CN106124428A - A kind of Determination of Polyphenols detection method - Google Patents
A kind of Determination of Polyphenols detection method Download PDFInfo
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- CN106124428A CN106124428A CN201610421197.5A CN201610421197A CN106124428A CN 106124428 A CN106124428 A CN 106124428A CN 201610421197 A CN201610421197 A CN 201610421197A CN 106124428 A CN106124428 A CN 106124428A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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Abstract
A kind of Determination of Polyphenols detection method, belong to food technology field, by making in sample polyphenol compound in alkaline solution, under the conditions of lucifuge, quantitative oxidation reaction is carried out with the tungsten acid in forint phenol reagent, tungsten acid self is then reduced generation blue compound, the depth degree of described blue compound is directly proportional to the number rolled into a ball containing phenolic group, and reacted sample is carried out absorbance measurement, calculates Determination of Polyphenols in sample according to gallic acid standard curve.Determination of Polyphenols detection method of the present invention carries out absorbance measurement by spectrophotometer, calculates according to gallic acid standard curve, shortens the detection time, reduces testing requirement, solves other detection method instrument requirements high, the problem that sample treatment is loaded down with trivial details.
Description
Technical field
The invention belongs to food technology field, be specifically related to a kind of Determination of Polyphenols detection method.
Background technology
Free radical be produced by oxidation reaction to health harmful substance.It almost attacks internal all cells.Make
Adult body tissue, organ are destroyed, and accelerate body aging and cause a disease.Free radical too much can cause the heart, brain, angiopathy, lures
Swell tumor, cause alzheimer disease, cataract, diabetes, hepatopathy, parkinson, hypoimmunity etc..Polyphenol is plant
Property food in find, there is the compound of potential promotion health benefit.Currently it is referred to as " the 7th class nutrient ".Due to polyphenol
Structure itself has multiple conjugated double bond, even if the electronics of some position is snatched away by free radical, by conjugation molecular structure stabilized
Thus protect cell, there is the multiple free radical function of removing.Therefore the detection of Determination of Polyphenols all has weight to food production exploitation
Want meaning.
The assay method of Determination of Polyphenols have permanganimetric method, tartaric acid spectrophotography, amino-antipyrine colorimetry,
Ultraviolet spectrophotometry etc..Wherein permanganimetric method is while oxidation aldehydes matter, some phenol derivatives and non-phenolic material
Matter can also be by potassium permanganate oxidation, the poor repeatability of measurement result；Be used for of tartaric acid spectrophotography detects phenol in Nicotiana tabacum L. more
Class content of material, the scope of application is little；Amino-antipyrine colorimetry usually applies in the mensuration of volatile phenol, therefore the method phenol
Extraction step is cumbersome, and experimentation is numerous and diverse, time-consuming.
Summary of the invention
According to above the deficiencies in the prior art, the technical problem to be solved is to propose a kind of Determination of Polyphenols inspection
Survey method, carries out absorbance measurement by spectrophotometer, carries out calculating sample total polyphenols according to gallic acid standard curve
Content, shortens the detection time, reduces testing requirement, solves other detection method instrument requirements high, and sample treatment is loaded down with trivial details
Problem.
In order to solve above-mentioned technical problem, the technical solution used in the present invention is:
A kind of Determination of Polyphenols detection method, by making in sample, polyphenol compound is in alkaline solution, with forint phenol
Tungsten acid in reagent carries out quantitative oxidation reaction under the conditions of lucifuge, and tungsten acid self is then reduced generation blue coloured compound
Thing, the depth degree of described blue compound is directly proportional to the number rolled into a ball containing phenolic group, and reacted sample is carried out absorbance
Measure, calculate Determination of Polyphenols in sample according to gallic acid standard curve.
Preferably, described alkaline solution is Na2CO3Solution, wherein Na2CO3It is 12:88 with the mass ratio of distilled water.
Preferably, the described concentration that gallic acid standard curve is gallic acid standard solution is closed with the linear of absorbance
It it is curve.
Preferably, the response time of described quantitative oxidation reaction is 2h.
