CN109503675A - The method of rubusoside and ursolic acid is extracted from Sweet tea - Google Patents
The method of rubusoside and ursolic acid is extracted from Sweet tea Download PDFInfo
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- CN109503675A CN109503675A CN201811491679.3A CN201811491679A CN109503675A CN 109503675 A CN109503675 A CN 109503675A CN 201811491679 A CN201811491679 A CN 201811491679A CN 109503675 A CN109503675 A CN 109503675A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
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- 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
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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
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/256—Polyterpene radicals
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Abstract
The present invention provides a kind of method that rubusoside and ursolic acid are extracted from Sweet tea, including crushing, alcohol takes, filters, is concentrated, water-soluble, alkali soluble, filtering, filter residue is through fermentation of Aspergillus niger, alcohol takes, filters, is concentrated, water-soluble, alkali soluble again, Column chromatography respectively, elution is concentrated under reduced pressure, and crystallization recrystallization is primary to obtain rubusoside sterling and ursolic acid sterling.The present invention improves dissolution and the transformation efficiency of its functional component by microorganism conversion, while neutralizing alkali process object using Aspergillus Niger acid, is also beneficial to aspergillus niger fast-growth；In addition rubusoside and ursolic acid are separated with water-soluble and alkali soluble mode using alcohol extracting, again, acquisition high purity product is chromatographed and crystallized through column, effectively prevent rubusoside to destroy under base reagent, improves its yield.
Description
Technical field
The invention belongs to the technical fields of natural product chemistry, relate generally to one kind and extract rubusoside and bear from sweet tea
The method of tartaric acid.
Background technique
Sweet tea is the perennial rattan wood of rosaceae, is a kind of distinctive wild sweet-tasting plant in Guangxi, main product in my area Dayao Mountain,
Since its is sweet, therefore named Sweet tea.With Siraitia grosvenorii, STEVIA REBAUDIANA and claim Guangxi three big sweet-tasting plant, my area it is civil have it is long
Applicating history, local people are also used to generation sugar processed food, can also be used as medicine always when tea-drinking use for a long time.Guangxi in 1979
Analysis testing research center has carried out Chemical bath deposition with regard to Sweet tea, determines its chemical structure and rebaudioside containing sweet substance
Unanimously, sugariness is 300 times of sucrose, and calorific value is only the 5% of sucrose.Identified, rubusoside (Sweet tea glucoside) content is up to
6-7%.Through pharmacological testing, have hypoglycemic, blood pressure lowering, the gastric acid inhibitory that boosts metabolism excessive, clearing heat and moistening lung, solution of promoting the production of body fluid
Yearningly and there is Gao Tiandu, the characteristic of low heat value can make diabetes, the adjuvant treatment of obesity.Have to thermal sensation, heat-syndrome cough preferable
Curative effect has no toxic side effect through toxicological test, is a kind of novel advanced sweet taste health beverages in the world.Mouthfeel sweetness is tasty and refreshing, always
It is few all suitable.It often drinks, can keep fit and healthy.
The separation and Extraction of rubusoside and terpene substances, functional component etc. are studied both at home and abroad, to small-scale
It extracts the rubusoside of higher degree and is also had been reported that with terpene substances, the Introduction To Cn Patent of 102239983 A of Publication No. CN
A kind of method for extracting rubusoside from sweet tea using column chromatography, crystallization and the means such as recrystallizes and is on a large scale made
Standby rubusoside sterling, is added as food additives, is widely used in the trouble such as obesity, hypertension, diabetes and heart disease
The health food of person.Detailed process is extracted, filters, is concentrated under reduced pressure with ethanol solution using the sweet tea after dried, so
It is concentrated under reduced pressure, decolourizes with resin adsorption, ethanol elution, by eluent afterwards, rubusoside sterling is made in crystallization and recrystallization.Publication number
For a kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea of Introduction To Cn Patent of CN107513094A,
Including using dehydrated alcohol ultrasonic wave extraction, suction filtration；Extracting solution after concentration merges obtains medicinal extract, and resulting medicinal extract chloroform is molten
It solves, brown medicinal extract is obtained after concentration and recovery chloroform；With 5% sodium hydroxide alkali soluble acid, heavy, sediment is separated medicinal extract with silica gel column chromatography
The methods of obtain the oleanolic acid and ursolic acid of high-purity, using high performance liquid chromatography carry out assay, oleanolic acid and
The purity of ursolic acid is 90% or more, and the yield of oleanolic acid is in 20mg/100g or more, and the yield of ursolic acid is in 90mg/
100g or more.
