CN107198708A - A kind of sunflower disk extract and its production and use - Google Patents
A kind of sunflower disk extract and its production and use Download PDFInfo
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- CN107198708A CN107198708A CN201710352702.XA CN201710352702A CN107198708A CN 107198708 A CN107198708 A CN 107198708A CN 201710352702 A CN201710352702 A CN 201710352702A CN 107198708 A CN107198708 A CN 107198708A
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- butanol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The present invention relates to medicine, more particularly to a kind of sunflower disk extract and its production and use.The sunflower disk extract of the present invention contains 17.1 ± 0.2wt% of general flavone, 6.7 ± 0.1wt% of chlorogenic acid.The sunflower disk extract of the present invention can be used for preparing medicament for expanding vascellum.
Description
Technical field
The present invention relates to medicine, more particularly to Plant Extracts and its production and use.
Background technology
Sunflower disk is the sunflower (Helianthus annuus L.) for sloughing sunflower seeds, and Helianthus is in composite family sunflower
Category, is annual herb plant, alias sunflower, and passionflower, zhang chrysanthemum are called on China's ancient books, sun flower etc. is met.China was cultivated to day
Certain herbaceous plants with big flowers at least has the history of nearly 400 years, and passing people can fall the sunflower disk for sloughing sunflower seeds as waste combustion, in recent years
Carrying out sunflower disk due to containing being made as feed or for extracting pectin multiple nutritional components more.Sunflower disk be also in fact simply in
Medicinal material, sunflower disk is sweet, cold in nature, nontoxic, return liver warp.Function wind-dispelling, soothing the liver, clearing away damp-heat dissipates stagnant gas, invigorating Qi and tonifying kidney.《China
Book on Chinese herbal medicine》Described in its play the role for the treatment of hypertension, headache, dizziness, the symptom such as tinnitus.
Sunflower is as the important oil crops of China, and extensively, its seed is used as oil expression to cultivated area, and floral disc is not yet opened
Hair is utilized, and is simply used as medicine in folk remedy.Research shows that sunflower disk has the effect reduced blood pressure, but right at present
Relevant report is had no in its vasodilatory effect.
The content of the invention
The purpose of the present invention aims to provide a kind of sunflower disk extract and preparation method thereof and vasodilator effect.
Specifically, the first aspect of the present invention there is provided a kind of sunflower disk extract, and the sunflower disk extract contains
16.9~17.3wt% of general flavone, 6.6~6.8wt% of chlorogenic acid.
In a preference, the sunflower disk extract is comprised the following steps：Sunflower disk is crushed, passes through reflux
Extracted with 75% ethanol；Extract is rotated, centrifuges and goes precipitation；Petroleum ether-ethyl acetate-n-butanol is extracted successively, described
Petroleum ether retains lower floor's liquid with ethyl acetate extraction step, is eventually adding after n-butanol, standing and is layered, and retains the extraction of its upper strata
Liquid；Precipitation is gone into gained upper strata extract solution revolving, centrifugation, n-butanol phase extract is obtained.
The second aspect of the present invention there is provided a kind of preparation method of above-mentioned sunflower disk extract, methods described bag
Include：Sunflower disk is crushed, extracted by reflux with 75% ethanol；Extract is rotated, centrifuges and goes precipitation；Petroleum ether-second
Acetoacetic ester-n-butanol is extracted successively, and the petroleum ether retains lower floor's liquid with ethyl acetate extraction step, is eventually adding positive fourth
It is layered after alcohol, standing, retains its upper strata extract solution；Precipitation is gone into gained upper strata extract solution revolving, centrifugation, n-butanol is obtained and mutually carries
Take thing.
Present invention also offers purposes of the above-mentioned sunflower disk extract in medicament for expanding vascellum is prepared.
The details of various aspects of the present invention will be able to detailed description in subsequent chapters and sections.By hereafter and claim
Description, the features of the present invention, purpose and advantage will become apparent from.
Brief description of the drawings
The rutin standard curve of the sunflower disk n-butanol phase extract of Fig. 1 present invention, illustrates rutin concentration in 0~640ug/
It is linearly good with absorbance in the range of ml；
The chlorogenic acid standard curve of the sunflower disk n-butanol phase extract of Fig. 2 present invention, illustrate chlorogenic acid concentration 0~
It is linearly good with absorbance in the range of 128ug/ml；
The sunflower disk n-butanol phase extract of Fig. 3 present invention has diastole effect, and its diastole to rat chest aorta
Concentration dependent is presented in blood vessel function, and diastole action effect is strong compared with rutin and chlorogenic acid；
The vasodilator of the sunflower disk n-butanol phase extract of Fig. 4 present invention mainly passes through NO approach.
