CN107513094B - A kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea - Google Patents
A kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea Download PDFInfo
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- CN107513094B CN107513094B CN201710817875.4A CN201710817875A CN107513094B CN 107513094 B CN107513094 B CN 107513094B CN 201710817875 A CN201710817875 A CN 201710817875A CN 107513094 B CN107513094 B CN 107513094B
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- acid
- oleanolic acid
- ursolic acid
- medicinal extract
- oleanolic
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- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 title claims abstract description 56
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 title claims abstract description 56
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 title claims abstract description 56
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 title claims abstract description 56
- 229940100243 oleanolic acid Drugs 0.000 title claims abstract description 56
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 title claims abstract description 56
- WCGUUGGRBIKTOS-GPOJBZKASA-N (3beta)-3-hydroxyurs-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C WCGUUGGRBIKTOS-GPOJBZKASA-N 0.000 title claims abstract description 54
- 229940096998 ursolic acid Drugs 0.000 title claims abstract description 54
- PLSAJKYPRJGMHO-UHFFFAOYSA-N ursolic acid Natural products CC1CCC2(CCC3(C)C(C=CC4C5(C)CCC(O)C(C)(C)C5CCC34C)C2C1C)C(=O)O PLSAJKYPRJGMHO-UHFFFAOYSA-N 0.000 title claims abstract description 54
- 238000000605 extraction Methods 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241001122767 Theaceae Species 0.000 title claims abstract description 22
- 235000009508 confectionery Nutrition 0.000 title claims abstract description 21
- 238000000746 purification Methods 0.000 title claims abstract description 15
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 44
- 239000000284 extract Substances 0.000 claims abstract description 27
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 18
- 239000013049 sediment Substances 0.000 claims abstract description 10
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 9
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 9
- 229960000935 dehydrated alcohol Drugs 0.000 claims abstract description 6
- 238000011084 recovery Methods 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 238000001514 detection method Methods 0.000 claims description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- 239000000706 filtrate Substances 0.000 claims description 9
- 239000012141 concentrate Substances 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 7
- 239000003480 eluent Substances 0.000 claims description 6
- 229960004756 ethanol Drugs 0.000 claims description 6
- 239000003208 petroleum Substances 0.000 claims description 5
- 238000001556 precipitation Methods 0.000 claims description 5
- 238000005057 refrigeration Methods 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 239000000779 smoke Substances 0.000 claims description 4
- 238000009835 boiling Methods 0.000 claims description 3
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 claims 1
- 239000000460 chlorine Substances 0.000 claims 1
- 229910052801 chlorine Inorganic materials 0.000 claims 1
- 238000005259 measurement Methods 0.000 claims 1
- 238000003556 assay Methods 0.000 abstract description 4
- 238000000926 separation method Methods 0.000 abstract description 4
- 241000196324 Embryophyta Species 0.000 abstract description 3
- 239000002253 acid Substances 0.000 abstract description 3
- 239000003513 alkali Substances 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 18
- 235000019441 ethanol Nutrition 0.000 description 6
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 235000002906 tartaric acid Nutrition 0.000 description 3
- 239000011975 tartaric acid Substances 0.000 description 3
- 241000830535 Ligustrum lucidum Species 0.000 description 2
- 241001092459 Rubus Species 0.000 description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 241001656831 Arctous alpina Species 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- YWPVROCHNBYFTP-UHFFFAOYSA-N Rubusoside Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC1OC(CO)C(O)C(O)C1O YWPVROCHNBYFTP-UHFFFAOYSA-N 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000767 anti-ulcer Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- QEVHRUUCFGRFIF-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C(C5=CC=C(OC)C=C5N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 QEVHRUUCFGRFIF-MDEJGZGSSA-N 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- YWPVROCHNBYFTP-OSHKXICASA-N rubusoside Chemical compound O([C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YWPVROCHNBYFTP-OSHKXICASA-N 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The process of the invention discloses a kind of from Sweet tea extraction purification oleanolic acid and ursolic acid, including using dehydrated alcohol ultrasonic wave extraction, filter；Extracting solution after concentration merges obtains medicinal extract, and resulting medicinal extract is dissolved with chloroform, and brown medicinal extract is obtained after concentration and recovery chloroform；Medicinal extract is heavy with 5% sodium hydroxide alkali soluble acid, the methods of sediment silica gel column chromatography separation obtains the oleanolic acid and ursolic acid of high-purity, assay is carried out using high performance liquid chromatography, the purity of oleanolic acid and ursolic acid is 90% or more, the yield of oleanolic acid is in 20mg/100g or more, and the yield of ursolic acid is in 90mg/100g or more.Present invention process method, required equipment is simple and convenient to operate, extraction time is short, extraction efficiency is high, energy saving, saving raw material, mild condition, not the features such as not destroying active component, a kind of feasibility source is provided to extract oleanolic acid and ursolic acid from natural plants, the added value of Sweet tea is improved, there is important application value and economic and social benefits.
