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CN104473977A - Cordyceps militaris water extract as well as preparation method and application thereof - Google Patents

Cordyceps militaris water extract as well as preparation method and application thereof Download PDF

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Publication number
CN104473977A
CN104473977A CN201410660797.8A CN201410660797A CN104473977A CN 104473977 A CN104473977 A CN 104473977A CN 201410660797 A CN201410660797 A CN 201410660797A CN 104473977 A CN104473977 A CN 104473977A
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Prior art keywords
cordyceps militaris
link
water extract
preparation
extract according
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CN201410660797.8A
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CN104473977B (en
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雍天乔
张命龙
谢意珍
焦春伟
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Institute of Microbiology of Guangdong Academy of Sciences
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Guangdong Institute of Microbiology
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A61K36/066—Clavicipitaceae
    • A61K36/068—Cordyceps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Mycology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention provides a method for preparing a cordyceps militaris water extract. Water which is rich in resources and low in price serves as a solvent, and the aims of reducing the production cost and avoiding pollution of organic solvents are achieved. The cordyceps militaris water extract prepared by the method can be used for preparing medicines, health care products or auxiliary agents for reducing the uric acid level and improving the gout diseases.

Description

A kind of Cordyceps militaris (L.) Link. water extract, preparation method and application thereof
[technical field]
The present invention relates to a kind of Cordyceps militaris (L.) Link. water extract, preparation method and application thereof, belong to health product and the field of Chinese medicines.
[background technology]
Goat seriously perplexs human lives, and the epidemiological study of New Zealand, the U.S. and China shows that gout disease becomes more and more general, and sickness rate is about 2%, and constantly raises.Wherein 65 years old and above crowd's incidence rate the highest, the sickness rate of male is also higher than women's sickness rate twice.Goat is the result of hyperuricemia development, because uric acid concentration in body is higher than blood dissolves ability (>360 μm of ol/L), cause sodium urate crystals in soft tissue, joint deposition, these depositing crystallines cause soft tissue, joint and osseous tissue repeatedly inflammatory have an intense pain, seriously reduce patients ' life quality.
Goat is divided into acute and chronic two kinds.Comprise NSAID (non-steroidal anti-inflammatory drug), Colchicine, thyroliberin or steroid for acute pain syndrome caused by wind pathogen medicine in the market, if these drug mains act on inflammatory factor, reduce patient suffering.For chronic pain syndrome caused by wind pathogen, management blood uric acid levels is crucial.At present, the medicine being used for reducing uric acid concentration in human body mainly contains: allopurinol, medicine for improving uric acid excretion and diuretic, comprise the uricase etc. of benzbromarone, uricase, PEG modification, these medicines are because have stronger side effect to make the patient suffering from hepatopathy and nephropathy not use to liver and kidney metabolic organ, and these medicines easily cause gastrointestinal tract and cardiovascular diseases simultaneously.This make to suffer from hypertension, congestive heart failure, gastritis/ulcer or renal insufficiency patient with gout take said medicine and usually there is higher risk.Chinese Herbs is good, side effect is low, and its advantage manifests gradually, thus widely uses clinically.
For goat, there is literary composition to report Cordyceps militaris (L.) Link. ethanol extraction and there is the effect reducing uric acid, alleviate goat.But its preparation method is complicated, cost is high, and all adopts organic solvent extraction, there is the problem that organic solvent pollution is large.
[summary of the invention]
For the problems referred to above, be of the present inventionly the preparation method that a kind of Cordyceps militaris (L.) Link. water extract is provided, use aboundresources, low-cost water as solvent, reach the object reducing production cost, avoid organic solvent pollution.Specifically comprise the following steps:
