CN106589016A - Method for extracting syringin compound from tobaccos - Google Patents
Method for extracting syringin compound from tobaccos Download PDFInfo
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- CN106589016A CN106589016A CN201611104589.5A CN201611104589A CN106589016A CN 106589016 A CN106589016 A CN 106589016A CN 201611104589 A CN201611104589 A CN 201611104589A CN 106589016 A CN106589016 A CN 106589016A
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- extract
- reduced pressure
- chromatography
- chloroform
- syringoside
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
- A24B15/26—Use of organic solvents for extraction
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
Abstract
The invention discloses a method for extracting a syringin compound from tobaccos. The method is characterized by comprising the following steps: preparing a tobacco extract; dissolving the tobacco extract into water, and extracting; eluting an extraction solution with ethanol in a macroporous resin column; collecting eluate, performing reduced pressure concentration, performing capillary sampling thin-layer chromatography, and combining eluate containing the similar components; performing reduced pressure concentration till a solid state to obtain a polar tobacco extract; performing thin-layer chromatography, combining samples with amaranth spots, and performing reduced pressure concentration to obtain a crude extract; uniformly stirring the crude extract and silica gel, separating by using a normal phase silica gel chromatographic column, and eluting; through the chromatographic column, combining fragrance-forming substances according to different Rf values and different polarities to obtain a fine extract; performing normal phase column chromatography on the fine extract, and eluting, segmenting into 5 parts A-E during TLC detection, performing gel column chromatography on a segment B, feeding the segment B into a semi-preparative chromatography system, and separating; under chromatographic conditions, retaining for 14.32 minutes, and separating to obtain the monomer compound.
Description
Technical field
The present invention relates to a kind of method that syringoside compound is extracted from Nicotiana tabacum L., belongs to technical field of tobacco chemistry.
Background technology
Syringoside is Phenylpropanoid Glycosides glycosides compound.Syringoside monomer be often used as investigate Radix Et Caulis Acanthopanacis Senticosi, Daphne giraldii Nitsche and
The quality control index of the Chinese herbal medicine such as Cortex Ilicis Rotundae.And about from plant extract separate obtain syringoside report it is little.From
So there are many plants to contain syringoside in boundary, but because its syringoside content is relatively low, the preparation of the syringoside standard substance for being
It is relatively costly.Therefore, simple efficient extraction prepares syringoside for the quality control and syringoside of Chinese crude drug are front
The new drug development of body is significant.
Syringoside is a kind of strong anti-liver cytotoxic drug, with the enzymatic activity and suppression lipid mistake of recovering microsomal enzyme system
Oxidation, can promote hepatotoxicant metabolism, and improve liver function, be allowed to normalization.
Meanwhile, it has been found that existing syringoside does not have antioxidant activity, purple fourth is extracted particularly from Nicotiana tabacum L.
The method of fragrant glycosides, yet there are no report, thus define its range of application, and especially syringoside is used as essence spice for cigarette
Addition.
The content of the invention
The technical problem to be solved in the present invention is：A kind of method that syringoside compound is extracted from Nicotiana tabacum L. is provided, it is purple
Ligustrin has preferable antioxidant activity, can expand the range of application of syringoside compound, to overcome prior art not
Foot.
Technical scheme：A kind of method that syringoside compound is extracted from Nicotiana tabacum L., the method is using following
Step：
(1), after with ethanol water extraction tobacco sample, tobacco extract is obtained by concentrating under reduced pressure；
(2) after will be tobacco extract water-soluble, using organic solvent extraction after, concentrating under reduced pressure extract to 1/10th volumes,
Obtain extract；
(3) after by the activation of D101 macroporous resins, load in chromatographic column, pure water is rinsed to without alcohol taste, by previous step
In macroporous resin chromatographic column, then the extract for obtaining is poured into from top to down, with the ethanol of variable concentrations gradient to macroporous resin
Post carries out eluting；
(4), after collecting eluent, concentrating under reduced pressure is carried out to eluent, capillary tube is carried out after being concentrated into 1/10th volumes and is taken
Sample thin layer chromatography, development system are chloroform：Methanol, developer are 5% ethanol solution of sulfuric acid, and display packing, will to heat colour developing
Eluent containing similar component is merged；Concentrating under reduced pressure eluent to obtain Nicotiana tabacum L. polar extract to dry；
(5) thin layer chromatography is adopted, developing solvent is chloroform：Acetone, developer be 5% ethanol solution of alpha-Naphthol, colour developing side
Method is heating colour developing, obtains glucosides class crude extract extractum after merging the sample concentrating under reduced pressure for showing aubergine speckle；
(6), after glucosides class crude extract extractum is stirred with silica gel, separated using normal phase silica gel chromatography post, mobile phase
For chloroform：Methanol elution gradient, eluting liquid proportional are 100:0-0:100；Collect each several part eluent；
(7) by thin layer chromatography, developing solvent is chloroform：Acetone, developer be 5% ethanol solution of alpha-Naphthol, colour developing side
Method is heating colour developing, according to different Rf value, merges glucosides class material according to opposed polarity, obtains the thin extract of glucosides class；
(8) normal phase column chromatography is carried out to the thin extract of Nicotiana tabacum L., with methanol：Chloroform be eluent gradient eluting, TLC detection segmentation
For A-E5 part, by B sections therein after gel filtration chromatography, separated in sample introduction to half preparing chromatography system；In color
Spectral condition：Mobile phase is acetonitrile：Water, volume ratio 18:82, retention time 14.32 minutes, the isolated monomeric compound.
