CN108096376A - Fermentate of the bulk pharmaceutical chemicals containing Fructus Corni and its preparation method and application - Google Patents
Fermentate of the bulk pharmaceutical chemicals containing Fructus Corni and its preparation method and application Download PDFInfo
- Publication number
- CN108096376A CN108096376A CN201810086333.9A CN201810086333A CN108096376A CN 108096376 A CN108096376 A CN 108096376A CN 201810086333 A CN201810086333 A CN 201810086333A CN 108096376 A CN108096376 A CN 108096376A
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- fermentation
- fructus corni
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- 238000000855 fermentation Methods 0.000 claims abstract description 54
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- 244000241838 Lycium barbarum Species 0.000 claims abstract description 44
- 230000004151 fermentation Effects 0.000 claims abstract description 44
- 235000015459 Lycium barbarum Nutrition 0.000 claims abstract description 43
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
- 239000007788 liquid Substances 0.000 claims abstract description 29
- 230000000694 effects Effects 0.000 claims abstract description 28
- 239000000725 suspension Substances 0.000 claims abstract description 23
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 20
- 241000894006 Bacteria Species 0.000 claims abstract description 19
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 18
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- 239000004310 lactic acid Substances 0.000 claims abstract description 10
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- 238000011081 inoculation Methods 0.000 claims abstract description 7
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- 229910019142 PO4 Inorganic materials 0.000 claims description 5
- 150000003863 ammonium salts Chemical class 0.000 claims description 5
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 10
- 239000002609 medium Substances 0.000 description 10
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- ZUKLFFYDSALIQW-MSUKCBDUSA-N Iridoid glycoside Chemical compound [H][C@]12CC[C@H](C(O)=O)[C@@]1([H])[C@H](OC1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)OC=C2 ZUKLFFYDSALIQW-MSUKCBDUSA-N 0.000 description 4
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- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 3
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
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- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
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- 230000000052 comparative effect Effects 0.000 description 2
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
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- 239000001103 potassium chloride Substances 0.000 description 2
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- 229910000160 potassium phosphate Inorganic materials 0.000 description 2
- 235000011009 potassium phosphates Nutrition 0.000 description 2
- 235000011121 sodium hydroxide Nutrition 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- ONDPHDOFVYQSGI-UHFFFAOYSA-N zinc nitrate Chemical compound [Zn+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ONDPHDOFVYQSGI-UHFFFAOYSA-N 0.000 description 2
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- DTHRMEYEJGYKKG-UHFFFAOYSA-N Ajugoside Natural products CC(=O)OC1(C)CC(O)C2C=COC(OC3OC(CO)C(O)C(O)C3O)C12C DTHRMEYEJGYKKG-UHFFFAOYSA-N 0.000 description 1
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 description 1
- 241000131317 Capitulum Species 0.000 description 1
- 235000008495 Chrysanthemum leucanthemum Nutrition 0.000 description 1
- 244000192528 Chrysanthemum parthenium Species 0.000 description 1
- 235000000604 Chrysanthemum parthenium Nutrition 0.000 description 1
- 241000142975 Cornaceae Species 0.000 description 1
- 241000759833 Cornus officinalis Species 0.000 description 1
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- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
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- 235000008708 Morus alba Nutrition 0.000 description 1
- WVVOBOZHTQJXPB-UHFFFAOYSA-N N-anilino-N-nitronitramide Chemical compound [N+](=O)([O-])N(NC1=CC=CC=C1)[N+](=O)[O-] WVVOBOZHTQJXPB-UHFFFAOYSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- 240000004371 Panax ginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 241000241413 Propolis Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
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- 244000269722 Thea sinensis Species 0.000 description 1
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- CAFTUQNGDROXEZ-XBDCZORHSA-N [(1s,4as,5r,7s,7as)-4a,5-dihydroxy-7-methyl-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,5,6,7a-tetrahydrocyclopenta[c]pyran-7-yl] acetate Chemical compound O([C@@H]1OC=C[C@@]2(O)[C@H](O)C[C@]([C@@H]12)(C)OC(=O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O CAFTUQNGDROXEZ-XBDCZORHSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
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- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 description 1
- CAFTUQNGDROXEZ-UHFFFAOYSA-N acetyl harpagide Natural products C12C(OC(=O)C)(C)CC(O)C2(O)C=COC1OC1OC(CO)C(O)C(O)C1O CAFTUQNGDROXEZ-UHFFFAOYSA-N 0.000 description 1
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid group Chemical class C(CC(O)(C(=O)O)CC(=O)O)(=O)O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 1
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- 239000008367 deionised water Substances 0.000 description 1
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- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
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- 235000019634 flavors Nutrition 0.000 description 1
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- 230000002496 gastric effect Effects 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- -1 iridoid glycoside glycosides Chemical class 0.000 description 1
- 239000008297 liquid dosage form Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- GVALZJMUIHGIMD-UHFFFAOYSA-H magnesium phosphate Chemical compound [Mg+2].[Mg+2].[Mg+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O GVALZJMUIHGIMD-UHFFFAOYSA-H 0.000 description 1
- 239000004137 magnesium phosphate Substances 0.000 description 1
- 229910000157 magnesium phosphate Inorganic materials 0.000 description 1
- 229960002261 magnesium phosphate Drugs 0.000 description 1
- 235000010994 magnesium phosphates Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 229940069949 propolis Drugs 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 239000007909 solid dosage form Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 235000020097 white wine Nutrition 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
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Classifications
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- 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)
- A61K36/40—Cornaceae (Dogwood family)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
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- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/287—Chrysanthemum, e.g. daisy
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
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- 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
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- 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
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Abstract
Fermentate the invention discloses a kind of bulk pharmaceutical chemicals containing Fructus Corni and its preparation method and application.The preparation method includes the following steps：(1) bulk pharmaceutical chemicals being made of Fructus Corni, matrimony vine and chrysanthemum are crushed, 40~65 parts by weight of Fructus Corni, 20~40 parts by weight of matrimony vine and 10~35 parts by weight of chrysanthemum is taken to be mixed with water respectively, obtain suspension；(2) suspension is digested using pectase, cellulase, zytase and trypsase, obtains enzymolysis liquid；(3) nutriment is added in into enzymolysis liquid, sterilizes, obtain fermentation medium；(4) into the fermentation medium, inoculation yeast bacterium carries out anaerobic fermentation 8~20h of culture, and then inoculating lactic acid bacterium carries out standing for fermentation 25~100h of culture, obtains cultured products；(5) cultured products are subjected to separation of solid and liquid, obtain fermentate.The method of the present invention can reduce fermentation time, and the antifatigue effect of gained fermentate is notable.
