CN106153618A - A kind of test kit measuring oxidation resistance and application thereof - Google Patents
A kind of test kit measuring oxidation resistance and application thereof Download PDFInfo
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- CN106153618A CN106153618A CN201610854490.0A CN201610854490A CN106153618A CN 106153618 A CN106153618 A CN 106153618A CN 201610854490 A CN201610854490 A CN 201610854490A CN 106153618 A CN106153618 A CN 106153618A
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
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Abstract
The invention provides a kind of test kit measuring oxidation resistance, including iodine liquid, starch solution and coagulant solution.The test kit that the present invention provides, operating procedure is simple and convenient, quick, can obtain measurement result in 1h.And by with the naked eye can the power of direct judgement sample solution oxidation resistance, and can oxidation resistance in quantitative analysis sample solution, the detectable safety non-toxic of the test kit of present invention offer, low cost, effect is obvious.
Description
Technical field
The present invention relates to the technical field of determination oxidative, particularly relate to a kind of reagent measuring oxidation resistance
Box.
Background technology
Oxyradical theory is thought, active oxygen (ROS) and active nitrogen (RNS) can generate free radical, thus cause thin
There is oxidative damage in born of the same parents' lipid, cell membrane, albumen and DNA.As time goes on, oxygen-derived free radicals produces and increases, and body
Scavenging activity declines, and SOD, GSH-PX, CAT enzyme isoreactivity reduces simultaneously.And the free radical of surplus can with body cell in not
Satisfied fatty acid combines hazardous compounds such as producing MDA.
In body, the summation of the oxidation resistance of all antioxidation bases is referred to as " total antioxidant capacity ", total antioxidant capacity
It is usually used to evaluate the overall capacity of body antioxidant system, is also used to evaluate some health foods and natural plants simultaneously
Antioxygenic property, discloses the power of the biological ability resisting intracellular oxidative stress induced damage.Resist energy of oxidation at present
Power detection is substantially to be detected body sample by purchase test kit.But test kit disclosed in prior art,
Complex operation step, complexity.
Summary of the invention
In view of this, present invention aims to the deficiencies in the prior art, and provide a kind of and measure oxidation resistance
Test kit.
In order to realize foregoing invention purpose, the present invention provides techniques below scheme:
The invention provides a kind of test kit measuring oxidation resistance, including following reagent: iodine liquid, starch solution and solidifying
Gu agent solution.
Preferably, described iodine liquid is formulated by elemental iodine, iodide and water.
Preferably, in described iodine liquid, the concentration of elemental iodine is (6.9～8.5) × 10-7mol/ml。
Preferably, one during described iodide are potassium iodide, sodium iodide, silver iodide, carbon tetraiodide and triiodide nitrogen or
Several.
Preferably, described iodide are potassium iodide.
Preferably, in described starch solution, the concentration of starch is (3.0～4.5) g/ml.
Preferably, in described coagulant solution, the concentration of coagulator is (1.0～1.4) g/ml.
Preferably, one or more during described coagulator is agar, gelatin and pectin.
Further, described test kit application in terms of measuring oxidation resistance.
Present invention also offers the application of mentioned reagent box, the process of described mensuration comprises the following steps:
1) described iodine liquid, starch solution and coagulant solution are mixed, obtain the pretreatment iodamylum mixed liquor of blueness；
2) by described step 1) cooling of the pretreatment iodamylum mixed liquor that obtains, obtain solidifying the iodamylum of state；
3) testing sample solution is placed in described step 2) on the iodamylum of solidification state that obtains, changing according to color
Measurement result to oxidability.
