CN104048866B - The extracting method of polyphenols in a kind of tobacco waste - Google Patents
The extracting method of polyphenols in a kind of tobacco waste Download PDFInfo
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- CN104048866B CN104048866B CN201410289090.0A CN201410289090A CN104048866B CN 104048866 B CN104048866 B CN 104048866B CN 201410289090 A CN201410289090 A CN 201410289090A CN 104048866 B CN104048866 B CN 104048866B
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Abstract
The invention discloses a kind of extracting method of polyphenols in tobacco waste.This method utilizes the polyphenols in organic solvent ultrasonic extraction tobacco waste, and nonionic surfactant is added in extract and auxiliary extraction agent is vortexed, centrifuged.Using chlorogenic acid as standard items, the content of the polyphenol extracted in ultraviolet spectrophotometry detection tobacco waste.The features such as compared with the extracting method of existing polyphenol substance, this method extraction conditions is gentle, and organic solvent amount used is greatly decreased, and extractant is environmentally friendly, extraction cost is low.
Description
Technical field
The invention belongs to analytical chemistry pre-treatment and preenrichment technical field, is related to polyphenols in a kind of tobacco waste
Extraction and assay method.
Background technology
The tobacco production in China accounts for the 45% of world wide production, and tobacco industry has substantial amounts of offal, the tobacco that goes mouldy etc. every year
Material rich in cellulose is thrown aside, and not only waste of resource but also has polluted environment.It is it that tobacco waste is handled by microorganism conversion
One of comprehensive Utilization Ways, but the polyphenols and nicotine etc. such as chlorogenic acid contained in tobacco have suppression to make to many microorganisms
With, it is therefore necessary to extracted and put forward, and the polyphenols such as chlorogenic acid has important application value in itself, extraction
It is organic fertilizer ground substance that residue can be handled with microorganism conversion.The polyphenols such as chlorogenic acid generally existing in plant, traditional work
Skill is extracted from honeysuckle, folium cortex eucommiae mostly, and it is necessary that polyphenols is extracted from Discarded Tobacco.
At present, in tobacco mainly there is the extracting method of polyphenol substance, and organic solvent extraction, ultrasonic wave extraction, microwave carry
The methods of taking.The cloud point extraction method that the present invention uses be using nonionic surfactant in aqueous with auxiliary extraction agent
It is miscible to change mutual polarity, it is had more preferable affinity, effect of extracting enhancing to polyphenols；And it has
At normal temperatures or relatively low temperature is with regard to operable, environmentally friendly, low cost and other advantages, suitable for low content polyphenols
Extraction.
The content of the invention
It is an object of the invention to for insufficient existing for present cloud point extraction method, there is provided a kind of new extraction cigarette
The method of polyphenols in careless waste material, there is directive significance for polyphenol substance extraction.
The purpose of the present invention is realized by following scheme：
The extracting method of polyphenols comprises the following steps in a kind of tobacco waste：
(1) tobacco waste of certain mass is taken to be placed in the organic solvent of certain volume, at a certain temperature ultrasonic extraction,
Filtering, filtrate are stand-by；
(2) above-mentioned sample solution is taken, adds a certain amount of deionized water and nonionic surfactant and auxiliary extraction agent,
It is vortexed, centrifugation, obtains surfactant phase and aqueous phase；Polyphenols is extracted to surfactant phase；
(3) content of polyphenols in determined by ultraviolet spectrophotometry surfactant phase is used.
The heretofore described w/v of tobacco waste and organic solvent that takes is 1:5-15, organic solvent are first
Alcohol, ethanol, one kind of acetone, its concentration are 30%-100%.
Heretofore described nonionic surfactant includes polyoxyethylene fatty alcohol Genapol X-080, fatty alcohol
APEO AEO-9 one kind, dosage are to add 0.1~0.5mL per the 5mL aqueous solution；Help extraction agent include n-hexyl alcohol, n-octyl alcohol,
One kind of isooctanol, n-amyl alcohol, dosage are to add 0.1~0.5mL per the 5mL aqueous solution.
Heretofore described Extracting temperature is 40-50 DEG C, ultrasonic time 10-30min, vortex time 1-3min,
Centrifugal speed is 3000-6000r/min, centrifugation time 3-15min.