Description
Technical field
The present invention relates to a kind of fermentate and its preparation method and application, the hair of especially a kind of bulk pharmaceutical chemicals containing Fructus Corni
Ferment composition and its preparation method and application.
Background technology
The Chinese medicine class health products with antifatigue effect are numerous at present, and mostly containing ginseng, cordyceps sinensis, ganoderma lucidum, red scape
My god, the rare drug such as Radix Astragali, pilose antler, propolis, it is expensive；Preparation method is mostly using conventional method, such as direct comminuting method, water
Extraction method, ethanol extraction method, water extraction and alcohol precipitation method, extraction volatile oil etc..Other Chinese medicine class health products with antifatigue effect
Use said extracted preparation method more.
For example, CN104663973A discloses a kind of health protection tea for enhancing body immunity, keeping fit and healthy, with the fleece-flower root
For primary raw material, specifically it is made of the fleece-flower root, Fructus Corni, matrimony vine, Radix Salviae Miltiorrhizae and chrysanthemum；Preparation method be by the fleece-flower root of processing with
Fructus Corni, the fruit of Chinese wolfberry, Radix Salviae Miltiorrhizae, chrysanthemum mixing, are directly crushed to 40~50 mesh and obtain.CN102132919A, which discloses one kind, to be had
Beverage that is antifatigue, adjusting immune function and preparation method thereof, is made of following methods：By the fruit of Chinese wolfberry, Fructus Corni, mulberry fruit,
Wolfberry leaf, dark plum, wild jujube, Gorgon fruit, chrysanthemum are after cleaning, and twice, for the first time plus the water of 10 times of weight, decoction 2 are small for extracting in water
When, second of the water for adding 8 times of weight when decoction 1.5 is small, decocts extracting solution respectively with the filtering of 120~200 mesh numbers, 11000 twice
It rev/min is centrifuged at a high speed, is concentrated into 1.25~1.28g/cm of density3, extraction cream is made, according to conventional beverage processing technology
Beverage is made, wherein the weight of extraction cream, which accounts for, is made beverage weight percentage as 2.7~3.0%.
The flavour of a drug that the above-mentioned product with antifatigue effect uses are numerous, and preparation method is conventional method.
The content of the invention
It is an object of the invention to provide a kind of preparation method of the fermentate based on the bulk pharmaceutical chemicals containing Fructus Corni, this method
Bulk pharmaceutical chemicals can be simplified, and shorten fermentation time.Another object of the present invention is to provide a kind of fermentate, have antifatigue
Effect.It is still another object of the present invention to provide a kind of purposes of fermentate.The purpose of the present invention is by following technical solution
It realizes.
The present invention provides a kind of preparation method of fermentate, includes the following steps：
(1) bulk pharmaceutical chemicals being made of Fructus Corni, matrimony vine and chrysanthemum are crushed, takes 40~65 parts by weight of Fructus Corni, Chinese holly respectively
20~40 parts by weight of Qi and 10~35 parts by weight of chrysanthemum are mixed with water, obtain suspension；
(2) suspension is digested using pectase, cellulase, zytase and trypsase, obtains enzyme
Solve liquid；
(3) nutriment is added in into the enzymolysis liquid, sterilizes, obtain fermentation medium；
(4) into the fermentation medium, inoculation yeast bacterium carries out anaerobic fermentation 8~20h of culture, then inoculating lactic acid bacterium
Standing for fermentation 25~100h of culture is carried out, obtains cultured products；
(5) cultured products are subjected to separation of solid and liquid, obtain the fermentate.
Preparation in accordance with the present invention, it is preferable that Fructus Corni is 45~50 parts by weight, and matrimony vine is 20~30 parts by weight,
And chrysanthemum is 10~30 parts by weight.
Preparation in accordance with the present invention, it is preferable that in step (1), the dosage of water is the 6~16 of bulk pharmaceutical chemicals total weight
Times.
Preparation in accordance with the present invention, it is preferable that in step (2), the total weight based on suspension, the dosage of pectase
For 0.05~0.25wt%, the dosage of cellulase is 0.03~0.2wt%, the dosage of zytase for 0.05~
0.25wt%, the dosage of trypsase is 0.03~0.2wt%.
Preparation in accordance with the present invention, it is preferable that in step (2), the pH value of enzymolysis is 6.0~7.5, and hydrolysis temperature is
45~65 DEG C, enzymolysis time is 2~5h.
