CN110314177B - Radix Puerariae rich in isoflavone and its preparation method - Google Patents
Radix Puerariae rich in isoflavone and its preparation method Download PDFInfo
- Publication number
- CN110314177B CN110314177B CN201910527719.3A CN201910527719A CN110314177B CN 110314177 B CN110314177 B CN 110314177B CN 201910527719 A CN201910527719 A CN 201910527719A CN 110314177 B CN110314177 B CN 110314177B
- Authority
- CN
- China
- Prior art keywords
- aspergillus niger
- isoflavone
- radix puerariae
- pachyrhizua angulatus
- pueraria thomsonii
- 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.)
- Active
Links
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 235000008696 isoflavones Nutrition 0.000 title claims abstract description 43
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims description 9
- 241000228245 Aspergillus niger Species 0.000 claims abstract description 35
- 239000003814 drug Substances 0.000 claims abstract description 27
- 241000731396 Pueraria montana var. thomsonii Species 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 21
- 238000000855 fermentation Methods 0.000 claims abstract description 20
- 230000004151 fermentation Effects 0.000 claims abstract description 20
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 15
- 229940079593 drug Drugs 0.000 claims description 7
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 239000000725 suspension Substances 0.000 claims description 5
- 241000220485 Fabaceae Species 0.000 claims description 3
- 239000003963 antioxidant agent Substances 0.000 claims description 3
- 235000006708 antioxidants Nutrition 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 210000000748 cardiovascular system Anatomy 0.000 abstract description 8
- 206010028980 Neoplasm Diseases 0.000 abstract description 7
- 239000000126 substance Substances 0.000 abstract description 7
- 230000032683 aging Effects 0.000 abstract description 6
- 229920002472 Starch Polymers 0.000 abstract description 4
- 150000004676 glycans Chemical class 0.000 abstract description 4
- 239000001963 growth medium Substances 0.000 abstract description 4
- 229920001282 polysaccharide Polymers 0.000 abstract description 4
- 239000005017 polysaccharide Substances 0.000 abstract description 4
- 235000019698 starch Nutrition 0.000 abstract description 4
- 239000008107 starch Substances 0.000 abstract description 4
- 235000015097 nutrients Nutrition 0.000 abstract description 3
- 230000007721 medicinal effect Effects 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 238000002835 absorbance Methods 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 239000012085 test solution Substances 0.000 description 10
- 230000009466 transformation Effects 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- RXUWDKBZZLIASQ-UHFFFAOYSA-N Puerarin Natural products OCC1OC(Oc2c(O)cc(O)c3C(=O)C(=COc23)c4ccc(O)cc4)C(O)C(O)C1O RXUWDKBZZLIASQ-UHFFFAOYSA-N 0.000 description 8
- 239000000843 powder Substances 0.000 description 8
- HKEAFJYKMMKDOR-VPRICQMDSA-N puerarin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C1=C(O)C=CC(C2=O)=C1OC=C2C1=CC=C(O)C=C1 HKEAFJYKMMKDOR-VPRICQMDSA-N 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 238000005303 weighing Methods 0.000 description 7
- 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 6
- 230000000694 effects Effects 0.000 description 5
- 239000013558 reference substance Substances 0.000 description 5
- 235000010575 Pueraria lobata Nutrition 0.000 description 4
- 241000219781 Pueraria montana var. lobata Species 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 4
- 150000002515 isoflavone derivatives Chemical class 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- -1 DPPH free radical Chemical class 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 230000002292 Radical scavenging effect Effects 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 210000004165 myocardium Anatomy 0.000 description 3
- 230000000144 pharmacologic effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- 241000222383 Polyporales Species 0.000 description 2
- 230000006793 arrhythmia Effects 0.000 description 2
- 206010003119 arrhythmia Diseases 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 230000036772 blood pressure Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000000302 ischemic effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 150000002894 organic compounds Chemical class 0.000 description 2
- 230000036284 oxygen consumption Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 230000002000 scavenging effect Effects 0.000 description 2
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- 238000003892 spreading Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010056771 Glucosidases Proteins 0.000 description 1