The test kit of the mensuration oxidation resistance that the present invention provides, including iodine liquid, starch solution and coagulant solution.At this
In invention, after iodine liquid, starch solution and coagulator mixing, obtaining the mixed solution of blueness, principle is the starch in starch solution
Become blue after running into the elemental iodine in iodine liquid, after blue mixed solution cooling, obtain solidifying the iodamylum of state, and sample
In reducing substances the elemental iodine being combined with starch is reduced to iodide ion, make blueness become white, the change according to color is fast
Speed differentiates the power of oxidation resistance.The test kit that the present invention provides, operating procedure is simple and convenient, quick, can obtain in 1h
Measurement result.And by with the naked eye can the power of direct judgement sample solution oxidation resistance, and energy quantitative analysis sample is molten
Oxidation resistance in liquid, the detectable safety non-toxic of the test kit that the present invention provides, low cost, effect is obvious.
Accompanying drawing explanation
Fig. 1 is the structural representation of test kit of the present invention；
Fig. 2 is the structural representation of test kit preferred version of the present invention；
The connection diagram of the mensuration oxidation resistance instrument that Fig. 3 provides for the embodiment of the present invention.
Detailed description of the invention
The invention provides a kind of test kit measuring oxidation resistance, including iodine liquid, starch solution and coagulant solution.
The test kit that the present invention provides includes iodine liquid.In the present invention, described iodine liquid is preferably by elemental iodine, iodide and water
Formulated.Described iodide are preferably the one or several in potassium iodide, sodium iodide, silver iodide, carbon tetraiodide and triiodide nitrogen
Kind.The present invention does not has special restriction to the source of described iodide, uses the medicine that those skilled in the art's routine is selected i.e.
Can.In the present invention, what what described iodide were concrete was preferably in above-mentioned medicine is the most several, more preferably two kinds.Work as institute
State iodide and be preferably potassium iodide and sodium iodide, the mass ratio of described potassium iodide and sodium iodide be preferably (0.5～2): (0.5～
2), more preferably (0.8～1.5): (0.8～1.5), most preferably 1:1.In the present invention, the optimum that described iodide are concrete
Elect the potassium iodide in above-mentioned medicine as.
In the present invention, in described iodine liquid, the concentration of elemental iodine is preferably (6.9～8.5) × 10-7Mol/ml, more preferably
(7.4～8.2) × 10-7Mol/ml, most preferably 7.9 × 10-7mol/ml.In the present invention, the iodine liquid in described test kit is excellent
Choosing uses the vial of brown to hold, and the capacity of described Brown Glass Brown glass bottles and jars only is preferably 10～100ml, more preferably 30～70ml,
Most preferably 50ml.
Described starch solution preferably employs vial or plastic bottle holds, and the capacity of described vial or plastic bottle is preferably
1～10ml, more preferably 3～7ml, most preferably 5ml.
The test kit that the present invention provides includes starch solution.In the present invention, in described starch solution, the concentration of starch is excellent
Elect (3.0～4.5) g/ml as, more preferably (3.5～4.0) g/ml, most preferably 3.75g/ml.The present invention is to described starch
Source is not particularly limited, and uses the commercial goods that those skilled in the art's routine is selected.In the present invention, described starch
Concrete is preferably soluble starch.In the present invention, described starch solution preferably employs vial or plastic bottle holds, institute
The capacity stating starch solution is preferably 1～10ml, more preferably 3～7ml, most preferably 5ml.
The test kit that the present invention provides includes coagulant solution.In the present invention, coagulator in described coagulant solution
Concentration is preferably (1.0～1.4) g/ml, more preferably (1.1～1.3) g/ml, most preferably 1.2g/ml.In the present invention, institute
State one or more that coagulator is preferably in agar, gelatin and pectin, more preferably two kinds.When described coagulator is more preferably
During in above-mentioned medicine two kinds, the mass ratio of described coagulator is preferably (0.5～2): (0.5～2), more preferably (0.8～
1.5): (0.8～1.5), optimum is 1:1.In the present invention, the most preferably agar that described coagulator is concrete.The present invention is to institute
The source stating coagulator is not particularly limited, and uses commercial goods well known to those skilled in the art.
In the present invention, the solvent in each reagent above-mentioned is water, and the present invention does not has special limit to the kind of described water
Fixed, use the water needed for preparation solution well known to those skilled in the art；In the present invention, described water is preferably distillation
Water, more preferably distilled water.