Preparation in accordance with the present invention, it is preferable that in step (3), the nutriment includes 0.1~0.5wt% ferment
Female cream, 1~5wt% ammonium salts, 0.02~0.2wt% zinc salts, 0.02~0.2wt% magnesium salts and 0.2~0.8wt% phosphate；With
Upper weight percent is all based on the total weight of the enzymolysis liquid.
Preparation in accordance with the present invention, it is preferable that in the anaerobic fermentation culture of step (4), the inoculum concentration of saccharomycete is
0.2~1.2wt% of the fermentation medium, fermentation temperature are 26~32 DEG C, and fermentation time is 10~18h.
Preparation in accordance with the present invention, it is preferable that in the standing for fermentation culture of step (4), the inoculum concentration of lactic acid bacteria is
0.2~1.0wt% of the fermentation medium, 24~36h of fermented and cultured at 35~40 DEG C, then ferments at 60~75 DEG C
Cultivate 18~35h.
The present invention also provides the fermentates that the preparation method of any of the above-described obtains.
The present invention also provides above-mentioned fermentate in the drug with antifatigue effect, feed addictive or health products are prepared
Purposes.
The present invention is first digested by using certain enzyme, after sequentially add saccharomycete and mode that lactic acid bacteria is fermented
Fructus Corni, matrimony vine and chrysanthemum bulk pharmaceutical chemicals are handled, obtain the fermentate with notable antifatigue effect；This method is fermented
Time is short, so as to be more advantageous to industrialized production.Customary preparation methods are decocted compared with the water of traditional Chinese medicine combination, it is of the invention
The active ingredient of original Chinese medicine is rich in fermentate；Meanwhile compared with Fructus Corni, matrimony vine, each medicinal material of chrysanthemum list, by the present invention
The anti-fatigue health efficacy of product has been obviously improved after method processing.
Specific embodiment
With reference to specific embodiment, the present invention is further illustrated, but protection scope of the present invention is not limited to
This.
The bulk pharmaceutical chemicals for fermentation of the present invention are made of Fructus Corni, matrimony vine and chrysanthemum.Fructus Corni is Cornaceae plant
The drying and ripening pulp of Fructus Corni Cornus officinalis Sieb.et Zucc..Matrimony vine is plant of Solanaceae lycium barbarum
The dry mature fruit of Lycium barbarum L..Chrysanthemum is feverfew chrysanthemum Chrysanthemum morifolium
Ramat. dry capitulum.
The preparation method of the fermentate of the bulk pharmaceutical chemicals containing Fructus Corni of the present invention includes the following steps：(1) grinding dispersion step；
(2) enzymolysis step；(3) culture medium preparation steps；(4) fermentation step：(5) separating step.It optionally, can also be dense including (6)
Contracting drying steps.It introduces in detail below.
In the grinding dispersion step (1) of the present invention, the bulk pharmaceutical chemicals being made of Fructus Corni, matrimony vine and chrysanthemum are crushed, point
40~65 parts by weight of Fructus Corni, 20~40 parts by weight of matrimony vine and 10~35 parts by weight of chrysanthemum and water is not taken to be mixed to get suspension.
Each bulk pharmaceutical chemicals can be crushed respectively, then weigh the amount of corresponding proportion, then mixed with water；Each raw material can also first be weighed
Then medicine crushes together, then mixed with water.The bulk pharmaceutical chemicals can be crushed to below 10 mesh, be preferably below 24 mesh.According to this
A preferred embodiment is invented, Fructus Corni is crushed to below 10 mesh, and matrimony vine and chrysanthemum are crushed to respectively below 24 mesh.At this
In invention, it is preferable that Fructus Corni is 45~60 parts by weight, matrimony vine is 20~30 parts by weight, and chrysanthemum is 10~30 parts by weight, example
Such as 25~30 parts by weight.According to embodiment of the present invention, Fructus Corni is 45 parts by weight, matrimony vine is 30 parts by weight, and chrysanthemum
Flower is 25 parts by weight.According to another implementation of the invention, Fructus Corni is 57 parts by weight, matrimony vine is 29 parts by weight, and chrysanthemum
Flower is 14 parts by weight.According to another implementation of the invention, Fructus Corni is 60 parts by weight, matrimony vine is 25 parts by weight, and chrysanthemum
Flower is 15 parts by weight.Such combination can so that the antifatigue effect of fermentate is more notable.
In the grinding dispersion step (1) of the present invention, the dosage of water is 6~16 times of bulk pharmaceutical chemicals total weight；Preferably 8~
12 times.Using such proportioning, be conducive to subsequent enzymolysis and fermentation process.Water can be distilled water or deionized water etc., only
Otherwise influence enzymolysis and fermentation process.
In the enzymolysis step of the present invention, using pectase, cellulase, zytase and trypsase to the suspension
Liquid is digested, and obtains enzymolysis liquid.Total weight based on suspension, the dosage of pectase can be 0.05~0.25wt%, excellent
Elect 0.06~0.2wt% as, more preferably 0.1~0.15wt%.Total weight based on suspension, the dosage of cellulase can be with
It is preferably 0.05~0.2wt% for 0.03~0.25wt%, more preferably 0.1~0.15wt%.Gross weight based on suspension
Amount, the dosage of zytase can be 0.05~0.25wt%, be preferably 0.06~0.2wt%, more preferably 0.1~
0.15wt%.Total weight based on suspension, the dosage of trypsase can be 0.03~0.2wt%, preferably 0.05~
0.15wt%, more preferably 0.1~0.15wt%.According to embodiment of the present invention, the dosage of pectase for 0.1~
0.15wt%, the dosage of cellulase are 0.1~0.15wt%, and the dosage of zytase is 0.1~0.15wt%, trypsase
Dosage be 0.1~0.15wt%.A specific embodiment according to the present invention, the dosage of pectase are 0.1wt%, fiber
The dosage of plain enzyme is 0.1wt%, and the dosage of zytase is 0.1wt%.Another embodiment according to the present invention,
The dosage of pectase is 0.15wt%, and the dosage of cellulase is 0.1wt%, and the dosage of zytase is 0.1wt%, pancreas egg
The dosage of white enzyme is 0.1wt%.
