CN108982640A - A kind of evaluation method of the total antioxidant capacity based on cross-linked hydrogel - Google Patents
A kind of evaluation method of the total antioxidant capacity based on cross-linked hydrogel Download PDFInfo
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Abstract
The invention discloses a kind of evaluation methods of total antioxidant capacity based on cross-linked hydrogel.The evaluation method passes through the preparation of polysaccharide-metal ion hydrogel；The preparation of lower concentrations of active oxygen cluster；Metal ion electrochemical redox in depolymehzation process measures；Analyze the relationship between metal probe signal and total antioxidation performance；It finally realizes and evaluation can be carried out to the total economic value of example.This method is easy, cheap, can be realized the efficient detection analysis to antioxidant total antioxidant capacity.
Description
Technical field
The invention belongs to technical field of electrochemical detection, and in particular to a kind of total antioxidant capacity based on cross-linked hydrogel
Evaluation method.
Background technique
The physical chemist Denham Harman of the forties in last century, Shell Co. Ltd attempt to begin look for mankind aging's
Mechanism.In 1956, he delivered famous paper " Aging:a theory based on free radical and
Radiation chemistry ", it is believed that the aging course of organism is the reactive oxygen species accumulation institute that histocyte constantly generates
It causes.Later, he further proposes that antioxidant can be with the theory of delaying senility.Itself have in organism a set of by anti-oxidant
Metabolite and enzyme constitute complication system, pass through cooperateing between the Metabolic Intermediate and product and enzyme by antioxidation
It acts on so that important cell component such as DNA, protein and lipid is from oxidative damage.If however active oxygen in organism
Dysequilibrium between cluster ingredient and antioxidant system can then cause oxidative stress, and oxidative stress is considered as causing a variety of diseases
One of the reason of case such as rheumatoid arthritis and Parkinson's disease.Therefore, in various hygienic measures, antioxidant is taken
It is considered the effect of possessing anti-aging with the edible vegetable and fruit rich in natural.Wherein total antioxidant capacity had been already
As an important indicator for measuring food, fruit juice and additive oxidation resistance.But in recent years, many researchs have also discovered
The opinion completely contradicted, excessive intake antioxidant may bring the risk of health.Such as Medical College of Zhejiang Univ. is immune
Research institute Lu Linrong professor seminar is learned from immunologic angle, discovery supplement intake antioxidant, which exists, promotes itself property
The risk of the occurrence and development of immunological diseases.Although academia always exists dispute for the effect of antioxidant, no matter its is right
It is healthy and beneficial or harmful, a kind of method of efficient detection total antioxidant capacity is established in nutrition, pharmacology, physiology and food
Product processing and other fields all there has been extensive demand.
It now, include the detection of oxygen radical absorbability, total oxidation for the common detection methods of total antioxidant capacity
Agent understands that ability detection, ROO cause beta carotene Fading Spectrophotometic Method, the measurement of iron ion reducing power and Folin-Ciocalteu ratio
Color method etc..These detection methods based on optical instrument generally possess it is some be difficult to the shortcomings that overcoming, for example need longer
Detection time, the deficiency of decolorization and anti-interference ability need to be carried out in advance to test sample.Enzyme process detection and latest model
DNA damage method can efficiently solve the influence of chaff interferent, and provide more sensitive model, but enzyme and DNA are easier to
Inactivation, price also costly, and are not easy to stablize immobilized in electrode surface.
In contrast, total antioxidant capacity detection is carried out since its detecting instrument is cheap, detection is believed using electrochemical method
It is number sensitive, can be achieved to carry detection, become the alternative by expectation.Wherein direct electrode detection method and chemical modification electricity
Pole detection method is easy the interference by other compositions in complex sample, relatively is suitable for assessing the oxidation resistance of single-oxidizer.
Such as Chinese patent application CN201210098387.X discloses a kind of electrochemistry of the antioxygenic property of quickly detection bee product
Method determines that its is anti-oxidant by the size of the redox current peak value of solution to be measured using gallic acid as standard items
Content of material.
Summary of the invention
The purpose of the present invention is directed to the insufficient status of Electrochemical Detection total antioxidant capacity practicability, provides a kind of based on friendship
Join the evaluation method of the total antioxidant capacity of hydrogel, this method is easy, cheap, can be realized to antioxidant total antioxidation energy
The efficient detection of power is analyzed.
