CN106265846A - A kind of fermentation process improving Radix Rhodiolae oxidation resistance - Google Patents
A kind of fermentation process improving Radix Rhodiolae oxidation resistance Download PDFInfo
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- 238000000855 fermentation Methods 0.000 title claims abstract description 48
- 230000004151 fermentation Effects 0.000 title claims abstract description 48
- 230000003647 oxidation Effects 0.000 title claims abstract description 10
- 238000007254 oxidation reaction Methods 0.000 title claims abstract description 10
- 239000000243 solution Substances 0.000 claims abstract description 23
- 239000000843 powder Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000002609 medium Substances 0.000 claims abstract description 19
- 238000002360 preparation method Methods 0.000 claims abstract description 18
- 239000000284 extract Substances 0.000 claims abstract description 14
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 238000011218 seed culture Methods 0.000 claims abstract description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 11
- 239000008103 glucose Substances 0.000 claims abstract description 11
- 241000228245 Aspergillus niger Species 0.000 claims abstract description 10
- 239000012535 impurity Substances 0.000 claims abstract description 10
- 239000012153 distilled water Substances 0.000 claims abstract description 7
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 6
- 238000004062 sedimentation Methods 0.000 claims abstract description 6
- 239000006228 supernatant Substances 0.000 claims abstract description 6
- 238000010790 dilution Methods 0.000 claims abstract description 5
- 239000012895 dilution Substances 0.000 claims abstract description 5
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 10
- 239000002054 inoculum Substances 0.000 claims description 9
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 5
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 5
- 235000010344 sodium nitrate Nutrition 0.000 claims description 5
- 239000004317 sodium nitrate Substances 0.000 claims description 5
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 3
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 3
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 3
- 239000011591 potassium Substances 0.000 claims description 3
- 229910052700 potassium Inorganic materials 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 claims 1
- 230000000857 drug effect Effects 0.000 abstract description 6
- 239000012467 final product Substances 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 230000003064 anti-oxidating effect Effects 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 231100000753 hepatic injury Toxicity 0.000 description 2
- 241000411851 herbal medicine Species 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 206010002660 Anoxia Diseases 0.000 description 1
- 241000976983 Anoxia Species 0.000 description 1
- 208000019901 Anxiety disease Diseases 0.000 description 1
- 241000220284 Crassulaceae Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 241001165494 Rhodiola Species 0.000 description 1
- 241001170117 Rhodiola dumulosa Species 0.000 description 1
- 241000130930 Rhodiola henryi Species 0.000 description 1
- 241001170115 Rhodiola kirilowii Species 0.000 description 1
- 244000042430 Rhodiola rosea Species 0.000 description 1
- 235000003713 Rhodiola rosea Nutrition 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000007953 anoxia Effects 0.000 description 1
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- 230000003026 anti-oxygenic effect Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N benzo-alpha-pyrone Natural products C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000001671 coumarin Nutrition 0.000 description 1
- 150000004775 coumarins Chemical class 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 229940079593 drug Drugs 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
- 235000008434 ginseng Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000002443 hepatoprotective effect Effects 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000009979 protective mechanism Effects 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/41—Crassulaceae (Stonecrop family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
The invention discloses a kind of fermentation process improving Radix Rhodiolae oxidation resistance, comprise the steps: the pretreatment of (1) Radix Rhodiolae: Radix Rhodiolae section is pulverized and is dried；(2) preparation of Radix Rhodiolae extracting solution: step (1) gained Radix Rhodiolae powder and distilled water are mixed, extract, mixed extract after eight layers of filtered through gauze, overnight, centrifugal remove impurity, supernatant is used for preparing fermentation medium in sedimentation；(3) preparation of seed culture medium；(4) preparation of fermentation medium: the Radix Rhodiolae extracting solution of step (2) gained added in the glucose of 0.5% 2% and get final product；(5) fermentation process: use physiological saline solution to clean the spore on the growth aspergillus niger inclined-plane of 7 days, 4 layers of filtered through gauze remove impurity, in succession cultivate in seed culture medium and fermentation medium through dilution, utilize fermentable pharmacy to be effectively improved the oxidation resistance of Radix Rhodiolae extracting solution, strengthen its drug effect.
Description
Technical field
The invention belongs to biological fermentation field, be specifically related to a kind of fermentation process improving Radix Rhodiolae oxidation resistance.