The polyphenol of the invention using chlorogenic acid as standard items, extracted in ultraviolet spectrophotometry detection tobacco waste
Content, measure wavelength are 540nm.
Relative to prior art, the present invention has following remarkable advantage:
1. nonionic surfactant in aqueous with auxiliary extraction agent is miscible changes mutual polarity, make it to more
Aldehydes matter has more preferable affinity, and effect of extracting is remarkably reinforced.
2. Cloud Points of Nonionic Surfactants is low used by, can be carried out extracting under normal temperature or slightly heat condition,
Operating condition is gentle.
3. compared with the extracting method of existing polyphenol substance, organic solvent amount used is greatly decreased in extraction, extractant pair
Environment-friendly, the features such as extraction cost is low.
Embodiment
The present invention is further described with reference to embodiment, but protection scope of the present invention is not limited to this.
Polyphenol substance detection method：
(1) making of working curve：The polyphenol standard liquid that concentration is 10mg/mL is taken respectively in 10mL graduated centrifuge tube
In, with deionized water be configured to concentration be respectively 1,2,4,6,8,10mg/mL polyphenol standard liquid, standard is used as using chlorogenic acid
Product are detected with ultraviolet spectrophotometry, wavelength 540nm, obtain equation of linear regression y=0.01435x-0.008, coefficient correlation
R2=0.9960.
(2) sample polyphenol content determines：The sample solution handled well is taken, determines its absorbance, according to working curve in (1)
Calculate polyphenol substance content in sample.
Embodiment 1
The polyphenol substance in tobacco waste is extracted using the present invention and determines its content, and implementation process comprises the following steps：
(1) pretreatment of sample：
Take waste material offal 10g to add 95% ethanol solution 90mL, transfer for 50 DEG C and 15min is extracted in ultrasound, filter, will filter
Liquid is put into standby in sample bottle.
(2) sample extraction：
Take sample solution 2mL in step (1) in 10mL graduated centrifuge tubes, to add 3mL deionized waters, add
0.5mLAEO-9 and 0.3mL n-hexyl alcohols are in centrifuge tube, vortex 2min, 3000r/min centrifugation 15min, object be extracted in
Centrifuge tube upper strata.
(3) detected by above-mentioned polyphenol substance detection method with ultraviolet spectrophotometry, calculate polyphenols
Measure as 2.13mg/mL.
Embodiment 2
The polyphenol substance in tobacco waste is extracted using the present invention and determines its content, and implementation process comprises the following steps：
(1) sample pretreatment：
Take waste material offal 10g to add 75% methanol 150mL solution, transfer for 45 DEG C and 20min is extracted in ultrasound, filter, will
Filtrate is put into standby in sample bottle.
(2) sample extraction：
Sample solution 2mL in step (1) is taken in 10mL graduated centrifuge tubes, to add 3mL deionized waters, take 0.2mLAEO-9
With 0.2mL n-octyl alcohols in centrifuge tube, vortex 3min, 4000r/min centrifugation 10min, object is extracted in centrifuge tube upper strata.
(3) detected by above-mentioned polyphenol substance detection method with ultraviolet spectrophotometry, calculate polyphenols
Measure as 5.65mg/mL.
Embodiment 3
The polyphenol substance in tobacco waste is extracted using the present invention and determines its content, and implementation process comprises the following steps：
(1) sample pretreatment：
Take waste material offal 10g to add 50mL acetone, transfer for 50 DEG C and 10min is extracted in ultrasound, filter, filtrate is put into sample
It is standby in product bottle.
(2) sample determines：
Sample solution 2mL in step (1) is taken in 10mL graduated centrifuge tubes, to add 3mL deionized waters, add 0.5mLX-080
With 0.1mL n-amyl alcohols in centrifuge tube, vortex 1min, 5000r/min centrifugation 6min, object is extracted in centrifuge tube upper strata.
(3) detected by above-mentioned polyphenol substance detection method with ultraviolet spectrophotometry, calculate polyphenols
Measure as 3.78mg/mL.