Preferably, in described gallic acid standard solution, gallic acid standard substance are 1 with the mass volume ratio of distilled water:
10mg/mL。
A kind of Determination of Polyphenols detection method, comprises the steps:
Step one: detection uses ultraviolet spectrophotometer, and condition determination is set to: wavelength 765nm, start preheating before detection
30min；
Step 2: the preparation of blank solution: add forint phenol reagent 1mL in test tube and mass percent is the carbon of 12%
Acid sodium solution 2mL, and it is settled to 25ml with water, obtaining blank solution, at room temperature lucifuge reaction 2h, reacted blank solution exists
At 765nm wavelength, spectrophotometer is returned to zero；
Step 3: the preparation of gallic acid standard solution: weigh gallic acid standard substance 25mg, with constant volume after water dissolution in
In 250mL volumetric flask, obtain the gallic acid standard solution of 0.1mg/mL；
Step 4: the formulation of standard curve: pipette respectively gallic acid standard solution 0.00,0.25,0.50,0.75,
1.00,1.25,1.50mL be placed in test tube and be separately added into forint phenol reagent 1mL, be separately added into mass percent after shaking up again
It is the Na of 12%2CO3Solution 2mL, is settled to 25mL with water, shakes up, and under room temperature after lucifuge reaction 2h, measures under 765nm wavelength
The absorbance of reactant；
Step 5: the drafting of standard curve: with the concentration of gallic acid standard solution as abscissa, with the extinction of reactant
Degree is vertical coordinate, makes standard curve, calculates standard curve formula；
Step 6: the mensuration of sample: take 1mL sample in test tube, and add forint phenol reagent 1mL and mass percent is
The sodium carbonate liquor 2mL of 12%, is settled to 25mL with water, measures the absorbance of sample after at room temperature lucifuge reaction 2h；
Step 7: the calculating of Determination of Polyphenols in sample: the absorbance of sample is brought in standard curve formula and calculates
Determination of Polyphenols in sample.
Preferably, the test tube described in step 2, step 4, step 6 is tool plug test tube.
Preferably, the amount of 1mL sample described in step 6 is to measure with pipet.
The medicine have the advantages that Determination of Polyphenols detection method of the present invention carries out absorbance survey by spectrophotometer
Fixed, calculate according to gallic acid standard curve, shorten the detection time, reduce testing requirement, solve other detections
Method instrument requirements is high, the problem that sample treatment is loaded down with trivial details.
Accompanying drawing explanation
Fig. 1 is the concentration graph of relation with absorbance of gallic acid standard solution.
Detailed description of the invention
Below by the description to embodiment, the present invention is further detailed explanation, to help people in the art
Member has more complete, accurate and deep understanding to inventive concept, the technical scheme of the present invention.
A kind of Determination of Polyphenols detection method, comprises the steps:
Step one: detection uses ultraviolet spectrophotometer, and condition determination is set to: wavelength 765nm, start preheating before detection
30min；
Step 2: the preparation of blank solution: add forint phenol reagent 1mL in tool plug test tube and mass percent is 12%
Sodium carbonate liquor 2mL, and be settled to 25ml with water, obtain blank solution, at room temperature lucifuge reaction 2h, reacted blank is molten
Spectrophotometer is returned to zero at 765nm wavelength by liquid；
Step 3: the preparation of gallic acid standard solution: weigh gallic acid standard substance 25mg, with constant volume after water dissolution in
In 250mL volumetric flask, obtain the gallic acid standard solution of 0.1mg/mL；
Step 4: the formulation of standard curve: pipette respectively gallic acid standard solution 0.00,0.25,0.50,0.75,
1.00,1.25,1.50mL be placed in tool plug test tube and be separately added into forint phenol reagent 1mL, be separately added into quality hundred after shaking up again
Mark is the Na of 12%2CO3Solution 2mL, is settled to 25mL with water, shakes up, under room temperature after lucifuge reaction 2h, under 765nm wavelength
Measure the absorbance of reactant；
Step 5: the drafting of standard curve: with the concentration of gallic acid standard solution as abscissa, with the extinction of reactant
Degree is vertical coordinate, makes standard curve, calculates standard curve formula；
Step 6: the mensuration of sample: carry out measuring 1mL sample in tool plug test tube with pipet, and add the examination of forint phenol
Agent 1mL and the sodium carbonate liquor 2mL that mass percent is 12%, is settled to 25mL with water, surveys after at room temperature lucifuge reaction 2h
The absorbance of random sample product；
Step 7: the calculating of Determination of Polyphenols in sample: the absorbance of sample is brought in standard curve formula and calculates
Determination of Polyphenols in sample.
Table 1 below is the concentration value with absorbance of the gallic acid standard solution using above-mentioned determination of experimental method:
Table 1
| Standard curve concentration | Absorbance |
| 0.00 | 0.000 |
| 0.25 | 0.134 |
| 0.50 | 0.263 |
| 0.75 | 0.380 |
| 1.00 | 0.484 |
| 1.25 | 0.586 |
| 1.50 | 0.680 |
Claims (8)
1. a Determination of Polyphenols detection method, it is characterised in that: by make in sample polyphenol compound in alkaline solution,
Carrying out quantitative oxidation reaction under the conditions of lucifuge with the tungsten acid in forint phenol reagent, tungsten acid self is then reduced generation indigo plant
Color compound, the depth degree of described blue compound is directly proportional to the number rolled into a ball containing phenolic group, carries out reacted sample
Absorbance measurement, calculates Determination of Polyphenols in sample according to gallic acid standard curve.