In the present invention, described coagulant solution is preferably obtained with water heating for dissolving by described coagulator.In the present invention,
The temperature of described heating for dissolving is preferably 90～110 DEG C, more preferably 95～105 DEG C, most preferably 100 DEG C；Described add thermosol
The time solved is preferably 3～8min, more preferably 4～6min, most preferably 5min.The present invention instrument to described heating for dissolving
It is not particularly limited, uses the instrument that those skilled in the art's conventional heating dissolves, such as microwave oven.
In the present invention, described coagulant solution preferably employs vial or plastic bottle holds, described vial or plastics
The capacity of bottle is 10～100ml, more preferably 30～70ml, most preferably 50ml.
The source of the described vial holding reagent or plastic bottle is not particularly limited by the present invention, uses commercial goods
?.
In the present invention, described test kit be preferably shaped to cuboid, upper end open, excellent in the bottom of test kit internal layer
Choosing has three grooves, is used for fixing the reagent bottle holding reagent, and the order of putting of the described reagent bottle holding reagent is not had spy
Different restriction.
The invention provides the application in detection oxidation resistance of the test kit described in technique scheme, in the present invention
In, described application preferably includes following steps:
1) described iodine liquid, starch solution and coagulant solution are mixed, obtain pretreatment iodamylum mixed liquor；
2) by described step 1) cooling of the pretreatment iodamylum mixed liquor that obtains, obtain solidifying the iodamylum of state；
3) testing sample solution is immersed described step 2) in the iodamylum of solidification state that obtains, changing according to color
Measurement result to oxidability.
In the present invention, the mixing of described iodine liquid, starch solution and coagulant solution is preferably: by iodine liquid and starch solution
After mixing, obtain iodamylum solution, then mix with coagulant solution.In the present invention, described coagulant solution is molten with iodamylum
Liquid mixing is more preferably by coagulator and the dissolving of water mixing post-heating, when coagulant solution is cooled to 45～60 DEG C, then with iodine
Starch solution mixes.In the present invention, described coagulant solution is more preferably cooled to 47～55 DEG C, most preferably 50 DEG C.This
The bright instrument heating described coagulant solution does not has special restriction, uses the instrument that those skilled in the art's routine is selected i.e.
Can, such as microwave oven.
In the present invention, for mixing described iodine liquid, starch solution and coagulant solution volume ratio be preferably (1～
3): (0.1～0.3): (5～15), more preferably (1.5～2.5): (0.15:0.25): (7～10), most preferably 2:0.2:8.
In the present invention, after mixing, starch becomes blue after running into elemental iodine, so that the pretreatment iodamylum mixed liquor arrived is indigo plant
Color.
After obtaining pretreatment iodamylum mixed liquor, described pretreatment iodamylum mixed liquor is cooled down by the present invention, is solidified
State iodamylum.In the present invention, the type of cooling of described pretreatment iodamylum mixed liquor is not particularly limited, uses this area
The mode of the cooling solution that technical staff's routine is selected, as being placed in solution on ice.
After obtaining solidifying state iodamylum, testing sample solution is immersed in the iodamylum of described solidification state by the present invention, root
The measurement result of oxidability is obtained according to the change of color.In the present invention, the kind of described sample includes food, plant and blood
Clearly.In the present invention, the process step of the testing sample of described serum preferably includes non-anticoagulant process whole blood is placed in centrifuge tube
In, after solidification, 3000rpm is centrifuged 10min, is placed in by supernatant in another centrifuge tube, and 3000rpm is centrifuged 10min again, by upper
Clear liquid is placed in another centrifuge tube and get final product.The instrument that described testing sample solution is placed on the iodamylum of solidification state by the present invention does not has
Particular determination, uses the instrument that those skilled in the art's routine is selected, such as syringe.In the present invention, described color
Change obtains the measurement result specially color of oxidation resistance and is become white from blueness.In the present invention, described color becomes
The principle of white is to become blue after the starch in starch solution runs into the elemental iodine in iodine liquid, and the reducing substances in sample will be with
The elemental iodine that starch combines is reduced to iodide ion, makes blueness become white, and then according to the change fast resolution antioxidation of color
The power of ability.