- 102000004366 Glucosidases Human genes 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- 241000219780 Pueraria Species 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 238000004820 blood count Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000004526 pharmaceutical effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- 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)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/488—Pueraria (kudzu)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/06—Antiarrhythmics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Diabetes (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Heart & Thoracic Surgery (AREA)
- Cardiology (AREA)
- Hematology (AREA)
- Mycology (AREA)
- Obesity (AREA)
- Nutrition Science (AREA)
- Toxicology (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Urology & Nephrology (AREA)
- Vascular Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides a method for preparing isoflavone-rich pachyrhizua angulatus, which comprises the following steps: inoculating Aspergillus niger to radix Puerariae, and fermenting. The invention utilizes the fermentation of aspergillus niger, takes rich polysaccharide and starch contained in the pueraria thomsonii as a culture medium, does not need to add other nutrient substances, can obviously improve the content of the total isoflavone in the pueraria thomsonii in a short time, further improves the antioxidant activity of the pueraria thomsonii by improving the content of the total isoflavone, and has wide application prospect in preparing medicaments with obvious medicinal effect, such as cardiovascular system protection, tumor resistance, aging resistance and the like.
Description
Technical Field
The invention relates to the field of microbial fermentation, in particular to isoflavone-rich pachyrhizua angulatus and a preparation method thereof.
Background
The conversion of chemical components of traditional Chinese medicine by utilizing microbial fermentation technology is one of the important branches of traditional Chinese medicine biotechnology, and the essence of the conversion is that a series of enzymes generated by microorganisms act on the chemical components in the traditional Chinese medicine, the structure and/or content of the chemical components are changed through the catalytic action of the enzymes, so that the activity and/or toxicity of the traditional Chinese medicine are changed. Aspergillus niger is a fungus commonly used in transforming traditional Chinese medicines by microbial fermentation technology, and belongs to Aspergillus of Aphyllophorales, Moniliaceae, Aphyllophorales, Deuteromycotina. The microbial organic compound fermentation medium contains abundant enzyme systems such as amylase, pectinase and glucosidase, has the advantages of high safety, short fermentation and transformation time and the like, and is widely applied to the fields of food processing, organic compound transformation, traditional Chinese medicine effective component transformation and the like. Researches show that the aspergillus niger is used for fermenting and converting the traditional Chinese medicine, so that the content of active ingredients in the traditional Chinese medicine can be improved, and the structure of the traditional Chinese medicine can be changed, thereby improving the pharmacological activity of the traditional Chinese medicine.
Radix Puerariae is derived from dried root of Pueraria thomsonii Benth of Leguminosae, is one of the traditional Chinese medicines commonly used in traditional Chinese medicine, has effects of relieving muscles and reducing fever, promoting fluid production, promoting eruption, invigorating yang and relieving diarrhea, and has high safety. In addition to medical use, pachyrhizua angulatus is widely used for preparing food, health products and the like, such as kudzu powder, fine dried noodles, vermicelli, oral liquid, beverage, wine and the like. Researches show that the pueraria lobata contains isoflavone, polysaccharide, starch and other chemical components, wherein isoflavone components represented by puerarin are main active components of the pueraria lobata, and the pueraria lobata has the cardiovascular system protection effects of reducing blood pressure, slowing down heart rate, reducing myocardial oxygen consumption, increasing coronary blood flow, improving metabolism of normal and ischemic myocardium, inhibiting platelet aggregation, resisting arrhythmia, reducing blood sugar, reducing blood fat and the like, and also has the activities of resisting oxidation, resisting tumors, resisting aging and the like. The antioxidation is one of important pharmacological actions of the pachyrhizua angulatus, and is one of partial mechanisms of the pharmacological actions of protecting the cardiovascular system, resisting aging and the like.
