glucose dehydrogenase, content 0.5 in the reagent of detection urine glucose ~5wt%, in a particular embodiment, the content of the glucose oxidase or glucose dehydrogenase are 0.8~2wt%.It is above-mentioned The content of glucose oxidase or glucose dehydrogenase will affect glucose in urine detection range and response current size.
During actually detected urine specific gravity of urine or urine glucose, mentioned reagent is not to be used alone, and is needed Cooperate with sensor, is just able to achieve the detection of specific gravity of urine or glucose in urine.One kind is additionally provided in one embodiment of the application as a result, Electrochemical sensor comprising substrate, electrode and reagent layer, the corresponding reagent of the reagent layer are examination described in above scheme Agent.
More specifically, during detecting urine specific gravity of urine, it is only necessary to specific gravity of urine can be realized using a sensor Detection, the corresponding reagent of the reagent layer of the sensor include electron mediator described in above scheme, stabilizer, anti-agglomerating agent, Surfactant, pH buffer and water, can be referred to as no enzyme sensor.
It is found by the applicant that: since different people or same people are in the differences such as different moments amount of drinking water and dietary intake, urine urine Specific gravity variation is obvious, and glucose detection is influenced very greatly by specific gravity of urine, if therefore only with a sensor containing enzymatic reagent The accuracy that is bound to of measurement glucose is not high, because sensor background electric current and sensitivity change with urine specific gravity of urine, to protect Measurement accuracy is demonstrate,proved, when detecting urine glucose, two kinds of sensors are used in the embodiment of the present application, one kind is that above-mentioned detection is urinated For the sensor-of specific gravity without enzyme sensor, a kind of sensor to detect glucose (contains glucose oxidase or grape in reagent Glucocorticoid dehydrogenase) thus-enzyme sensor according to the relationship of specific gravity of urine and glucose obtains the concentration of glucose.
When detecting urine specific gravity of urine or detection urine glucose, the sensor used can also be those skilled in the art Well known sensor is not particularly limited this application.
More specifically, this application provides a kind of specific sensor, structural schematic diagram is as shown in Figure 1 or 2, in figure 1 be substrate, 2 be working electrode, 3 be trigger electrode to electrode, 4,5 be insulating film, 6 be reagent layer, 7 be hydrophilic film, 8 be viscous Layer, 9 are closed to lead sample slot, 10 being gas outlet, specifically, the sensor include substrate, it is electrode, insulating film, reagent layer, hydrophilic Film, adhesive layer lead sample slot and gas outlet;Specifically,
Multiple electrodes are provided in substrate;
At least one electrode surface is provided with insulating film, and the insulating film offers opening, and reagent layer is set to described open In mouthful;
Adhesive layer and hydrophilic film are being disposed with along far from the insulating film surface on the substrate direction;
The adhesive layer, which offers, leads sample slot, and the hydrophilic film offers gas outlet, described to lead some or all of sample slot Reagent layer described in face leads sample slot described in the gas outlet face.
Shape the application of the electrode is without particularly limiting, according to shape well known to those skilled in the art It can.In a specific embodiment, the electrode includes trigger electrode, working electrode with to electrode, and the working electrode is L Type, described is L-type to electrode;The end L of the working electrode and one end that neighbouring reagent layer is all set in the end L of electrode, institute Trigger electrode is stated to be set in the working electrode and the space for forming electrode.
Insulating film is set on the surface at the end L of the working electrode, for limiting the area of reagent layer.It can be in insulating film Multiple openings, such as two or three openings are set, a reagent layer is provided in the opening at the end L for closing on working electrode. If opening is two, the end L of one of opening face working electrode, another opening face is to the end electrode L;If opening is three A, the end L of one of opening face working electrode, other two opening difference face is to the end L of electrode, the T of trigger electrode End.The hydrophilic film, the adhesive layer are and insulating film is in the same side.The hydrophilic film is complete PET film, and itself and bonding Layer forms one and leads sample slot, i.e. adhesive layer offers one and leads sample slot, leads hydrophilic film within the scope of sample slot at this and offers gas outlet, makees For the gas outlet of siphon sample introduction.Hydrophilic film or the length of adhesive layer can be equal, for example, the one third of trigger electrode length Or half, guarantee three electrodes some be located at the underface of adhesive layer and hydrophilic film.Liquid sample such as urine From leading after sample slot enters sensor, and electrode, trigger electrode and reagent layer are in contact.In this application, trigger electrode The end T represent T-type top end, working electrode represents one end that L-type has bending with to the end L of electrode.