In the present invention, described pretreatment iodamylum mixed liquor capillary tube is filled, and quickly after cooling, obtains solidifying state iodine
Starch, then one end of capillary tube is sealed.In the present invention in the present invention, kind and source to capillary tube are the most special
Limit, use commercial goods.In the present invention, the specification of described capillary tube preferably capacity is 5～10 μ l, more preferably
6～9 μ l, most preferably 8 μ l；The length of described capillary tube is preferably 5～15cm；More preferably 8～12cm, most preferably
10cm。
In the present invention, what the connected mode of described capillary tube was concrete be preferably one end connection syringe of silica gel tube,
The other end of silica gel tube connects the capillary tube not having sealing.The present invention is the most special to kind and the source of silica gel tube and syringe
Limit, use commercial goods.
In the present invention, the detecting step of described testing sample includes taking the capillary tube that one end has been sealed, and syringe takes to be treated
Testing sample is also placed in the unsealed one end of capillary tube by test sample product, is vertically placed by capillary tube, then do not seal after injection
Mouth one end connects one end of silica gel tube, and the other end of silica gel tube connects syringe, and after connection, syringe injection air applies
Pressure, obtains measurement result in 1h.
In the present invention, described test kit the most also includes above-mentioned capillary tube, syringe and silica gel tube, and described test kit is excellent
Choosing also includes three grooves, is used for placing above-mentioned capillary tube, syringe and silica gel tube, to described capillary tube, syringe and silica gel
The order of putting of pipe does not has special restriction.
Below in conjunction with the embodiment in the present invention, the technical scheme in the present invention is clearly and completely described.Aobvious
So, described embodiment is only a part of embodiment of the present invention rather than whole embodiments.Based on the reality in the present invention
Execute example, the every other embodiment that those of ordinary skill in the art are obtained under not making creative work premise, all belong to
In the scope of protection of the invention.
Embodiment 1
Take non-anticoagulant process Mouse whole blood 2ml in centrifuge tube, after solidification, 3000rpm is centrifuged 10min, take supernatant in
In another centrifuge tube, then 3000rpm is centrifuged 10min, takes supernatant and is placed in clean centrifuge tube, i.e. obtains testing sample molten
Liquid.Taking 2ml iodine liquid to mix with 0.2ml soluble starch solution, obtain iodamylum solution, in iodine liquid, the concentration of elemental iodine is 7.9
×10-7Mol/ml, in starch solution, the concentration of starch is 3.75g/ml.Taking 8ml agar solution, 100 DEG C are heated 5min, to be cooled
Mixing with iodamylum solution during to 50 DEG C, obtain pretreatment iodamylum mixed liquor, in agar solution, the concentration of agar is 1.2g/
ml.Taking specification is 8 μ l, and the capillary tube of a length of 10cm is filled pretreatment iodamylum solution, quickly the solution in cooling capillary tube
Until solidification, then by an end closure of capillary tube, take syringe and testing sample solution is placed in above capillary tube without coagulum
Place, and vertically place, then syringe is connected by silica gel tube with capillary tube, applies pressure by syringe air and makes it quick
Reaction, 1h measurement result.Measurement result is 1.4cm for the partial-length that bleaches, and oxidation resistance is equivalent to consume 3.36 × 10- 10The amount of mol iodide ion.
