CN102229638A - Method for extracting oleanolic acid from chaenomeles fruit and preparing oleanolic acid standard - Google Patents
Method for extracting oleanolic acid from chaenomeles fruit and preparing oleanolic acid standard Download PDFInfo
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- CN102229638A CN102229638A CN 201110104760 CN201110104760A CN102229638A CN 102229638 A CN102229638 A CN 102229638A CN 201110104760 CN201110104760 CN 201110104760 CN 201110104760 A CN201110104760 A CN 201110104760A CN 102229638 A CN102229638 A CN 102229638A
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- oleanolic acid
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- fructus chaenomelis
- methanol
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- 229940100243 oleanolic acid Drugs 0.000 title claims abstract description 45
- 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 44
- 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 44
- 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 44
- 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 44
- 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 44
- 235000013399 edible fruits Nutrition 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 22
- 241001507936 Chaenomeles Species 0.000 title abstract 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 13
- 238000000605 extraction Methods 0.000 claims abstract description 12
- 239000000126 substance Substances 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000000284 extract Substances 0.000 claims abstract description 10
- 239000000843 powder Substances 0.000 claims abstract description 10
- 238000000746 purification Methods 0.000 claims abstract description 9
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000000287 crude extract Substances 0.000 claims abstract description 7
- 239000011347 resin Substances 0.000 claims abstract description 6
- 229920005989 resin Polymers 0.000 claims abstract description 6
- 239000007864 aqueous solution Substances 0.000 claims abstract description 4
- 229920006395 saturated elastomer Polymers 0.000 claims abstract description 4
- 238000004140 cleaning Methods 0.000 claims abstract description 3
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 42
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 238000004237 preparative chromatography Methods 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 239000012141 concentrate Substances 0.000 claims description 6
- 239000012043 crude product Substances 0.000 claims description 6
- 239000013078 crystal Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 5
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical group O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 5
- 238000000926 separation method Methods 0.000 claims description 5
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- 239000012535 impurity Substances 0.000 claims description 4
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 4
- 229930182490 saponin Natural products 0.000 claims description 4
- 150000007949 saponins Chemical class 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 239000012044 organic layer Substances 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 230000003068 static effect Effects 0.000 claims description 3
- 238000002137 ultrasound extraction Methods 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 238000005374 membrane filtration Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 abstract description 2
- 239000012488 sample solution Substances 0.000 abstract 2
- 238000013375 chromatographic separation Methods 0.000 abstract 1
- 238000001728 nano-filtration Methods 0.000 abstract 1
- 238000007670 refining Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 238000011161 development Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- -1 (SOD) Substances 0.000 description 1
- 244000189799 Asimina triloba Species 0.000 description 1
- 235000006264 Asimina triloba Nutrition 0.000 description 1
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 235000009467 Carica papaya Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- JLPDBLFIVFSOCC-UHFFFAOYSA-N Oleandrin Natural products O1C(C)C(O)C(OC)CC1OC1CC(CCC2C3(CC(C(C3(C)CCC32)C=2COC(=O)C=2)OC(C)=O)O)C3(C)CC1 JLPDBLFIVFSOCC-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003260 anti-sepsis Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000000496 cardiotonic agent Substances 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 1
- 238000001914 filtration 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
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 208000021822 hypotensive Diseases 0.000 description 1
- 230000001077 hypotensive effect Effects 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- JLPDBLFIVFSOCC-XYXFTTADSA-N oleandrin Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1C[C@@H](CC[C@H]2[C@]3(C[C@@H]([C@@H]([C@@]3(C)CC[C@H]32)C=2COC(=O)C=2)OC(C)=O)O)[C@]3(C)CC1 JLPDBLFIVFSOCC-XYXFTTADSA-N 0.000 description 1
- 229950010050 oleandrin Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 229940032362 superoxide dismutase Drugs 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Landscapes
- Steroid Compounds (AREA)
Abstract
The invention discloses a method for extracting oleanolic acid from chaenomeles fruit and preparing an oleanolic acid standard. The method comprises the following steps of: cleaning the chaenomeles fruit, drying the chaenomeles fruit in the shade, crushing the chaenomeles fruit into powder, dissolving the powder by using ethanol, performing ultrasonic aided extraction to obtain a crude extract, adding the crude extract into water for dissolving, adding saturated aqueous solution of n-butanol, performing extraction for many times, concentrating the extract to obtain an extract substance, dissolving the extract substance, performing nano-filtration to obtain sample solution of the oleanolic acid, and performing preliminary purification of macroporous resin, silica gel column chromatography, high performance liquid chromatography (HPLC), chromatographic separation and refining on the obtained sample solution to obtain the oleanolic acid standard. The method is simple and convenient in operation, and the oleanolic acid has high yield and high purity and can be made into the standard.