During actually detected urine specific gravity of urine, urine is dropped in the sensor containing reagent layer, while to biography Sensor applies a voltage such as+0.4V voltage, measures after a period of time response current when such as 30s, can according to response electric current and Corresponding relationship between specific gravity of urine directly obtains specific gravity of urine value.When specific implementation, it can be realized by a measuring instrument, or It is realized by the test circuit and related testing tool of setting.
For measuring instrument, the corresponding relationship formula being previously provided between urine specific gravity of urine and response current, one In a embodiment, the first relational expression of urine specific gravity of urine and response current be may be expressed as:
Y=A* | X |+B;
Wherein Y is urine specific gravity of urine, | X | the examination for the absolute value of response current value, in the occurrence and reagent layer of A and B Agent is related.In a specific embodiment, the corresponding reagent of reagent layer by weight percentage, the wherein potassium ferricyanide 6.45%, Trehalose 1.8%, hydrophilic silicon dioxide 0.9%, Tween-20 0.45%, phosphate buffer solution 45.2%, water 45.2%, thus The first obtained relational expression are as follows: Y=0.00148*X+0.99922;
In a specific embodiment, by weight percentage, wherein the potassium ferricyanide 4%, extra large for the corresponding reagent of reagent layer Thus algae sugar 3%, hydrophilic silicon dioxide 1.5%, Tween-20 0.3%, phosphate buffer solution 45.6%, water 45.6% obtain The first relational expression are as follows: Y=0.00183*X+1.00162.It will be understood by those skilled in the art that above are only specific example, if Its weight percent is accordingly adjusted within the scope of each component, coefficient A and B also can accordingly change.
Therefore, the first relational expression that urine specific gravity of urine and response current can be preset in test equipment, passes through electrification It learns sensor test urine and obtains electric current (when+0.4V) when 30s, directly obtain urine specific gravity of urine, such as straight at measuring instrument end Connect output urine specific gravity of urine.
In practical application, urine, which drops to first sensor, (for convenience of description, measures the sensor of specific gravity of urine in the application Referred to as first sensor or without enzyme sensor, the scales for measuring glucose in urine are second sensor or enzyme sensor), measuring instrument Measurement instrument port can directly obtain specific gravity of urine value after applying voltage.
During actually detected urine glucose, urine is added to the first sensor containing reagent layer and respectively In two sensors, and from measuring instrument into two sensors application+0.4V voltage, when measuring instrument measures 30s first sensing The first response current (no enzyme electrode electric current) of device and the second response current (enzyme electrode electric current) of second sensor, measuring instrument End directly exports urine glucose according to the corresponding relationship between urine glucose value and the first response current, the second response current Value.
It is specific: to be obtained after being corrected using no enzyme electrode current value to enzyme electrode urine glucose test job curve One working curve, working curve formula (the second relational expression) are as follows: I=(A*i+B) * Cglu+ (C*i+D), I=enzyme electrode electricity Stream, for i=without enzyme electrode electric current, Cglu is urine glucose value, the value and first sensor of coefficient A, B, C and D, the second sensing The reagent of reagent layer is related in device, i.e., the concrete composition of reagent has determined in reagent layer, and above-mentioned coefficient A, B, C and D's specifically takes Value is also just fixed, the occurrence of the concrete composition of each reagent corresponding specific A, B, C and a D.Thus it may make survey The concentration of glucose of examination is more acurrate;As shown in Figure 5 and Figure 6, Fig. 5 is that glucose in urine testing background electric current and no enzyme electrode current relationship are bent Line, Fig. 6 are glucose in urine measurement sensitivity and no enzyme electrode current relationship.