In the present invention, the enzyme activity of pectase can be 15~550,000 U/g, be preferably 25~500,000 U/g, more preferably
35~500,000 U/g.The enzyme activity of cellulase can be 20~550,000 U/g, be preferably 25~550,000 U/g, more preferably 35~
500000 U/g.The enzyme activity of zytase can be 15~600,000 U/g, be preferably 25~500,000 U/g, more preferably 35~500,000
U/g, the enzyme activity of trypsase can be 15~550,000 U/g, be preferably 25~500,000 U/g, more preferably 35~500,000 U/g.
A specific embodiment according to the present invention, the enzyme activity of pectase is 500,000 U/g, and the enzyme activity of cellulase is 500,000 U/
G, the enzyme activity of zytase is 500,000 U/g, and the enzyme activity of trypsase is 500,000 U/g.
During using mentioned kind and the biological enzyme of dosage, bulk pharmaceutical chemicals can fully be digested, amino is provided for fermented and cultured
The raw materials such as acid, monose, oligosaccharides, so as to reduce fermentation time.
In the enzymolysis step (2) of the present invention, the pH value of enzymolysis can be 6.0~7.5, be preferably 6.0~7.0；Enzymolysis
Temperature can be 45~65 DEG C, be preferably 50~60 DEG C, more preferably 52~58 DEG C；Enzymolysis time can be 2~5h, be preferably
2.5~4.5h, more preferably 3~4h.A specific embodiment according to the present invention, the pH value of enzymolysis is 6.5, hydrolysis temperature
For 55 DEG C, enzymolysis time 3.5h.Enzymatic hydrolysis condition using the present invention can make above-mentioned three kinds of biological enzyme fill bulk pharmaceutical chemicals
Divide enzymolysis, so as to reduce fermentation time.
In the culture medium preparation steps (3) of the present invention, nutriment is added in into the enzymolysis liquid, sterilizes, is fermented
Culture medium.Methods known in the art may be employed to sterilize, which is not described herein again.Based on the total weight of the enzymolysis liquid,
The nutriment include 0.1~0.5wt% yeast extracts, 1~5wt% ammonium salts, 0.02~0.2wt% zinc salts, 0.02~
0.2wt% magnesium salts and 0.2~0.8wt% phosphate.Commercially available yeast extract may be employed.The dosage of yeast extract is preferably 0.2~
0.4wt%.Ammonium salt can be selected from least one of ammonium sulfate, ammonium nitrate or ammonium carbonate, be preferably ammonium sulfate.The dosage of ammonium salt
Preferably 1~3wt%, more preferably 1.5~2wt%.Zinc salt can be in zinc sulfate, zinc chloride, zinc nitrate at least one
Kind, and preferably zinc chloride.Based on the total weight of the enzymolysis liquid, the nutriment can include 0.02~0.2wt%, excellent
Elect the zinc salt of 0.04~0.1wt%, more preferably 0.05~0.08wt% as.Magnesium salts can be selected from magnesium sulfate, magnesium chloride, nitric acid
At least one of magnesium, magnesium carbonate are preferably magnesium sulfate.The dosage of magnesium salts is preferably 0.03~0.12wt%, more preferably
0.04~0.06wt%.Phosphate can be selected from sodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, dipotassium hydrogen phosphate,
At least one of potassium dihydrogen phosphate, calcium phosphate, magnesium phosphate are preferably potassium dihydrogen phosphate.Phosphatic dosage is preferably 0.25
~0.6wt%, more preferably 0.3~0.5wt%.Above-mentioned nutriment is added in enzymolysis liquid, can effectively meet saccharomycete, breast
The nutrient demand of the fermented and cultured of sour bacterium, and fermentation time can be reduced.
In the culture medium preparation steps (3) of the present invention, nutriment can also contain sylvite.The sylvite and phosphate are excellent
It elects same substance, such as potassium phosphate, dipotassium hydrogen phosphate or potassium dihydrogen phosphate as, is preferably potassium dihydrogen phosphate.
In the fermentation step (4) of the present invention, into the fermentation medium, inoculation yeast bacterium carries out anaerobic fermentation culture 8
~20h, then inoculating lactic acid bacterium, which be left to ferment, cultivates 25~100h, obtains cultured products.The present invention fermentation time compared with
It is short.The example of the saccharomycete of the present invention includes but not limited to the high activity dried yeast of Angel Yeast Co., Ltd's production.This
The lactic acid bacteria of invention can be selected from the one or more of lactobacillus bulgaricus and streptococcus thermophilus；Preferably bulgarian milk bar
Bacterium and the mixture of streptococcus thermophilus.The weight ratio of lactobacillus bulgaricus and streptococcus thermophilus can be 1:10~10:1, it is excellent
Elect 1 as:5~5:1, more preferably 1:1.