However, the total isoflavone content in the pueraria thomsonii is low, so that the further development and application of the pueraria thomsonii are limited. Therefore, a method is found for improving the content of isoflavone in the pueraria thomsonii, and the pueraria thomsonii has a very great application prospect in preparing medicaments for protecting cardiovascular systems, resisting tumors and resisting aging.
Disclosure of Invention
The invention aims to provide isoflavone-rich pachyrhizua angulatus and a preparation method thereof.
The invention provides a method for preparing isoflavone-rich pachyrhizua angulatus, which comprises the following steps:
inoculating Aspergillus niger to radix Puerariae, and fermenting.
Further, the fermentation time is 7 days or more, preferably 9 days or more, and more preferably 11 days or more.
Further, the pachyrhizua angulatus is sterilized pachyrhizua angulatus;
inoculated with a suspension of aspergillus niger spores;
the ratio of the pachyrhizua angulatus to the aspergillus niger spores is (8-12) g: 2X 108CFU；
The fermentation condition is an aseptic environment with the temperature of 25-35 ℃ and the humidity of 95-99%.
Further, the mode of the mould sterilization treatment is as follows: sterilizing radix puerariae under irradiation of an ultraviolet lamp for 30-60 min;
the Aspergillus niger is Aspergillus niger spore suspension with concentration of 1 × 108CFU/ml；
The ratio of the pachyrhizua angulatus to the aspergillus niger spores is 10 g: 2X 108CFU；
The fermentation conditions were an aseptic environment with a temperature of 30 ℃ and a humidity of 99%.
The invention also provides isoflavone-rich pachyrhizua angulatus, and the weight percentage of the isoflavone in the isoflavone-rich pachyrhizua angulatus is more than 1.651%.
Further, the isoflavone-rich pachyrhizua angulatus has the weight percentage of the isoflavone of more than 2.019%, preferably more than 2.562%.
Further, the isoflavone-rich pachyrhizua angulatus is prepared according to the method.
The invention also provides a pharmaceutical composition which is prepared by taking the pueraria thomsonii rich in isoflavone as an active ingredient and adding pharmaceutically acceptable auxiliary materials.
The invention also provides the application of the isoflavone-rich pachyrhizua angulatus in preparing pachyrhizua angulatus products;
preferably, the radix puerariae product is a medicament for protecting cardiovascular system, an antioxidant medicament, an anti-tumor medicament or an anti-aging medicament.
Furthermore, the medicine for protecting the cardiovascular system comprises medicines for reducing blood pressure, slowing heart rate, reducing oxygen consumption of cardiac muscle, increasing coronary blood flow, improving metabolism of normal and ischemic cardiac muscle, inhibiting platelet aggregation, resisting arrhythmia, reducing blood sugar and reducing blood fat;
the tumor is a tumor associated with estrogen abnormality, preferably breast cancer.
CFU/mL represents the number of total colonies per mL.
The radix Puerariae product is medicine, food or health product containing radix Puerariae.
Experiments prove that the method for treating the pachyrhizua angulatus by adopting the Aspergillus niger fermentation takes rich polysaccharide and starch contained in the pachyrhizua angulatus as culture media, does not need to add other nutrient substances, can obviously improve the content of the total isoflavone in the pachyrhizua angulatus in a short time, further improves the antioxidant activity of the pachyrhizua angulatus by improving the content of the total isoflavone, and has wide application prospect in preparing medicaments with obvious pharmaceutical effect, such as cardiovascular system protection, aging resistance, tumor resistance and the like, and pharmaceutical and edible pachyrhizua angulatus products.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
The raw materials and instruments used in the present invention are known products, and are obtained by purchasing commercially available products.
Example 1A. niger fermentatively transformed Pueraria thomsonii sample
1. Preparation of aspergillus niger spores: collecting mature Aspergillus niger grown on PDA culture medium, gently scraping spores with inoculating loop, suspending in sterile ultrapure water, counting under microscope by using blood count plate method, and adjusting to 1 × 10 concentration with sterile ultrapure water8CFU/ml Aspergillus niger spore suspension.