In a specific embodiment, the corresponding reagent of the reagent layer of no enzyme sensor by weight percentage, wherein iron Potassium cyanide 6.45%, trehalose 1.8%, hydrophilic silicon dioxide 0.9%, Tween-20 0.45%, phosphate buffer solution 45.2%, Water 45.2%, the corresponding reagent of the reagent layer of enzyme sensor by weight percentage, wherein glucose oxidase 0.9%, iron cyanogen Change potassium 6.3%, trehalose 1.8%, hydrophilic silicon dioxide 0.9%, Tween-20 0.45%, phosphate buffer solution 44.825%, Water 44.825%, thus obtained second relational expression are as follows: I=(- 0.02241*i+0.82955) * Cglu+ (0.84547*i- 0.18419);
In a specific embodiment, the corresponding reagent of the reagent layer of no enzyme sensor by weight percentage, wherein iron Potassium cyanide 5.4%, trehalose 1.8%, hydrophilic silicon dioxide 1.8%, Tween-20 0.9%, phosphate buffer solution 45.05%, Water 45.05%, the corresponding reagent of the reagent layer of enzyme sensor by weight percentage, wherein glucose oxidase 1.8%, iron cyanogen Change potassium 5.3%, trehalose 1.8%, hydrophilic silicon dioxide 1.8%, Tween-20 0.9%, phosphate buffer solution 44.2%, water 44.2%, thus obtained second relational expression are as follows: I=(- 0.02852*i+0.67899) * Cglu+ (0.85354*i- 0.18966)。
It will be understood by those skilled in the art that above are only specific example, if accordingly adjusting its weight within the scope of each component Percentage, coefficient A, B, C and D also can accordingly change.
During atual detection, the second relational expression is previously provided in test equipment (to be set according to reagent concrete composition Set), after being respectively dropped into same urine sample in enzyme sensor and no enzyme sensor, in measuring instrument to enzyme sensor and nothing After enzyme sensor applies voltage, urine dextrose equivalent directly can be exported at measuring instrument end.
In the reagent of detection urine specific gravity of urine provided by the present application, due to including electron mediator, surfactant, stabilization Agent, anti-agglomerating agent, buffer and water, and by the content of control mentioned component, so that it is applied to test urine specific gravity of urine has Preferable accuracy;Equally, in the reagent of detection urine glucose, due to including glucose oxidase or glucose dehydrogenation Enzyme, electron mediator, surfactant, stabilizer, anti-agglomerating agent, buffer and water, and the content by controlling mentioned component, So that it is applied to test urine glucose has preferable accuracy.
For a further understanding of the present invention, below with reference to embodiment to detection urine specific gravity of urine provided by the invention and detection Urine glucose is described in detail, and protection scope of the present invention is not limited by the following examples.
Embodiment 1
As depicted in figs. 1 and 2, Fig. 1 and Fig. 2 is a kind of electrochemical sensor used in the embodiment of the present invention, and 1 is base in figure Material, 2 be working electrode, 3 be trigger electrode to electrode, 4,5 be insulating film, 6 be reagent layer, 7 be hydrophilic film, 8 be adhesive layer Sample slot is is led, 10 is gas outlet in (such as layers of two-sided), 9;
The surface screen-printed of substrate goes out electrode shape, is followed successively by L-type working electrode 2, L-type and triggers electricity to electrode 3 and T-type Pole 4, trigger electrode 4 are set in working electrode 2 and the space formed to electrode 3;
The end side the L surface of working electrode 2 is provided with insulating film 5, and insulating film 5 offers an opening, and reagent layer 6 is set to In opening;Preferably, the surrounding of reagent layer 6 is fitted closely with insulating film 5.
Adhesive layer 8 and hydrophilic film 7 are being disposed with along far from 1 direction upper nonconductive Film of substrate, 5 surface;
Bonding glue-line 8, which offers, leads sample slot 9, and hydrophilic film 7 offers gas outlet 10, leads some or all of sample slot 9 face Reagent layer 6 leads sample slot 9 described in 10 face of gas outlet.