Description
Technical field
The present invention relates to effective ingredients in plant method for extraction and purification, specifically a kind of extraction purification oleanolic acid from Sweet tea
With the process of ursolic acid.
Background technique
Guangxi Folium hydrangeae strigosae (Rubus Suavissimus S.Lee) is rose family rubus, is that the distinctive sweet taste in Guangxi is planted
Object is often used as tea-drinking civil, containing main actives such as rubusoside, flavones, tea polyphenols, has relieving heat and thirst, kidney tonifying, drop
Blood pressure and treatment diabetes and other effects.Document report is identified in Sweet tea by extracting separation and method of spectroscopy and contains oleanolic acid
And ursolic acid.Modern research shows that oleanolic acid has AntiHIV1 RT activity, antibacterial, anticancer, antiulcer, treatment osteoporosis etc. extensively
Pharmacological action and bioactivity, have been used to clinic as hepatic.Ursolic acid has antitumor, protect liver, angiocarpy, sugar
The multiple biological activities such as sick, anti-inflammatory, antiviral are urinated, extensive research is at home and abroad received.Oleanolic acid and ursolic acid market
Demand is big, artificial synthesized more complex, still relies on to extract from natural plants at present and obtains.Therefore, to oleanolic acid in Sweet tea
And the research of ursolic acid extraction and purification process has great importance.Through Literature Consult, oleanolic acid and ursolic acid are mentioned in Sweet tea
The research of purifying process is taken to have not been reported.The present invention is using ultrasonic wave extraction and silica gel column chromatography separation method to pier neat in Sweet tea
Tartaric acid and ursolic acid extract and purifying process is studied, and lays the foundation for the comprehensive utilization of Sweet tea.
Summary of the invention
The work of the technical problem to be solved in the present invention is to provide a kind of from Sweet tea extraction purification oleanolic acid and ursolic acid
Process.
Present invention provide the technical scheme that
A kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea, includes the following steps:
(1) take Sweet tea sample to smash to pieces, weigh the sample after smashing to pieces, be added the dehydrated alcohols of 2~4 times of its weight amounts 25~
10~20min of ultrasonic wave extraction at 40 DEG C, extracting solution filter；
The filtered residue of ethanol washing is used again, then is filtered, and smoke filtrate merges with the extracting solution of first time, after concentration merges
Extracting solution obtain medicinal extract, resulting medicinal extract is dissolved with chloroform, after concentration and recovery chloroform brown medicinal extract；
(2) medicinal extract that step (1) obtains is boiled with 5% sodium hydroxide solution, is filtered while hot, filtrate is placed in refrigerator 5
~10 DEG C of refrigeration 1h are filtered, are washed with water to neutrality；
Filtered residue adds boiling to boil, and being adjusted to pH with hydrochloric acid is 1~2, lets cool precipitation, filters, wash precipitating with hot water
Object；
Sediment is separated with silica gel column chromatography, with petroleum ether: chloroform=1:2 is eluted, and TLC detection discards eluent；Again
With chloroform: ethyl acetate=18:1 elution, every 100mL collect a, 30 parts of collection, and TLC detection merges 5-15 parts, concentration elution
Liquid obtains high purity oleanolic acid sample；
Merge 17-30 parts, concentrate eluant obtains high-purity ursolic acid sample；
By resulting oleanolic acid and ursolic acid sample, dry at 101 DEG C~105 DEG C to constant weight respectively；
(3) oleanolic acid and ursolic acid sample by step (2) purifying after dry, is contained using high performance liquid chromatography
It is fixed to measure, and the purity of oleanolic acid and ursolic acid is 90% or more, and the yield of oleanolic acid is in 20mg/100g or more, ursolic acid
Yield in 90mg/100g or more.