From the point of view of existing literature data, the functional component utilization rate of Sweet tea is not high, and is handled as waste, waste
A large amount of Sweet tea resource can substantially reduce the production cost of its effective component and to ring if Sweet tea functional component can be made full use of
The protection in border.
Summary of the invention
Not high for Sweet tea utilization rate, the present invention provides that a kind of Sweet tea utilization rate is high, method is simple, at low cost, function produces
The rubusoside of product purity is high and the preparation method of ursolic acid.
Therefore, the present invention provides a kind of method that rubusoside and ursolic acid are extracted from Sweet tea, and step includes:
(1) dry Sweet tea is crushed, 70~85% ethanol waters are added, in 70~80 DEG C of progress 1~3h of refluxing extraction, mistake
Filter, obtains filtrate I and filter residue I, and wherein ethanol water volume L and Sweet tea weight kg is 2~4:1；
(2) filtrate I is concentrated under reduced pressure into medicinal extract, and water dissolution is added, and filtering obtains filtrate II and filter residue II, wherein water body is added
Product L and I volume L ratio of filtrate are 1~2:10；
(3) 0.2~0.8mol/L sodium hydroxide solution is added to 7.2~7.8 in filter residue II, is stirred dissolution, filters, obtain
Filtrate III and filter residue III；
(4) filter residue III is mixed with filter residue I, 4~10% fermentation of Aspergillus niger liquid of addition, and normal temperature fermentation 4~6 days, then plus
Enter 70~85% ethanol waters, in 70~80 DEG C of progress 1~3h of refluxing extraction, filtering obtains filtrate IV and filter residue IV, wherein second
Alcohol solution volume L and Sweet tea weight kg is 2~4:1；
(5) filtrate IV is concentrated under reduced pressure into medicinal extract, and water dissolution is added, and filtering obtains filtrate V and filter residue V, wherein water body is added
Product L and IV volume L ratio of filtrate are 1~2:10；
(6) 0.2~0.8mol/L sodium hydroxide solution is added to 7.2~7.8 in filter residue V, is stirred dissolution, filters, obtain
Filtrate VI；
(7) filtrate II is merged with filtrate V, upper XD-5 resin, 40%~60% ethyl alcohol of three to five times of column volumes of use into
Row elution, collects eluent I, is directly added into 95% ethyl alcohol to final concentration of 75%~85%, refrigerator is stood overnight, and crystallization is tied again
Crystalline substance is primary to obtain rubusoside sterling；
(8) filtrate III is merged with filtrate VI, is adjusted to 5.5~6.5, added two to four times of volume butanol and extracted
It takes, obtains butanol, before immunoassay liquid, be concentrated under reduced pressure into the 1/5~1/15 of original volume, directly upper D4006 macroporous resin column, use four to six
80%~85% ethyl alcohol of times column volume is eluted, and the 1/5~1/10 of original volume is concentrated under reduced pressure into, and stands 15~30min,
It filters 1 time while hot, gained filtrate refrigerator is stood overnight, and is crystallized and is recrystallized primary ursolic acid sterling.
Above-mentioned filtering refers to ceramic membrane filter 1 time.
The temperature of above-mentioned reduced pressure is 40~60 DEG C, is concentrated under reduced pressure using rotary evaporator.
Above-mentioned fermentation of Aspergillus niger liquid can be bought, or self-control obtains -- use common fungus fluid nutrient medium inoculated aspergillus niger
Strain carries out shaking table culture 2~3 days at 28~30 DEG C, wherein the amount L and Sweet tea weight kg of fermentation of Aspergillus niger liquid is added
Than for 4~10:100.
Technical effect
1, the present invention improves dissolution and the transformation efficiency of its functional component by microorganism conversion, while utilizing aspergillus niger
It produces acid and neutralizes alkali process object, be also beneficial to aspergillus niger fast-growth.
2, the present invention is separated rubusoside and ursolic acid using alcohol extracting, again with water-soluble and alkali soluble mode, is chromatographed and is tied through column
Crystalline substance obtains high purity product, effectively prevent rubusoside to destroy under base reagent, improves its yield.
3, it is purified using crystallization and recrystallization, simple process is easily operated, and purification efficiency is high.
Specific embodiment
In the following, the present invention will be further detailed with embodiment, but its any for being not limited to these examples
A or similar example.