In the anaerobic fermentation culture of step (4), the inoculum concentration of saccharomycete for the fermentation medium 0.2~
1.2wt% is preferably 0.5~1.0wt%, more preferably 0.7~0.9wt%.Fermentation temperature is 26~32 DEG C, preferably 28~
30 DEG C, more preferably 28 DEG C.Fermentation time is 10~18h, is preferably 12~16h, more preferably 16h.Under these conditions, may be used
So that the effect of saccharomycete gives full play to and saves the time.
In the standing for fermentation culture of step (4), the inoculum concentration of lactic acid bacteria for the fermentation medium 0.2~
1.0wt% is preferably 0.2~0.8wt%, more preferably 0.4~0.6wt%.35~40 DEG C, be preferably 36~38 DEG C, more
25~36h of fermented and cultured, preferably 28~34h, more preferably 32h at preferably 37 DEG C；Then 60~75 DEG C, be preferably 62
18~35h of fermented and cultured, preferably 25~32h, more preferably 30h at~68 DEG C, more preferably 65 DEG C.
In the separating step (5) of the present invention, the cultured products are subjected to separation of solid and liquid, obtain the fermentate.Gu
The isolated zymotic fluid of liquid and solid residue, the method for separation of solid and liquid can be centrifugation or filtering, and be preferably to centrifuge.
In the concentrate drying step (6) of the present invention, by zymotic fluid concentration, drying, yeast powder is obtained.It can subtract
It is concentrated and is dried under the conditions of pressure, which is not described herein again.
Fermentate is obtained using above-mentioned preparation method.The fermentate of the present invention can be above-mentioned zymotic fluid, or hair
Ferment powder.Fermentate is sometimes referred to as ferment, fermentation composition etc..According to embodiment of the present invention, the yeast powder
Water content less than 5wt%, the vigor of superoxide dismutase SOD is more than or equal to 420 ± 5U/g, and viable count of lactobacillus is more than etc.
In 3.0 ± 0.5 × 108CFU/g。
The fermentate of the present invention is notable with antifatigue effect under antifatigue effect, particularly low pressure, thus can be used in
It prepares in the drug with antifatigue effect, feed addictive or health products.Health products can be food with health role,
Such as candy, beverage, drinks, bread, biscuit etc..Drug can be a variety of conventional solid dosage forms or liquid dosage form, no longer go to live in the household of one's in-laws on getting married
It states.Feed addictive refers in Feed Manufacturing processing, a small amount of or micro substance added during use, and dosage is very in feed
Less but effect is notable.
The raw material of following embodiment and comparative example is described as follows：
Saccharomycete：The high activity dried yeast of Angel Yeast Co., Ltd's production；
Lactic acid bacteria：Weight ratio is 1:1 lactobacillus bulgaricus and streptococcus thermophilus.
Following embodiment and the test method of comparative example are described as follows：
<The measure of iridoid glycoside content before and after fermentation>
The preparation of reference substance solution：It is appropriate that precision weighs 8-O-Acetylharpagide reference substance, and methanol dissolving is configured to every 1mL about
Reference substance solution containing 0.4mg.
The preparation of test solution：
Fermentate sample：The yeast powder 0.25g of embodiment 2 is weighed (compared with Fructus Corni 1.5g crude drugs, matrimony vine 1.0g crude drugs
With chrysanthemum 0.83g crude drugs), it is accurately weighed, it is placed in conical flask with cover, precision adds in methanol 25mL, and weighed weight is ultrasonically treated
(power 250W, frequency 40KHz) 30min, lets cool, then weighed weight, and the weight of less loss is supplied with methanol, is shaken up, and filtration takes continuous
Filtrate is as test solution.
Aqueous extracts sample：Fructus Corni 1.5g crude drugs, matrimony vine 1.0g crude drugs and chrysanthemum 0.83g crude drugs are taken, water impregnates 30min,
Decoct extraction 2 times, 10 times of amounts of each amount of water, when each extraction time 2 is small, filtering, filtrate merges, and is concentrated into dry powder, is placed in
In conical flask with cover, precision adds in methanol 25mL, and weighed weight is ultrasonically treated (power 250W, frequency 40KHz) 30min, puts
It is cold, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, filters, takes subsequent filtrate as test solution.
<Absorbance measurement>
Precision measures reference substance solution, test solution and each 0.4mL of methanol, is placed in 10mL measuring bottles, adds 1.0mol/L
Hydrochloric acid solution 3mL puts in 90 degree of water-baths and keeps the temperature 15min, lets cool, and adds dinitrophenylhydrazine ethyl alcohol test solution 0.5mL, puts 90 degree of water-baths and protects
Warm 30min, lets cool, and adds 1moL/L sodium hydroxides methanol aqueous solution (4g sodium hydroxides, 20mL water, 80mL methanol) 4mL, adds methanol
Scale is diluted to, is shaken up, after room temperature 2h, using above-mentioned methanol reaction solution as blank, spectrophotometric determination is used at 586nm
Absorbance A.Iridoid glycoside content calculation formula is as follows：
CSample=ASample×CMark/AMark
Table 1, fermentate sample and the measurement result of Aqueous extracts sample iridoid glycoside glycosides content
Embodiment 1
1) Fructus Corni, matrimony vine and chrysanthemum are crushed respectively, and is sieved using No. 2 sieves (24 mesh) of pharmacopeia, obtain raw material
Medicinal powder end；Then be 45 parts by weight according to Fructus Corni, matrimony vine is 30 parts by weight and chrysanthemum is that 25 parts by weight weigh bulk pharmaceutical chemicals powder,
Mixing, and the pure water of 10 times of amounts (weight ratio) of bulk pharmaceutical chemicals is added in, suspension is made.
