CN117679448A - Method for extracting and purifying total saponins of panax notoginseng - Google Patents
Method for extracting and purifying total saponins of panax notoginseng Download PDFInfo
- Publication number
- CN117679448A CN117679448A CN202311565440.7A CN202311565440A CN117679448A CN 117679448 A CN117679448 A CN 117679448A CN 202311565440 A CN202311565440 A CN 202311565440A CN 117679448 A CN117679448 A CN 117679448A
- Authority
- CN
- China
- Prior art keywords
- extracting
- total saponins
- panax notoginseng
- enzymolysis
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229930182490 saponin Natural products 0.000 title claims abstract description 75
- 150000007949 saponins Chemical class 0.000 title claims abstract description 75
- 235000017709 saponins Nutrition 0.000 title claims abstract description 75
- 241000180649 Panax notoginseng Species 0.000 title claims abstract description 65
- 235000003143 Panax notoginseng Nutrition 0.000 title claims abstract description 65
- 238000000034 method Methods 0.000 title claims abstract description 48
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 60
- 244000131316 Panax pseudoginseng Species 0.000 claims abstract description 38
- 235000003181 Panax pseudoginseng Nutrition 0.000 claims abstract description 38
- 239000011347 resin Substances 0.000 claims abstract description 27
- 229920005989 resin Polymers 0.000 claims abstract description 27
- 239000000463 material Substances 0.000 claims abstract description 17
- 238000000605 extraction Methods 0.000 claims abstract description 12
- 239000000843 powder Substances 0.000 claims abstract description 12
- 238000010992 reflux Methods 0.000 claims abstract description 8
- 239000004094 surface-active agent Substances 0.000 claims abstract description 7
- 238000000746 purification Methods 0.000 claims abstract description 6
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 43
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 32
- 108010019077 beta-Amylase Proteins 0.000 claims description 17
- 108010038658 exo-1,4-beta-D-xylosidase Proteins 0.000 claims description 17
- 238000005406 washing Methods 0.000 claims description 12
- 108010059892 Cellulase Proteins 0.000 claims description 11
- 229940106157 cellulase Drugs 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 8
- PYIDGJJWBIBVIA-UYTYNIKBSA-N lauryl glucoside Chemical compound CCCCCCCCCCCCO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O PYIDGJJWBIBVIA-UYTYNIKBSA-N 0.000 claims description 7
- 238000010828 elution Methods 0.000 claims description 6
- 239000012535 impurity Substances 0.000 claims description 5
- 238000012856 packing Methods 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 abstract description 5
- 238000009776 industrial production Methods 0.000 abstract description 4
- 230000008569 process Effects 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 9
- 238000001035 drying Methods 0.000 description 6
- 230000007071 enzymatic hydrolysis Effects 0.000 description 5
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 238000001179 sorption measurement Methods 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 238000004440 column chromatography Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- TYLVGQKNNUHXIP-MHHARFCSSA-N 10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)C=4C=CC=CC=4)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 TYLVGQKNNUHXIP-MHHARFCSSA-N 0.000 description 2
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 2
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000000916 dilatatory effect Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000005057 refrigeration Methods 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 description 2
- RAQNTCRNSXYLAH-RFCGZQMISA-N (20S)-ginsenoside Rh1 Chemical compound O([C@@H]1[C@H]2C(C)(C)[C@@H](O)CC[C@]2(C)[C@H]2C[C@@H](O)[C@H]3[C@@]([C@@]2(C1)C)(C)CC[C@@H]3[C@@](C)(O)CCC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RAQNTCRNSXYLAH-RFCGZQMISA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 241000208340 Araliaceae Species 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 235000002791 Panax Nutrition 0.000 description 1
- 241000208343 Panax Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- 229930002877 anthocyanin Natural products 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect 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
- 235000013405 beer Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- MRUAUOIMASANKQ-UHFFFAOYSA-O carboxymethyl-[3-(dodecanoylamino)propyl]-dimethylazanium Chemical compound CCCCCCCCCCCC(=O)NCCC[N+](C)(C)CC(O)=O MRUAUOIMASANKQ-UHFFFAOYSA-O 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 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
- 235000015203 fruit juice Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- YURJSTAIMNSZAE-HHNZYBFYSA-N ginsenoside Rg1 Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(C[C@@H]([C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YURJSTAIMNSZAE-HHNZYBFYSA-N 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- -1 glycoside compounds Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940075468 lauramidopropyl betaine Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000006993 memory improvement Effects 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- LLPWNQMSUYAGQI-OOSPGMBYSA-N notoginsenoside R1 Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(C[C@@H]([C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O[C@H]2[C@@H]([C@@H](O)[C@H](O)CO2)O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O LLPWNQMSUYAGQI-OOSPGMBYSA-N 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Chemical class 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of extraction and purification of total saponins, and particularly relates to a method for extracting and purifying total saponins of panax notoginseng. The method comprises the following steps: taking pseudo-ginseng coarse powder, and carrying out enzymolysis to obtain an enzymolysis material; (2) Adding ethanol solution and surfactant into the enzymolysis material, and performing reflux extraction to obtain an extracting solution; (3) Refrigerating the extractive solution, decolorizing, and concentrating to obtain decolorized solution; (4) Purifying the decolorized solution with macroporous resin to obtain Notoginseng radix total saponin. The preparation process is simple, and is suitable for industrial production, and the total saponins of the final pseudo-ginseng can reach about 97%.