" one kind extracts oleanolic acid side from the fruit of glossy privet for the present invention and the patent of invention of 105348364 B of Publication No. CN
Method " and master's thesis " research of the chemical component of Guangxi Folium hydrangeae strigosae leaf " compare, have the advantages that certain.It is " a kind of from glossy privet
Oleanolic acid method is extracted in son " it is slightly purified after ultrasonic wave extraction using the separation elution of D101 macroreticular resin in patent, then
Be further purified using Sephadex LH-20 gel column, purification process is complicated for operation, time-consuming, consumption reagent, and because with the present invention
Raw material is different, can only extraction purification oleanolic acid, do not include ursolic acid.Master's thesis " chemical component of Guangxi Folium hydrangeae strigosae leaf
Research " in be heated to reflux mode using chloroform and extract oleanolic acid and ursolic acid, extraction time is long, and energy consumption is high, low efficiency.It mentions
It is not purified slightly using straightforward procedure after taking, and uses silica gel column chromatography repeatedly with different organic solvent mixing eluent systems
It is isolated and purified, it is same complicated for operation, it is time-consuming, and consume a large amount of organic solvents, uneconomical environmental protection.The unjustified pier tartaric acid of the method
And the purity and extraction yield of ursolic acid are examined, the oleanolic acid and ursolic acid of purifying are only used for Components identification.The present invention
It is extracted using ultrasonic wave, is slightly purified using straightforward procedures such as the heavy, water washing and precipitatings of alkali soluble acid, pass through silica gel column chromatography point
It is studied from the oleanolic acid and ursolic acid for respectively obtaining high-purity, and to purity and extraction yield, oleanolic acid and bear
The purity of tartaric acid 90% or more, the yield of oleanolic acid in 20mg/100g or more, the yield of ursolic acid 90mg/100g with
On.Present invention process is simple, easy to operate, purification process is simple, and extraction time is short, extraction efficiency is high, mild condition, does not destroy
Active component, it is easily operated.
Detailed description of the invention
Fig. 1 is the HPLC figure of embodiment oleanolic acid, ursolic acid standard solution；
Fig. 2 is the HPLC figure of oleanolic acid sample after embodiment purifying is dry；
Fig. 3 is the HPLC figure of ursolic acid sample after embodiment purifying is dry.
In figure, 1. be oleanolic acid 2. be ursolic acid.
Specific embodiment
The content of present invention is further elaborated below with reference to embodiment and attached drawing, but not as to limit of the invention
It is fixed.
Embodiment 1
The process of extraction purification oleanolic acid and ursolic acid, includes the following steps: from Sweet tea
(1) it takes Sweet tea sample to be smashed to pieces with bruisher, weighs sample 150g, the dehydrated alcohol of 2 times of its weight amounts is added 25
Ultrasonic wave extraction 10min at DEG C, extracting solution filter；It again with the filtered residue of 300 mL ethanol washings 1 time, then filters, filters
Liquid merges with the extracting solution of first time, and the extracting solution after concentration merges obtains medicinal extract, and resulting medicinal extract 30mL chloroform dissolves, concentration
Brown medicinal extract is obtained after recycling chloroform；
(2) medicinal extract obtained adds 30mL5% sodium hydroxide solution to boil 10min, filters while hot, and filtrate is placed in refrigerator and is existed
In 5~10 DEG C of refrigeration 1h, filter, be washed with water to neutrality；Filtered residue adds 30 mL boilings to boil 10min, is adjusted to pH with hydrochloric acid
It is 1, lets cool precipitation, filters, with hot water washing sediment 3 times；Sediment is separated with silica gel column chromatography, with petroleum ether: chloroform=
1:2 elutes 1000mL, and TLC detection discards eluent；Use chloroform again: ethyl acetate=18:1 elution, every 100mL collect portion,
30 parts are collected, TLC detection merges 5-15 parts, concentrate eluant obtains high purity oleanolic acid sample；Merge 17-30 parts, concentration is washed
De- liquid obtains high-purity ursolic acid sample；By resulting oleanolic acid and ursolic acid sample, dry at 101 DEG C to constant weight respectively；
(3) oleanolic acid and ursolic acid sample after purifying is dry carry out assay using high performance liquid chromatography.