Embodiment 2
Urine specific gravity of urine is detected, agent formulations such as 1 institute of table using the sensor of embodiment 1, in sensor in reagent layer Show;
The reagent data table of 1 embodiment of table, 2 reagent layer
Substance Mass ratio
The potassium ferricyanide 6.45%
Trehalose 1.8%
Hydrophilic silicon dioxide 0.9%
Tween-20 0.45%
Phosphate buffer solution 45.2%
Water 45.2%
10 parts of urine sample are taken, respectively number 1-10, using measuring instrument to sensor application+0.4V voltage, tests 30s response Electric current, measurement instrument side directly obtain specific gravity of urine, on the basis of mentioned reagent composition, the calculating of specific gravity of urine and electrode current Formula is Y=0.00148*X+0.99922.Fig. 3 is that urine specific gravity of urine and electrochemical sensor test electric current and be fitted Line Chart.The public affairs Formula is pre-set inside measuring instrument, and when measuring, after obtaining response current, measuring instrument can directly exist after calculating Output end exports specific gravity of urine value.Every part of urine sample specific gravity of urine is tested using refractometer simultaneously, by what is be calculated by electrode current It calculates specific gravity of urine and is compared with the specific gravity of urine tested using refractometer, error is calculated, the results are shown in Table 2.
2 the present embodiment of table obtains specific gravity of urine and refractometer tests specific gravity of urine error information table
Test result (the urine test paper specific gravity of urine neighboring gradient phase that specific gravity of urine result is better than urine test paper is calculated using electrode current It is poor 0.005), compared with the test result of more accurate refractometer, as a result also have very high comparativity.It is provided in this embodiment Reagent can guarantee to obtain preferable accuracy when testing specific gravity of urine.
Embodiment 3
The process for testing specific gravity of urine is same as Example 2, and difference is that the reagent composition of reagent layer is different, as shown in table 3; Test result is as shown in table 4 with comparison result;Fig. 4 is urine specific gravity of urine provided in this embodiment and electrochemical sensor test electricity Stream fitting Line Chart;
The reagent data table of 3 embodiment of table, 3 reagent layer
4 the present embodiment of table obtains specific gravity of urine and refractometer tests specific gravity of urine error information table
Embodiment 4
It provides without enzyme sensor and enzyme sensor, wherein being the biography that embodiment 1 provides without enzyme sensor and enzyme sensor Sensor, the reagent in reagent layer in no enzyme sensor is as shown in table 5, such as 6 institute of table of the reagent in reagent layer in enzyme sensor Show;
Composition table of the table 5 without reagent in enzyme sensor reagent layer
Substance Mass ratio
The potassium ferricyanide 6.45%
Trehalose 1.8%
Hydrophilic silicon dioxide 0.9%
Tween-20 0.45%
Phosphate buffer solution 45.2%
Water 45.2%
The composition table of reagent in 6 enzyme sensor reagent layer of table
The urine various concentration value of three parts of different specifics gravity of urine is taken to pass through survey using above-mentioned no enzyme sensor and enzyme sensor Test instrument is respectively to two sensor application+0.4V voltages, the first response current and the second response current when measuring 30s, and leads to It crosses test equipment and directly obtains urine dextrose equivalent, according to the composition of mentioned reagent, obtaining the formula being arranged inside measuring instrument is I =(- 0.02241*i+0.82955) * Cglu+ (0.84547*i-0.18419).When measuring, it is only necessary to obtain the first sound Electric current and the second response current are answered, urine dextrose equivalent can be obtained by above-mentioned calculating formula pre-set inside measuring instrument; A urine sample is separately taken, tests specific gravity of urine using refractometer, and prepare the glucose of various concentration with it;By grape obtained above The concentration of sugar and the contrasting glucose concentrations of preparation, as shown in Figure 7.
As shown in Figure 7, directly being measured by measuring instrument to substantially conform to blood glucose with the glucose actually prepared error Strip test national standard (when grape concentration of glucose≤4.2mM, within ± 0.83mM, when concentration of glucose > 4.2mM, ± 15% Within).
Embodiment 5
The method of detection urine glucose is same as Example 4, and difference is: the reagent in sensor reagent layer is different, tool Body is as shown in table 7 and table 8, in the present embodiment, the calculation formula for the glucose being arranged inside measuring instrument are as follows: I=(- 0.02852*i+0.67899)*Cglu+(0.85354*i-0.18966);
Composition table of the table 7 without reagent in enzyme sensor reagent layer
Substance Mass ratio
The potassium ferricyanide 5.4%
Trehalose 1.8%
Hydrophilic silicon dioxide 1.8%
Tween-20 0.9%
Phosphate buffer solution 45.05%
Water 45.05%
The composition table of reagent in 8 enzyme sensor reagent layer of table
The concentration value that five kinds of different specific gravity of urine urine samples are measured is compared with compound concentration value, and error accords with substantially after discovery correction Close the test of blood glucose strip national standard (when grape concentration of glucose≤4.2mM, within ± 0.83mM, when concentration of glucose > 4.2mM, Within ± 15%), as shown in Figure 8.
Embodiment 6
The method of detection urine glucose is same as Example 5, and difference is: sensor and embodiment 5 in the present embodiment Sensor is different, and the sensor that the present embodiment uses is as shown in figure 9,1 for substrate, 2 being wherein working electrode, 3 being to electrode, 4 It is insulating film for trigger electrode, 5,6 be reagent layer, 7 be hydrophilic film, 8 is layers of two-sided, 9 to lead sample slot, 10 be gas outlet;Absolutely The opening of velum is multiple such as two, and reagent layer 6 is set in the opening of face working electrode.
The concentration value that five kinds of different specific gravity of urine urine samples are measured is compared with compound concentration value, and error accords with substantially after discovery correction Close the test of blood glucose strip national standard (when concentration of glucose≤4.2mM, within ± 0.83mM, when concentration of glucose > 4.2mM, ± Within 15%), as shown in Figure 10.
Reagent provided in this embodiment is also applied for the electrochemical detection test strip that existing sensor for example measures glucose in urine, only needs The composition of reagent layer therein and ratio are replaced in such a way that the present embodiment provides.
The above description of the embodiment is only used to help understand the method for the present invention and its core ideas.It should be pointed out that pair For those skilled in the art, without departing from the principle of the present invention, the present invention can also be carried out Some improvements and modifications, these improvements and modifications also fall within the scope of protection of the claims of the present invention.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one The widest scope of cause.

Claims (10)

1. a kind of reagent for detecting urine specific gravity of urine, comprising:
Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1.
2. a kind of reagent for detecting urine glucose, comprising:
Surplus is pH buffer and water, and the mass ratio of pH buffer and water is 1:1.
3. reagent according to claim 1 or 2, which is characterized in that the electron mediator is the iron cyanide, ferrocyanide Object, ferrocene or derivatives thereof;The stabilizer is selected from one or both of trehalose and chitosan;The anti-agglomerating agent choosing From one or both of hydrophilic silicon dioxide and xanthan gum;The pH buffer be selected from phosphate buffer, MES buffer and One of citrate buffer solution is a variety of;The surfactant is in Tween-20, Tween-80 and Qula logical -100 It is one or more.
4. reagent according to claim 1 or 2, which is characterized in that the content of the electron mediator is 4~7wt%.
5. reagent according to claim 2, which is characterized in that the content of the glucose oxidase or glucose dehydrogenase For 0.8~3.2wt%.
6. a kind of electrochemical sensor for urine detection, including substrate, electrode and reagent layer, which is characterized in that the examination The corresponding reagent of oxidant layer is the described in any item reagents of Claims 1 to 5.
7. electrochemical sensor according to claim 6, which is characterized in that the electrochemical sensor further includes insulation Film, adhesive layer, leads sample slot and gas outlet at hydrophilic film;
Multiple electrodes are provided in substrate;
At least one electrode surface is provided with insulating film, and the insulating film offers opening, and reagent layer is set in the opening;
Adhesive layer and hydrophilic film are being disposed with along far from the insulating film surface on the substrate direction;
The adhesive layer, which offers, leads sample slot, and the hydrophilic film offers gas outlet, described to lead some or all of sample slot face The reagent layer leads sample slot described in the gas outlet face.
8. electrochemical sensor according to claim 7, which is characterized in that the electrode include working electrode with to electricity Pole, the working electrode are L-type, and described is L-type to electrode;The end L of the working electrode and base is all set in the end L of electrode The same end at bottom.
9. electrochemical sensor according to claim 8, which is characterized in that it is described opening be it is multiple, the reagent layer is set It is placed in proper pair of L of an opening of the working electrode.
10. a kind of kit for urine detection, which is characterized in that including use described in any one of Claims 1 to 5 Reagent in urine detection or the reagent layer that is formed by the described in any item reagents of Claims 1 to 5.
CN201810063224.5A 2018-01-19 2018-01-19 A kind of reagent and electrochemical sensor Pending CN109085218A (en)

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