Background technology
Radix Rhodiolae, belongs to Crassulaceae perennial herb or shrub plant, is to be grown in the rare wild plant of more than 1700 meters,
The growing environment of severe cold region gives the vitality that it is extremely indomitable, is described as " plateau Radix Ginseng ".The Radix Rhodiolae being grown in China has
72 kinds, about more than 90, whole world kind, the principal item of China have Radix Rhodiolae, Radix Rhodiolae, Radix Rhodiolae,
Rhodiola kirilowii (Regel) Maxim., Rhodiola dumulosa, Rhodiola henryi, Rhodiola phariensis, Rhodiola rosea L. etc..Radix Rhodiolae contains multiple chemistry
Composition, the common macromolecular complex such as including flavonoid, saponins, amino acids, phenols, Coumarins, volatile oil, and polysaccharide
Matter.Experimental studies have found that Radix Rhodiolae can improve anoxia endurance, resisting fatigue, antioxidation, defying age, raising immunity, anticancer, guarantor
Hepatoprotective dirty and have anti-good fortune penetrate, the effect such as anxiety.Radix Rhodiolae extract has antioxidation, can be obviously improved body
Anaerobic environment.Alcoholic liver injury and the hepatic injury that stress cause are had protective effect, and mechanism is due to Radix Rhodiolae extract
There is oxidation resistance, can be by removing free radical and metabolite thereof, anti-lipid peroxidation reacts.
Fermentation is an importance of Chinese medicine processing, by means of enzyme and the effect of microorganism, under the conditions of certain environment
(such as temperature, humidity, air, moisture etc.), make medicine pass through Fermentation, and its effective ingredient is sufficiently separated, extracts, and changes it
Original performance, strengthens or produces new effect, expands medicine variety, to adapt to the needs of clinical application.After bioconversion, in
Medicine has more biological activity.At present, the microbial fermentation technology of Chinese herbal medicine has become raising Chinese herbal medicine active component, has reduced pair work
With, strengthen drug effect important means.Radix Rhodiolae has antioxidant activity and is widely recognized as, but the main work of its antioxidation
Property composition is also still not clear.
Summary of the invention
For the problems referred to above, the present invention utilizes fermentable pharmaceutical technology to improve the antioxidation energy of Radix Rhodiolae extracting solution
Power, strengthens its drug effect.Radix Rhodiolae is after fermentation of Aspergillus niger, and its chemical composition can change, thus changes oxidation resistance.
The present invention adopts the following technical scheme that realization:
A kind of fermentation process improving Radix Rhodiolae oxidation resistance, comprises the steps:
(1) pretreatment of Radix Rhodiolae: Radix Rhodiolae section is crushed to 40-60 mesh；Being dried and making water content is 3%-5%, institute
State baking temperature and be 60-80 DEG C, 2-3 hour drying time, obtain Radix Rhodiolae powder, standby；
(2) preparation of Radix Rhodiolae extracting solution: by step (1) gained Radix Rhodiolae powder and distilled water in the ratio of material-water ratio 1:25
Mixing, 100 DEG C extract twice, each one hour, mixed extract after eight layers of filtered through gauze, supply moisture, by extracting solution at 4 DEG C
Overnight, 5000rpm is centrifuged remove impurity in 20-30 minute, and supernatant is used for preparing fermentation medium in sedimentation；
(3) preparation of seed culture medium: consist of 10g/L glucose, 5g/L yeast powder, 3g/L sodium nitrate, 1.5g/L phosphorus
Acid dihydride potassium, 2g/L Magnesium sulfate heptahydrate, 0.01g/L ferrous sulfate heptahydrate, 121 DEG C of sterilizing 20min；
(4) preparation of fermentation medium: the Radix Rhodiolae extracting solution of step (2) gained is added the glucose of 0.5%-2%
In, 121 DEG C of sterilizing 20min；
(5) fermentation process: use physiological saline solution to clean the spore on the growth aspergillus niger inclined-plane of 7 days, 4 layers of gauze mistake
Filter miscellaneous, use blood counting chamber to count under the microscope, then to be diluted to spore count be 107Individual/ml, by the inoculation of 2%-5%
Amount is seeded in seed culture medium, incubated overnight, then is seeded in fermentation medium by the inoculum concentration of 5%-10%, 28-30 DEG C
200rpm cultivates 2-3 days.
The present invention is compared with prior art had the advantages that to be fermented altogether by microorganism and Chinese medicine and carries out Chinese medicine
Concocting, the secondary metabolites of fermentable and ingredient generation synergism can strengthen drug effect so that Radix Rhodiolae extracting solution
Oxidation resistance improve, DPPH clearance rate by 35% improve to 75%.The decomposition of microorganism can poisonous by medical material
Substance decomposition, or it is carried out modification make its toxicity reduce.The medicine effective active can not being directly utilized after entering people's human body
Component, available fermentation technique completes in vitro to convert and make it to be directly utilized by body, plays rapidly proper efficiency.Fermentation
Protection active ingredient of Chinese herbs exempts from destruction, makes the active component of medicine, active substance be extracted to greatest extent, utilize.Make
Using fermentation of Aspergillus niger Radix Rhodiolae, save herb resource, protect environment, method is simple to operate, low cost, and the cycle is short, is in gentleness
Under the conditions of improve the method for Radix Rhodiolae drug effect, be to realize the modernization of Chinese medicine, there is the new technique of high-tech level.