Chloroform in described step (5) developing solvent：Acetone volume ratio is 5:1.
Chloroform in described step (7) developing solvent：Acetone volume ratio is 10:1.
In described step (5) and (7), developer is 5% ethanol solution of alpha-Naphthol, and coloration method is heating colour developing.
The invention has the beneficial effects as follows：In present invention process, the extraction means of each step are especially right in step 8
In TLC detection segmentations, B sections are separated in sample introduction to half preparing chromatography system after reversed-phase silica gel column chromatography；In each chromatograph
Under the conditions of, retention time obtains syringoside for 14.32 minutes is and its important step, especially retention time therein, protects
Stay the length of time, even 0.1-0.2 point of difference can all cause the change of separation product, this be inventor in an experiment
Constantly grope what repetition test was obtained.
The present invention is extracted from Nicotiana tabacum L. by separation and extraction technology and obtains syringoside, and syringoside is used as cigaret additive
The gaseous phase free radical of Medicated cigarette can be reduced, its possible mechanism is that the effective ingredient in Chinese herbal medicine has blocked Medicated cigarette free radical in burning
Generation or to generate free radical be rapidly quenched.Simultaneously as be the material of Nicotiana tabacum L. itself, in addition to Medicated cigarette
After in product, the harmful effects such as bad abnormal smells from the patient will not be produced to cigarette product due to adding non-tobacco product class material.
Description of the drawings
Proton nmr spectras (1H-NMR) of the Fig. 1 for the compounds of this invention；
Carbon-13 nmr spectras (13C-NMR) of the Fig. 2 for the compounds of this invention；
Fig. 3 is composed for the heteronuclear Multiple-Quantum Coherences of the compounds of this invention；
Fig. 4 is the 1H 1H COSY spectrums that the double quantum of the compounds of this invention are filtered；
Fig. 5 is the compounds of this invention heteronuclear polysaccharide Correlated Spectroscopy；
Antioxidation activity in vitro figures of the Fig. 6 for YC-7 (syringoside) and VC in the present invention；
Fig. 7 is that half preparative hplc of the compounds of this invention analyzes testing result figure.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, the present invention is made into one below in conjunction with accompanying drawing
Step ground is described in detail.
A kind of syringoside compound with antioxidant activity, the compound have following structural formula：
White crystal (CD3OD), Chemical Formula:C17H24O9, Exact Mass:372.14, Molecular
Weight:372.37m/z:372.14 (100.0%), 373.15 (19.0%), 374.15 (3.6%), Elemental
Analysis:C, 54.83；H, 6.50；O, 38.67.