(1) Cordyceps militaris (L.) Link.sporophore of drying is pulverized;
(2) Cordyceps militaris (L.) Link.sporophore after pulverizing is placed in the water of 5-25 times of weight, at 60-100 DEG C, extracts 1-2 hour, filter with 400 mesh filter screens;
(3) repeat to extract the nutrient substance in gained filtering residue by the condition identical with step (2).
(4) the rear distilling under reduced pressure of filtrate merging of step (2) and (3) gained is concentrated into 250-350mL;
(5) the concentrated solution lyophilizing of step (4) gained is obtained Cordyceps militaris (L.) Link. water extract.
Preferably, the leaching process in above-mentioned steps (2) is supersound extraction, and temperature is preferably 80 DEG C.
Cordyceps militaris (L.) Link. water extract obtained according to the method described above, it is the nutritional labeling extracted from Cordyceps militaris (L.) Link.sporophore, containing crude protein, crude fat, carbohydrate, mannitol, Cordyceps oil, cordycepin, sterols, ucleosides, aminoacid, peptide class, polysaccharide, monosaccharide, organic acid, vitamin and inorganic elements, wherein, mannitol content is: 4-7%, polyoses content is 20-25%, and nucleoside content is 0.25-0.35%.
Get the obtained Cordyceps militaris (L.) Link. water extract of said method to carry out being tested by uric acid in body, result shows to use the continuous gavage of Cordyceps militaris (L.) Link. water extract 7 days, the blood uric acid concentration of metabolic arthritis mice can be reduced to normal mouse uric acid level.
Therefore, another object of the present invention is to provide the application of above-mentioned Cordyceps militaris (L.) Link. water extract in preparation reduction uric acid concentration, the medicine improving gout disease, health product or assistant medicament.
Another object of the present invention is to provide a kind of medicine for the treatment of goat, it is characterized in that, this medicine contains above-mentioned Cordyceps militaris (L.) Link. water extract.
Beneficial effect of the present invention is:
1, the preparation technology of Cordyceps militaris (L.) Link. water extract of the present invention is simple, and cost is low;
2, the present invention adopts water as solvent extraction Cordyceps militaris (L.) Link. water extract, there is not the problem of organic solvent pollution, safety and environmental protection;
3, Cordyceps militaris (L.) Link. water extract of the present invention has significant uric acid resisting effect, can be used in preparing the medicine alleviating gout disease.
[accompanying drawing explanation]
Fig. 1: uric acid concentration in each group mice serum
[detailed description of the invention]
Explain the present invention further below in conjunction with embodiment, but embodiment does not limit in any form to the present invention.
Embodiment 1
One, the preparation of Cordyceps militaris (L.) Link. water extract (ETCM)
Get Cordyceps militaris (L.) Link.sporophore 100g, add in conical flask after using pulverizer to pulverize, and add 2L distilled water, mixture water-bath at 80 DEG C used 400 mesh filter screens to filter after extracting 2 hours.Extracting experiment repeats once, and after gained filtrate being merged, distilling under reduced pressure is concentrated into 300mL, and lyophilizing obtains Cordyceps militaris (L.) Link. water extract 53g, and productive rate is 53%.
Two, in Cordyceps militaris (L.) Link. water extract, each constituent content measures
Mannitol measures: utilize the chromogenic reaction of mannitol and sodium metaperiodate and Nash reagent to measure mannitol content.Wherein being formulated as follows of agents useful for same: Nash reagent: get ammonium acetate 150g, after dissolving, adds 2mL glacial acetic acid and 2mL acetylacetone,2,4-pentanedione, is dissolved to 1000mL with distilled water with appropriate distilled water; 0.015mol/L sodium metaperiodate: get sodium metaperiodate 3.2g and be dissolved in the 0.12mol/L HCI of 1000mL; 0.1%L-rhamnose: get L-rhamnose 0.1g, be dissolved in 100mL distilled water.Mannitol standard solution: get mannitol standard substance 100mg, be dissolved in 100mL distilled water.
Testing sample solution 0.03mL is added in test tube, mend distilled water to 1mL, distilled water 1mL is added in blank pipe, add mannitol standard solution 1mL in titer control tube, then in experiment tube, blank pipe and titer control tube, add 0.015mol/L sodium metaperiodate liquid 1mL respectively, after mixing, room temperature places 10 minutes, each test tube adds 0.1% rhamnose 2mL, after abundant mixing, add Nash reagent 4mL, 53 DEG C of water bath heat preservation 15min.Ultraviolet-uisible spectrophotometer is used to measure its absorption value at 413nm place, according to the mannitol content in the absorbance value calculation sample of mannitol standard solution after cooling.It is 4% that this method records mannitol content in embodiment 1 sample.