Embodiment 2
Take non-anticoagulant process Mouse whole blood 2ml in centrifuge tube, after solidification, 3000rpm is centrifuged 10min, take supernatant in
In another centrifuge tube, then 3000rpm is centrifuged 10min, takes supernatant and is placed in clean centrifuge tube, i.e. obtains testing sample molten
Liquid.Taking 1ml iodine liquid to mix with 0.1ml starch solution, obtain iodamylum solution, in iodine liquid, the concentration of iodide ion is 6.9 × 10- 7Mol/ml, in starch solution, the concentration of starch is 3.0g/ml.Take 5ml agar solution, 100 DEG C of heating 5min, be cooled to 60
DEG C time mix with iodamylum solution, obtain pretreatment iodamylum mixed liquor, in agar solution, the concentration of agar is 1.0g/ml.Take
Specification is 5 μ l, and the capillary tube of a length of 5cm is filled pretreatment iodamylum solution, and quickly the solution in cooling capillary tube is until coagulating
Gu, then by an end closure of capillary tube, take syringe and testing sample solution be placed in above capillary tube at without coagulum, and erect
Straight placement, then syringe is connected by silica gel tube with capillary tube, applies pressure by syringe air and makes its fast reaction,
45min measurement result.Measurement result is 2.0cm for the partial-length that bleaches, and oxidation resistance is equivalent to consume 4.4 × 10-10mol
The amount of iodide ion.
Embodiment 3
Take non-anticoagulant process Mouse whole blood 2ml in centrifuge tube, after solidification, 3000rpm is centrifuged 10min, take supernatant in
In another centrifuge tube, then 3000rpm is centrifuged 10min, takes supernatant and is placed in clean centrifuge tube, i.e. obtains testing sample molten
Liquid.Taking 3ml iodine liquid to mix with 0.3ml starch solution, obtain iodamylum solution, in iodine liquid, the concentration of iodide ion is 8.5 × 10- 7Mol/ml, in starch solution, the concentration of starch is 4.5g/ml.Take 15ml agar solution, 90 DEG C of heating 6min, be cooled to 45
DEG C time mix with iodamylum solution, obtain pretreatment iodamylum mixed liquor, in agar solution, the concentration of agar is 1.4g/ml.Take
Specification is 10 μ l, and the capillary tube of a length of 15cm is filled pretreatment iodamylum solution, quickly the solution in cooling capillary tube until
Solidification, then by an end closure of capillary tube, takes syringe and is placed in by testing sample solution above capillary tube at without coagulum, and
Vertically placing, then syringe is connected by silica gel tube with capillary tube, applies pressure by syringe air and makes its fast reaction,
50min measurement result.Measurement result is 2.6cm for the partial-length that bleaches, and oxidation resistance is equivalent to consume 4.68 × 10-10mol
The amount of iodide ion.
Embodiment 4
Take ultra-pure water and repeatedly rinse young leaflet tablet or stem section or sarcocarp, weigh 1-5g blade or sarcocarp, by material in mortar
The ratio of liquor ratio 1:9 adds ultra-pure water, grinds to form homogenate, the material in mortar is fully enclosed centrifuge tube, 10000r/min from
Heart 10min, Aspirate supernatant, obtain testing sample solution.Take 1ml iodine liquid to mix with 0.1ml starch solution, obtain iodamylum molten
Liquid, in iodine liquid, the concentration of iodide ion is 6.9 × 10-7Mol/ml, in starch solution, the concentration of starch is 3.0g/ml.Take 5ml agar
Solution, 100 DEG C of heating 4min, mix with iodamylum solution when being cooled to 55 DEG C, obtain pretreatment iodamylum mixed liquor, agar
In solution, the concentration of agar is 1.0g/ml.Taking specification is 5 μ l, and the capillary tube of a length of 5cm is filled pretreatment iodamylum solution,
Quickly the solution in cooling capillary tube is until solidification, then by an end closure of capillary tube, takes syringe by testing sample solution
Being placed in above capillary tube at without coagulum, and vertically place, then syringe is connected by silica gel tube with capillary tube, by injection
Device air applies pressure makes its fast reaction, 55min measurement result.Measurement result is 2.0cm for the partial-length that bleaches, antioxidation
Ability is equivalent to consume 4.4 × 10-10The amount of mol iodide ion.
