CN110314174A - A kind of red ginseng extract and preparation method with anti-aging effects - Google Patents
A kind of red ginseng extract and preparation method with anti-aging effects Download PDFInfo
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- A61K36/185—Magnoliopsida (dicotyledons)
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- 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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Abstract
The invention belongs to a kind of red ginseng extract and preparation method with anti-aging effects.It is characterized in that steaming at a certain temperature using fresh ginseng as raw material, processing is steamed and after drying obtains red ginseng, by 50%-80% ethyl alcohol heating and refluxing extraction, extracting solution concentration, freeze-drying obtain red ginseng crude extract.It after water dissolves concentrate, by C18 Solid Phase Extraction, isolates and purifies, collects 80% ethanol elution component.80% ethanol elution component is dissolved with methanol, acetone precipitation is added, is separated by solvent deposition, is filtered, filtrate concentration, step of freeze drying composition.Definite ingredients are prepared, stablize red ginseng extract.It is characterized in that red ginseng extract substance group includes 20 (S/R)-Rh1、Rg9、Rg8、F2、Rg6、Rg4、Rk3、Rh4、20(S/R)‑Rg3、20(S/R)‑Rs3、Rk1、Rg5、20(S/R)‑Rh2、Rs5、Rs4、Rk2、Rh3.Content is 80%, and yield is 4.68% or more.This method is at low cost, simple and efficient, efficient, easy to operate, extract obtained definite ingredients, and activity of fighting against senium is good.
Description
Technical field
The invention belongs to a kind of red ginseng extract and preparation method with anti-aging effects.
Background technique
Ginseng (panaxGinseng C. A. Meyer) be rare Chinese medicine, referred to as " King of Herbs ", from ancient times to the present by
People be widely applied, it is with a history of thousands of years, have extensive pharmacological action (Liyang etc., Chinese herbal medicine: Vol. 40 (1),
2009,164-166).Red ginseng is fresh ginseng by the product for steaming and being dried into, and is widely used, has anti-aging, antioxygen
Change and waits pharmacological activity (Zhou Qiuqiu etc., Shanghai Chinese magazine: Vol. 50(2) with antifatigue, 2016,97-100).In recent years,
Document also reports effect of the red ginseng in terms of anti-aging.The accumulation that red ginseng is able to suppress fat is related to specific fat cell
The expression of gene (PPAR γ, C/EBP α, aP2, leptin), while the level and 3T3-L1 cell of mRNA can also be reduced
Middle MMP-2 and MMP-9 activity (Oh J, etc., Evid Based Complement Alternat Med. Epub 2012,
2012(6): 265023.).Research finds that red ginseng can reduce (the Park B.J, etc. BMB of the DNA damage as caused by oxygen radical
Rep 2009;42 (8): 500-505.).Kim's etc. research shows that red ginseng can reduce leaching by up-regulation activities of antioxidant enzymes
Bar DNA damage of cell and oxidation (Kim J.Y. etc., the Nutr J. 2012 of low-density lipoprotein;11 (1): 1-9.).It is red
The pharmacological basis research of ginseng anti-aging effects has shown that red ginseng is a kind of good antidotal agent.
Document report, the complex chemical composition in red ginseng, main active is ginsenoside, mainly includes protoplast's ginseng two
Alcohol type saponin(e (such as Rb1, R2, Rc, Rd, Rg3, Rh2), protopanaxatriol type saponin(e (such as Re, Rf, Rg1, Rg2, Rh1) and processing
The Δ 20 (21)-and Δ 20 (22)-ginsenoside, C-24 generated in the process, 25 hydrations-ginsenoside (grain husk etc., Chinese experimental
Pharmacology of traditional Chinese medical formulae magazine: Vol. 19 (20), 2013,16-19；).Due to the complexity of its chemical component, the anti-aging chemicals of red ginseng
Matter basis is not also very clear.This just promotes us to use modern analysis and screening technique to the anti-aging of red ginseng anti-aging
Material base is screened.It there is no the report of red ginseng extract and preparation method with anti-aging effects at present.In aging
In occurrence and development Mechanism Study, free radical theory is always generally acknowledged one of key factor.This patent is anti-oxidant by red ginseng
Effect is to screen and determine a kind of red ginseng extract and preparation method with anti-aging effects.It is characterized in that being with fresh ginseng
Raw material steams processing at a certain temperature and steams and after drying obtains red ginseng, pass through 50%-80% ethyl alcohol heating and refluxing extraction, extract
Liquid concentration, freeze-drying, obtain red ginseng crude extract.It after water dissolves concentrate, by C18 Solid Phase Extraction, isolates and purifies, collects 80% second
Alcohol elution fraction.80% ethanol elution component is dissolved with methanol, acetone precipitation is added, is separated by solvent deposition, is filtered, filtrate
Concentration, step of freeze drying composition.Definite ingredients are prepared, stablize red ginseng extract.It is characterized in that red ginseng extract substance group
Including 20 (S/R)-Rh1、Rg9、Rg8、F2、Rg6、Rg4、Rk3、Rh4、20(S/R)-Rg3、20(S/R)-Rs3、Rk1、Rg5、20(S/R)-
Rh2、Rs5、Rs4、Rk2、Rh3.Content is 80%, and yield is 4.68% or more.This method is at low cost, simple and efficient, efficient, easy to operate,
Extract obtained definite ingredients, activity of fighting against senium are good.
Shared following several: Jin Shanshi [the patent public affairs close about the patent of red ginseng extract and preparation method in document
The number of opening: CN105614885A] it is extracted using red ginseng as raw material using alkali ion water and ethyl alcohol, process is static, precipitating, concentration, divides
From and obtain Radix Ginseng Rubra total saponins extract.Jin Hengtai [patent publication No.: CN10294024A] invent one kind be able to ascend it is specific
The red ginseng extract preparation method of saponin content, using be tied be provided with the pipe of coil of power supply who, extract.These red ginsengs
The extracting method and the present invention of extract are different, and maximum difference ingredient is not known.Wei Xuening [patent publication No.:
CN106890204A] using rare saponin(e in superhigh pressure technique acquisition ginseng, Rg5 and Rk1 total recovery is 5-10%.With the present invention
Preparation method and gained ingredient are different.Battle array Rong Yu [patent publication No.: CN107073055A] is contained by cell culture preparation
There is the extract of rare ginsenoside.Different using medicinal material extract with this method, active constituent is different.
Summary of the invention
It is an object of the present invention to provide a kind of red ginseng extract and preparation method with anti-aging effects.
One, prepared by red ginseng extract
1. utilizing the red ginseng after -5 years 4 years fresh ginseng steamings and drying.- 5 years 4 years fresh ginsengs are 110-125 DEG C in temperature
Under conditions of, steam ginseng case in steam processing 9.5 hours, naturally cool to temperature be 70 DEG C when, by ginseng taking-up be put into baking oven
80 DEG C dry 72 hours.
2. 50-80% ethyl alcohol heating and refluxing extraction is measured 2 times with 8-10 times, 1.5 hours every time.Ethyl alcohol is recycled to after doing, is frozen
It is dry to obtain red ginseng crude extract.
3. dissolving crude extract with 5 times of water, then isolated and purified by C18 solid phase mixing, first with the water of 5-8 times of column volume
Wash, after again respectively with 50% ethyl alcohol of 8 times of volumes, 80% ethanol elution, collect 80% ethanol elution component.
4. pair the obtained further solvent deposition purifying of component.After collected 80% ethanol elution component solvent is evaporated,
With methanol sample dissolution, 20 times of amount acetone precipitations are added, staticly settle 2-4 hours, filters to take supernatant, solvent evaporated, freeze-drying
Up to the red ginseng extract of stable components.
5. definite ingredients stablize red ginseng extract, tested and analyzed through RRLC-Q-TOF-MS, includes the rare saponin(e of ginseng
20(S/R)-Rh1、Rg9、Rg8、F2、Rg6、Rg4、Rk3、Rh4、20(S/R)-Rg3、20(S/R)-Rs3、Rk1、Rg5、20(S/
R)-Rh2,Rs5,Rs4,Rk2,Rh3.Content is 80%, and yield is 4.68% or more.
Two, red ginseng extract activity of fighting against senium result
The specified monthly age is respectively reached using 4 groups of rats (every group 15), i.e. when 4 monthly ages, 12 monthly ages, 18 monthly ages and 24 monthly age, uses
Glass capillary carries out inner canthus and takes blood.After taking blood, every group of Oral Administration in Rats give red ginseng extract aqueous solution (16 mg/100g,
It is equivalent to crude drug 40mg/100g), dosage period is 2 weeks, is taken again respectively after administration 1 week, after 2 weeks and after drug withdrawal 1 week
Blood, method are same as above.Blood sample after collection carries out refrigerated centrifuge, 3000 turns/10 minutes, takes supernatant superoxide dismutase
Enzyme (SOD) and malonaldehyde (MDA) kit are measured.Gained SOD content and MDA content will be calculated with SPSS13.0 software
Result carry out statistical analysis, comparison among groups use one-way analysis of variance, data with mean plus-minus standard deviation (± S)
It indicates.
Experimental result is shown in such as Tables 1 and 2s.
The statistical result of 1 each age group rat of table administration front and back SOD
Note: each group is compared with age group in April before being administered: P < 0.05 *
Group compares before its corresponding administration after each group administration: P < 0.05 #
The statistical result of 2 each age group rat of table administration front and back MDA
Note: each group is compared with age group in April before being administered: P < 0.01 * P < 0.05, * *
Group compares before its corresponding administration after each group administration: P < 0.01 # P < 0.05, ##
SOD detection the results show that the intracorporal SOD activity of 4 different age group rats occurs obviously with advancing age
Variation, the SOD activity in especially 24 months age group rat plasmas is significantly lower than age group in April (P < 0.05), December age group and
18 months age groups have no significant difference compared with age group in April, but downward trend then occur in 18 months age groups, show the increasing with the age
Long, the activity of internal SOD is being gradually reduced.After giving red ginseng extract, April age group and December age group rat plasma in
SOD activity does not change significantly, and increasing significantly occurs in the SOD activity of 18 months age groups and 24 months age groups, meanwhile,
It is discontinued after 1 week, the equal decrease to some degree of activity of SOD illustrates that this raised variation of SOD activity is by red ginseng extract
It is caused.This phenomenon explanation is in the normal situation of SOD Activity of Rats, and red ginseng extract has no apparent influence, but in SOD
In the senile rat of activity decline, red ginseng extract is capable of the activity of apparent increase SOD, and the result shows red ginseng extracts pair
The activity of SOD has good adjustment effect.
And MDA detection the results show that the MDA content in 4 different age group rat bloods occurred significantly with year
Age increases and increased trend, although the MDA content at 4 monthly ages and 12 monthly ages is almost the same, from 18 months age groups and 24 months age groups
From the point of view of the result of (especially 24 months age groups) in contrast, the increasing rheological properties of MDA content increases clearly, illustrates with the age
Growth, the degree of body lipid peroxidating aggravates significantly, in conjunction with SOD's as a result, the raising of MDA to be likely due to SOD living
Property decline caused by.After giving red ginseng extract and intervening, it may be seen that similar with SOD as a result, i.e. 4 monthly ages and 12
The MDA content at monthly age does not change substantially, and significantly reducing then occur in 24 months age groups, and the knot that MDA content is gone up after being discontinued
Fruit, which has also confirmed this variation, to be generated after being interfered by TGRG；Though the MDA content of 18 months age groups is after red ginseng extract intervention
So there is no significantly changing, but there is certain downward trend.
Detection of the experiment by SOD and MDA content in blood plasma after intervening different age group rat and red ginseng extract, table
It is bright with advancing age, especially into senescence phase (24 monthly age), body weakens the Scavenging activity of oxygen radical, intracorporal
Level of lipid peroxidation aggravates, and the red ginseng extract that we prepare can lipid peroxidation in effective delaying senility course
Process.
After ginseng is simply steamed and dried by this method, separated by the extraction of 80% ethyl alcohol, C18 solid phase mixing pure
Change, solvent separation, has isolated and purified out the red ginseng extract group of stable components, yield is 4.68% or more.This method is at low cost,
It is simple and efficient, efficient, easy to operate, anti-aging effects are good.
Detailed description of the invention
The RRLC-Q-TOF-MS total ion current figure of 1 red ginseng extract group of attached drawing
Attached drawing 1 is the RRLC-Q-TOF-MS total ion current figure of red ginseng extract group after separation.It is analyzed, including 20 (S/
R)-Rh1、Rg9、Rg8、F2、Rg6、Rg4、Rk3、Rh4、20(S/R)-Rg3、20(S/R)-Rs3、Rk1、Rg5、20(S/R)-
The mixture of the rare saponin(e of Rh2, Rs5, Rs4, Rk2, Rh3, does not include common ginsenoside, and the ginseng for obtaining stable components is rare
Saponin(e group, yield are higher.
Specific embodiment
Embodiment 1:
One, prepared by red ginseng extract
By 1500 grams of fresh ginsengs, under conditions of temperature is 125 DEG C, processing 9.5 hours is steamed in steaming ginseng case, is naturally cooled to
When temperature is 70 DEG C, ginseng taking-up is put into 80 DEG C of baking oven and is dried 72 hours, is crushed spare.Added with 10 times of amount (W/V) 80% ethyl alcohol
Circumfluence distillation 2 times, every time 1.5 hours.Ethyl alcohol is recycled to after doing, freeze-drying obtains the thick total saposins of red ginseng.5 times of water dissolutions, then
It is isolated and purified by C18 solid phase mixing, first with the washing of 8 times of column volumes, then respectively with 50% ethyl alcohol of 8 times of volumes, 80% ethyl alcohol
80% ethanol elution component is collected in elution.It is small using rare saponin(e polarity later, the dissolution sex differernce in different solvents, to institute
Component is obtained to be further purified.After collected 80% ethanol elution component solvent is evaporated, with methanol sample dissolution, it is added 20 times
Measure acetone precipitation, staticly settle 2-4 hours, filter to take supernatant, solvent evaporated to get stable components red ginseng extract 0.7g
(yield 4.68%).It is tested and analyzed through RRLC-Q-TOF-MS, red ginseng extract includes 20 (S/ rich in the rare saponin(e group of ginseng
R)-Rh1、Rg9、Rg8、F2、Rg6、Rg4、Rk3、Rh4、20(S/R)-Rg3、20(S/R)-Rs3、Rk1、Rg5、20(S/R)-
Rh2,Rs5,Rs4,Rk2,Rh3.The rare saponin(e group ingredient of ginseng as shown in Fig. 1, is free of common ginsenoside.
Two, red ginseng extract activity of fighting against senium is tested
2.1 administrations and the acquisition and pre-treatment of blood sample
At 4 groups of rats (every group 15) each arrival specified monthly age, i.e. 4 monthly ages, 12 monthly ages, 18 monthly ages and 24 monthly age, using hair
Thin glass tube carries out inner canthus and takes blood.After taking blood, every group of Oral Administration in Rats gives red ginseng extract aqueous solution (16 mg/100g, phase
When in crude drug 40mg/100g), dosage period is 2 weeks, blood is taken again respectively after administration 1 week, after 2 weeks and after drug withdrawal 1 week,
Method is same as above.Blood sample after collection carries out refrigerated centrifuge, 3000 turns/10 minutes, takes supernatant superoxide dismutase
(SOD) it is measured with malonaldehyde (MDA) kit.
2.2 the assay of SOD and MDA
The content of SOD or IC50 (50% inhibition concentration) and MDA are measured by colorimetric method.
2.3 statistical analysis
The result for calculating gained SOD content and MDA content is subjected to statistical analysis with SPSS13.0 software, comparison among groups are adopted
With one-way analysis of variance, data with mean plus-minus standard deviation (± S) it indicates.
2.4 result
Experimental result is shown in Table 3 and table 4.
The statistical result of 3 each age group rat of table administration front and back SOD
Note: each group is compared with age group in April before being administered: P < 0.05 *
Group compares before its corresponding administration after each group administration: P < 0.05 #
The statistical result of 4 each age group rat of table administration front and back MDA
Note: each group is compared with age group in April before being administered: P < 0.01 * P < 0.05, * *
Group compares before its corresponding administration after each group administration: P < 0.01 # P < 0.05, ##
Experiment by the detection of SOD and MDA content in blood plasma after intervening different age group rat and red ginseng extract, show with
The growth at age, especially into senescence phase (24 monthly age), body weakens the Scavenging activity of oxygen radical, intracorporal lipid
Extent of peroxidation aggravates, and the red ginseng extract that we prepare can lipid peroxidation in effective delaying senility course into
Journey.
Claims (6)
1. one kind with fresh ginseng after steaming and drying for raw material, prepare red ginseng by refluxing extraction and Solid phase extraction separation and mention
The method for taking object, it is characterised in that using fresh ginseng be raw material after steaming and drying obtains red ginseng, based on 50%-80% ethyl alcohol
Extraction solvent is wanted to extract, extracting solution concentration, freeze-drying, after water dissolves concentrate, by Solid Phase Extraction, solvent deposition separation is filtered,
Filtrate concentration, step of freeze drying composition are prepared definite ingredients, stablize red ginseng extract.
2. raw material feature described in claim 1 is: red ginseng of the raw material for -5 years 4 years fresh ginseng steamings and after drying, 4
- 5 years fresh ginsengs steamed processing 9.5 hours in steaming ginseng case, naturally cooled under conditions of temperature is 110-125 DEG C year
When temperature is 70 DEG C, ginseng taking-up is put into 80 DEG C of baking oven and is dried 72 hours.
3. method according to claim 1, it is characterised in that: measure 50-80% ethyl alcohol heating and refluxing extraction 2 with 8-10 times
Secondary, 1.5 hours every time, for recycling ethyl alcohol to after doing, freeze-drying obtained red ginseng crude extract.
4. method according to claim 1, it is characterised in that: 5 times of water dissolve crude extract, then mixed by C18 solid phase
Conjunction isolates and purifies, first with the washing of 5-8 times of column volume, after again respectively with 50% ethyl alcohol of 8 times of volumes, 80% ethanol elution, collect
80% ethanol elution component.
5. method according to claim 1, it is characterised in that: the further solvent deposition of obtained component is purified, it will
After collected 80% ethanol elution component solvent is evaporated, with methanol sample dissolution, 20 times of amount acetone precipitations is added, staticly settle 2-4
Hour, supernatant is filtered to take, the red ginseng extract up to stable components is lyophilized in solvent evaporated.
6. method according to claim 1, definite ingredients stablize red ginseng extract it is characterized by: dilute comprising ginseng
Have saponin(e 20 (S/R)-Rh1, Rg9, Rg8, F2, Rg6, Rg4, Rk3, Rh4,20 (S/R)-Rg3,20 (S/R)-Rs3, Rk1, Rg5,
20 (S/R)-Rh2, Rs5, Rs4, Rk2, Rh3, content 80%, yield is 4.68% or more.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101244103A (en) * | 2008-03-24 | 2008-08-20 | 中国科学院长春应用化学研究所 | Method for processing Chinese medicine red ginseng |
| CN102133251A (en) * | 2011-03-15 | 2011-07-27 | 李仁� | Processing method for improving ginsenosides content in radix ginseng rubra |
| CN102973623A (en) * | 2012-12-20 | 2013-03-20 | 刘淑莹 | Separation method of ginseng rare saponins in ginseng fibrils |
-
2018
- 2018-03-28 CN CN201810263608.1A patent/CN110314174A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101244103A (en) * | 2008-03-24 | 2008-08-20 | 中国科学院长春应用化学研究所 | Method for processing Chinese medicine red ginseng |
| CN102133251A (en) * | 2011-03-15 | 2011-07-27 | 李仁� | Processing method for improving ginsenosides content in radix ginseng rubra |
| CN102973623A (en) * | 2012-12-20 | 2013-03-20 | 刘淑莹 | Separation method of ginseng rare saponins in ginseng fibrils |
Non-Patent Citations (2)
| Title |
|---|
| 姜大成: "《中药鉴定学》", 31 January 2016, 中国农业大学出版社 * |
| 张乐等: "人参稀有皂苷的研究", 《长春中医药大学学报》 * |
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