Embodiment 1
The dry Sweet tea of 100kg is crushed, 80% ethanol water 300L is added, in 80 DEG C of progress refluxing extraction 2h, filtering is obtained
260L filtrate I and filter residue I；Filtrate I is concentrated under reduced pressure into medicinal extract, and the dissolution of 30L water is added, and filtering obtains filtrate II and filter residue II；Filter residue
II is added 0.5mol/L sodium hydroxide solution to 7.5, is stirred dissolution, filters, obtain filtrate III and filter residue III.
Filter residue III is mixed with filter residue I, and 8L fermentation of Aspergillus niger liquid is added, normal temperature fermentation 4~6 days, adds 85% second
Alcohol solution 300L, in 80 DEG C of progress refluxing extraction 2h, filtering obtains 280L filtrate IV and filter residue IV；Filtrate IV is concentrated under reduced pressure into
Medicinal extract, is added the dissolution of 35L water, and filtering obtains filtrate V and filter residue V；0.5mol/L sodium hydroxide solution is added to 7.5 in filter residue V,
It is stirred dissolution, filters, obtains filtrate VI.
Filtrate II is merged with filtrate V, upper XD-5 resin is eluted with 55% ethyl alcohol of five times of column volumes, and collection is washed
De- liquid I, is directly added into 95% ethyl alcohol, until ethyl alcohol final concentration of 80% in solution, refrigerator is stood overnight, and crystallization recrystallization is primary to be obtained
3.26kg rubusoside sterling；Filtrate III is merged with filtrate VI, is adjusted to 6, four times of volume butanol is added and is extracted, obtain fourth
Alcohol extract liquor is concentrated under reduced pressure into the 1/5~1/15 of original volume, directly upper D4006 macroporous resin column (the erudite beautification industry science in Tianjin
Skill Co., Ltd), it is eluted with 80%~85% ethyl alcohol of six times of column volumes, is concentrated under reduced pressure into the 1/5~1/ of original volume
10,15~30min is stood, is filtered 1 time while hot, gained filtrate refrigerator is stood overnight, and is crystallized and is recrystallized primary 1.83kg bear
Tartaric acid sterling.It is detected through HPLC method, Sweet tea cellulose content is 98.68% in obtained rubusoside sterling, black bearberry in ursolic acid sterling
Acid content is 98.71%.
Comparative examples 1
The dry Sweet tea of 100kg is crushed, 80% ethanol water 300L is added, in 80 DEG C of progress refluxing extraction 2h, filtering is obtained
260L filtrate I and filter residue I；Filter residue I adds 85% ethanol water 300L, and in 80 DEG C of progress refluxing extraction 2h, filtering is obtained
280L filtrate II.
Filtrate I and filtrate II merge, and are concentrated under reduced pressure into medicinal extract, and the dissolution of 60L water is added, and filtering obtains filtrate III and filter residue III；
0.5mol/L sodium hydroxide solution is added to 7.5 in filter residue II, is stirred dissolution, filters, obtain filtrate IV and filter residue IV；Filter residue IV
0.5mol/L sodium hydroxide solution is added to 7.5, is stirred dissolution, filters, obtain filtrate V.
By filtrate III, upper XD-5 resin is eluted with 55% ethyl alcohol of five times of column volumes, collects eluent I, is directly added
Enter 95% ethyl alcohol, until ethyl alcohol final concentration of 80% in solution, refrigerator is stood overnight, and crystallization recrystallization is primary to obtain 2.75kg rubusoside
Sterling；Filtrate V is merged, is adjusted to 6, four times of volume butanol is added and is extracted, obtain butanol, before immunoassay liquid, be concentrated under reduced pressure into
The 1/5~1/15 of original volume, directly upper D4006 macroporous resin column, is washed with 80%~85% ethyl alcohol of six times of column volumes
It is de-, it is concentrated under reduced pressure into the 1/5~1/10 of original volume, stands 15~30min, is filtered 1 time while hot, gained filtrate refrigerator was placed
Night, crystallization with recrystallize primary 1.53kg ursolic acid sterling.It is detected through HPLC method, rubusoside contains in obtained rubusoside sterling
Amount is 98.52%, and ursolic acid content is 98.48% in ursolic acid sterling.