2. Preparation of aspergillus niger fermentation transformed pachyrhizua angulatus sample:
(1) collecting coarse powder of radix Puerariae (dried root of Pueraria thomsonii Benth of Leguminosae, identified by professor Chenolo, university of Chinese medicine and pharmacy) 10g, spreading on the bottom of culture dish, sterilizing with ultraviolet irradiation on ultra-clean bench for 30min (ultraviolet irradiation intensity: 90 μ W/cm)2Irradiation distance: 25-35cm), turning over once, and continuing to irradiate for 30 min.
(2) The sterilized radix puerariae samples are divided into a blank control group and an aspergillus niger fermentation transformation group (each group is in parallel with 3 parts). Adding Aspergillus niger fermentation conversion group with concentration of 1 × 1082ml of Aspergillus niger spore suspension of CFU/ml, mixing, adding equal volume of sterile ultrapure water into blank control group, culturing in artificial climate box (temperature: 30 deg.C, humidity: 99%), sampling at 7 th, 9 th and 11 th days, spreading the sample on the bottom of culture dish, sterilizing on ultra-clean bench under ultraviolet lamp for 30min (ultraviolet irradiation intensity: 90 μ W/cm)2Irradiation distance: 25-35cm), turning over once, continuously irradiating for 30min for sterilization, placing in a reduced pressure drying oven, and drying under reduced pressure at 60 ℃ for 24 h. Obtaining the radix puerariae samples obtained by fermenting the aspergillus niger for different time (7 days, 9 days and 11 days). And blank control samples obtained at different times (7, 9 and 11 days).
Example 2 Aspergillus niger fermentatively transformed Pueraria thomsonii sample
The same method as that of example 1 is adopted, the pachyrhizua angulatus crude powder in the step 2 is increased from 10g to 12g, and the pachyrhizua angulatus crude powder is cultured under the following conditions of an artificial climate box at constant temperature and humidity respectively: (a) temperature: 25 ℃, humidity: 95%, (b) temperature: 35 ℃, humidity: 99%, (c) temperature: 30 ℃, humidity: 99 percent; culturing for 11 days to obtain a pachyrhizua angulatus sample.
Example 3 Aspergillus niger fermentatively transformed Pueraria thomsonii sample
The same method as that of example 1 is adopted, the pachyrhizua angulatus crude powder in the step 2 is reduced from 10g to 8g, and the pachyrhizua angulatus crude powder is cultured under the following conditions of an artificial climate box at constant temperature and constant humidity respectively: (a) temperature: 25 ℃, humidity: 95%, (b) temperature: 35 ℃, humidity: 99%, (c) temperature: 30 ℃, humidity: 99 percent; culturing for 11 days to obtain a pachyrhizua angulatus sample.
The beneficial effects of the present invention are demonstrated by the following experimental examples.
Experimental example 1 measurement of Total isoflavone content in Pueraria thomsonii sample
1. Preparation of control solutions
A proper amount of puerarin contrast (Kagaku Biotech Co., Ltd., Sichuan, batch No. wkq18040806, purity: HPLC ≥ 98%) is precisely weighed, and 70% ethanol is added to make into contrast stock solution containing 1mg per ml for use.
2. Preparation of test solution
Taking about 0.5g of a sample to be detected, precisely weighing, placing in a 100mL triangular flask, adding 20mL of 70% ethanol, weighing, ultrasonically extracting for 30min, cooling, weighing again, complementing the loss weight with 70% ethanol, shaking up, tightly sucking 0.1mL of supernatant into a 10mL measuring flask, adding distilled water to dilute to scale, and shaking up to obtain the product.