Description
Technical Field
The invention belongs to the technical field of extraction and purification of total saponins, and particularly relates to a method for extracting and purifying total saponins of panax notoginseng.
Background
Notoginseng radix is a plant of Panax of Araliaceae, also called Notoginseng radix, and is a traditional precious medicinal material. At present, at least two hundred active compounds including saponin, polysaccharide, flavonoid, volatile oil, amino acid and the like are found, roots, stems, leaves and flowers of the active compounds can be used as medicines, and at least twenty preparations in the national basic medicine catalogue and the national Chinese medicine protection variety catalogue contain pseudo-ginseng components. Modern pharmacology shows that the pseudo-ginseng has the pharmacological effects of stopping bleeding, dilating blood vessels, promoting blood circulation and removing blood stasis, and has the activities of reducing myocardial oxygen consumption, inhibiting platelet aggregation, dilating coronary artery, resisting atherosclerosis, resisting thrombosis and the like. Wherein, the total saponins of Notoginseng radix (PNS) has good effects of promoting blood circulation, removing blood stasis, relieving inflammation, and protecting myocardium.
The existing production and purification process of the pseudo-ginseng total saponins mainly adopts a column chromatography and a solvent extraction method. Column chromatography commonly used at present is silica gel column chromatography, gel column chromatography and macroporous adsorption resin method. The silica gel column chromatography and the gel column chromatography can purify to obtain the high-purity saponin component, but the operation is complex, the cost is high, and the method is not suitable for technological production; macroporous adsorption resin is a common method for industrially producing and purifying the total saponins of panax notoginseng at present.
The Chinese patent application CN109700841A discloses a method for producing and preparing high-purity panax notoginseng saponins, which comprises the following steps: (1) crushing and sieving pseudo-ginseng, extracting with alcohol, and concentrating; (2) Extracting with acetone, removing acetone, and dissolving precipitate with water; or extracting with n-butanol, retaining n-butanol, concentrating, and dissolving in water; (3) Loading onto macroporous adsorbent resin, washing with alkali water, purifying, eluting with ethanol, collecting eluate, and concentrating; purifying the refined resin and washing with water; (5) adding ethanol into the water washing liquid, and then, decoloring the resin; (6) decolorizing with active carbon, filtering, concentrating and drying. The total content of the final pseudo-ginseng total saponins reaches more than 90 percent, but the preparation method is more complex.
The Chinese patent application CN1919222A discloses a preparation method of total saponins of panax notoginseng, which comprises the following preparation steps: soaking Notoginseng radix in water, adding cellulase for enzymolysis, extracting the residue with water or 10% -90% ethanol, concentrating the extractive solution, sequentially loading onto anion exchange resin column and macroporous resin column, concentrating the eluate under reduced pressure to obtain extract, and drying. The method can remove most of water-soluble impurities and fat-soluble impurities such as saccharide and pigment, has the advantages of less impurities, high purity and stable quality, and can be used alone or with other active ingredients to prepare medicine with good therapeutic effect and high quality. Although the content of the total saponins finally obtained by the invention meets the requirement of Chinese pharmacopoeia (namely, the oral administration is not lower than 75%), the content is in the range of 81.3-85.7%, and is relatively less.