2) by suspension and the pectase (500,000 U/g) based on suspension total weight 0.1wt%, the cellulose of 0.1wt%
Enzyme (500,000 U/g), the zytase (500,000 U/g) of 0.1wt%, the trypsase (500,000 U/g) of 0.1wt% mix and carry out enzyme
Solution, obtains enzymolysis liquid；Enzymatic hydrolysis condition is：PH is 6.5, and temperature is 55 DEG C, enzymolysis time 3.5h.
3) total weight based on the enzymolysis liquid, adds following nutriment：Yeast extract 0.3wt%, ammonium sulfate
1.5wt%, magnesium sulfate 0.05wt%, potassium dihydrogen phosphate 0.4wt% and zinc chloride 0.05wt%, fermented and cultured is obtained after sterilizing
Base.
4) to the saccharomycete of fermentation medium inoculation 0.8wt%, anaerobic fermentation 16h at 28 DEG C；Then 0.5wt% breasts are inoculated with
Sour bacterium is left to ferment 32h at 37 DEG C, is then left to ferment 30h at 65 DEG C.
5) centrifugal treating after fermentation, is carried out, isolated zymotic fluid and solid residue, the zymotic fluid are this implementation
The fermentate of example.
Embodiment 2
The zymotic fluid of embodiment 1 is concentrated under reduced pressure, is dried under reduced pressure to obtain yeast powder.The water content of gained yeast powder is less than
5wt%；The vigor of superoxide dismutase (SOD) is more than (420 ± 5) U/g；Viable count of lactobacillus be more than (3.0 ± 0.5) ×
108CFU/g；Iridoid glycoside is more than 5wt%.
Embodiment 3
According to weight ratio it is 1 by the zymotic fluid of embodiment 1 and water:6 are mixed to get mixture, are then based on the total of mixture
Weight addition 15wt% sucrose, 0.5wt% citric acids ,-filling-cooling of progress homogeneous-degassing-pasteurize and etc., it obtains
Enzyme beverage.
Embodiment 4
By the zymotic fluid of embodiment 1 and 45 ° of white wine and drinking water according to 1:8:4 ratio mixing, carries out homogeneous-Pasteur and kills
Bacterium-filling step, obtains ferment wine.
Embodiment 5
1) Fructus Corni, matrimony vine and chrysanthemum are crushed respectively, Fructus Corni crosses No. 1 sieve (10 mesh) of pharmacopeia, and matrimony vine and chrysanthemum cross medicine
No. 2 sieves (24 mesh sieve) of allusion quotation, obtain bulk pharmaceutical chemicals powder；Then be 57 parts by weight according to Fructus Corni, matrimony vine is 29 parts by weight and chrysanthemum
Bulk pharmaceutical chemicals powder is weighed for 14 parts by weight, is mixed, and adds in the pure water of 10 times of amounts (weight ratio) of bulk pharmaceutical chemicals, suspension is made.
2) by suspension and the pectase (500,000 U/g) based on suspension total weight 0.1wt%, the cellulose of 0.1wt%
Enzyme (500,000 U/g), the zytase (500,000 U/g) of 0.1wt%, the trypsase (500,000 U/g) of 0.1wt% mix and carry out enzyme
Solution, obtains enzymolysis liquid；Enzymatic hydrolysis condition is：PH is 6.5, and temperature is 55 DEG C, enzymolysis time 3.5h.
3) total weight based on the enzymolysis liquid, adds following nutriment：Yeast extract 0.3wt%, ammonium sulfate
1.5wt%, zinc chloride 0.05wt%, magnesium sulfate 0.05wt% and potassium dihydrogen phosphate 0.4wt%, fermented and cultured is obtained after sterilizing
Base.
4) to the saccharomycete of fermentation medium inoculation 0.8wt%, anaerobic fermentation 16h at 28 DEG C；Then 0.5wt% breasts are inoculated with
Sour bacterium is left to ferment 32h at 37 DEG C, is then left to ferment 30h at 65 DEG C.
5) centrifugal treating after fermentation, is carried out, isolated zymotic fluid and solid residue, the zymotic fluid are this implementation
The fermentate of example.
Embodiment 6
The zymotic fluid of embodiment 5 is concentrated under reduced pressure, is dried under reduced pressure to obtain yeast powder.The water content of gained yeast powder is less than
5wt%；The vigor of superoxide dismutase (SOD) is more than (420 ± 5) U/g；Viable count of lactobacillus be more than (3.0 ± 0.5) ×
108CFU/g。
Embodiment 7
1) Fructus Corni, matrimony vine and chrysanthemum are crushed respectively, Fructus Corni crosses No. 1 sieve (10 mesh) of pharmacopeia, and matrimony vine and chrysanthemum cross medicine
No. 2 sieves (24 mesh sieve) of allusion quotation, obtain bulk pharmaceutical chemicals powder；Then be 60 parts by weight according to Fructus Corni, matrimony vine is 30 parts by weight and chrysanthemum
Bulk pharmaceutical chemicals powder is weighed for 10 parts by weight, is mixed, and adds in the pure water of 12 times of amounts (weight ratio) of bulk pharmaceutical chemicals, suspension is made.
2) by suspension and the pectase (500,000 U/g) based on suspension total weight 0.15wt%, the cellulose of 0.1wt%
Enzyme (500,000 U/g), the zytase (500,000 U/g) of 0.1wt%, the trypsase (500,000 U/g) of 0.1wt% mix and carry out enzyme
Solution, obtains enzymolysis liquid；Enzymatic hydrolysis condition is：PH is 6.5, and temperature is 55 DEG C, enzymolysis time 3.5h.
3) total weight based on the enzymolysis liquid, adds following nutriment：Yeast extract 0.3wt%, ammonium sulfate
2.0wt%, magnesium sulfate 0.05wt%, potassium dihydrogen phosphate 0.4wt% and zinc chloride 0.05wt%, fermented and cultured is obtained after sterilizing
Base.
4) to the saccharomycete of fermentation medium inoculation 0.8wt%, anaerobic fermentation 16h at 28 DEG C；Then 0.5wt% breasts are inoculated with
Sour bacterium is left to ferment 32h at 37 DEG C, is then left to ferment 30h at 65 DEG C.
5) centrifugal treating after fermentation, is carried out, isolated zymotic fluid and solid residue, the zymotic fluid are this implementation
The fermentate of example.
Embodiment 8
The zymotic fluid of embodiment 7 is concentrated under reduced pressure, is dried under reduced pressure to obtain yeast powder.The water content of gained yeast powder is less than
5wt%；The vigor of superoxide dismutase (SOD) is more than (420 ± 5) U/g；Viable count of lactobacillus be more than (3.0 ± 0.5) ×
108CFU/g；Iridoid glycoside is more than 5wt%.
Experimental example 1- effect experiments
<The preparation of drug>
(yeast powder group) of the invention：The yeast powder 2.5g of embodiment 2 is weighed (compared with Fructus Corni 15g crude drugs, matrimony vine 10g
Crude drug and chrysanthemum 8.3g crude drugs), add in distilled water be settled to 120ml to get.
Aqueous extract group：Fructus Corni 15g, matrimony vine 10g and chrysanthemum 8.3g are taken, water impregnates 30min, decocts extraction 2 times, every time
When 10 times of amount of water amount, each extraction time 2 are small, filtering, filtrate merges, and rotary evaporation concentration is settled to distilled water
120ml to get.
Fructus Corni extracting solution group：Fructus Corni 33.3g is taken, water impregnates 30min, decocts extraction 2 times, each 10 times of amount of water
When amount, each extraction time 2 are small, filtering, filtrate merges, rotary evaporation concentration, with distilled water be settled to 120ml to get.
Fructus lycii extracted solution group：Matrimony vine 33.3g is taken, water impregnates 30min, decocts extraction 2 times, and each 10 times of amount of water is measured, often
When secondary extraction time 2 is small, filtering, filtrate merges, rotary evaporation concentration, with distilled water be settled to 120ml to get.
Chrysanthemum extract liquid group：Chrysanthemum 33.3g is taken, water impregnates 30min, decocts extraction 2 times, and each 10 times of amount of water is measured, often
When secondary extraction time 2 is small, filtering, filtrate merges, rotary evaporation concentration, with distilled water be settled to 120ml to get.
<Experimental method>
Kunming mice 60 is taken, is randomly divided into 6 groups, every group 10, medication is as follows：
Blank control group：Daily gavage 20ml/kg physiological saline；
Ferment powder group：The yeast powder liquid of the present invention of daily gavage 20ml/kg weight；
Aqueous extracts group：The aqueous extract of daily gavage 20ml/kg weight；
Fructus Corni group：The Fructus Corni extracting solution of daily gavage 20ml/kg weight；
Matrimony vine group：The fructus lycii extracted solution of daily gavage 20ml/kg weight；
Chrysanthemum group：The chrysanthemum extract liquid of daily gavage 20ml/kg weight.
Each group presses above-mentioned dosage gastric infusion 7 days, after last dose 60min, mouse is placed in environment under low pressure and fills water
1000ml beakers in, the lead of heavy burden own body weight 5wt%, water temperature is room temperature, carry out swimming test.It is all sunk to head
It cannot emerge within 10 seconds in water and exhaust state for power, the time that record each group exhausts state from swim to power exhausts as mouse power
Swimming time takes average value in each group.Solution takes mouse thymus and spleen and weighs, as dirty with the ratio of Organs Weight and weight
Device index.
<Experimental result>
Mouse low pressure swimming with a load attached to the body experimental result is shown in Table 2.
Table 2, mouse low pressure swimming with a load attached to the body experimental result
| Group | Animal number of elements (n) | Dosage (g/kg) | Swimming time (s) |
| Blank control group | 10 | -- | 859.72±88.21△△ |
| Ferment powder group | 10 | 5.5 | 1108.74±136.80** |
| Aqueous extracts group | 10 | 5.5 | 1005.53±77.46**△ |
| Fructus Corni group | 10 | 5.5 | 954.73±91.01*△△ |
| Matrimony vine group | 10 | 5.5 | 958.66±72.00*△△ |
| Chrysanthemum group | 10 | 5.5 | 876.15±60.41△△ |
Note：Compared with blank control group,*P ＜ 0.05,**P ＜ 0.01；
Compared with ferment powder group,ΔP ＜ 0.05,ΔΔP ＜ 0.01.
From experimental data statistical result, compared with blank control group, ferment powder group, Aqueous extracts group, Fructus Corni group, Chinese holly
The more equal conspicuousness of mouse swimming time of Qi group improves (P ＜ 0.05~0.01)；Compared with ferment powder group, Aqueous extracts group, mountain
Fruit of medicinal cornel group, matrimony vine group and chrysanthemum group swimming time significantly reduce (P ＜ 0.05~0.01), this shows that ferment powder group promotes mouse low
The extension effect of pressure walking weight load is significantly better than Aqueous extracts group, Fructus Corni group, matrimony vine group and chrysanthemum group.