Embodiment 5
Weigh the food to be measured of 1-5g, first with liquid nitrogen by mortar pre-cooling, add food liquid nitrogen grinding to be measured and become powder,
Adding ultra-pure water in the ratio of solid-liquid ratio 1:9 in mortar, mixing is made homogenate, the material in mortar is fully enclosed centrifuge tube,
10000r/min is centrifuged 10min, Aspirate supernatant, obtains testing sample solution.Take 2ml iodine liquid molten with 0.2ml soluble starch
Liquid mixes, and obtains iodamylum solution, and in iodine liquid, the concentration of elemental iodine is 7.9 × 10-7Mol/ml, the concentration of starch in starch solution
For 3.75g/ml.Take 8ml agar solution, 110 DEG C of heating 6min, mix with iodamylum solution when being cooled to 50 DEG C, obtain pre-
Processing iodamylum mixed liquor, in agar solution, the concentration of agar is 1.2g/ml.Taking specification is 8 μ l, the capillary tube of a length of 10cm
Being filled pretreatment iodamylum solution, quickly the solution in cooling capillary tube is until solidification, then by an end closure of capillary tube, takes
Testing sample solution is placed in above capillary tube at without coagulum by syringe, and vertically places, and then syringe leads to capillary tube
Cross silica gel tube to connect, apply pressure by syringe air and make its fast reaction, 1h measurement result.Measurement result is the part that bleaches
A length of 1.4cm, oxidation resistance is equivalent to consume 3.36 × 10-10The amount of mol iodide ion.
As seen from the above embodiment, the test kit that the present invention provides, operating procedure is simple and convenient, quick, can obtain in 1h
To measurement result.And by with the naked eye can the power of direct judgement sample solution oxidation resistance, and energy quantitative analysis sample
Oxidation resistance in solution, the detectable safety non-toxic of the test kit that the present invention provides, low cost, effect is obvious.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. the test kit measuring oxidation resistance, it is characterised in that include following reagent: iodine liquid, starch solution and solidification
Agent solution.
Test kit the most according to claim 1, it is characterised in that described iodine liquid is to be prepared by elemental iodine, iodide and water
Form.
Test kit the most according to claim 2, it is characterised in that in described iodine liquid, the concentration of elemental iodine is (6.9～8.5)
×10-7mol/ml。
4. according to the test kit described in Claims 2 or 3, it is characterised in that described iodide are potassium iodide, sodium iodide, iodate
One or more in silver, carbon tetraiodide and triiodide nitrogen.
Test kit the most according to claim 4, it is characterised in that described iodide are potassium iodide.
Test kit the most according to claim 1, it is characterised in that in described starch solution the concentration of starch be (3.0～
4.5)g/ml。
Test kit the most according to claim 1, it is characterised in that in described coagulant solution, the concentration of coagulator is (1.0
～1.4) g/ml.
8. according to the test kit described in claim 1 or 7, it is characterised in that described coagulator is in agar, gelatin and pectin
One or more.
9. the application in terms of measuring oxidation resistance of the test kit described in claim 1～8 any one.
Application the most according to claim 9, it is characterised in that the process of described mensuration comprises the following steps:
1) described iodine liquid, starch solution and coagulant solution are mixed, obtain pretreatment iodamylum mixed liquor；
2) by described step 1) cooling of the pretreatment iodamylum mixed liquor that obtains, obtain solidifying the iodamylum of state；
3) testing sample solution is placed in described step 2) on the iodamylum of solidification state that obtains, obtain oxygen according to the change of color
The measurement result of change ability.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109521068A (en) * | 2018-12-06 | 2019-03-26 | 杭州电子科技大学 | A method of detection honey oxidation resistance |
| CN111505002A (en) * | 2020-04-29 | 2020-08-07 | 北京福纳康生物技术有限公司 | Method for testing in-vitro oxidation resistance of antioxidant and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109521068A (en) * | 2018-12-06 | 2019-03-26 | 杭州电子科技大学 | A method of detection honey oxidation resistance |
| CN111505002A (en) * | 2020-04-29 | 2020-08-07 | 北京福纳康生物技术有限公司 | Method for testing in-vitro oxidation resistance of antioxidant and application thereof |
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