To achieve the above object, the present invention takes following technical scheme:
A kind of evaluation method of the total antioxidant capacity based on cross-linked hydrogel, includes the following steps:
Step (1): the preparation of polysaccharide-metal ion hydrogel:
Polysaccharide is dispersed in solvent, certain density metal precursor is then added into polysaccharide dispersion liquid
Salting liquid, stirring, obtains polysaccharide-metal ion crosslinked hydrogel；
Step (2): lower concentrations of active oxygen cluster technology of preparing:
Mixing hydrogen peroxide and ferrous sulfate solution prepare lower concentrations of active oxygen cluster；
Step (3): metal ion electrochemical redox in depolymehzation process measures:
Utilize metal ion redox current in cyclic voltammetry scan measurement polysaccharide-metal ion crosslinked hydrogel
Value；
Certain time is waited after reactive oxygen species solution is added in polysaccharide-metal ion crosslinked hydrogel, measures metal
Ionic redox current value；
When waiting identical after addition reactive oxygen species solution and antioxidant in polysaccharide-metal ion crosslinked hydrogel
Between, measure metal ion redox current value.
Step (4): analysis metal probe signal and total antioxidation performance between relationship: cross-linked hydrogel whether be added it is anti-
In the case of oxygen agent can depolymerization, compare whether there is or not antioxidant exist in the case of metal ion electrochemical redox signal difference,
Realize the evaluation to the total antioxidant capacity of antioxidant.
Step (5): detection can be carried out to the total economic value of example: the antioxidant in step (4) is changed to reality
Example sample, the example includes serum, blood plasma, urine or plant extraction liquid, corresponding by metal ion probe electrochemistry
Difference evaluation example sample total antioxidant capacity.
Preferably, the polysaccharide in the step (1) is selected from chitosan, sodium alginate, carragheen, gelatin, carrageenan
One of or it is a variety of.
Preferably, the solvent is 0.1-1M acetic acid or hydrochloric acid solution.
Preferably, the metal precursor salt is the acetate or chloride of silver, copper, nickel, zinc, cadmium, iron, cobalt or palladium, institute
The solution concentration of configuration is 0.001-0.5M.
Preferably, the volume ratio of the polysaccharide dispersion liquid and metal precursor salting liquid is 100:1-10:1.
Preferably, the hydrogen peroxide mass concentration of lower concentrations of active oxygen cluster is 1%-10%, ferrous iron in the step (2)
Ion concentration is 0.001-0.01M.
Preferably, the section of cyclic voltammetry scan is -1.5-1.5V in the step (3)；Scanning speed be 20~
100mV/s。
Preferably, the evaluation method in the step (4) specifically: take percentage system, polysaccharide-metal ion is taken to hand over
Joining the current value of metal ion oxidation or reduction peak in hydrogel is background current, in polysaccharide-metal ion crosslinked hydrogel
It is the upper limit after the current value of measured metal ion oxidation or reduction peak subtracts background current value after addition reactive oxygen species solution
Value, calculate metal measured after reactive oxygen species solution and antioxidant are added in polysaccharide-metal ion crosslinked hydrogel from
The current value of son oxidation or reduction peak is experimental group data.
Preferably, example is fresh sample in the step (5), or one month saved under -20 DEG C of environment
The three months samples saved under sample, or -80 DEG C of environment.
Method of the invention is caused using polysaccharide-metal ion crosslinked hydrogel as detection platform based on reactive oxygen species
The scavenging effect to reactive oxygen species of hydrogel depolymerization reaction and antioxidant, utilize the electrochemistry letter of metal ion in hydrogel
Number variation, realize to antioxidant total economic value can efficient detection analyze.The project is it is intended that nutrition, pharmacology, life
Increasing demand is evaluated to total antioxidation performance in the fields such as Neo-Confucianism, food processing, provides a kind of strong operability, without electrode
Surface modification and low-cost analysis means.
The invention has the advantages that:
1. entire analytic process proposed by the present invention is not necessarily to biological reagent, the evaluation of electrochemistry total antioxidation performance is substantially reduced
The cost of mode.
2. evaluating total antioxidation performance as probe using metal ion, Direct Electrochemistry detection antioxidant is avoided
Shi Rongyi by chaff interferent influenced.
3. total antioxidation performance evaluation method proposed by the present invention potential can be fabricated to mini kit product, have real
The potentiality of ground detection.
Specific embodiment
Following specific embodiments are the further explanations to method provided by the invention and technical solution, but are not construed as
Limitation of the present invention.
The total antioxidant capacity of 1. rutin and luteolin mixed solution of embodiment detects
(1) it disperses low-molecular weight chitoglycan in 0.5M acetum, is configured to 10mL, add into chitosan dispersion
Enter the 0.05M zinc acetate of 0.1mL.Mixed liquor carries out 5min with magnetic stirrer and quickly stirs, and forms chitosan-metal zinc ion
Cross-linked hydrogel.