3. Methodology investigation
(1) And (3) linear relation investigation: precisely absorbing 0.1mL, 0.2 mL, 0.3 mL, 0.40 mL, 0.50 mL and 0.6mL of the puerarin reference substance stock solution respectively, placing the puerarin reference substance stock solution in a 50mL measuring flask, dissolving the puerarin reference substance stock solution to the scale with 70 percent ethanol, shaking the solution evenly to prepare 0.002, 0.004, 0.006, 0.008, 0.010 and 0.012mg/mL of reference substance solution respectively, measuring the absorption value under the condition of 250nm of an ultraviolet spectrophotometer, drawing a standard curve by taking the puerarin concentration (mg/mL) as an abscissa (X) and the absorbance as an ordinate (Y), and obtaining a linear regression equation as follows: y is 71.17X +0.012, r is 0.9990, which shows that the concentration and the absorbability of the total isoflavone are in good linear relationship in the range of 0.002-0.012 mg/mL.
(2) And (3) precision test: taking puerarin control solution with concentration of 0.006mg/ml, measuring absorbance at 250nm of an ultraviolet spectrophotometer, and repeatedly measuring for 6 times, wherein RSD value of the absorbance is 0.03%, which indicates that the precision of the instrument is good.
(3) And (3) stability test: taking the powder of the pueraria thomsonii before conversion, preparing a test solution according to the same method in the step 2, and measuring the absorbance at 250nm of an ultraviolet spectrophotometer at 5 time points of 0 hour, 2 hours, 4 hours, 6 hours and 8 hours respectively, wherein the average value of the absorbance is 0.2515, and the RSD value is 1.95%, which shows that the test solution is basically stable within 8 hours.
(4) And (3) repeatability experiment: taking 6 parts of the pueraria thomsonii medicinal material before conversion, wherein each part is about 0.5g, precisely weighing, preparing a sample solution according to the same method in the step 2, and measuring the absorbance at 250nm of an ultraviolet spectrophotometer. As a result, the average content of total isoflavones is 12.71mg/g, and the RSD value is 2.00%, which shows that the method has good repeatability.
(5) Sample recovery rate test: taking 6 parts of same sample powder with known content, each part is about 0.25g, adding 3.1mg puerarin reference substance into each sample, preparing test solution according to the same method of the step 2, measuring absorbance at 250nm, calculating recovery rate, and indicating that the average value of the recovery rate is 99.61%, and the RSD is 3.79%, the recovery rate of the experimental method is good.
4. Determination of total isoflavone content in sample
Separately, each sample (Aspergillus niger fermentation group: radix Puerariae sample obtained in example 1; blank control group: blank control group sample obtained in example 1) was prepared in the same manner as in step 2, and the absorbance at 250nm in an ultraviolet spectrophotometer was measured and the total isoflavone content was calculated, the results are shown in Table 1.
It can be seen that after the transformation of aspergillus niger for 7, 9 and 11 days, the total isoflavone content of the sample after the transformation of pueraria thomsonii is respectively 1.46, 1.70 and 1.91 times of the total isoflavone content of the sample which is not transformed in the corresponding time. The results show that under the action of Aspergillus niger, the content of the total isoflavone which is the main active ingredient of the pueraria thomsonii is obviously increased, and the longer the conversion time is, the more the content of the total isoflavone is increased.
TABLE 1 Total isoflavone content determination results of each Pueraria thomsonii sample
Experimental example 2 comparison of antioxidant Activity of Pueraria thomsonii samples before and after fermentation and transformation of Aspergillus niger
The research adopts a DPPH free radical removal method to evaluate the antioxidant activity of the pachyrhizua angulatus samples before and after the fermentation and transformation of Aspergillus niger.
1. Preparing a DPPH solution: DPPH (1, 1-diphenyl-2-trinitrophenylhydrazine) is precisely weighed and added with methanol to prepare 160 mu mol/L solution which is stored in dark place.
2. Preparing a test solution: taking about 0.5g of each sample (Aspergillus niger fermentation conversion group: radix Puerariae sample prepared in example 1; blank control group: blank control group sample prepared in example 1), precisely weighing, placing in a 100mL triangular flask, adding 20mL of 70% ethanol, weighing, ultrasonically extracting for 30min, cooling, weighing again, complementing the weight loss with 70% ethanol, and shaking up to obtain the product.