After the coarse powder of pseudo-ginseng is subjected to enzymolysis in the Chinese patent application CN107669721A, the enzymolysis product is subjected to water extraction and multistage countercurrent alcohol extraction, and then macroporous adsorption resin is adopted for adsorption treatment, so that the yield of the total saponins of pseudo-ginseng is about 80%. Pulverizing Notoginseng radix into coarse powder, adding 6-8 times of water into multifunctional extraction tank, adding emulsifier, mixing, and reflux extracting; then the extract is passed through macroporous adsorption resin, and the purity is not obviously improved although the extract yield is improved by 30 percent.
Therefore, the process for extracting and purifying the total saponins of panax notoginseng needs to be further researched, and the process parameters of enriching and purifying the total saponins of panax notoginseng by optimizing macroporous resin are optimized, so that the purposes of simplicity, rapidness, suitability for industrial production and high content are achieved.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for purifying total saponins of panax notoginseng. The high-content total saponins of panax notoginseng is prepared by purifying LXD-100 macroporous resin, and the method is simple and quick and is suitable for industrial production.
In order to achieve the above purpose, the invention adopts the following specific technical scheme:
a method for extracting and purifying total saponins of panax notoginseng comprises the following steps:
(1) Taking pseudo-ginseng coarse powder, and carrying out enzymolysis to obtain an enzymolysis material;
(2) Adding ethanol solution and surfactant into the enzymolysis material, and performing reflux extraction to obtain an extracting solution;
(3) Refrigerating the extractive solution, decolorizing, and concentrating to obtain decolorized solution;
(4) Purifying the decolorized solution with macroporous resin to obtain Notoginseng radix total saponin.
Preferably, the enzymatic hydrolysis in step (1) comprises two enzymatic hydrolysis steps, the first enzymatic hydrolysis adding cellulase and the second enzymatic hydrolysis adding β -xylosidase and β -amylase.
Further preferably, the addition amount of the cellulase is 1% -1.5% of the weight of the pseudo-ginseng coarse powder, and the total addition amount of the beta-xylosidase and the beta-amylase is 0.5% -0.8% of the weight of the pseudo-ginseng coarse powder.
Further preferably, the mass ratio of the beta-xylosidase to the beta-amylase is 2-3:1.
Further preferably, the temperature of the first enzymolysis is 40-45 ℃, and the enzymolysis time is 1-1.5h; the temperature of the second enzymolysis is 50-60 ℃, and the enzymolysis time is 0.5-1h.
Beta-xylosidase belongs to glycoside hydrolase (glycoside hydrolase, GH), is one of main xylan degrading enzymes, and can be applied to the fields of food, feed, papermaking, energy sources and the like. Beta-xylosidase has special hydrolysis function, and can cut off notoginsenoside R 1 And R is 2 And xylose groups on ASI, XDT and anthocyanin to respectively form ginsenoside Rg 1 And Rh 1 10-Deacetylpaclitaxel (DT), cycloastragal (CA) and anthocyanidin, and these products with removed xylosyl have important biological activities such as anticancer, anti-inflammatory, antioxidant and memory improvement.
Beta-amylase generally plays an important role in industries such as food processing, grain processing, fermentation, brewing, textiles and the like, the beta-amylase serving as a saccharifying agent can be applied to industrial production such as beer, maltose, beverages and the like, is an important enzyme source in the food processing and brewing industries, is commonly used for starch decomposition and filtration speed improvement in fruit juice processing, and is commonly used for conversion of glycoside compounds.
Preferably, the mass concentration of the ethanol solution in the step (2) is 85% -95%, and the addition amount of the ethanol solution is 2-4 times of the weight of the enzymolysis materials.
Preferably, the surfactant in the step (2) is dodecyl glucoside, and the addition amount of the surfactant is 5% -10%, preferably 5% -8% of the weight of the enzymolysis material.
Preferably, the number of times of reflux extraction in the step (2) is 2-4, and the temperature of the reflux extraction is 75-85 ℃.
Preferably, the temperature of the refrigeration in the step (3) is 0-10 ℃, the time of the refrigeration is 24-48 hours, the time of the decolorization is 20-30 minutes, and the concentration is carried out until no ethanol smell exists.
Preferably, in the step (4), the macroporous resin is LXD-100, a wet method column packing is adopted, and the diameter-to-height ratio of the macroporous resin column is 1:3-5.
Preferably, the purification in step (4) comprises water washing to remove impurities and solution a elution; the volume of the water washing is 5-7BV, and the flow rate of the water washing is 2-3BV/h.
Further preferably, the solution a is a mixed solution of ethanol, n-butanol and water.