Claims (3)
1. a kind of method for extracting rubusoside and ursolic acid from Sweet tea, step include the following:
(1) dry Sweet tea is crushed, 70~85% ethanol waters are added, in 70~80 DEG C of progress 1~3h of refluxing extraction, is filtered,
Filtrate I and filter residue I are obtained, wherein ethanol water volume L and Sweet tea weight kg is 2~4:1；
(2) filtrate I is concentrated under reduced pressure into medicinal extract, and water dissolution is added, and filtering obtains filtrate II and filter residue II, wherein be added water volume L with
I volume L ratio of filtrate is 1~2:10；
(3) 0.2~0.8mol/L sodium hydroxide solution is added to 7.2~7.8 in filter residue II, is stirred dissolution, filters, obtain filtrate
III and filter residue III；
(4) filter residue III is mixed with filter residue I, and 4~10% fermentation of Aspergillus niger liquid are added, normal temperature fermentation 4~6 days, add 70
~85% ethanol water, in 70~80 DEG C of progress 1~3h of refluxing extraction, filtering obtains filtrate IV and filter residue IV, wherein ethanol water
Liquor capacity L and Sweet tea weight kg is 2~4:1；
(5) filtrate IV is concentrated under reduced pressure into medicinal extract, and water dissolution is added, and filtering obtains filtrate V and filter residue V, wherein water volume L is added
It is 1~2:10 with IV volume L ratio of filtrate；
(6) 0.2~0.8mol/L sodium hydroxide solution is added to 7.2~7.8 in filter residue V, is stirred dissolution, filters, obtain filtrate
Ⅵ；
(7) filtrate II is merged with filtrate V, upper XD-5 resin is washed with 40%~60% ethyl alcohol of three to five times of column volumes
It is de-, eluent I is collected, is directly added into 95% ethyl alcohol to final concentration of 75%~85%, refrigerator is stood overnight, crystallization recrystallization one
It is secondary to obtain rubusoside sterling；
(8) filtrate III is merged with filtrate VI, is adjusted to 5.5~6.5, added two to four times of volume butanol and extracted, obtained
Butanol, before immunoassay liquid is concentrated under reduced pressure into the 1/5~1/15 of original volume, directly upper D4006 macroporous resin column, four to six times of cylinders of use
80%~85% long-pending ethyl alcohol is eluted, and the 1/5~1/10 of original volume is concentrated under reduced pressure into, and stands 15~30min, while hot mistake
Filter 1 time, gained filtrate refrigerator is stood overnight, and is crystallized and is recrystallized primary ursolic acid sterling.
2. the method according to claim 1, the filtering refers to ceramic membrane filter 1 time.
3. the method according to claim 1, the temperature of the reduced pressure is 40~60 DEG C, is depressurized using rotary evaporator
Concentration.
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| CN201811491679.3A CN109503675B (en) | 2018-12-07 | 2018-12-07 | Method for extracting rubusoside and ursolic acid from sweet tea |
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| CN201811491679.3A CN109503675B (en) | 2018-12-07 | 2018-12-07 | Method for extracting rubusoside and ursolic acid from sweet tea |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239983A (en) * | 2011-05-16 | 2011-11-16 | 向华 | Method for extracting rubusoside from sweet tea leaves |
| CN104370981A (en) * | 2014-09-30 | 2015-02-25 | 桂林市一峰食品有限公司 | Method for extraction of rubusoside from sweet tea |
| CN107513094A (en) * | 2017-09-12 | 2017-12-26 | 桂林市产品质量检验所 | A kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea |
| CN108178775A (en) * | 2017-12-28 | 2018-06-19 | 长沙湘资生物科技有限公司 | The method that cape jasmine extracts gardenoside and ursolic acid |
-
2018
- 2018-12-07 CN CN201811491679.3A patent/CN109503675B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239983A (en) * | 2011-05-16 | 2011-11-16 | 向华 | Method for extracting rubusoside from sweet tea leaves |
| CN104370981A (en) * | 2014-09-30 | 2015-02-25 | 桂林市一峰食品有限公司 | Method for extraction of rubusoside from sweet tea |
| CN107513094A (en) * | 2017-09-12 | 2017-12-26 | 桂林市产品质量检验所 | A kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea |
| CN108178775A (en) * | 2017-12-28 | 2018-06-19 | 长沙湘资生物科技有限公司 | The method that cape jasmine extracts gardenoside and ursolic acid |
Non-Patent Citations (4)
| Title |
|---|
| CAMILLE S. BOWEN-FORBES，等: "Ursolic acid analogues: non-phenolic functional food components in Jamaican raspberry fruits", 《FOOD CHEMISTRY》 * |
| YOON SOO CHOI,等: "Triterpenoids from the Roots of Rubus parvifolius", 《ARCH.PHARM.RES.》 * |
| 刘敬，等: "近5年悬钩子属植物化学成分研究进展", 《时珍国医国药》 * |
| 谭冬明,等: "广西甜茶化学成分、提取工艺及生物活性研究进展", 《中成药》 * |
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