Adopting sulfuric acid-phynol method to record polyoses content in embodiment 1 sample is 22%.
Utilizing HPLC to record nucleoside content in embodiment 1 sample is 0.29%.
Three, Cordyceps militaris (L.) Link. water extract uric acid resisting experiment
Get 60 male SPF kunming mices (20 ± 2g), be divided into 6 groups at random: Normal group, hyperuricemia model matched group, positive controls 1, positive controls 2, ETCM low dose group, ETCM high dose group.Except Normal group lumbar injection and gavage normal saline, other groups according to the dosage lumbar injection oxonic acid potassium salt of 100mg/kg/d, the simultaneously hypoxanthine modeling of gavage 600mg/kg/d dosage.The previous h fast of modeling can't help water, after modeling 1 hour, the allopurinol (AL) of positive controls gavage 5mg/kg/d dosage, the allopurinol (AL) of positive controls 2 gavage 10mg/kg/d dosage, the Cordyceps militaris (L.) Link. water extract (ETCM) of Cordyceps militaris (L.) Link. water extract low dose group gavage 50mg/kg/d dosage, the Cordyceps militaris (L.) Link. water extract (ETCM) of Cordyceps militaris (L.) Link. water extract high dose group gavage 200mg/kg/d dosage, the normal saline of Normal group and hyperuricemia model matched group then gavage same volume, continuous 7 days.Within 7th day, gastric infusion is after 1 hour, anesthesia is plucked eyeball and is got blood, use centrifuge centrifugal 10min under 3500r/min speed to be separated and obtain serum, detect uric acid in serum concentration, (* represents compared with Normal group result, significant difference (P<0.01) as shown in Figure 1; # represents compared with model control group, significant difference (P<0.01)).
As can be seen from Figure 1, Cordyceps militaris (L.) Link. water extract of the present invention has good uric acid resisting effect, and its uric acid resisting effect can make metabolic arthritis model group blood uric acid concentration be reduced to normal mouse uric acid level.In addition, Cordyceps militaris (L.) Link. is as the fungus of integration of edible and medicinal herbs, and to person not damaged, its water extract may be used in medicine, health product or the assistant medicament prepared uric acid resisting, improve gout.
Embodiment 2
Get Cordyceps militaris (L.) Link.sporophore 100g, add in conical flask after using pulverizer to pulverize, and add 500mL distilled water, mixture supersound extraction at 60 DEG C used 400 mesh filter screens to filter after 2 hours.Extracting experiment repeats once, and after gained filtrate being merged, distilling under reduced pressure is concentrated into 250mL, and lyophilizing obtains Cordyceps militaris (L.) Link. water extract 39g, and productive rate is 39%.
Recording mannitol content in this Cordyceps militaris (L.) Link. water extract according to the detection method described in embodiment 1 is 7%, and polyoses content is 20%, and nucleoside content is 0.35%.
Embodiment 3
Get Cordyceps militaris (L.) Link.sporophore 100g, add in conical flask after using pulverizer to pulverize, and add 2.5L distilled water, mixture is decocted at 100 DEG C extraction and after 2 hours, use 400 mesh filter screens to filter.Extracting experiment repeats once, and after gained filtrate being merged, distilling under reduced pressure is concentrated into 350mL, and lyophilizing obtains Cordyceps militaris (L.) Link. water extract 45g, and productive rate is 45%.
Recording mannitol content in this Cordyceps militaris (L.) Link. water extract according to the detection method described in embodiment 1 is 5.4%, and polyoses content is 25%, and nucleoside content is 0.25%.
Embodiment 4
Get Cordyceps militaris (L.) Link.sporophore 100g, add in conical flask after using pulverizer to pulverize, and add 1.5L distilled water, mixture water-bath at 90 DEG C used 400 mesh filter screens to filter after extracting 2 hours.Extracting experiment repeats once, and after gained filtrate being merged, distilling under reduced pressure is concentrated into 300mL, and lyophilizing obtains Cordyceps militaris (L.) Link. water extract 49g, and productive rate is 49%.
Recording mannitol content in this Cordyceps militaris (L.) Link. water extract according to the detection method described in embodiment 1 is 6.2%, and polyoses content is 24%, and nucleoside content is 0.33%.

Claims (7)

1. a preparation method for Cordyceps militaris (L.) Link. water extract, is characterized in that, comprises the following steps:
(1) Cordyceps militaris (L.) Link.sporophore of drying is pulverized;
(2) Cordyceps militaris (L.) Link.sporophore after pulverizing is placed in the water of 5-25 times of weight, at 60-100 DEG C, extracts 1-2 hour, filter with 400 mesh filter screens;
(3) repeat to extract the nutrient substance in gained filtering residue by the condition identical with step (2).
(4) the rear distilling under reduced pressure of filtrate merging of step (2) and (3) gained is concentrated into 250-350mL;
(5) by the concentrated solution lyophilizing of step (4) gained.
2. the preparation method of Cordyceps militaris (L.) Link. water extract according to claim 1, is characterized in that, the Extracting temperature in described step (2) is 80 DEG C.
3. the preparation method of Cordyceps militaris (L.) Link. water extract according to claim 1, is characterized in that, the leaching process in described step (2) is supersound extraction.
4. the Cordyceps militaris (L.) Link. water extract that the preparation method of the Cordyceps militaris (L.) Link. water extract according to any one of claim 1-3 is obtained.
5. Cordyceps militaris (L.) Link. water extract according to claim 4, is characterized in that, in described Cordyceps militaris (L.) Link. water extract, mannitol content is 4-7%, and polyoses content is 20-25%, and nucleoside content is 0.25-0.35%.
6. Cordyceps militaris (L.) Link. water extract according to claim 4 reduces the application in uric acid concentration, the medicine improving gout disease, health product or assistant medicament in preparation.
7. treat a medicine for goat, it is characterized in that, described medicine contains Cordyceps militaris (L.) Link. water extract according to claim 4.
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Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105087332A (en) * 2015-08-31 2015-11-25 遵义鸿霖生物技术有限公司 Preparing process of soy sauce flavor type cordyceps militaris wine
CN105876777A (en) * 2016-04-12 2016-08-24 周安琪 Food having effect of reducing uric acid and preparation technology thereof
CN105943958A (en) * 2016-06-03 2016-09-21 黑龙江省玄鸟生物科技股份有限公司 Inonotus obliquus composite solid particles for treating gout and preparing method thereof
CN106692158A (en) * 2017-01-18 2017-05-24 广东省微生物研究所 Application of vitamin B1, cordycepic acid and riboflavin and composition thereof in uric acid-lowering drugs
CN107050067A (en) * 2016-11-02 2017-08-18 仪征康海农业发展有限公司 A kind of industrialized process for preparing of Cordyceps militaris water soluble powder
CN108042627A (en) * 2017-12-28 2018-05-18 广东粤微食用菌技术有限公司 A kind of composition for dropping hyperuricemia and its preparation method and application
CN108403729A (en) * 2018-05-14 2018-08-17 广东省微生物研究所（广东省微生物分析检测中心） The preparation method of agrocybe extract and its in the application for preparing anti-trioxypurine drug
CN108992471A (en) * 2018-08-28 2018-12-14 广东粤微生物科技有限公司 The preparation method of selenium-enriched cordceps militaris extract and its application in preparation inhibiting hyperuricemia and/or oxidation resistant drug or food
CN109170424A (en) * 2018-08-22 2019-01-11 江苏国桢生物科技有限公司 A kind of cordycepin beverage and preparation method thereof
CN109329671A (en) * 2018-11-15 2019-02-15 杨齐辉 A kind of edible drink product and preparation method thereof for treating gout
CN109553647A (en) * 2018-12-26 2019-04-02 中国科学院西北高原生物研究所 The extracting method of metabolite, additive
CN109875031A (en) * 2019-01-25 2019-06-14 常德炎帝生物科技有限公司 A kind of complex method for improveing Cordyceps militaris effect component extract
CN110448579A (en) * 2019-09-10 2019-11-15 北京汉藏金生物科技有限公司 A kind of quick anti-trioxypurine formula
CN110916200A (en) * 2019-12-02 2020-03-27 广东微量元素生物科技有限公司 Uric acid reducing composition and preparation method thereof
CN114224957A (en) * 2021-12-30 2022-03-25 仙乐健康科技股份有限公司 Composition with effect of inhibiting uric acid generation and preparation method thereof

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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105087332A (en) * 2015-08-31 2015-11-25 遵义鸿霖生物技术有限公司 Preparing process of soy sauce flavor type cordyceps militaris wine
CN105876777A (en) * 2016-04-12 2016-08-24 周安琪 Food having effect of reducing uric acid and preparation technology thereof
CN105943958A (en) * 2016-06-03 2016-09-21 黑龙江省玄鸟生物科技股份有限公司 Inonotus obliquus composite solid particles for treating gout and preparing method thereof
CN107050067A (en) * 2016-11-02 2017-08-18 仪征康海农业发展有限公司 A kind of industrialized process for preparing of Cordyceps militaris water soluble powder
CN106692158A (en) * 2017-01-18 2017-05-24 广东省微生物研究所 Application of vitamin B1, cordycepic acid and riboflavin and composition thereof in uric acid-lowering drugs
CN108042627B (en) * 2017-12-28 2021-03-26 广东粤微食用菌技术有限公司 Composition for treating hyperuricemia and preparation method and application thereof
CN108042627A (en) * 2017-12-28 2018-05-18 广东粤微食用菌技术有限公司 A kind of composition for dropping hyperuricemia and its preparation method and application
CN108403729A (en) * 2018-05-14 2018-08-17 广东省微生物研究所（广东省微生物分析检测中心） The preparation method of agrocybe extract and its in the application for preparing anti-trioxypurine drug
CN108403729B (en) * 2018-05-14 2021-09-14 广东省微生物研究所（广东省微生物分析检测中心） Preparation method of agrocybe cylindracea extract and application of agrocybe cylindracea extract in preparation of uric acid reducing medicine
CN109170424A (en) * 2018-08-22 2019-01-11 江苏国桢生物科技有限公司 A kind of cordycepin beverage and preparation method thereof
CN108992471A (en) * 2018-08-28 2018-12-14 广东粤微生物科技有限公司 The preparation method of selenium-enriched cordceps militaris extract and its application in preparation inhibiting hyperuricemia and/or oxidation resistant drug or food
CN109329671A (en) * 2018-11-15 2019-02-15 杨齐辉 A kind of edible drink product and preparation method thereof for treating gout
CN109553647A (en) * 2018-12-26 2019-04-02 中国科学院西北高原生物研究所 The extracting method of metabolite, additive
CN109875031A (en) * 2019-01-25 2019-06-14 常德炎帝生物科技有限公司 A kind of complex method for improveing Cordyceps militaris effect component extract
CN110448579A (en) * 2019-09-10 2019-11-15 北京汉藏金生物科技有限公司 A kind of quick anti-trioxypurine formula
CN110916200A (en) * 2019-12-02 2020-03-27 广东微量元素生物科技有限公司 Uric acid reducing composition and preparation method thereof
CN114224957A (en) * 2021-12-30 2022-03-25 仙乐健康科技股份有限公司 Composition with effect of inhibiting uric acid generation and preparation method thereof

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Application publication date: 20150401

Assignee: Guangdong Meili engine Brand Management Co.,Ltd.

Assignor: GUANGDONG INSTITUTE OF MICROBIOLOGY

Contract record no.: X2020980006416

Denomination of invention: Water extract, preparation method and application of Cordyceps militaris

Granted publication date: 20170912

License type: Common License

Record date: 20200924

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Address after: No. 56, courtyard, No. 100, Xianlie Middle Road, Guangzhou, Guangdong 510070

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Patentee after: Institute of Microbiology, Guangdong Academy of Sciences

Address before: No. 56, courtyard, No. 100, Xianlie Middle Road, Guangzhou, Guangdong 510070

Patentee before: GUANGDONG INSTITUTE OF MICROBIOLOGY (GUANGDONG DETECTION CENTER OF MICROBIOLOGY)