Embodiment 4
The polyphenol substance in tobacco waste is extracted using the present invention and determines its content, and implementation process comprises the following steps：
(1) sample pretreatment：
Take waste material offal 10g to add 30% aqueous acetone solution 90mL, transfer for 40 DEG C and 30min is extracted in ultrasound, filter, will
Filtrate is put into standby in sample bottle.
(2) sample determines：
Sample solution 5mL is taken in step (1) in 10mL graduated centrifuge tubes, add 0.1mLX-080 and 0.3mL isooctanol in
In centrifuge tube, vortex 2min, 6000r/min centrifugation 3min, object is extracted in centrifuge tube upper strata.
(3) detected by above-mentioned polyphenol substance detection method with ultraviolet spectrophotometry, calculate polyphenols
Measure as 3.56mg/mL.
Claims (1)
1. the detection method of polyphenols in a kind of tobacco waste, it is characterised in that comprise the following steps：
(1) tobacco waste of certain mass is taken to be placed in the organic solvent of certain volume, ultrasonic extraction filters, and filtrate is stand-by；
(2) above-mentioned sample solution is taken, adds a certain amount of deionized water and nonionic surfactant and auxiliary extraction agent, whirlpool
Rotation, centrifugation, obtains surfactant phase and aqueous phase；Polyphenols is extracted to surfactant phase；
(3) content of polyphenols in determined by ultraviolet spectrophotometry surfactant phase is used；
The weight of tobacco waste described in step (1) and the volume ratio of organic solvent are 1:5-1:15, wherein, tobacco waste
Unit of weight for gram, the volume unit of organic solvent is milliliter；Organic solvent is methanol, ethanol, one kind of acetone；
Extracting temperature is 40-50 DEG C in step (1), ultrasonic time 10-30min；
Nonionic surfactant described in step (2) includes polyoxyethylene fatty alcohol Genapol X-080, fatty alcohol polyoxy
Vinethene AEO-9 one kind, dosage are to add 0.1~0.5mL per the 5mL aqueous solution；
Auxiliary extraction agent described in step (2) includes n-hexyl alcohol, n-octyl alcohol, isooctanol, one kind of n-amyl alcohol, and dosage is per 5mL
The aqueous solution adds 0.1~0.5mL；
Vortex time described in step (2) is 1-3min, centrifugal speed 3000-6000r/min, centrifugation time 3-15min；
The content of polyphenols described in step (3), is using chlorogenic acid as standard items, is detected using ultraviolet spectrophotometry
The content of the polyphenol extracted in tobacco waste, measure wavelength are 540nm.
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103830430A (en) * | 2014-02-21 | 2014-06-04 | 湖北中烟工业有限责任公司 | Method for extracting polyphenol from tobacco through double-water-phase system |
| CN105997632A (en) * | 2016-05-30 | 2016-10-12 | 江南大学 | Preparation method of pomegranate peel antioxidant substance |
| CN106723289A (en) * | 2017-03-31 | 2017-05-31 | 江苏中烟工业有限责任公司 | A kind of method for lifting papermaking-method reconstituted tobaccos recovery rate |
| CN109111364B (en) * | 2017-06-22 | 2021-06-22 | 中国烟草总公司广东省公司 | Method for extracting chlorogenic acid from tobacco waste by combining high-voltage pulsed electric field with microwaves and ultrasonic waves |
| CN108283329B (en) * | 2018-01-25 | 2021-02-02 | 云南中烟工业有限责任公司 | Method for extracting polyphenol compounds from tobacco waste and application |
| CN108904607B (en) * | 2018-08-13 | 2021-03-16 | 郑州轻工业学院 | Method for extracting polyphenols from oil peony seed shells |
| CN112961136A (en) * | 2021-02-06 | 2021-06-15 | 云南省烟草公司玉溪市公司 | Method for extracting polyphenol from tobacco leaves |
| CN113975308A (en) * | 2021-10-20 | 2022-01-28 | 广东省科学院测试分析研究所(中国广州分析测试中心) | Method for extracting total polyphenol from folium cortex eucommiae by using surfactant and microwave |
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| CN101126752B (en) * | 2007-09-06 | 2010-12-15 | 复旦大学 | Method for simultaneously determining multiple polyphenolic compound content of fruit |
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