Description
Technical background
The invention belongs to Separation of Natural Products purifying field, be specifically related to a kind of method of from the Fructus Chaenomelis fruit, extracting Oleanolic Acid and preparing the Oleanolic Acid standard substance.
Background technology
Fructus Chaenomelis is the famous special product in Anhui, is celebrated with " hundred benefits are fruit ", is one of 30 dietotherapeutic kinds of Ministry of Health's announcement in 2003.Fructus Chaenomelis is rich in flavonoid, saponin, amino acid, superoxide-dismutase multiple biologically active substances such as (SOD), and various trace elements such as potassium, calcium, magnesium, zinc, iron, all have wide practical use in fields such as medicochemistry, vegetable chemistry, bio-pharmaceuticals, health care and green foods.
(oleanolic acid, OA) another name oleanolic acid, celebrating four elements etc. are pentacyclic triterpene acid to Oleanolic Acid, belong to oleandrin, are physiologically active substances main in the Fructus Chaenomelis.Correlative study at present concentrates on the extraction of rutin in the Fructus Chaenomelis, flavones, trace element mostly and measures.Oleanolic Acid is complicated because of structure, and synthetic is loaded down with trivial details, and main at present the dependence extracted from plant.Therefore, research separates Oleanolic Acid in the Fructus Chaenomelis, and important application value is arranged.
Oleanolic Acid has and protects the liver, protects outside stomach, cardiac stimulant, hypoglycemic, lipopenicillinase, the hypotensive biological activity, also have effects such as antisepsis and anti-inflammation, anti-oxidant, immunomodulatory, inhibition platelet aggregation, has multiple pharmacological effect, and certain antitumor, antitumour activity are arranged, be a kind of natural active matter that exploitation is worth that has.
Technology and the research approaching with the present invention are pawpaw (with the Fructus Chaenomelis composition, the purposes difference) mostly.Having only one piece about the Oleanolic Acid assay aspect the research of Fructus Chaenomelis Oleanolic Acid, leaching process is wherein arranged, but mainly be needed step in the mensuration process, not to extract and produce product, so, there is essential difference, and the extraction process complex steps, products obtained therefrom---Oleanolic Acid purity is not high.
Summary of the invention
The invention discloses a kind of method of from the Fructus Chaenomelis fruit, extracting Oleanolic Acid and preparing the Oleanolic Acid standard substance.The inventive method is extracted Oleanolic Acid, and technology is simple, the yield height of Oleanolic Acid, good stability; Oleanolic Acid purity behind the purifying is very high, and the finished product purity reaches more than 97.50%.
Purpose of the present invention can realize by following technical measures:
A kind of method of extracting Oleanolic Acid from the Fructus Chaenomelis fruit is characterized in that: specifically may further comprise the steps:
(1) will 40 ℃ of-60 ℃ of following vacuum-dryings, and then pulverize through after Fructus Chaenomelis fruit, the cleaning of screening, cutting into slices, dry in the shade, cross the 40-45 mesh sieve, Fructus Chaenomelis fruit powder;
(2) Fructus Chaenomelis fruit powder is joined in the ethanolic soln, under 30 ℃-50 ℃, ultrasonic-assisted extraction 30-50min, after multiple extraction, with the extracting solution vacuum concentration, get crude extract, and reclaim solvent, the weight ratio of Fructus Chaenomelis fruit powder and ethanolic soln is 1: (10-15), the concentration of ethanolic soln is 60-85%;
(3) the gained crude extract is added water miscible after, the saturated aqueous solution extraction that adds the equivalent propyl carbinol more repeatedly, collected organic layer and vacuum concentration, medicinal extract shape material, reclaim propyl carbinol simultaneously;
(4) step (3) gained is got medicinal extract shape material dissolve with methanol,, promptly get the Oleanolic Acid sample liquid through membrane filtration.