3. Evaluation of antioxidant Activity
And (3) taking the test solution prepared in the step (2), and diluting the test solution into test solutions with different concentrations by using methanol. Placing 0.12mL of test solution with different concentrations and 0.18mL of DPPH solution in a 96-well plate, uniformly mixing, placing for 1h at room temperature in a dark place, measuring absorbance Ai at 517nm of an enzyme-labeling instrument, simultaneously measuring absorbance A0 of 0.12mL of methanol +0.18mL of DPPH solution, and measuring absorbance Aj of 0.12mL of test solution +0.18mL of methanol solution, wherein each sample is paralleled for three times, and calculating the clearance rate according to a formula: DPPH radical clearance ═ 1- (Ai-Aj)/a0) × 100%. The clearance rate (Y) is used to perform logarithmic function equation regression on the concentration (X) of the drug, and the concentration (IC) of the drug at 50% clearance is obtained from the logarithmic function equation50)，IC50The smaller the value, theThe stronger the antioxidant activity of the drug, the results are shown in table 2.
It can be seen that samples after Pueraria thomsonii transformation cleared the average IC of DPPH radicals after 7, 9 and 11 days of Aspergillus niger transformation50The values are respectively the corresponding time unconverted sample IC50Values 30.09%, 10.23% and 5.61%, IC50Is significantly reduced. The result shows that the antioxidant activity of the pachyrhizua angulatus is obviously increased under the action of the aspergillus niger, and the longer the transformation time of the aspergillus niger is, the stronger the antioxidant activity is.
TABLE 2 measurement results of DPPH radical scavenging of radix Puerariae samples
Note: IC (integrated circuit)50The smaller the value, the stronger the antioxidant activity of the drug.
Experimental example 3 correlation study of Total isoflavone content and DPPH radical scavenging
The correlation between the content of total isoflavones and the activity of eliminating DPPH free radicals was analyzed by using software SPSS13.0, Pearson. As shown in Table 3, it can be seen that the total content of total isoflavones and DPPH radical scavenging activity (IC) in Pueraria thomsonii50Value), i.e. the higher the total isoflavone content in the sample, the stronger the antioxidant activity (IC for scavenging DPPH radicals)50The smaller the value).
Therefore, the content of the total isoflavone in the radix puerariae is closely related to the antioxidant activity of the radix puerariae, and the total isoflavone plays an important role in the antioxidant process of the radix puerariae.
TABLE 3 analysis results of Pearson correlation between the content of total isoflavones of Pueraria thomsonii and the scavenging of DPPH free radicals
Note: denotes P <0.05
In conclusion, the method for treating the pachyrhizua angulatus by adopting the aspergillus niger fermentation adopts rich polysaccharide and starch contained in the pachyrhizua angulatus as the culture medium, does not need to add other nutrient substances, can obviously improve the content of the total isoflavone in the pachyrhizua angulatus in a short time, further improves the antioxidant activity of the pachyrhizua angulatus by improving the content of the total isoflavone, and has wide application prospect in preparing medicines with obvious medicinal effect, such as protecting cardiovascular systems, resisting aging, resisting tumors and the like, and medicine-food homologous pachyrhizua angulatus products.
Claims (4)
1. A method for preparing isoflavone-rich pachyrhizua angulatus is characterized in that: the method comprises the following steps: inoculating Aspergillus niger to radix Puerariae, and fermenting; the radix Puerariae is dried root of Pueraria thomsonii Benth of Leguminosae;
the fermentation time is more than 11 days;
the Aspergillus niger is Aspergillus niger spore suspension with concentration of 1 × 108CFU/ml；
The ratio of the pachyrhizua angulatus to the aspergillus niger spores is 10 g: 2X 108 CFU；
The fermentation conditions were an aseptic environment with a temperature of 30 ℃ and a humidity of 99%.
2. The method of claim 1, wherein: the radix Puerariae is sterilized.
3. The method of claim 2, wherein: the sterilization treatment mode is as follows: and (3) irradiating and sterilizing the pachyrhizua angulatus under an ultraviolet lamp for 30-60 min.