A kind of method that syringoside compound is extracted from Nicotiana tabacum L., comprises the steps of：
(1), after with 75% ethanol water extraction tobacco sample, tobacco extract is obtained by concentrating under reduced pressure；
(2) by tobacco extract water, after ultrasound wave hydrotropy, after ethyl acetate or petroleum ether extraction, concentrating under reduced pressure extraction
Liquid is taken to 1/10th volumes, extract is obtained；
(3) during D101 macroporous resins to be required chromatographic column to specifications that load after activation appropriate size, pure water punching
It is washed till without alcohol taste, in macroporous resin chromatographic column, then the extract obtained in previous step is poured into from top to down, it is dense with difference
The ethanol (10%-100%) of degree gradient carries out eluting to macroporous resin column；
(4), after collecting many bottles of eluents, concentrating under reduced pressure is carried out to each bottle of eluent, 1/10th volumes is concentrated into laggard
Row capillary tube samples thin layer chromatography, and development system is chloroform：Methanol, developer are 5% ethanol solution of sulfuric acid, and display packing is to add
Heat colour developing.Eluent containing similar component is merged；；Concentrating under reduced pressure eluent to obtain Nicotiana tabacum L. polar extract to dry；
(5) thin layer chromatography is adopted, developing solvent is chloroform：Acetone (5:1), developer is 5% ethanol solution of alpha-Naphthol, is shown
Color method is heating colour developing, obtains glucosides class crude extract extractum after merging the sample concentrating under reduced pressure for showing aubergine speckle；
(7), after glucosides class crude extract extractum is stirred with silica gel, separated using normal phase silica gel chromatography post, mobile phase
For chloroform：Methanol elution gradient, eluting liquid proportional are 100:0-0:100；Collect each several part eluent；
(8) by thin layer chromatography, developing solvent is chloroform：Acetone (10:1), developer is 5% ethanol solution of alpha-Naphthol,
Coloration method is heating colour developing, according to different Rf value, merges glucosides class material according to opposed polarity, obtains glucosides class and carefully carry
Thing；
(9) normal phase column chromatography is carried out to the thin extract of Nicotiana tabacum L., with methanol：Chloroform be eluent gradient eluting, TLC detection segmentation
For A-E5 part, by B sections therein after gel filtration chromatography, separated in sample introduction to half preparing chromatography system；In color
Under spectral condition, (mobile phase is acetonitrile：Water, 18:82), retention time 14.32 minutes, the isolated monomeric compound.
Antioxidant activity is studied
(1) preparation of DPPH storing solutions accurately weighs 1,1- diphenyl -2- trinitrophenyl-hydrazines (DPPH) 0.078g, with anhydrous
Ethanol dissolving is settled in 100ml brown volumetric flasks, is shaken up, and obtains concentration for 2mmol/L mother solutions, 4 DEG C of preservations.It is female that used time takes 10ml
Liquid dilutes constant volume in 100ml volumetric flasks, obtains concentration for 0.2mmol/L.
(2) the preparation precision of VC solution weighs VC 0.25g, and water dissolution obtains mother liquid concentration 2.5mg/ml, takes mother solution respectively
0.01ml, 0.02ml, 0.03ml, 0.05ml, 0.08ml, 0.1ml, 0.15ml are settled in 25ml volumetric flasks, obtain finite concentration
Gradient 0.001mg/L, 0.002mg/L, 0.003mg/L, 0.005mg/L, 0.008mg/L, 0.01mg/L, 0.015mg/L shake up,
Room temperature is placed.Draw 2ml sample solutions respectively to determine according to method under " 1.1.4 " item.
(3) need testing solution, according to finite concentration gradient dilution then divides with a certain amount of separated monomeric compound is produced
2ml test solutions are not taken, is determined according to method under " 1.1.4 " item.
(4) to take 2ml variable concentrations need testing solution molten with 2ml 0.2mmol/L DPPH for free radical scavenging activity assay method
During liquid adds color comparison tube, after mixing, room temperature places 30min, and A values are determined at 517nm, and parallel assay three times is right as the positive with VC
According to.Using the absorption of the feature aubergine group of DPPH solution, determine reacted for examination with ultraviolet visible spectrophotometry
Solution represents its Scavenging activity to organic free radical in the decline degree that 517nm absorbs.
Sample understands that to DPPH rate is calculated according to below equation：
DPPH clearance rate=1- (Ai- Aj)/A0
AiFor the meansigma methodss of sample+DPPH absorbances；
AjFor the meansigma methodss of sample solution absorbance；
A0For the meansigma methodss of DPPH absorbances.
As a result
The antioxidation activity in vitro of table 1YC-7 (syringoside) and VC
| Sample | Linear regression | R2 | IC50/(mg·mL-1) |
| YC-7 (syringoside) | Y=118.05x-1.5872 | 0.9986 | 0.4370 |
| VC | Y=6573.2x-3.1645 | 0.9996 | 0.0081 |
There is certain antioxidant activity to syringoside.
Claims (4)
1. it is a kind of from Nicotiana tabacum L. extract syringoside compound method, it is characterised in that：The method includes following step：
(1), after with ethanol water extraction tobacco sample, tobacco extract is obtained by concentrating under reduced pressure；
(2), after will be tobacco extract water-soluble, after being extracted using organic solvent, concentrating under reduced pressure extract be obtained to 1/10th volumes
Extract；
(3) after by the activation of D101 macroporous resins, load in chromatographic column, pure water is rinsed to without alcohol taste, will be obtained in previous step
Extract pour into from top to down in macroporous resin chromatographic column, then, macroporous resin column is entered with the ethanol of variable concentrations gradient
Row eluting；
(4) after collecting eluent, concentrating under reduced pressure is carried out to eluent, capillary tube sampling is carried out after being concentrated into 1/10th volumes thin
Analyse layer by layer, development system is chloroform：Methanol, developer are 5% ethanol solution of sulfuric acid, and display packing is heating colour developing, will be contained
The eluent of similar component is merged；Concentrating under reduced pressure eluent to obtain Nicotiana tabacum L. polar extract to dry；
(5) thin layer chromatography is adopted, developing solvent is chloroform：Acetone, after merging shows the sample concentrating under reduced pressure of aubergine speckle
Obtain glucosides class crude extract extractum；
(6) after glucosides class crude extract extractum is stirred with silica gel, separated using normal phase silica gel chromatography post, mobile phase is chlorine
It is imitative：Methanol elution gradient, collects each several part eluent；
(7) by thin layer chromatography, developing solvent is chloroform：Acetone, according to different Rf value, merges glucosides class according to opposed polarity
Material, obtains the thin extract of glucosides class；
(8) normal phase column chromatography is carried out to the thin extract of Nicotiana tabacum L., with methanol：Chloroform be eluent gradient eluting, TLC detection be segmented into A-
E5 part, by B sections therein after gel filtration chromatography, is separated in sample introduction to half preparing chromatography system；In chromatostrip
Part：Mobile phase is acetonitrile：Water, volume ratio 18:82, retention time 14.32 minutes, the isolated monomeric compound.
2. it is according to claim 1 from Nicotiana tabacum L. extract syringoside compound method, it is characterised in that：Described step
Suddenly chloroform in (5) developing solvent：Acetone volume ratio is 5:1.
3. it is according to claim 1 from Nicotiana tabacum L. extract syringoside compound method, it is characterised in that：Described step
Suddenly chloroform in (7) developing solvent：Acetone volume ratio is 10:1.
4. it is according to claim 1 from Nicotiana tabacum L. extract syringoside compound method, it is characterised in that：Described step
Suddenly in (5) and (7), developer is 5% ethanol solution of alpha-Naphthol, and coloration method is heating colour developing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611104589.5A CN106589016A (en) | 2016-12-05 | 2016-12-05 | Method for extracting syringin compound from tobaccos |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611104589.5A CN106589016A (en) | 2016-12-05 | 2016-12-05 | Method for extracting syringin compound from tobaccos |
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| CN106589016A true CN106589016A (en) | 2017-04-26 |
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| CN201611104589.5A Pending CN106589016A (en) | 2016-12-05 | 2016-12-05 | Method for extracting syringin compound from tobaccos |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113951534A (en) * | 2021-11-05 | 2022-01-21 | 云南中烟工业有限责任公司 | Jasmine extract, preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3001530B1 (en) * | 1998-09-30 | 2000-01-24 | 日本たばこ産業株式会社 | Tobacco flavor enhancer and tobacco products containing it |
| CN101328198A (en) * | 2008-08-01 | 2008-12-24 | 陕西科技大学 | Extraction and separation method of syringin |
| CN102643322A (en) * | 2012-03-29 | 2012-08-22 | 中国中医科学院中药研究所 | Method for simultaneously preparing pedunculoside and syringin |
-
2016
- 2016-12-05 CN CN201611104589.5A patent/CN106589016A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3001530B1 (en) * | 1998-09-30 | 2000-01-24 | 日本たばこ産業株式会社 | Tobacco flavor enhancer and tobacco products containing it |
| CN101328198A (en) * | 2008-08-01 | 2008-12-24 | 陕西科技大学 | Extraction and separation method of syringin |
| CN102643322A (en) * | 2012-03-29 | 2012-08-22 | 中国中医科学院中药研究所 | Method for simultaneously preparing pedunculoside and syringin |
Non-Patent Citations (2)
| Title |
|---|
| KENJI ITO，ET AL.: ""Glycosidic Fraction of Flue-curred Tobacco Leaves: Its Separation and Component Analysis"", 《BIOSCI. BIOTECHNOL. BIOCHEM.》 * |
| 郭丽娜等: "《常用中药材快速鉴定》", 30 June 2006, 黑龙江科学技术出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113951534A (en) * | 2021-11-05 | 2022-01-21 | 云南中烟工业有限责任公司 | Jasmine extract, preparation method and application thereof |
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