Embodiment
The extraction and quantitative analysis of the sunflower disk n-butanol phase of embodiment one
1. materials and methods
1.1 material
Sunflower disk, the purchase place of production is the cultivar of Hebei Guoan, can swallow through Shanghai Univ. of Traditional Chinese Medicine's Experimental Teaching Centre
Professor is accredited as composite family Helianthus sunflower (Helianthus annuus L.)；Rutin, chlorogenic acid standard items (Shanghai source leaf
Bio tech ltd)；75% ethanol, n-butanol (Chemical Reagent Co., Ltd., Sinopharm Group)；722 visible light light-splitting luminosity
Count (Shanghai precision instrument Co., Ltd)；Air dry oven (the upper grand experimental facilities Co., Ltd of Nereid).
1.2 method
1.2.1 the extraction of sunflower disk n-butanol phase
The fresh sunflower disk that collection comes 60 DEG C of drying into air dry oven, direct pulverizer is crushed；Take 1kg sunflower disk powder
End, sunflower disk is crushed, and is extracted by reflux with 75% ethanol；Extract is rotated, centrifuges and goes precipitation；Petroleum ether-second
Acetoacetic ester-n-butanol is extracted successively, and petroleum ether and ethyl acetate extraction reservation are lower floor's liquid, are eventually adding n-butanol, quiet
Layering is postponed, retains its upper strata extract solution；Precipitation is gone into gained upper strata extract solution revolving, centrifugation, its n-butanol is obtained and mutually extracts
1.024 grams of thing, is configured to 1mg/mL, the quantitative analysis for general flavone and chlorogenic acid in sunflower disk n-butanol phase extract.
1.2.2 the general flavone quantitative analysis in sunflower disk n-butanol phase extract
1.2.2.1 the making of rutin standard curve
Rutin standard items 10mg accurately is weighed, is dissolved with 90% ethanol and is settled to 10m L, 4 DEG C of guarantors in brown bottle are placed in
Deposit standby (1000 μ g/m L).Standard solution 0.1,0.2,0.4,0.8,0.16,0.32,0.64mL are drawn respectively in 10m L
In volumetric flask, 0.3mL5%NaNO is added2Solution, is well mixed, and 0.3mL10%Al (NO3) is added after 6min3, it is quiet after being well mixed
6min is put, 4mL 4%NaOH solution is then added, 10mL is settled to 90% ethanol solution after standing 15min, is mixed, 15min
Light absorption value is determined at 510nm.Linear regression analysis is carried out to absorbance (Y) with rutin concentration (X), rutin standard is drawn bent
Line.
1.2.2.2 the general flavone quantitative analysis in sunflower disk n-butanol phase extract
In above-mentioned 1.2.1 after extract solution dilution, the preparation method of reference standard curve joins by ratio 5%NaNO respectively2It is molten
Liquid, 10%Al (NO3)3With 4%NaOH solution, sample liquid is made in constant volume, and its absorbance is surveyed at 510nm.Recovery rate calculates public
Formula is：
W=C × V × n/m
Wherein：The concentration of C- samples, is calculated according to standard curve；Volume after the dilution of V- samples；
The multiple of n- dilutions；The quality of m- samples.
1.2.3 the chlorogenic acid quantitative analysis in sunflower disk n-butanol phase extract
1.2.3.1 the making of chlorogenic acid standard curve
Precision weighs chlorogenic acid reference substance 5.12mg and is placed in 10ml brown volumetric flasks, adds 50% methanol solution and is settled to
Scale, is made the chlorogenic acid reference substance solution (less than 10 DEG C preservations) that concentration is 512 μ g/ml.Take chlorogenic acid standard items molten respectively
Liquid 0.039ml, 0.078ml, 0.156ml, 0.313ml, 0.625ml, 1.25ml, 2.5m, l5ml in 10ml brown volumetric flasks,
Plus 50% methanol constant volume, it is made into the chlorogenic acid standard solution that concentration is 2,4,8,16,32,64,128,256 μ g/ml, 50% first
Alcohol is blank control.The absorbance of various concentrations standard solution is surveyed under 327nm wavelength, with chlorogenic acid concentration (X) to extinction
Spend (Y) and carry out linear regression analysis, draw chlorogenic acid standard curve.
1.2.3.2 the chlorogenic acid quantitative analysis in sunflower disk n-butanol phase extract
In above-mentioned 1.2.1 after extract solution dilution, precision weighs sunflower disk n-butanol phase extract sample 5mg and is placed in 10ml palm fibres
In color tolerance measuring bottle, add 50% methanol solution and be settled to scale, the sample solution that concentration is 500 μ g/ml is made.At 327nm
Survey its absorbance.Recovery rate calculation formula is：
W=C × V × n/m
Wherein：The concentration of C- samples, is calculated according to standard curve；Volume after the dilution of V- samples；
The multiple of n- dilutions；The quality of m- samples.
2. result
2.1 rutin concentration are linearly good with absorbance in the range of 0~640ug/ml.Standard curve is y=0.0013x+
0.0429.It is shown in Table 1 and Fig. 1.
The rutin standard curve of table 1
2.2 according to 2.1 standard curves, substitute into general flavone content=17.1 in sunflower disk n-butanol phase extract ±
0.2wt%.
2.3 chlorogenic acid concentrations are linearly good with absorbance in the range of 0~128ug/ml.Standard curve is y=0.0177x
+0.1827.It is shown in Table 2 and Fig. 2.
The chlorogenic acid standard curve of table 2
2.4 according to 2.3 standard curves, substitute into chlorogenic acid content=6.7 in sunflower disk n-butanol phase extract ±
0.1wt%.
The vasodilator effect research of the sunflower disk n-butanol phase extract of embodiment two
1. materials and methods
1.1 material
Acetylcholine (Acetylcholine, ACh), phyenlephrinium (Phenylephrine, PE), sodium nitroprussiate
(Sodium Nitroprusside, SNP) etc. is purchased from Sigma companies, and the inorganic reagent such as NaCl is that domestic analysis is pure.Kreb-
Henseleit (K-H) liquid (mmolL-1)：NaCl 118,KCl 4.7,KH2PO4 1.2,MgCl2 1.2,NaHCO3 25,
CaCl2 2.5, Glucose 11, PH=7.4).Animal：200 ± 20g, rat 30 is bought in Shanghai Si Laike experimental animals
Co., Ltd, raises in Shanghai Univ. of Traditional Chinese Medicine's animal experimental center.Separatory funnel, dissection plate, tack, dissecting scissors, tweezers,
Cotton balls (absorbent cotton), culture dish etc..(AD Instruments companies, Australia is big for the channel multi-functional physiographs of PowerLab eight
Leah).
1.2 method
1.2.1 the preparation of thoracic aortic ring
Rat anesthesia is put to death, and the rapid aorta pectoralis that takes out is placed in 4 DEG C of K-H liquid, divests and 2- is made after surrounding connective tissue
3mm vascular circle.In the bath that vascular circle is hung on to 4mL K-H liquid, 37 DEG C of constant temperature, and it is continually fed into 95%O2+ 5%CO2's
Mixed gas.
Each vascular circle gives 2.0g pulling force, and K-H liquid is changed once per 15min, balances 60min.With 60mmolL-1's
KCl shrinks twice in advance, to induce maximum contraction, after K-H liquid elution pulling force stabilization, adds 10-6mol·L-1PE shrink,
After pulling force reaches plateau value, 10 are added-5Mol/L Ach, if vasodilation is up to more than 80%, it is believed that endothelium is complete, under can be used for
One step is tested.
1.2.2 effect of the sunflower disk n-butanol phase extract to rat chest aorta endothelium-dependent relaxation
The complete rat chest aorta vascular circle of endothelium is made, isodose PE shrinks, and is adding various concentrations sunflower disk just
Butanol phase extract (is prepared) by embodiment one, effect of the observation various concentrations gradient group to aorta pectoralis endothelium-dependent relaxation.
Carried out repeating experiment more than 3 times according to above experimental procedure.
1.2.3 mechanism of the sunflower disk n-butanol phase extract to rat chest aorta endothelium-dependent relaxation
The complete rat chest aorta vascular circle of endothelium is made, respectively with L-NAME (0.1mmolL-1), Indomethacin
(0.1mmol·L-1)、TEA(5mmol·L-1) pretreatment 20min, shunk in advance with PE, add sunflower disk n-butanol phase extract
(being prepared by embodiment one) observes its effect, and control group adds same dose K-H liquid.
2 results
The sunflower disk n-butanol phase extract of 2.1 present invention has diastole effect, and its diastole to rat chest aorta
Concentration dependent is presented in blood vessel function, and action effect is better than rutin and the chlorogenic acid (being shown in Table 3 and Fig. 3) that it contains.
The various concentrations sunflower disk n-butanol phase extract of table 3 is to vasodilative effect
Compared with 0.1mg/ μ L sunflower disk n-butanols,**P<0.01
The diastole effect that 2.2 sunflower disk n-butanols extract relative rat chest aorta mainly (is shown in Table 4 and figure by NO approach
4)。
The various concentrations sunflower disk n-butanol phase extract of table 4 is to vasodilative effect
Compared with 0.1mg/ μ L sunflower disk n-butanols,**P<0.01
Many aspects involved in the present invention have been explained as above.However, it should be understood that without departing from spirit of the invention
Under the premise of, those skilled in the art can carry out equivalent change and modification to it, and the change and modification equally fall into the application
The coverage of appended claims.
Claims (4)
1. a kind of sunflower disk extract, it is characterised in that the sunflower disk extract contains 16.9~17.3wt% of general flavone, green original
6.6~6.8wt% of acid.
2. sunflower disk extract as claimed in claim 1, it is characterised in that the sunflower disk extract is made by the following method
：Sunflower disk is crushed, extracted by reflux with 75% ethanol；Extract is rotated, centrifuges and goes precipitation；Petroleum ether-second
Acetoacetic ester-n-butanol is extracted successively, and the petroleum ether retains lower floor's liquid with ethyl acetate extraction step, is eventually adding positive fourth
It is layered after alcohol, standing, retains its upper strata extract solution；Precipitation is gone into gained upper strata extract solution revolving, centrifugation, n-butanol is obtained and mutually carries
Take thing.
3. the preparation method of sunflower disk extract as claimed in claim 1 or 2, it is characterised in that methods described includes：By certain herbaceous plants with big flowers
Floral disc is crushed, and is extracted by reflux with 75% ethanol；Extract is rotated, centrifuges and goes precipitation；Petroleum ether-acetic acid second
Ester-n-butanol is extracted successively, and the petroleum ether retains lower floor's liquid with ethyl acetate extraction step, is eventually adding n-butanol, quiet
Layering is postponed, retains its upper strata extract solution；Precipitation is gone into gained upper strata extract solution revolving, centrifugation, n-butanol phase extract is obtained.
4. application of the sunflower disk extract as claimed in claim 1 or 2 in medicament for expanding vascellum is prepared.
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| CN201710352702.XA CN107198708B (en) | 2017-05-18 | 2017-05-18 | Sunflower disc extract and preparation method and application thereof |
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| CN201710352702.XA CN107198708B (en) | 2017-05-18 | 2017-05-18 | Sunflower disc extract and preparation method and application thereof |
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| CN107198708A true CN107198708A (en) | 2017-09-26 |
| CN107198708B CN107198708B (en) | 2020-10-16 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6790465B2 (en) * | 2000-12-01 | 2004-09-14 | Snore-Fix, Inc. | Composition and method for treating snoring |
| US20080206161A1 (en) * | 2002-10-25 | 2008-08-28 | Dov Tamarkin | Quiescent foamable compositions, steroids, kits and uses thereof |
| CN103641717A (en) * | 2013-12-18 | 2014-03-19 | 中国农业科学院兰州畜牧与兽药研究所 | Method for extracting and separating chlorogenic acid from flower disk of sunflower in florescence |
| CN105920067A (en) * | 2016-05-06 | 2016-09-07 | 洮南市金塔生物科技有限公司 | Sunflower calathide extract containing polysaccharide, flavonoid and alkaloid and preparation method of sunflower calathide extract |
-
2017
- 2017-05-18 CN CN201710352702.XA patent/CN107198708B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6790465B2 (en) * | 2000-12-01 | 2004-09-14 | Snore-Fix, Inc. | Composition and method for treating snoring |
| US20080206161A1 (en) * | 2002-10-25 | 2008-08-28 | Dov Tamarkin | Quiescent foamable compositions, steroids, kits and uses thereof |
| CN103641717A (en) * | 2013-12-18 | 2014-03-19 | 中国农业科学院兰州畜牧与兽药研究所 | Method for extracting and separating chlorogenic acid from flower disk of sunflower in florescence |
| CN105920067A (en) * | 2016-05-06 | 2016-09-07 | 洮南市金塔生物科技有限公司 | Sunflower calathide extract containing polysaccharide, flavonoid and alkaloid and preparation method of sunflower calathide extract |
Non-Patent Citations (2)
| Title |
|---|
| 叶橘泉编: "《叶橘泉食物中药与便方》", 30 November 2013, 中国中医药出版社 * |
| 索金玲: "向日葵花盘中有效化学成分分析及抗氧化性测定", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
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