Determination of Polyphenols detection method the most according to claim 1, it is characterised in that: described alkaline solution is Na2CO3
Solution, wherein Na2CO3It is 12:88 with the mass ratio of distilled water.
Determination of Polyphenols detection method the most according to claim 1, it is characterised in that: described gallic acid standard curve
Concentration and the linear relationship curve of absorbance for gallic acid standard solution.
Determination of Polyphenols detection method the most according to claim 1, it is characterised in that: described quantitative oxidation reaction anti-
It is 2h between Ying Shi.
Determination of Polyphenols detection method the most according to claim 1, it is characterised in that: described gallic acid standard solution
Middle gallic acid standard substance are 1:10mg/mL with the mass volume ratio of distilled water.
6. according to the Determination of Polyphenols detection method described in any one of claim 1-5, it is characterised in that comprise the steps:
Step one: detection uses ultraviolet spectrophotometer, and condition determination is set to: wavelength 765nm, start preheating 30min before detection；
Step 2: the preparation of blank solution: add forint phenol reagent 1mL in test tube and mass percent is the sodium carbonate of 12%
Solution 2mL, and it is settled to 25ml with water, obtaining blank solution, at room temperature lucifuge reaction 2h, reacted blank solution exists
At 765nm wavelength, spectrophotometer is returned to zero；
Step 3: the preparation of gallic acid standard solution: weigh gallic acid standard substance 25mg, with constant volume after water dissolution in
In 250mL volumetric flask, obtain the gallic acid standard solution of 0.1mg/mL；
Step 4: the formulation of standard curve: pipette respectively gallic acid standard solution 0.00,0.25,0.50,0.75,1.00,
1.25, during 1.50mL is placed in test tube and be separately added into forint phenol reagent 1mL, being separately added into mass percent after shaking up again is 12%
Na2CO3Solution 2mL, is settled to 25mL with water, shakes up, and under room temperature after lucifuge reaction 2h, measures reactant under 765nm wavelength
Absorbance；
Step 5: the drafting of standard curve: with the concentration of gallic acid standard solution as abscissa, with the absorbance of reactant be
Vertical coordinate, makes standard curve, calculates standard curve formula；
Step 6: the mensuration of sample: take 1mL sample in test tube, and add forint phenol reagent 1mL and mass percent is 12%
Sodium carbonate liquor 2mL, be settled to 25mL with water, after at room temperature lucifuge reaction 2h, measure the absorbance of sample；
Step 7: the calculating of Determination of Polyphenols in sample: the absorbance of sample is brought in standard curve formula and calculates sample
Middle Determination of Polyphenols.
Determination of Polyphenols detection method the most according to claim 6, it is characterised in that: in step 2, step 4, step 6
Described test tube is tool plug test tube.
Determination of Polyphenols detection method the most according to claim 6, it is characterised in that: 1mL's sample described in step 6
Amount is to measure with pipet.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107307416A (en) * | 2017-05-23 | 2017-11-03 | 集美大学 | A kind of method that low temperature ultrasonic extracts Moringa polyphenol |
| CN107621456A (en) * | 2017-09-12 | 2018-01-23 | 合肥师范学院 | A kind of kiwi polyphenol content assaying method |
| CN108169146A (en) * | 2017-11-20 | 2018-06-15 | 湖北大学 | A kind of qualitative in aryloxy group phosphorus-containing compound is prepared and quantitative determination phenol method |
| CN111044465A (en) * | 2019-12-20 | 2020-04-21 | 山西省农业科学院农产品加工研究所 | Physiological marking method for evaluating storage capacity of buckwheat rice |
| CN113109277A (en) * | 2021-03-15 | 2021-07-13 | 杭州融凯盛科技有限公司 | Method for determining propyl gallate in Lyocell production system |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107307416A (en) * | 2017-05-23 | 2017-11-03 | 集美大学 | A kind of method that low temperature ultrasonic extracts Moringa polyphenol |
| CN107621456A (en) * | 2017-09-12 | 2018-01-23 | 合肥师范学院 | A kind of kiwi polyphenol content assaying method |
| CN108169146A (en) * | 2017-11-20 | 2018-06-15 | 湖北大学 | A kind of qualitative in aryloxy group phosphorus-containing compound is prepared and quantitative determination phenol method |
| CN111044465A (en) * | 2019-12-20 | 2020-04-21 | 山西省农业科学院农产品加工研究所 | Physiological marking method for evaluating storage capacity of buckwheat rice |
| CN113109277A (en) * | 2021-03-15 | 2021-07-13 | 杭州融凯盛科技有限公司 | Method for determining propyl gallate in Lyocell production system |
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