Preferably, the mass concentration of the ethanol in the solution A is 60% -70%, and the mass concentration of the n-butanol is 0.5% -2%.
Preferably, the flow rate of the elution of the solution A is 2-4BV/h, preferably 2-3BV/h, and the volume of the elution is 3-6BV.
Compared with the prior art, the invention has the following beneficial effects:
(1) The extraction and purification method is simple and efficient, and the total saponins of the pseudo-ginseng finally extracted has high content and high yield, wherein the content of the total saponins of the pseudo-ginseng can reach about 97 percent, and the yield can reach about 30 percent;
(2) According to the invention, different types of enzymes are used for enzymolysis, the cellulose firstly promotes the dissolution of plant cell walls to dissolve out more plant cell inner solubles, then the dissolved substances are further subjected to enzymolysis by adopting beta-xylosidase and beta-amylase at the same time, so that the enzymolysis efficiency is fully improved, and the content and the yield of the finally obtained total saponins of panax notoginseng are further improved;
(3) Compared with other macroporous resins, the LXD-100 macroporous resin is more suitable for purifying the total saponins of the pseudo-ginseng, and the elution is carried out by the mixed solution, so that the elution effect is obviously improved.
Detailed Description
The technical solutions of the embodiments of the present invention are further clearly described, and the described embodiments are only a part of the present invention, which are used to explain the present invention, but not to limit the present invention, so that other embodiments obtained by other persons skilled in the art without creative efforts fall within the protection scope of the present invention.
LXD-100 macroporous resin, available from New Material Co., ltd.
Example 1
A method for extracting and purifying total saponins of Notoginseng radix comprises the following steps:
(1) Taking pseudo-ginseng coarse powder, adding cellulase accounting for 1.2% of the weight of pseudo-ginseng, carrying out enzymolysis for 1h under the water bath condition of 45 ℃, adding beta-xylosidase accounting for 0.6% of the weight of pseudo-ginseng and beta-amylase (the mass ratio of the beta-xylosidase to the beta-amylase is 2:1), and carrying out enzymolysis for 1h under the water bath condition of 55 ℃ to obtain an enzymolysis material;
(2) Adding 3 times of ethanol solution with the mass concentration of 90% and dodecyl glucoside with the mass concentration of 6% into the enzymolysis material, reflux-extracting for 3 times at 80 ℃, and mixing the extracting solutions to obtain pseudo-ginseng extracting solution;
(3) Refrigerating Notoginseng radix extractive solution at 5deg.C for 30 hr, filtering, adding active carbon into the filtrate, stirring, adsorbing for 30min, and concentrating to obtain decolorized solution;
(4) Passing the decolorized solution through LXD-100 macroporous resin, wet packing (column volume is 50mL, diameter-to-height ratio is 1:4), washing water with 6BV ultrapure water at 3BV/h flow rate, eluting with mixed solution of ethanol, n-butanol and water (wherein the mass concentration of ethanol is 70% and the mass concentration of n-butanol is 1%), co-eluting at 2BV/h for 6BV, collecting eluate, concentrating and drying to obtain Notoginseng radix total saponin with yield of 21.7%.
According to the method for detecting the content of the total saponins of pseudo-ginseng in the Chinese pharmacopoeia of 2020 edition, the content of the total saponins of pseudo-ginseng is detected to be 97.4 percent.
Example 2
A method for extracting and purifying total saponins of Notoginseng radix comprises the following steps:
(1) Taking pseudo-ginseng coarse powder, adding cellulase accounting for 1.5% of the weight of pseudo-ginseng, carrying out enzymolysis for 1.5 hours under the water bath condition of 40 ℃, adding beta-xylosidase accounting for 0.5% of the weight of pseudo-ginseng and beta-amylase (the mass ratio of the beta-xylosidase to the beta-amylase is 3:1), and carrying out enzymolysis for 0.5 hour under the water bath condition of 60 ℃ to obtain an enzymolysis material;
(2) Adding 2 times of ethanol solution with the mass concentration of 95% and dodecyl glucoside with the mass concentration of 5% into the enzymolysis material, reflux-extracting for 4 times at 75 ℃, and mixing the extracting solutions to obtain pseudo-ginseng extracting solution;
(3) Refrigerating Notoginseng radix extractive solution at 10deg.C for 48 hr, filtering, adding active carbon into the filtrate, stirring, adsorbing for 30min, and concentrating to obtain decolorized solution;
(4) Passing the decolorized solution through LXD-100 macroporous resin, wet packing (column volume is 50mL, diameter-to-height ratio is 1:4), washing water with 6BV ultrapure water at 3BV/h flow rate, eluting with mixed solution of ethanol, n-butanol and water (wherein the mass concentration of ethanol is 70% and the mass concentration of n-butanol is 0.5%), co-eluting at 3BV/h flow rate for 5.5BV, collecting eluate, concentrating and drying to obtain Notoginseng radix total saponins with yield of 21.1%.