4. Use of isoflavone enriched pueraria thomsonii prepared according to the method of any one of claims 1 to 3 for the preparation of pueraria thomsonii products; the radix Puerariae product is an antioxidant drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910527719.3A CN110314177B (en) | 2019-06-18 | 2019-06-18 | Radix Puerariae rich in isoflavone and its preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910527719.3A CN110314177B (en) | 2019-06-18 | 2019-06-18 | Radix Puerariae rich in isoflavone and its preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110314177A CN110314177A (en) | 2019-10-11 |
| CN110314177B true CN110314177B (en) | 2021-10-26 |
Family
ID=68119748
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910527719.3A Active CN110314177B (en) | 2019-06-18 | 2019-06-18 | Radix Puerariae rich in isoflavone and its preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110314177B (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104844583B (en) * | 2015-03-25 | 2018-06-22 | 陕西鑫茂生物科技有限公司 | A kind of production method of Puerarin |
-
2019
- 2019-06-18 CN CN201910527719.3A patent/CN110314177B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110314177A (en) | 2019-10-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111249218B (en) | Saussurea involucrate fermentation stock solution and preparation method and application thereof | |
| CN110982868B (en) | Co-culture method for improving triterpene content of ganoderma lucidum and application thereof | |
| Huo et al. | A preliminary study on polysaccharide extraction, purification, and antioxidant properties of sugar-rich filamentous microalgae Tribonema minus | |
| CN106722943B (en) | Apple enzyme and preparation method thereof | |
| CN109970538A (en) | The Dimeric sesquiterpene compound in a kind of marine fungi source and preparation method thereof and application in preparing anti-inflammatory drugs | |
| CN109463744B (en) | Method for performing solid state fermentation on leucocarp powder by eurotium cristatum, product prepared by method and application of product | |
| JP2023548948A (en) | Use of hyaluronic acid or a salt thereof and/or trehalose in stabilizing ergothioneine and ergothioneine compositions containing hyaluronic acid or a salt thereof and/or trehalose | |
| CN112245343B (en) | Method for fermenting burdock root by lucid ganoderma, method for compositely fermenting burdock root, fermented product and application | |
| CN110314177B (en) | Radix Puerariae rich in isoflavone and its preparation method | |
| CN102250170A (en) | Preparation method and application of two active flavonoid glycosides in okra fruits | |
| CN108175040A (en) | A kind of preparation method of millet oat Antrodia camphorata mycoplasma health food | |
| CN109007790A (en) | A kind of preparation method of Chinese yam mycoplasma powder and its Chinese yam mycoplasma powder of preparation | |
| CN108901587A (en) | A kind of solid culture method of cicada fungus | |
| CN111449239A (en) | Functional food additive of ganoderma lucidum fermented sea buckthorn seed meal and preparation method thereof | |
| CN106176564B (en) | The method for preparing ginseng PORIA ALBA fermentation liquid using ginseng endogenetic fungus | |
| CN114940948B (en) | Tricholoma matsutake and tricholoma matsutake mycelium fermentation medium and preparation method of fermentation broth | |
| CN114304551B (en) | Application of lactobacillus plantarum P101 fermented towel gourd | |
| CN106617009A (en) | Antioxidant health care enzyme and preparation method thereof | |
| CN108841891B (en) | Method for reducing capacity of monascus to produce citrinin through liquid and solid state fermentation by rutin derivatives | |
| CN106434787A (en) | Corn peel polysaccharide compound, preparation method thereof and medicinal use thereof | |
| CN106922389A (en) | A kind of artificial culture method of cicada fungus | |
| CN114317348B (en) | Lactobacillus plantarum and application thereof | |
| CN109793699A (en) | A kind of preparation method of Rhizoma Gastrodiae facial mask cream | |
| CN113750018B (en) | Plant composition and preparation method and application thereof | |
| CN116270826B (en) | Method for improving activity of rose in reducing blood sugar and blood fat and application |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |