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 applicationThe 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 stabilizationAgent is selected from one or both of trehalose and chitosan；The anti-agglomerating agent in hydrophilic silicon dioxide and xanthan gum oneKind 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, institutesStating 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 openIn 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 slotReagent 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 electrodeType；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 electrodeIn.
Present invention also provides a kind of kits for urine detection, including being used for urine detection described in above schemeReagent 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 urineWith preferable accuracy；It further include Portugal in reagent meanwhile present invention also provides a kind of reagent for detecting urine glucoseGrape carbohydrate oxidase or glucose dehydrogenase, by with said components electron mediator, surfactant, stabilizer, anti-agglomerationAgent, buffer and water are used cooperatively, and so that it is applied to test urine glucose has preferable accuracy.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described below with reference to embodiment, stillIt 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 inventionLimitation.
For electrochemical sensor in addition to that can be used to test in urine other than glucose, present inventor passes through experiment hairIt 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 urineIt 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 oneThe 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 artSometimes 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, utilizeThe 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 inventionA 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 matchmakerMediator, 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 electronicsThe content of mediator is 2~10wt%, and in a particular embodiment, the content of the electron mediator is 4~7wt%；It is directlyInfluence 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 isTrehalose；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, instituteStating anti-agglomerating agent is hydrophilic silicon dioxide；Its content is 0.5~2wt%, in a particular embodiment, the content of the anti-agglomerating agentFor 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 realIt applies in example, the surfactant is Tween-20；Its content is 0.1~0.5wt%.The surfactant make aqueous solution withHydrophobic 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 detectingIn 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-mentionedThe 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 neededCooperate 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 schemeAgent.
More specifically, during detecting urine specific gravity of urine, it is only necessary to specific gravity of urine can be realized using a sensorDetection, 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 urineSpecific gravity variation is obvious, and glucose detection is influenced very greatly by specific gravity of urine, if therefore only with a sensor containing enzymatic reagentThe 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 protectMeasurement 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 urinatedFor the sensor-of specific gravity without enzyme sensor, a kind of sensor to detect glucose (contains glucose oxidase or grape in reagentGlucocorticoid 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 artWell 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 figure1 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 viscousLayer, 9 are closed to lead sample slot, 10 being gas outlet, specifically, the sensor include substrate, it is electrode, insulating film, reagent layer, hydrophilicFilm, 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 openIn 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 slotReagent 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 artIt can.In a specific embodiment, the electrode includes trigger electrode, working electrode with to electrode, and the working electrode is LType, 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, instituteTrigger 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 filmMultiple 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 threeA, 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 electrodeEnd.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 bondingLayer 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, makeesFor 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 lengthOr half, guarantee three electrodes some be located at the underface of adhesive layer and hydrophilic film.Liquid sample such as urineFrom leading after sample slot enters sensor, and electrode, trigger electrode and reagent layer are in contact.In this application, trigger electrodeThe 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 biographySensor 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 andCorresponding relationship between specific gravity of urine directly obtains specific gravity of urine value.When specific implementation, it can be realized by a measuring instrument, orIt 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, oneIn 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 BAgent 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%, thusThe 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 layerThus algae sugar 3%, hydrophilic silicon dioxide 1.5%, Tween-20 0.3%, phosphate buffer solution 45.6%, water 45.6% obtainThe 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, ifIts 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 electrificationIt 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 endConnect 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 applicationReferred to as first sensor or without enzyme sensor, the scales for measuring glucose in urine are second sensor or enzyme sensor), measuring instrumentMeasurement 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 respectivelyIn two sensors, and from measuring instrument into two sensors application+0.4V voltage, when measuring instrument measures 30s first sensingThe first response current (no enzyme electrode electric current) of device and the second response current (enzyme electrode electric current) of second sensor, measuring instrumentEnd directly exports urine glucose according to the corresponding relationship between urine glucose value and the first response current, the second response currentValue.
It is specific: to be obtained after being corrected using no enzyme electrode current value to enzyme electrode urine glucose test job curveOne working curve, working curve formula (the second relational expression) are as follows: I=(A*i+B) * Cglu+ (C*i+D), I=enzyme electrode electricityStream, 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 sensingThe 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 takesValue 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 surveyThe 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 bentLine, 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 ironPotassium 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 cyanogenChange 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 ironPotassium 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 cyanogenChange potassium 5.3%, trehalose 1.8%, hydrophilic silicon dioxide 1.8%, Tween-20 0.9%, phosphate buffer solution 44.2%, water44.2%, 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 componentPercentage, 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 compositionSet), after being respectively dropped into same urine sample in enzyme sensor and no enzyme sensor, in measuring instrument to enzyme sensor and nothingAfter 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, stabilizationAgent, 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 hasPreferable accuracy；Equally, in the reagent of detection urine glucose, due to including glucose oxidase or glucose dehydrogenationEnzyme, 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 detectionUrine glucose is described in detail, and protection scope of the present invention is not limited by the following examples.