CN108693287A - A kind of method that ABTS- Headspace solid-phase microextraction GC/MS method detects antioxidant content in white wine - Google Patents
A kind of method that ABTS- Headspace solid-phase microextraction GC/MS method detects antioxidant content in white wine Download PDFInfo
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- CN108693287A CN108693287A CN201810498901.6A CN201810498901A CN108693287A CN 108693287 A CN108693287 A CN 108693287A CN 201810498901 A CN201810498901 A CN 201810498901A CN 108693287 A CN108693287 A CN 108693287A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
Abstract
The invention discloses a kind of methods that ABTS- Headspace solid-phase microextraction GC/MS method detects antioxidant content in white wine, first by free radical 2'Bis- -3- ethyl benzo thiazole phenanthrolines -6- the sulfonic acid (ABTS) of hydrazine-are protected from light with white wine, white wine after being reacted;Respectively by unreacted white wine with react after white wine use Headspace solid phase microextractiom sample introduction, detected with gas chromatograph-mass spectrometer, the chromatogram of white wine after then obtaining the chromatogram of white wine and reacting;Finally, the peak area at the chromatogram of white wine retention time peak identical with the chromatogram of white wine after the reaction;The corresponding ingredient in the peak of peak area reduction in the chromatogram of white wine after reaction participates in the ingredient of radicals scavenging effect as in white wine.The present invention has the characteristics that simple to operate, influence factor is few, high sensitivity, precision are high, stability is good, and the evaluation to volatility constituent in white wine oxidation resistance may be implemented.
Description
Technical field
The present invention relates to a kind of detection method of white wine antioxidant content more particularly to a kind of micro- extractions of ABTS- head spaces solid phase
The method for taking gas chromatography mass spectrometry method to detect antioxidant content in white wine.
Background technology
White wine be using Cereals as primary raw material, with yeast, Chinese yeast or wheat bran and distiller's yeast etc. for saccharifying ferment, through boiling,
Saccharification, is distilled and manufactured beverage at fermentation.White wine has blood circulation and channel invigorating, helps the efficacy of a drug, improves a poor appetite, dispelling fatigue, cultivates one's taste,
The function of making one brisk and have resisting cold to refresh oneself has certain facilitation to health, is the traditional zymotic of the Chinese nation
Food has substance and spiritual double attribute.
Containing bioactive substances such as terpenes, Tetramethylpyrazine class and phenols in white wine, there is certain antioxygen to be turned into
With.Increase with people to drinking with healthy relationship attention rate, research antioxidative to white wine are on the increase, but at present
White wine anti-oxidation method mostly uses spectrophotometry and requires operating process to be protected from light more, and stability is bad, and testing result is not
System comprehensively analysis and classification can be carried out to white wine antioxidant content.There are no system, complete inspections for white wine antioxidant content
Survey method is unfavorable for the propulsion of the anti-oxidant research of white wine.
Invention content
In order to avoid above-mentioned the deficiencies in the prior art, the present invention is intended to provide a kind of ABTS- headspace solid-phase microextractions makings
The method that combination method detects antioxidant content in white wine.This method need to only compare the variation that chromatogram corresponds to the peak area at peak
Judge whether certain ingredient has antioxidation, while heterogeneity oxidation resistance may compare according to peak area amplitude of variation
Power has the characteristics that easy to operate, sensitivity precision is high and stability is good.
The method of antioxidant content in ABTS- Headspace solid-phase microextraction GC/MS method detection white wine of the present invention, including such as
Lower step:
Step 1:ABTS working solutions and white wine sample to be measured are pressed 2:1~10:1 volume ratio mixing is kept away at 15~30 DEG C
3~8h of light reaction, white wine after being reacted;
Step 2:Pure water and white wine sample to be measured are pressed 2:1~10:1 volume ratio mixing, it is protected from light 3 at 15~30 DEG C~
8h obtains white wine comparative sample;
Step 3:White wine and white wine comparative sample Headspace solid phase microextractiom sample introduction, use gas chromatograph-mass spectrometer after reacting respectively
Detection, the chromatogram of the chromatogram of white wine and white wine comparative sample after being reacted;Compare the chromatogram and white wine of white wine after reacting
The peak area at the chromatogram of comparative sample each peak accordingly, the corresponding substance in peak of peak area reduction have anti-as in white wine
The ingredient of oxidation, while different antioxidant content oxidation resistances in white wine are compared according to the percentage of peak area reduction
It is strong and weak.
In step 1, ABTS working solutions absorbance at 734nm is 0.7~2.0, and preparation method is as follows:
The ABTS storing solutions of 7mM are mixed in equal volume with the potassium peroxydisulfate storing solution of 2.45mM, are protected from light at 15~30 DEG C anti-
Answer 10~20h to get ABTS working stocks;ABTS working stocks described in 2~3mL are taken to be diluted to 10~30mL, at 15~30 DEG C
30min is protected from light to get ABTS working solutions.
Wherein ABTS storing solutions are obtained after 0.192g ABTS are settled to 50mL with pure water.
Wherein potassium peroxydisulfate storing solution is obtained after 0.066g potassium peroxydisulfates are settled to 100mL with pure water.
In step 3, the detection parameters setting of Headspace solid-phase microextraction GC/MS is as follows:
Chromatographic column is DB-wax capillary column chromatography columns, and injector temperature is 250 DEG C, 230 DEG C of mass detector temperature, just
40 DEG C of beginning column temperature keeps 2min, rises to 190 DEG C with 6 DEG C/min, keeps 2min, rises to 230 DEG C with 6 DEG C/min, keeps 15min;
The helium carrier gas of purity >=99.999%, flow velocity 1.6mL/min;Splitless injecting samples.
Compared with prior art, beneficial effects of the present invention are embodied in:
The present invention measures antioxidant content in white wine using the detection method of Headspace solid-phase microextraction GC/MS, in white wine
The 1%-2% of wine body total amount is accounted in fragrant, flavor and functional components, content is relatively fewer, is not easy to detect, traditional spectrophotometry
It indirect detection and can only learn total antioxidant capacity in wine sample by calculating, cannot learn it is which kind of ingredient is playing a role;
And the present invention directly can quickly detect the antioxidant content of wine sample, and can directly be found out specifically according to the amplitude that peak area changes
Certain single component oxidation resistance size, be white wine health Journal of Sex Research providing method.
Contain blue-green free radical ABTS in ABTS working solutions+, it is protected from light operation and oxidation resistance is detected to photometry
Accuracy is extremely important;The present invention is detected using the method for Headspace solid-phase microextraction GC/MS, principle:White wine and ABTS+A variety of antioxidant contents therein can be reduced after radical reaction, pass through the difference of white wine ingredient after comparing white wine and reacting
It can be learnt that in white wine the type and its oxidation resistance of antioxidant content size, method of the invention is direct detection white wine
The variation of middle ingredient relaxes operating condition, improves the precision of experiment so not being very harsh to the requirement being protected from light.
Since the present invention is using the antioxidant content of the method detection white wine of Headspace solid-phase microextraction GC/MS, method tool
Have the characteristics that easy to operate, sensitivity precision is high and stability is good, therefore suitable for detecting the lower ingredient of content in white wine,
And then realize the effective evaluation that healthy functions are carried out to white wine.
Since the present invention is using antioxidant content in the method detection white wine of Headspace solid-phase microextraction GC/MS, the method is only
The variation that the peak area that chromatogram corresponds to peak need to be compared can determine whether certain ingredient has antioxidation, while according to peak face
Product variation percentage compares the power of heterogeneity oxidation resistance.If Fig. 1 is the white wine chromatogram that ABTS working solutions are not added
(red) and white wine chromatogram (black) after the reaction of ABTS working solutions is added.Compare the peak area at each peak in chromatogram, peak
A, the peak area of B, C, D, E are reduced, and are illustrated that ingredient A, B, C, D, E have inoxidizability in white wine, are compared the hundred of peak area reduction
Divide ratio, it is known that the oxidation resistance size of the ingredient is followed successively by E>C>B>A>D.
The method of traditional detection white wine antioxidant content needs the ingredient of first screening white wine, then verifies each ingredient one by one again
Free radical scavenging ability and oxidation resistance, conventional method is there are heavy workload, test period is asynchronous, each experimental enviroment
The problems such as condition has differences, the present invention solve problem above well.
Description of the drawings
Fig. 1, Fig. 2 are respectively the detection gained chromatogram of embodiment 1,2, wherein red peak is the color that ABTS working solutions are not added
Spectrogram;Black peak is the chromatogram that ABTS working solutions are added.It can be seen from the figure that after ABTS working solutions are added, wine sample is a variety of
The content of ingredient is substantially reduced, i.e., the ingredient that content reduces takes part in Antioxidation reaction, and understands heterogeneity oxidation resistance
It is different.
Specific implementation mode
Technical solution of the present invention is further analyzed and described with specific embodiment below in conjunction with the accompanying drawings.
The absorbances at 734nm of ABTS working solutions described in following embodiment are 0.7~2.0, and preparation method is as follows:
The ABTS storing solutions of 7mM are mixed in equal volume with the potassium peroxydisulfate storing solution of 2.45mM, are protected from light at 15~30 DEG C anti-
Answer 10~20h to get ABTS working stocks;ABTS working stocks described in 2~3mL are taken to be diluted to 10~30mL, at 15~30 DEG C
30min is protected from light to get ABTS working solutions.
Wherein ABTS storing solutions are obtained after 0.192g ABTS are settled to 50mL with pure water;Potassium peroxydisulfate storing solution is
It is obtained after 0.066g potassium peroxydisulfates are settled to 100mL with pure water.
Embodiment 1:
ABTS- Headspace solid-phase microextraction GC/MS method detects the method for antioxidant content in white wine such as in the present embodiment
Under:
1, the ABTS working solutions that absorbance is 1.3 at 734nm are pressed 2.5 with white wine to be measured:1 volume ratio mixing, 20
4h is protected from light at DEG C, white wine after being reacted;
2, pure water and white wine to be measured are pressed 2.5:1 volume ratio mixing, 4h is protected from light at 20 DEG C, obtains white wine comparative sample;
3, white wine and white wine comparative sample Headspace solid phase microextractiom sample introduction after reacting respectively, are examined with gas chromatograph-mass spectrometer
It surveys, the chromatogram of the chromatogram of white wine and white wine comparative sample after being reacted;Compare the chromatogram of white wine and white wine pair after reacting
The peak area at each peak accordingly than the chromatogram of sample, the corresponding substance in peak of peak area reduction have antioxygen as in white wine
The ingredient of change effect, while the strong of different antioxidant content oxidation resistances in white wine may compare according to the amplitude of peak area reduction
It is weak.
In step 3, the detection parameters setting of Headspace solid-phase microextraction GC/MS is as follows:
Chromatographic column is DB-wax capillary column chromatography columns, and injector temperature is 250 DEG C, 230 DEG C of mass detector temperature, just
40 DEG C of beginning column temperature keeps 2min, rises to 190 DEG C with 6 DEG C/min, keeps 2min, rises to 230 DEG C with 6 DEG C/min, keeps 15min;
The helium carrier gas of purity >=99.999%, flow velocity 1.6mL/min;Splitless injecting samples.
Testing result is shown in Fig. 1, and ingredient A, B, C, D, E have inoxidizability in white wine as can be seen from Figure 1, compares peak area reduction
Percentage, it is known that the oxidation resistance size of the ingredient is followed successively by E>C>B>A>D.
Embodiment 2:
ABTS- Headspace solid-phase microextraction GC/MS method detects the method for antioxidant content in white wine such as in the present embodiment
Under:
1, the ABTS working solutions that absorbance is 1.8 at 734nm are pressed 3 with white wine to be measured:1 volume ratio mixing, 22 DEG C
Under be protected from light 5h, white wine after being reacted;
2, pure water and white wine to be measured are pressed 3:1 volume ratio mixing, 5h is protected from light at 22 DEG C, obtains white wine comparative sample;
3, white wine and white wine comparative sample Headspace solid phase microextractiom sample introduction after reacting respectively, are examined with gas chromatograph-mass spectrometer
It surveys, the chromatogram of the chromatogram of white wine and white wine comparative sample after being reacted;Compare the chromatogram of white wine and white wine pair after reacting
The peak area at each peak accordingly than the chromatogram of sample, the corresponding substance in peak of peak area reduction have antioxygen as in white wine
The ingredient of change effect, while the strong of different antioxidant content oxidation resistances in white wine may compare according to the amplitude of peak area reduction
It is weak.
In step 3, the detection parameters setting of Headspace solid-phase microextraction GC/MS is as follows:
Chromatographic column is DB-wax capillary column chromatography columns, and injector temperature is 250 DEG C, 230 DEG C of mass detector temperature, just
40 DEG C of beginning column temperature keeps 2min, rises to 190 DEG C with 6 DEG C/min, keeps 2min, rises to 230 DEG C with 6 DEG C/min, keeps 15min;
The helium carrier gas of purity >=99.999%, flow velocity 1.6mL/min;Splitless injecting samples.
Testing result is shown in Fig. 2, and ingredient A, B, C, D, E have inoxidizability in white wine as can be seen from Figure 2, compares peak area reduction
Percentage, it is known that the oxidation resistance size of the ingredient is followed successively by E>C>B>A>D.
Claims (4)
1. a kind of method of antioxidant content in ABTS- Headspace solid-phase microextraction GC/MS method detection white wine, it is characterised in that
Include the following steps:
Step 1:ABTS working solutions and white wine sample to be measured are pressed 2:1~10:1 volume ratio mixing is protected from light at 15~30 DEG C anti-
3~8h is answered, white wine after being reacted;
Step 2:Pure water and white wine sample to be measured are pressed 2:1~10:1 volume ratio mixing, 3~8h is protected from light at 15~30 DEG C,
Obtain white wine comparative sample;
Step 3:White wine and white wine comparative sample Headspace solid phase microextractiom sample introduction, are examined with gas chromatograph-mass spectrometer after reacting respectively
It surveys, the chromatogram of the chromatogram of white wine and white wine comparative sample after being reacted;Compare the chromatogram of white wine and white wine pair after reacting
The peak area at each peak accordingly than the chromatogram of sample, the corresponding substance in peak of peak area reduction have antioxygen as in white wine
The ingredient of change effect, while the strong of different antioxidant content oxidation resistances is compared in white wine according to the percentage of peak area reduction
It is weak.
2. according to the method described in claim 1, it is characterized in that:
In step 1, ABTS working solutions absorbance at 734nm is 0.7~2.0.
3. method according to claim 1 or 2, it is characterised in that the preparation method of the ABTS working solutions is as follows:
The ABTS storing solutions of 7mM are mixed in equal volume with the potassium peroxydisulfate storing solution of 2.45mM, 10 are protected from light at 15~30 DEG C
~20h is to get ABTS working stocks;It takes ABTS working stocks described in 2~3mL to be diluted to 10~30mL, is protected from light at 15~30 DEG C
30min is reacted to get ABTS working solutions.
4. according to the method described in claim 3, it is characterized in that:
In step 3, the detection parameters setting of Headspace solid-phase microextraction GC/MS is as follows:
Chromatographic column is DB-wax capillary column chromatography columns, and injector temperature is 250 DEG C, 230 DEG C of mass detector temperature, initial column
40 DEG C of temperature keeps 2min, rises to 190 DEG C with 6 DEG C/min, keeps 2min, rises to 230 DEG C with 6 DEG C/min, keeps 15min;Purity
>=99.999% helium carrier gas, flow velocity 1.6mL/min;Splitless injecting samples.
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| CN107144623A (en) * | 2017-06-29 | 2017-09-08 | 神威药业集团有限公司 | A kind of method of oxidation-resistant active ingredient in online quick screening and quantitative Chinese medicine |
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2018
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| CN1209833A (en) * | 1996-01-29 | 1999-03-03 | 诺沃挪第克公司 | Process for removing or bleaching soilng or stains from cellulosic fabric |
| CN101893615A (en) * | 2010-06-23 | 2010-11-24 | 江南大学 | Method for measuring off flavor compound geosmin not caused by hulls in Chinese white spirit |
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| CN107144623A (en) * | 2017-06-29 | 2017-09-08 | 神威药业集团有限公司 | A kind of method of oxidation-resistant active ingredient in online quick screening and quantitative Chinese medicine |
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