Detailed description of the invention
Embodiment 1
(1) pretreatment of Radix Rhodiolae: Radix Rhodiolae section is crushed to all cross 60 mesh sieves；It is dried again, described dry
Temperature is 80 DEG C, 3 hours drying times, makes water content 5%, obtains Radix Rhodiolae powder, standby.
(2) preparation of Radix Rhodiolae extracting solution: by step (1) gained Radix Rhodiolae powder and distilled water in the ratio of material-water ratio 1:25
Mixing, 100 DEG C extract twice, each one hour, mixed extract after eight layers of filtered through gauze, supply moisture.By extracting solution at 4 DEG C
Overnight, 5000rpm is centrifuged remove impurity in 30 minutes, and supernatant is used for preparing fermentation medium in sedimentation.
(3) preparation of seed culture medium: consist of 10g/L glucose, 5g/L yeast powder, 3g/L sodium nitrate, 1.5g/L phosphorus
Acid dihydride potassium, 2g/L Magnesium sulfate heptahydrate, 0.01g/L ferrous sulfate heptahydrate, 121 DEG C of sterilizing 20min；
(4) preparation of fermentation medium: the Radix Rhodiolae extracting solution of step (2) gained is added the glucose of 2%, is divided in
In the conical flask of 500ml, every bottled liquid measure 100ml, 121 DEG C of sterilizing 20min.
(5) fermentation process: use physiological saline solution to be washed down by the spore on the growth aspergillus niger inclined-plane of 7 days, 4 layers of gauze
Filtering and impurity removing, uses blood counting chamber to count under the microscope, then to be diluted to spore count is 107Individual/ml, by the inoculum concentration of 2%
It is seeded to seed culture medium, incubated overnight.Being seeded in fermentation medium by the inoculum concentration of 5%, 28 DEG C of 200rpm cultivate 3 again
My god.
Embodiment 2
(1) pretreatment of Radix Rhodiolae: 40 mesh are pulverized in Radix Rhodiolae section；Being dried, described baking temperature is 60 DEG C again,
3 hours drying times, make water content 3%, obtain Radix Rhodiolae powder, standby.
(2) preparation of Radix Rhodiolae extracting solution: by step (1) gained Radix Rhodiolae powder and distilled water in the ratio of material-water ratio 1:25
100 DEG C extract twice, each one hour, mixed extract after eight layers of filtered through gauze, supply moisture.By extracting solution 4 DEG C of sedimentations
Overnight, 5000r is centrifuged remove impurity in 30 minutes, and supernatant is used for preparing fermentation medium.
(3) preparation of seed culture medium: by 10g glucose, 5g yeast powder, 3g sodium nitrate, 1.5g potassium dihydrogen phosphate, 2g seven
Water magnesium sulfate, 0.01g ferrous sulfate heptahydrate are dissolved in water, are settled to 1L, are divided in the conical flask of 500ml, every bottled liquid measure
100ml, 121 DEG C of sterilizing 20min.
(4) preparation of fermentation medium: the Radix Rhodiolae extracting solution of step (2) gained is added the glucose of 1%, is divided in
In the conical flask of 500ml, every bottled liquid measure 100ml, 121 DEG C of sterilizing 20min.
(5) fermentation process: use physiological saline solution to be washed down by the spore on the growth aspergillus niger inclined-plane of 7 days, 4 layers of gauze
Filtering and impurity removing, uses blood counting chamber to count under the microscope, then to be diluted to spore count is 107Individual/ml, by the inoculum concentration of 2%
It is seeded to seed culture medium, incubated overnight.Being seeded in fermentation medium by the inoculum concentration of 5%, 28 DEG C of 200rpm cultivate 2 again
My god.
Embodiment 3
(1) pretreatment of Radix Rhodiolae: Radix Rhodiolae section being pulverized, cross 50 mesh sieves, middle powder and fine powder are continued to employ, coarse powder powder again
Broken, to whole mistake 50 mesh sieves；Being dried, described baking temperature is 60 DEG C again, 2 hours drying times, makes water content 5%, i.e.
Obtain Radix Rhodiolae powder, standby.
(2) preparation of Radix Rhodiolae extracting solution: by step (1) gained Radix Rhodiolae powder and distilled water in the ratio of material-water ratio 1:25
100 DEG C extract twice, each one hour, mixed extract after eight layers of filtered through gauze, supply moisture.By extracting solution 4 DEG C of sedimentations
Overnight, 5000rpm is centrifuged remove impurity in 20 minutes, and supernatant is used for preparing fermentation medium.