A kind of Oleanolic Acid that utilizes claim 1 to extract prepares the method for Oleanolic Acid standard substance, it is characterized in that: specifically may further comprise the steps:
(1) it is concentrated that the macroporous resin preliminary purification is handled the Oleanolic Acid sample liquid that claim 1 is made, use 95% dissolve with ethanol, with sample on the 0.9-1.1mL/min flow velocity, behind static 3～4h, earlier with distilled water flush away water-soluble impurity, use 20%, 50%, 95% ethanolic soln wash-out more respectively, elution flow rate is 1.5mL/min, collects 95% ethanol eluate, reclaims ethanol, evaporate to dryness promptly gets the saponin crude product;
(2) silica gel column chromatography will be after the crude product of macroporous resin chromatography preliminary purification be with dissolve with methanol upper prop, and with 50% and 95% methanol-eluted fractions, the Fractional Collections elutriant concentrates evaporate to dryness with 95% meoh eluate, promptly gets white, needle-shaped crystals;
(3) white, needle-shaped crystals is dissolved in the methyl alcohol, with HPLC preparative chromatography separation and purification, be in charge of the collection elutriant by preparative chromatography figure appearance time, HPLC is detected purity to be merged greater than stream part of 95%, nitrogen is blown away neat solvent, promptly get the Oleanolic Acid standard substance, the moving phase that HPLC detects purity is acetonitrile: water=95: 5 (V: V); Detect wavelength: 210nm; Column temperature: 25 ℃.
The described Oleanolic Acid that utilizes claim to extract prepares the method for Oleanolic Acid standard substance, and it is characterized in that: the condition of the described HPLC preparative chromatography of step (3) is: elutriant is a methanol-water, and the volume ratio of methyl alcohol and water is 85: 15 in the methanol-water; Volumetric flow rate: 10mL/min; Detect wavelength: 210nm; Column temperature: 25 ℃; Sample size: 1.5mL.
The invention has the advantages that:
(1) the inventive method, extraction and separation process is simple and tight, and method is reliable;
(2) the inventive method, the yield height of extraction Oleanolic Acid, good stability;
(3) the inventive method, the purity height of the Oleanolic Acid of extraction reaches more than 97.50%.
(4) utilize the Fructus Chaenomelis fruit to extract Oleanolic Acid for raw material, aboundresources can deep development Fructus Chaenomelis comprehensive utilization value.
The present invention is a raw material with the Fructus Chaenomelis fruit, and for comprehensive utilization Fructus Chaenomelis resource provides theoretical and practical application foundation, the present invention will be to promoting the Fructus Chaenomelis industry development, aspects such as Fructus Chaenomelis resource deep development being had positive effect behind industrializing implementation.