According to the method for detecting the content of the total saponins of pseudo-ginseng in the Chinese pharmacopoeia of 2020 edition, the content of the total saponins of pseudo-ginseng is detected to be 97.0 percent.
Example 3
A method for extracting and purifying total saponins of Notoginseng radix comprises the following steps:
(1) Taking pseudo-ginseng coarse powder, adding cellulase accounting for 1% of the weight of pseudo-ginseng, carrying out enzymolysis for 1.2 hours under the water bath condition of 45 ℃, adding beta-xylosidase accounting for 0.8% of the weight of pseudo-ginseng and beta-amylase (the mass ratio of the beta-xylosidase to the beta-amylase is 2:1), and carrying out enzymolysis for 1 hour under the water bath condition of 50 ℃ to obtain an enzymolysis material;
(2) Adding 2 times of ethanol solution with the mass concentration of 85% and dodecyl glucoside with the mass concentration of 8% into the enzymolysis material, reflux-extracting for 4 times at 85 ℃, and mixing the extracting solutions to obtain pseudo-ginseng extracting solution;
(3) Refrigerating Notoginseng radix extractive solution at 0deg.C for 24 hr, filtering, adding active carbon into the filtrate, stirring, adsorbing for 30min, and concentrating to obtain decolorized solution;
(4) Passing the decolorized solution through LXD-100 macroporous resin, loading into a column (column volume is 50mL, diameter-to-height ratio is 1:4) by wet method, washing with 5.5BV ultrapure water at a flow rate of 2.5BV/h, eluting with a mixed solution of ethanol, n-butanol and water (wherein the mass concentration of ethanol is 70% and the mass concentration of n-butanol is 2%), co-eluting at a flow rate of 2.5BV/h for 5BV, collecting the eluent, concentrating and drying to obtain Notoginseng radix total saponins with a yield of 22.4%.
According to the method for detecting the content of the total saponins of pseudo-ginseng in the Chinese pharmacopoeia of 2020 edition, the content of the total saponins of pseudo-ginseng is detected to be 97.1 percent.
Example 4
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: and (4) eluting with a mixed solution of ethanol, n-butanol and water (wherein the mass concentration of the ethanol is 70% and the mass concentration of the n-butanol is 1%), eluting at a speed of 4BV/h, eluting for 6BV altogether, collecting the eluent, concentrating and drying to obtain the total saponins of the pseudo-ginseng with the content of 93.2% and the yield of 20.1%.
Example 5
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: the temperature of the two enzymatic hydrolysis steps in step (1) was 45℃and the remaining steps were identical to those of example 1. The content of the total saponins of panax notoginseng is 95.1 percent, and the yield is 19.7 percent.
Comparative example 1
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: in the step (1), cellulase, beta-xylosidase and beta-amylase are added into pseudo-ginseng at the same time, and enzymolysis is carried out for 1h under the water bath condition of 45 ℃. The content of the total saponins of panax notoginseng is 89.2 percent, and the yield is 16.5 percent.
Comparative example 2
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: in the step (1), cellulase is added for enzymolysis for 1h, then beta-amylase accounting for 0.6 percent of the weight of the pseudo-ginseng is added, and the enzymolysis is carried out for 1h under the water bath condition of 55 ℃. The content of the total saponins of panax notoginseng is 91.3 percent, and the yield is 17.1 percent.
Comparative example 3
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: in the step (1), cellulase is added for enzymolysis for 1h, then beta-xylosidase accounting for 0.6% of the weight of the pseudo-ginseng is added, and the enzymolysis is carried out for 1h under the water bath condition of 55 ℃. The content of the total saponins of panax notoginseng is 90.9 percent and the yield is 17.3 percent.
Comparative example 4
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: in the step (2), dodecyl glucoside is not added. The content of the total saponins of panax notoginseng is 88.7 percent, and the yield is 16.9 percent.
Comparative example 5
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: the dodecyl glucoside is replaced by lauramidopropyl betaine in the step (2). The content of the total saponins of panax notoginseng is 89.8 percent, and the yield is 17.1 percent.
Comparative example 6
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: the eluent in the step (4) is ethanol solution with the mass concentration of 70 percent. The content of the total saponins of panax notoginseng is 90.2 percent, and the yield is 17.3 percent.
Comparative example 7
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: the eluent in the step (4) is a mixed solution of ethanol, n-butanol and water (wherein the mass concentration of the ethanol is 50% and the mass concentration of the n-butanol is 1%). The content of the total saponins of panax notoginseng is 91.4 percent, and the yield is 16.2 percent.
Comparative example 8
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: and (3) replacing LXD-100 macroporous resin with D101 macroporous resin in the step (4). The content of the total saponins of panax notoginseng is 89.2 percent, and the yield is 17.2 percent.
Comparative example 9
The process for extracting and purifying total saponins of panax notoginseng is different from example 1 only in that: and (3) replacing the LXD-100 macroporous resin with the LXD-200 macroporous resin in the step (4). The content of the total saponins of panax notoginseng is 90.5 percent, and the yield is 17.5 percent.
The foregoing detailed description is directed to one of the possible embodiments of the present invention, which is not intended to limit the scope of the invention, but is to be accorded the full scope of all such equivalents and modifications so as not to depart from the scope of the invention.
Claims (13)
1. The method for extracting and purifying the total saponins of panax notoginseng is characterized by comprising the following steps of:
(1) Taking pseudo-ginseng coarse powder, and carrying out enzymolysis to obtain an enzymolysis material;
(2) Adding ethanol solution and surfactant into the enzymolysis material, and performing reflux extraction to obtain an extracting solution;
(3) Refrigerating the extractive solution, decolorizing, and concentrating to obtain decolorized solution;
(4) Purifying the decolorized solution with macroporous resin to obtain Notoginseng radix total saponin.
2. The method for extracting and purifying total saponins of panax notoginseng according to claim 1, wherein the enzymolysis in the step (1) comprises two enzymolysis steps, wherein cellulase is added in the first enzymolysis step, and beta-xylosidase and beta-amylase are added in the second enzymolysis step.
3. The method for extracting and purifying total saponins of panax notoginseng according to claim 2, wherein the addition amount of cellulase is 1% -1.5% of the weight of the coarse powder of panax notoginseng, and the total addition amount of beta-xylosidase and beta-amylase is 0.5% -0.8% of the weight of the coarse powder of panax notoginseng.
4. The method for extracting and purifying total saponins of panax notoginseng as claimed in claim 3, wherein the mass ratio of the beta-xylosidase to the beta-amylase is 2-3:1.
5. The method for extracting and purifying total saponins of panax notoginseng according to claim 2, wherein the temperature of the first enzymolysis is 40-45 ℃, and the enzymolysis time is 1-1.5h; the temperature of the second enzymolysis is 50-60 ℃, and the enzymolysis time is 0.5-1h.
6. The method for extracting and purifying total saponins of panax notoginseng according to claim 1, wherein the mass concentration of the ethanol solution in the step (2) is 85% -95%, and the addition amount of the ethanol solution is 2-4 times of the weight of the enzymolysis material.
7. The method for extracting and purifying total saponins of panax notoginseng according to claim 1, wherein the surfactant in the step (2) is dodecyl glucoside, and the addition amount of the surfactant is 5% -10% of the weight of the enzymolysis material.
8. The method for extracting and purifying total saponins of panax notoginseng according to claim 1, wherein the number of times of reflux extraction in the step (2) is 2 to 4, and the temperature of reflux extraction is 75 ℃ to 85 ℃.
9. The method for extracting and purifying total saponins of panax notoginseng according to any one of claims 1 to 8, wherein the macroporous resin in the step (4) is LXD-100, wet packing is adopted, and the diameter-height ratio of the macroporous resin column is 1:3-5.
10. The method for extracting and purifying total saponins of panax notoginseng according to claim 1, wherein the purification in step (4) includes water washing to remove impurities and solution a elution; the volume of the water washing is 5-7BV, and the flow rate of the water washing is 2-3BV/h.
11. The method for extracting and purifying total saponins of panax notoginseng according to claim 10, wherein the solution a is a mixed solution of ethanol, n-butanol and water.