(3) preparation of seed culture medium: by 10g glucose, 5g yeast powder, 3g sodium nitrate, 1.5g potassium dihydrogen phosphate, 2g seven
Water magnesium sulfate, 0.01g ferrous sulfate heptahydrate are dissolved in water, are settled to 1L, are divided in the conical flask of 500ml, every bottled liquid measure
100ml, 121 DEG C of sterilizing 20min.
(4) preparation of fermentation medium: the Radix Rhodiolae extracting solution of step (2) gained is added the glucose of 0.5%, subpackage
In the conical flask of 500ml, every bottled liquid measure 100ml, 121 DEG C of sterilizing 20min.
(5) fermentation process: use physiological saline solution to be washed down by the spore on the growth aspergillus niger inclined-plane of 7 days, 4 layers of gauze
Filtering and impurity removing, uses blood counting chamber to count under the microscope, then to be diluted to spore count is 107Individual/ml, by the inoculum concentration of 2%
It is seeded to seed culture medium, incubated overnight.Being seeded in fermentation medium by the inoculum concentration of 5%, 28 DEG C of 200rpm cultivate 3 again
My god.
Embodiment 4
DPPH clearance rate measures: measure 2mL ethanol, takes 2mL DPPH solution (0.2mM is dissolved in ethanol), mixing shaking, room
After the lower lucifuge of temperature stands 30min, at 517mn, survey absorbance A c；500 times of precisions of fermentation liquid dilution measure 2mL, then take 2mL second
Alcohol, vibration, after lucifuge stands 30min under room temperature, at 517nm, survey absorbance A j；Precision measures the fermentation liquid of dilution 500 times
2mL, then take 2ml DPPH solution, vibration, at 517nm, survey absorbance A i after 30min.Calculate clearance rate: K=[l-(Ai-
Aj)/Ac] × l00%.The DPPH clearance rate of the Radix Rhodiolae fermentation liquid of embodiment 1-3 gained is shown in Table 1.
The DPPH clearance rate of table 1 embodiment 1-3 gained Radix Rhodiolae fermentation liquid
Radix Rhodiolae extracting solution DPPH clearance rate before fermentation is 35%, and through fermentation of Aspergillus niger, DPPH clearance rate is significantly
Improve, can reach 75%, improve 114%.Under the effect of aspergillus niger, the antioxygenic property of Radix Rhodiolae is strengthened,
Improve drug effect.
Claims (4)
1. improve a fermentation process for Radix Rhodiolae oxidation resistance, comprise the steps:
The pretreatment of Radix Rhodiolae: Radix Rhodiolae section is pulverized and is drying to obtain Radix Rhodiolae powder, standby；
The preparation of Radix Rhodiolae extracting solution: step (1) gained Radix Rhodiolae powder and distilled water are mixed, 100 DEG C are extracted twice, and each one
Hour, mixed extract after eight layers of filtered through gauze, supply the moisture of loss, overnight, centrifugal remove impurity, supernatant is used for preparing in sedimentation
Fermentation medium；
The preparation of seed culture medium: consist of 10g/L glucose, 5g/L yeast powder, 3g/L sodium nitrate, 1.5g/L biphosphate
Potassium, 2g/L Magnesium sulfate heptahydrate, 0.01g/L ferrous sulfate heptahydrate seed culture medium through 121 DEG C of sterilizing 20min；
The preparation of fermentation medium: being added by the Radix Rhodiolae extracting solution of step (2) gained in the glucose of 0.5%-2%, 121 DEG C go out
Bacterium 20min；
(5) fermentation process: using physiological saline solution to clean the spore on the growth aspergillus niger inclined-plane of 7 days, 4 layers of filtered through gauze are removed
Miscellaneous, after dilution, reach 1-3 × 107/ ml, cultivates in succession in culture medium and fermentation medium.
2. fermentation process as claimed in claim 1, in described step (1), Radix Rhodiolae is crushed to 40-60 mesh；Described baking temperature
For 60-80 DEG C, 2-3 hour drying time, described Radix Rhodiolae powder water content is 3%-5%.
3. fermentation process as claimed in claim 1, in described step (2), Radix Rhodiolae powder is 1:25-with the mass ratio of distilled water
1:50, described extracting solution settling temperature is 4 DEG C, and described centrifugal condition is 5000rpm, 20-30 minute.
4. fermentation process as claimed in claim 1, in described step (5), after dilution, spore presses 2%-5% inoculum concentration in seed training
Support incubated overnight in base, then be seeded in fermentation medium by the inoculum concentration of 5%-10%, cultivate 2-3 days for 28 DEG C-30 DEG C.
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| CN109528602B (en) * | 2019-01-22 | 2021-06-29 | 广州睿森生物科技有限公司 | Plant fermentation composition, preparation method and application thereof |
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