Embodiment
Be example with the laboratory implementation now, non-limiting examples is described below:
After screening, clean, cut into slices, drying in the shade, 50 ℃ of vacuum-dryings are pulverized, and cross 40 mesh sieves (aperture 0.425mm) with the Fructus Chaenomelis fruit; Add in the raw material powder and be equivalent to raw material powder weight 10-15 60-85% ethanol doubly, 30 ℃-50 ℃, ultrasonic-assisted extraction 30-50min under the 300W power condition after extracted twice, reclaims solvent with the extracting solution vacuum concentration and gets crude extract.Gained medicinal extract is added the saturated aqueous solution extraction three times that water adds the equivalent propyl carbinol after miscible again, organic layer is merged final vacuum concentrate, reclaim propyl carbinol, concentrate medicinal extract shape material dissolve with methanol, shake up, through 0.45 μ m filtering with microporous membrane, promptly get sample liquid.Sample liquid is used 95% dissolve with ethanol after concentrating, and with sample on the 1.0mL/min flow velocity, behind static 3～4h, with distilled water flush away water-soluble impurity, uses 20%, 50%, 95% ethanolic soln wash-out more respectively earlier, and elution flow rate is 1.5mL/min.Collect 95% ethanol eluate, reclaim ethanol, evaporate to dryness promptly gets the saponin crude product; With C
18After silica gel fully soaks the degassing with methyl alcohol, settling methods dress post.After fully washing out impurity with methyl alcohol, use 50-70% acetonitrile solution balance again.Will be through the crude product of macroporous resin chromatography preliminary purification, be dissolved in upper prop behind the methyl alcohol, and with 50% and 95% methanol-eluted fractions, the chromatography process is carried out at normal temperatures and pressures; The Fractional Collections elutriant concentrates evaporate to dryness with 95% meoh eluate, promptly gets white, needle-shaped crystals.White, needle-shaped crystals is dissolved in methyl alcohol, and (HPLC preparative chromatography condition is methanol-water (85: 15) with HPLC preparative chromatography separation and purification; Volumetric flow rate: 10mL/min; Detect wavelength: 210nm; Column temperature: 25 ℃; Sample size: 1.5mL), be in charge of the collection elutriant by preparative chromatography figure appearance time, (the HPLC testing conditions is an acetonitrile: water=95: 5 (V: V) be moving phase with the HPLC detection; Detect wavelength: 210nm; Column temperature: 25 ℃) purity merges greater than stream part of 95%, and nitrogen is blown away neat solvent, promptly gets the Oleanolic Acid standard substance, and recording purity is 97.50%, and productive rate is 1.23mg/g.
Claims (3)
1. method of extracting Oleanolic Acid from the Fructus Chaenomelis fruit is characterized in that: specifically may further comprise the steps:
(1) will 40 ℃ of-60 ℃ of following vacuum-dryings, and then pulverize through after Fructus Chaenomelis fruit, the cleaning of screening, cutting into slices, dry in the shade, cross the 40-45 mesh sieve, Fructus Chaenomelis fruit powder;
(2) Fructus Chaenomelis fruit powder is joined in the ethanolic soln, under 30 ℃-50 ℃, ultrasonic-assisted extraction 30-50min, after multiple extraction, with the extracting solution vacuum concentration, get crude extract, and reclaim solvent, the weight ratio of Fructus Chaenomelis fruit powder and ethanolic soln is 1: (10-15), the concentration of ethanolic soln is 60-85%;
(3) the gained crude extract is added water miscible after, the saturated aqueous solution extraction that adds the equivalent propyl carbinol more repeatedly, collected organic layer and vacuum concentration, medicinal extract shape material, reclaim propyl carbinol simultaneously;
(4) step (3) gained is got medicinal extract shape material dissolve with methanol,, promptly get the Oleanolic Acid sample liquid through membrane filtration.
2. an Oleanolic Acid that utilizes claim 1 to extract prepares the method for Oleanolic Acid standard substance, it is characterized in that: specifically may further comprise the steps:
(1) it is concentrated that the macroporous resin preliminary purification is handled the Oleanolic Acid sample liquid that claim 1 is made, use 95% dissolve with ethanol, with sample on the 0.9-1.1mL/min flow velocity, behind static 3～4h, earlier with distilled water flush away water-soluble impurity, use 20%, 50%, 95% ethanolic soln wash-out more respectively, elution flow rate is 1.5mL/min, collects 95% ethanol eluate, reclaims ethanol, evaporate to dryness promptly gets the saponin crude product;
(2) silica gel column chromatography will be after the crude product of macroporous resin chromatography preliminary purification be with dissolve with methanol upper prop, and with 50% and 95% methanol-eluted fractions, the Fractional Collections elutriant concentrates evaporate to dryness with 95% meoh eluate, promptly gets white, needle-shaped crystals;
(3) white, needle-shaped crystals is dissolved in the methyl alcohol, with HPLC preparative chromatography separation and purification, be in charge of the collection elutriant by preparative chromatography figure appearance time, HPLC is detected purity to be merged greater than stream part of 95%, nitrogen is blown away neat solvent, promptly get the Oleanolic Acid standard substance, the moving phase that HPLC detects purity is acetonitrile: water=95: 5 (V: V); Detect wavelength: 210nm; Column temperature: 25 ℃.