Mice organs index results are shown in Table 3.
Table 3, mice organs index results
| Group | Animal number of elements (n) | Dosage (g/kg) | Thymus index (%) | Index and spleen index (%) |
| Blank control group | 10 | -- | 0.24±0.08ΔΔ | 0.46±0.06ΔΔ |
| Ferment powder group | 10 | 5.5 | 0.46±0.07** | 0.65±0.07** |
| Aqueous extracts group | 10 | 5.5 | 0.40±0.04**Δ | 0.59±0.06**Δ |
| Fructus Corni group | 10 | 5.5 | 0.31±0.04ΔΔ | 0.53±0.06*ΔΔ |
| Matrimony vine group | 10 | 5.5 | 0.27±0.06ΔΔ | 0.49±0.07ΔΔ |
| Chrysanthemum group | 10 | 5.5 | 0.27±0.04ΔΔ | 0.44±0.07ΔΔ |
Note：Compared with blank control group,*P ＜ 0.05,**P ＜ 0.01；
Compared with ferment powder group,△P ＜ 0.05,△△P ＜ 0.01.
From experimental data statistical result, compared with blank control group, ferment powder group, the mouse thymus of Aqueous extracts group refer to
Number conspicuousness improves (P ＜ 0.01).Compared with ferment powder group, Aqueous extracts group, Fructus Corni group, matrimony vine group and chrysanthemum group thymus index
Conspicuousness reduces (P ＜ 0.05~0.01), shows that ferment powder group promotes mouse thymus index increase effect to be significantly better than Aqueous extracts
Group, Fructus Corni group, matrimony vine group and chrysanthemum group.Compared with blank control group, ferment powder group, Aqueous extracts group, the mouse of Fructus Corni group
Index and spleen index conspicuousness improves (P ＜ 0.05~0.01).Compared with ferment powder, Aqueous extracts group, Fructus Corni group, matrimony vine group and chrysanthemum
Group index and spleen index conspicuousness reduces (P ＜ 0.05~0.01).This shows that ferment powder group promotes mouse spleen index increase effect aobvious
It writes better than Aqueous extracts group, Fructus Corni group, matrimony vine group and chrysanthemum group.
Present invention is not limited to the embodiments described above, in the case of without departing substantially from the substantive content of the present invention, this field skill
Any deformation, improvement, the replacement that art personnel are contemplated that each fall within the scope of the present invention.
Claims (10)
1. a kind of preparation method of fermentate, which is characterized in that include the following steps：
(1) bulk pharmaceutical chemicals being made of Fructus Corni, matrimony vine and chrysanthemum are crushed, takes 40~65 parts by weight of Fructus Corni, matrimony vine 20 respectively
~40 parts by weight and 10~35 parts by weight of chrysanthemum are mixed with water, obtain suspension；
(2) suspension is digested using cellulase, pectase, zytase and trypsase, obtains enzymolysis liquid；
(3) nutriment is added in into the enzymolysis liquid, sterilizes, obtain fermentation medium；
(4) into the fermentation medium, inoculation yeast bacterium carries out anaerobic fermentation 8~20h of culture, and then inoculating lactic acid bacterium carries out
Standing for fermentation 25~100h of culture, obtains cultured products；
(5) cultured products are subjected to separation of solid and liquid, obtain the fermentate.
2. preparation method according to claim 1, which is characterized in that Fructus Corni is 45~50 parts by weight, matrimony vine for 20~
30 parts by weight, and chrysanthemum is 10~30 parts by weight.
3. preparation method according to claim 1, which is characterized in that in step (1), the dosage of water is bulk pharmaceutical chemicals total weight
6~16 times.
4. preparation method according to claim 1, which is characterized in that in step (2), the total weight based on suspension, fruit
The dosage of glue enzyme is 0.05~0.25wt%, and the dosage of cellulase is 0.03~0.2wt%, and the dosage of zytase is 0.05
~0.25wt%, the dosage of trypsase is 0.03~0.2wt%.
5. preparation method according to claim 4, which is characterized in that in step (2), the pH value of enzymolysis is 6.0~7.5,
Hydrolysis temperature is 45~65 DEG C, and enzymolysis time is 2~5h.
6. preparation method according to claim 5, which is characterized in that in step (3), the nutriment include 0.1~
0.5wt% yeast extracts, 1~5wt% ammonium salts, 0.02~0.2wt% zinc salts, 0.02~0.2wt% magnesium salts and 0.2~0.8wt%
Phosphate；More than weight percent is all based on the total weight of the enzymolysis liquid.
7. preparation method according to claim 6, which is characterized in that in the anaerobic fermentation culture of step (4), saccharomycete
Inoculum concentration is 0.2~1.2wt% of the fermentation medium, and fermentation temperature is 26~32 DEG C, and fermentation time is 10~18h.
8. preparation method according to claim 7, which is characterized in that in the standing for fermentation culture of step (4), lactic acid bacteria
Inoculum concentration is the 0.2~1.0wt%, 24~36h of fermented and cultured at 35~40 DEG C, then 60~75 of the fermentation medium
18~35h of fermented and cultured at DEG C.
9. the fermentate obtained according to claim 1~8 any one of them preparation method.
10. fermentate according to claim 9 is preparing drug, feed addictive or health products with antifatigue effect
In purposes.
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