Chromatographic condition is as follows:
Chromatographic column: the silent winged generation that Hypersil Gold C of match18(250 mm × 4.6 mm, 5 μm), mobile phase are methanol-
0.2% ammonium acetate solution (83:17), flow velocity: 0.6 mL/min, column temperature: 30 DEG C, Detection wavelength: 210 nm, sampling volume: 20
μ L, spectral scanning range: the nm of 190 nm~600.
0.1110 g of standard substance oleanolic acid is accurately weighed respectively, 0.0950 g of ursolic acid is placed in 50mL volumetric flask,
Scale is dissolved and is settled to ethyl alcohol, and concentration is respectively 2.220,1.900mg/mL.Accurate mark for measuring certain volume respectively
Quasi- mixed liquor with 70% ethanol water (volume ratio) be diluted to step by step various concentration series, oleanolic acid concentration be 2.22,4.44,
11.1,22.2,44.4,111 μ g/mL, black bearberry acid concentration are 1.90,3.80,9.50,19.0,38.0,95.0 μ g/mL, are drawn
Standard curve.Oleanolic acid and each 20mg of ursolic acid sample after accurately weighing the purifying drying of embodiment 1 respectively, use dehydrated alcohol
Dissolution is settled to 25.0 mL, is measured after taking the solution to dilute 20 times with 70% ethanol water according to above-mentioned liquid phase chromatogram condition,
Use external standard method with its purity of calculated by peak area.
Through detecting, oleanolic acid, the linear relationship of ursolic acid are good, and the purity of oleanolic acid is 93.3%, and yield is
23.3mg/100g；The purity of ursolic acid is 94.2%, yield 94.9mg/100g.
Embodiment 2
The process of extraction purification oleanolic acid and ursolic acid, includes the following steps: from Sweet tea
(1) it takes Sweet tea sample to be smashed to pieces with bruisher, weighs sample 150g, the dehydrated alcohol of 3 times of amount weight is added at 35 DEG C
Lower ultrasonic wave extraction 15min, extracting solution filter, then with the filtered residue of 300 mL ethanol washings 1 time, then filter, smoke filtrate
Merge with the extracting solution of first time, the extracting solution after concentration merges obtains medicinal extract, and resulting medicinal extract 30mL chloroform is dissolved, is concentrated back
Brown medicinal extract is obtained after receiving chloroform；
(2) medicinal extract obtained adds 30mL5% sodium hydroxide solution to boil 10min, filters while hot, and filtrate is placed in refrigerator and is existed
In 5~10 DEG C of refrigeration 1h, filter, be washed with water to neutrality；Filtered residue adds the heating of 30 mL water to boil 10min, with hydrochloric acid tune
It is 2 to pH, lets cool precipitation, filters, with hot water washing sediment 3 times；Sediment is separated with silica gel column chromatography, with petroleum ether:
Chloroform=1:2 elutes 1000mL, and TLC detection discards eluent；Use chloroform again: ethyl acetate=18:1 elution, every 100mL are collected
Portion, collects 30 parts, and TLC detection merges 5-15 parts, concentrate eluant obtains high purity oleanolic acid sample；Merge 17-30 parts,
Concentrate eluant obtains high-purity ursolic acid sample；By resulting oleanolic acid and ursolic acid sample, dried extremely at 103 DEG C respectively
Constant weight；
(3) oleanolic acid and ursolic acid sample after purifying is dry are carried out according to the high performance liquid chromatography in embodiment 1
Assay.
Through detecting, the purity of oleanolic acid is 93.1%, yield 24.1mg/100g；The purity of ursolic acid is 94.1%, is obtained
Rate is 94.0mg/100g.