3. the Oleanolic Acid that utilizes claim to extract according to claim 2 prepares the method for Oleanolic Acid standard substance, it is characterized in that: the condition of the described HPLC preparative chromatography of step (3) is: elutriant is a methanol-water, and the volume ratio of methyl alcohol and water is 85: 15 in the methanol-water; Volumetric flow rate: 10mL/min; Detect wavelength: 210nm; Column temperature: 25 ℃; Sample size: 1.5mL.
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| CN 201110104760 CN102229638B (en) | 2011-04-26 | 2011-04-26 | Method for extracting oleanolic acid from chaenomeles fruit and preparing oleanolic acid standard |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103054972A (en) * | 2012-12-20 | 2013-04-24 | 蚌埠丰原涂山制药有限公司 | Method of extracting oleanolic acid and ursolic acid from pawpaw |
| CN103059092A (en) * | 2012-12-25 | 2013-04-24 | 宁蒗女儿珍生物工程有限公司 | Method of extracting oleanolic acid from pawpaw |
| CN104004048A (en) * | 2014-05-27 | 2014-08-27 | 上海交通大学 | High-utilization-rate extraction and processing method of wrinkled papaya |
| CN105061544A (en) * | 2015-07-30 | 2015-11-18 | 利德健康科技（青岛）有限公司 | Method of extracting oleanolic acid from fresh pawpaw fruits |
| CN107125533A (en) * | 2017-06-09 | 2017-09-05 | 合肥工业大学 | A kind of preparation method of Xuancheng's pawpaw effervescent tablet |
| WO2024078517A1 (en) * | 2022-10-13 | 2024-04-18 | Hong Kong Baptist University | Identification of oleanolic acid and plant extract for glucose-6-phosphate dehydrogenase-related disorders including bag3opathy |
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2011
- 2011-04-26 CN CN 201110104760 patent/CN102229638B/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 20100715 寻华凤 宣木瓜中活性物质提取及含片与胶囊的研制 , 第7期 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103054972A (en) * | 2012-12-20 | 2013-04-24 | 蚌埠丰原涂山制药有限公司 | Method of extracting oleanolic acid and ursolic acid from pawpaw |
| CN103059092A (en) * | 2012-12-25 | 2013-04-24 | 宁蒗女儿珍生物工程有限公司 | Method of extracting oleanolic acid from pawpaw |
| CN103059092B (en) * | 2012-12-25 | 2016-03-23 | 宁蒗女儿珍生物工程有限公司 | A kind of method extracting Oleanolic Acid from pawpaw |
| CN104004048A (en) * | 2014-05-27 | 2014-08-27 | 上海交通大学 | High-utilization-rate extraction and processing method of wrinkled papaya |
| CN104004048B (en) * | 2014-05-27 | 2015-11-04 | 上海交通大学 | Cockle pawpaw high utilization rate extracts and working method |
| CN105061544A (en) * | 2015-07-30 | 2015-11-18 | 利德健康科技（青岛）有限公司 | Method of extracting oleanolic acid from fresh pawpaw fruits |
| CN107125533A (en) * | 2017-06-09 | 2017-09-05 | 合肥工业大学 | A kind of preparation method of Xuancheng's pawpaw effervescent tablet |
| WO2024078517A1 (en) * | 2022-10-13 | 2024-04-18 | Hong Kong Baptist University | Identification of oleanolic acid and plant extract for glucose-6-phosphate dehydrogenase-related disorders including bag3opathy |
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|---|---|
| CN102229638B (en) | 2013-05-08 |
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