(2) 13.9 μ g ferrous sulfate solutions are added in 2% hydrogen peroxide, prepare lower concentrations of active oxygen cluster.
(3) zinc ion restore current value is measured in chitosan-zinc ion cross-linked hydrogel as evaluating antioxygenic property
Background current.10min is waited after 0.1mL reactive oxygen species solution is added in chitosan-zinc ion cross-linked hydrogel, measures zinc
Belong to after ion reduction current value subtracts background current and is used as upper limit value.0.1mL is added in chitosan-zinc ion cross-linked hydrogel
The 0.01M rutin and luteolin mixed solution of reactive oxygen species solution and 0.1mL waits 10min, measures zinc ion reduction current
Value.The section of the scanning of cyclic voltammetric is -0.8~-0.8V；Scanning speed is 50mV/s.
(4) it zinc ion restore current value after rutin and luteolin will be added will subtract and be reduced compared with upper limit value after background current
78.4%, which is used to evaluate the total antioxidant capacity of 0.01M rutin and luteolin mixed solution.
The detection of the total antioxidant capacity of 2. ascorbic acid of embodiment and uric acid
Step (1), (2) are the same as embodiment 1.
It (3) is zinc ion restore current value in measurement chitosan-zinc ion cross-linked hydrogel as evaluation antioxygenic property
Background current.10min is waited after 0.1mL reactive oxygen species solution is added in chitosan-zinc ion cross-linked hydrogel, measures zinc
Belong to after ion reduction current value subtracts background current and is used as upper limit value.0.1mL is added in chitosan-zinc ion cross-linked hydrogel
The 0.01M ascorbic acid and 0.005M uric acid mixed solution of reactive oxygen species solution and 0.1mL wait 10min, measure zinc ion
Restore current value.The section of the scanning of cyclic voltammetric is -0.8~-0.8V；Scanning speed is 50mV/s.
(4) zinc ion restore current value after ascorbic acid and uric acid solution will be added to subtract after background current compared with upper limit value
Reduce 66.7%, which is used to evaluate the total antioxidant capacity of 0.01M ascorbic acid and 0.005M uric acid solution.
The total antioxidant capacity of 3. Lycoris Lycoris aurea bulb extracting solution of embodiment detects
(1) 10mL methanol solution is added after Lycoris aurea bulb tissue 1g pulverizes, carries out 30min ultrasonic extraction, filter paper filters,
1mL extracting solution is obtained after solvent concentration.
(2) it disperses low molecular weight seaweed acid sodium in 0.5M hydrochloric acid solution, is configured to 10mL, to sodium alginate dispersion liquid
The middle 0.01M silver nitrate that 0.1mL is added.Mixed liquor carries out 5min with magnetic stirrer and quickly stirs, and forms sodium alginate-metal
Silver ion cross-linked hydrogel.
(3) 13.9 μ g ferrous sulfate solutions are added in 1% hydrogen peroxide, prepare lower concentrations of active oxygen cluster.
(4) it measures silver ion reduction current value in sodium alginate-silver ion cross-linked hydrogel and is used as evaluation antioxygenic property
Background current.30min is waited after 0.1mL reactive oxygen species solution is added in sodium alginate-silver ion cross-linked hydrogel, is measured
Silver, which belongs to after ion reduction current value subtracts background current, is used as upper limit value.It is added in sodium alginate-silver ion cross-linked hydrogel
0.1mL reactive oxygen species solution and 0.1mL Lycoris aurea extracting solution wait 30min, measure zinc ion restore current value.Cyclic voltammetric
Scanning section be -0.5~-0.5V；Scanning speed is 100mV/s.
(5) silver ion reduction current value reduces after subtracting background current compared with upper limit value after addition Lycoris aurea extracting solution
32.3%, which is used to evaluate the total antioxidant capacity of Lycoris aurea extracting solution.
4. Lycoris of embodiment changes the total antioxidant capacity detection of brocade flower leaf extract
(1) it changing after brocade flower leaf tissue 1g pulverizes and 10mL methanol solution is added, carry out 30min ultrasonic extraction, filter paper filters,
1mL extracting solution is obtained after solvent concentration.
Step (2), (3) are the same as embodiment 3.
(4) it measures silver ion reduction current value in sodium alginate-silver ion cross-linked hydrogel and is used as evaluation antioxygenic property
Background current.30min is waited after 0.1mL reactive oxygen species solution is added in sodium alginate-silver ion cross-linked hydrogel, is measured
Silver, which belongs to after ion reduction current value subtracts background current, is used as upper limit value.It is added in sodium alginate-silver ion cross-linked hydrogel
0.1mL reactive oxygen species solution and 0.1mL change brocade flower extracting solution, wait 30min, measure zinc ion restore current value.Cyclic voltammetric
Scanning section be -0.5~-0.5V；Scanning speed is 100mV/s.
(5) silver ion reduction current value after changing brocade flower extracting solution, which is added, to be reduced after subtracting background current compared with upper limit value
11.4%, which is used to evaluate the total antioxidant capacity for changing brocade flower extracting solution.
The method of the present invention that the above embodiments are only used to help understand and its core concept.It should be pointed out that for
For those skilled in the art, without departing from the principle of the present invention, if can also be carried out to the present invention
Dry improvement and modification, these improvement and modification are also fallen into the claims in the present invention protection scope.
Claims (8)
1. a kind of evaluation method of the total antioxidant capacity based on cross-linked hydrogel, which comprises the steps of:
Step (1): the preparation of polysaccharide-metal ion hydrogel:
Polysaccharide is dispersed in solvent, it is molten that certain density metal precursor salt is then added into polysaccharide dispersion liquid
Liquid, stirring, obtains polysaccharide-metal ion crosslinked hydrogel；
Step (2): lower concentrations of active oxygen cluster technology of preparing:
Mixing hydrogen peroxide and ferrous sulfate solution prepare lower concentrations of active oxygen cluster；
Step (3): metal ion electrochemical redox in depolymehzation process measures:
Utilize metal ion redox current value in cyclic voltammetry scan measurement polysaccharide-metal ion crosslinked hydrogel；
Certain time is waited after reactive oxygen species solution is added in polysaccharide-metal ion crosslinked hydrogel, measures metal ion
Redox current value；
Same time is waited after reactive oxygen species solution and antioxidant is added in polysaccharide-metal ion crosslinked hydrogel, is surveyed
Complexing metal ion redox current value.
Step (4): the relationship between analysis metal probe signal and total antioxidation performance: whether cross-linked hydrogel is being added antioxidant
In the case of can depolymerization, compare whether there is or not antioxidant exist in the case of metal ion electrochemical redox signal difference, realize
Evaluation to the total antioxidant capacity of antioxidant.
Step (5): detection can be carried out to the total economic value of example: the antioxidant in step (4) is changed to example sample
Product, the example include serum, blood plasma, urine or plant extraction liquid, pass through the corresponding area of metal ion probe electrochemistry
The total antioxidant capacity of other evaluation example sample.
2. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In the polysaccharide in the step (1) is selected from one of chitosan, sodium alginate, carragheen, gelatin, carrageenan or more
Kind.
3. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In the solvent is 0.1-1M acetic acid or hydrochloric acid solution.
4. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In the metal precursor salt is the acetate or chloride of silver, copper, nickel, zinc, cadmium, iron, cobalt or palladium, and the solution configured is dense
Degree is 0.001-0.5M.
5. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In the volume ratio of the polysaccharide dispersion liquid and metal precursor salting liquid is 100:1-10:1.
6. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In the hydrogen peroxide mass concentration of lower concentrations of active oxygen cluster is 1%-10% in the step (2), and ferrous ion concentration is
0.001-0.01M。
7. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In the section of cyclic voltammetry scan is -1.5-1.5V in the step (3)；Scanning speed is 20~100mV/s.
8. a kind of evaluation method of total antioxidant capacity based on cross-linked hydrogel according to claim 1, feature exist
In evaluation method in the step (4) specifically: take percentage system, take golden in polysaccharide-metal ion crosslinked hydrogel
The current value for belonging to ionic oxide formation or reduction peak is background current, and reactive oxygen species are added in polysaccharide-metal ion crosslinked hydrogel
It is upper limit value after the current value of the oxidation of measured metal ion or reduction peak subtracts background current value after solution, calculates poly
Metal ion oxidation or reduction measured after reactive oxygen species solution and antioxidant are added in sugar-metal ion crosslinked hydrogel
The current value at peak is experimental group data.
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Cited By (2)
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| CN112280092A (en) * | 2020-11-05 | 2021-01-29 | 云南师范大学 | High-toughness porous composite hydrogel material and preparation and application thereof |
| CN116003827A (en) * | 2022-09-19 | 2023-04-25 | 中南大学 | Injectable hydrogel for promoting bone chitosan to complex zinc as well as preparation method and application thereof |
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