Embodiment 3
The process of extraction purification oleanolic acid and ursolic acid, includes the following steps: from Sweet tea
(1) it takes Sweet tea sample to be smashed to pieces with bruisher, weighs sample 150g, the dehydrated alcohol of 4 times of amount weight is added at 40 DEG C
Lower ultrasonic wave extraction 20min, extracting solution filter, then with the filtered residue of 300 mL ethanol washings 1 time, then filter, smoke filtrate
Merge with the extracting solution of first time, the extracting solution after concentration merges obtains medicinal extract, and resulting medicinal extract 30mL chloroform is dissolved, is concentrated back
Brown medicinal extract is obtained after receiving chloroform；·
(2) medicinal extract obtained adds 30mL5% sodium hydroxide solution to boil 10min, filters while hot, and filtrate is placed in refrigerator and is existed
In 5~10 DEG C of refrigeration 1h, filter, be washed with water to neutrality；Filtered residue adds the heating of 30 mL water to boil 10min, with hydrochloric acid tune
It is 2 to pH, lets cool precipitation, filters, with hot water washing sediment 3 times；Sediment is separated with silica gel column chromatography, with petroleum ether:
Chloroform=1:2 elutes 1000mL, and TLC detection discards eluent；Use chloroform again: ethyl acetate=18:1 elution, every 100mL are collected
Portion, collects 30 parts, and TLC detection merges 5-15 parts, concentrate eluant obtains high purity oleanolic acid sample；Merge 17-30 parts,
Concentrate eluant obtains high-purity ursolic acid sample；By resulting oleanolic acid and ursolic acid sample, done at 105 DEG C 1 respectively
It is dry to constant weight；
(3) oleanolic acid and ursolic acid sample after purifying is dry are carried out according to the high performance liquid chromatography in embodiment 1
Assay.
Through detecting, the purity of oleanolic acid is 93.0%, yield 22.8mg/100g；The purity of ursolic acid is 94.4%, is obtained
Rate is 93.6mg/100g.
Claims (1)
1. a kind of process of extraction purification oleanolic acid and ursolic acid from Sweet tea, which comprises the steps of:
(1) it takes Sweet tea sample to smash to pieces, weighs the sample after smashing to pieces, the dehydrated alcohol of 2~4 times of its weight amounts is added at 25~40 DEG C
10~20min of lower ultrasonic wave extraction, extracting solution filter；
The filtered residue of ethanol washing is used again, then is filtered, and smoke filtrate merges with the extracting solution of first time, mentioning after concentration merging
Liquid is taken to obtain medicinal extract, resulting medicinal extract is dissolved with chloroform, and brown medicinal extract is obtained after concentration and recovery chloroform；
(2) medicinal extract that step (1) obtains is boiled with 5% sodium hydroxide solution, is filtered while hot, filtrate is placed in refrigerator 5~10
DEG C refrigeration 1h, filter, be washed with water to neutrality；
Filtered residue adds boiling to boil, and being adjusted to pH with hydrochloric acid is 1~2, lets cool precipitation, filters, use hot water washing sediment；
Sediment is separated with silica gel column chromatography, with petroleum ether: chloroform=1:2 is eluted, and TLC detection discards eluent；Chlorine is used again
Imitative: ethyl acetate=18:1 elution, every 100mL collect a, 30 parts of collection, and TLC detection merges 5-15 parts, concentrate eluant obtains
Oleanolic acid sample；
Merge 17-30 parts, concentrate eluant obtains ursolic acid sample；
By resulting oleanolic acid and ursolic acid sample, dry at 101 DEG C~105 DEG C to constant weight respectively；
(3) oleanolic acid and ursolic acid sample by step (2) purifying after dry, is carried out using high performance liquid chromatography containing measurement
Fixed, the purity of oleanolic acid and ursolic acid is 90% or more, and in 20mg/100g or more, ursolic acid obtains the yield of oleanolic acid
Rate is in 90mg/100g or more.
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| CN105218616A (en) * | 2015-10-23 | 2016-01-06 | 山东省果树研究所 | A kind of method extracting ursolic acid and Oleanolic Acid from Blueberry |
| CN105232679A (en) * | 2015-11-11 | 2016-01-13 | 北京林业大学 | Method for simultaneously extracting ursolic acid and oleanolic acid in hawthorn by means of subcritical water |
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| CN105218616A (en) * | 2015-10-23 | 2016-01-06 | 山东省果树研究所 | A kind of method extracting ursolic acid and Oleanolic Acid from Blueberry |
| CN105232679A (en) * | 2015-11-11 | 2016-01-13 | 北京林业大学 | Method for simultaneously extracting ursolic acid and oleanolic acid in hawthorn by means of subcritical water |
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| Title |
|---|
| 广西甜茶叶的化学成分研究;谭冬明;《广西师范大学硕士学位论文》;20111215;全文 |
| 甜茶中齐墩果酸及熊果酸的检测方法的建立;谭冬明等;《食品研究与开发》;20170720;第38卷(第14期);全文 |
| 甜茶果实化学成分的研究;罗达伟;《广西师范大学硕士学位论文》;20110115;全文 |
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