12. The method for extracting and purifying total saponins of panax notoginseng according to claim 11, wherein the mass concentration of ethanol in the solution a is 60% -70%, and the mass concentration of n-butanol is 0.5% -2%.
13. The method for extracting and purifying total saponins of panax notoginseng according to claim 10, wherein the flow rate of the solution a is 2-4BV/h, and the eluting volume is 3-6BV.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311565440.7A CN117679448A (en) | 2023-11-22 | 2023-11-22 | Method for extracting and purifying total saponins of panax notoginseng |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311565440.7A CN117679448A (en) | 2023-11-22 | 2023-11-22 | Method for extracting and purifying total saponins of panax notoginseng |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117679448A true CN117679448A (en) | 2024-03-12 |
Family
ID=90125498
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311565440.7A Pending CN117679448A (en) | 2023-11-22 | 2023-11-22 | Method for extracting and purifying total saponins of panax notoginseng |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117679448A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104997009A (en) * | 2015-07-25 | 2015-10-28 | 云南蓝钻生物科技股份有限公司 | Health-care food for resisting aging and enhancing immunity and preparation method of health-care food |
| CN105147753A (en) * | 2015-10-23 | 2015-12-16 | 云南大学 | Application of panax notoginseng saponins to direct HCV resisting medicine preparing |
| CN116712469A (en) * | 2023-05-19 | 2023-09-08 | 长春医学高等专科学校(长春职工医科大学长春市医学情报所) | Traditional Chinese medicine extract and preparation method thereof |
-
2023
- 2023-11-22 CN CN202311565440.7A patent/CN117679448A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104997009A (en) * | 2015-07-25 | 2015-10-28 | 云南蓝钻生物科技股份有限公司 | Health-care food for resisting aging and enhancing immunity and preparation method of health-care food |
| CN105147753A (en) * | 2015-10-23 | 2015-12-16 | 云南大学 | Application of panax notoginseng saponins to direct HCV resisting medicine preparing |
| CN116712469A (en) * | 2023-05-19 | 2023-09-08 | 长春医学高等专科学校(长春职工医科大学长春市医学情报所) | Traditional Chinese medicine extract and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103933092B (en) | The method of Radix Notoginseng total arasaponins in the fresh Radix Notoginseng of a kind of multiplex-enzyme extraction | |
| CN103266154A (en) | Biological transformation method for preparing high-activity theasaponin | |
| CN111297928B (en) | Method for extracting panax notoginseng saponins | |
| CN114949915B (en) | Hericium erinaceus compound extract and preparation method thereof | |
| CN109369733B (en) | Method for simultaneously extracting multiple flavonoid compounds from tartary buckwheat leaves | |
| CN110917240B (en) | Continuous method for separating multiple effective components from cyclocarya paliurus | |
| CN111187328B (en) | Method for preparing mogrol | |
| CN116712469A (en) | Traditional Chinese medicine extract and preparation method thereof | |
| CN116987056A (en) | Method for extracting dihydroquercetin from larch | |
| CN102492667A (en) | Enzyme preparation, and application of same in extraction of phellodendron berberine and method thereof | |
| CN112870254A (en) | Method for separating flavone, saponin and polysaccharide from cyclocarya paliurus by continuous method | |
| CN117679448A (en) | Method for extracting and purifying total saponins of panax notoginseng | |
| CN107375356B (en) | Method for simultaneously preparing high-purity total flavonol glycosides and ginkgolides | |
| CN111393490B (en) | Method for extracting linarin from buddleja officinalis | |
| CN111349139B (en) | Method for extracting sapindoside | |
| CN115043889A (en) | Method for extracting synephrine, hesperidin and naringin from seville orange flower | |
| CN113662976A (en) | Extraction method of purslane extract and obtained product | |
| CN108558645B (en) | Method for extracting crocin from gardenia | |
| CN113754626A (en) | Method for preparing fisetin by enzyme method | |
| CN110484577B (en) | Method for extracting and preparing mannose from dragon fruit stems | |
| CN113024679A (en) | Method for extracting selenium polysaccharide and polyphenol from selenium-rich moringa seeds | |
| LU502945B1 (en) | Efficient preparation method of active substances in sanghuangporus vaninii | |
| CN108430596B (en) | Method for purifying ginsenoside Rh2 | |
| CN112194689B (en) | Method for extracting effective active ingredients of rhodiola rosea | |
| CN116370517B (en) | Extraction method for extracting flavonoid substances from cranberries |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |