CN105709031B - A kind of ganoderma lucidum polysaccharide compound and preparation method thereof for treating hand-foot-and-mouth disease - Google Patents
A kind of ganoderma lucidum polysaccharide compound and preparation method thereof for treating hand-foot-and-mouth disease Download PDFInfo
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Abstract
The present invention relates to a kind of ganoderma lucidum polysaccharide compounds and preparation method thereof for treating hand-foot-and-mouth disease, belong to medicines and health protection food technology field, it is made of following parts by weight of component: 20-30 parts of ganoderma lucidum polysaccharide, cordate houttuynia, Fructus Forsythiae, 3-5 parts of rhizoma anemarrhenae compound Chinese medicine extract, 5-10 parts of sucrose, 0.03-0.05 parts of Aspartame, 0.03-0.08 parts of essence, 200-300 parts of pure water, ganoderma lucidum removes oil-soluble impurities by alcohol reflux, hot water extracts and ethanol precipitation, ganoderma lucidum holosaccharide is obtained after column chromatographic purifying, with the Chinese Medicinal Houttuynia Cordata Thunb of preparation, Fructus Forsythiae and anemarrhena asphodefoides extract compound.Experiments have shown that compound has obvious inhibitory effect to enterovirns type 71.
Description
Technical field
The present invention relates to medicines and health protection food technology field, specially a kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease
And preparation method thereof.
Background technique
Hand-foot-and-mouth disease typically occurs in 3 years old or less children, with forerunners' disease such as low-heat, abdominal pain, general malaise, oral cavity, pharynx,
Soft palate etc. will appear painful blister, and severe cases often the clinics such as meningitis, encephalitis, encephalomyelitis occur in 1-5 days in morbidity
Performance.Enterovirns type 71 is to cause one of Children main pathogens.Up to the present, clinically there are no ten
Divide effective therapeutic scheme.
Ganoderma lucidum is precious medicinal fungi, has strengthen immunity, antitumor, anti-aging and other effects.Ganoderic acid is from spirit
The triterpene compound extracted in sesame has report in terms of antiphlogistic antibacterial, antitumor and anti HIV-1 virus and enterovirns type 71
Road.Ganoderma lucidum polysaccharide is active constituent important in ganoderma lucidum, there is not yet ganoderma lucidum polysaccharide prevention and treatment hand-foot-and-mouth disease and anti-Enterovirus 71
The correlative study of type.
Cordate houttuynia is the plant of dual-purpose of drug and food, has the function of the clearing heat and detoxicating, carbuncle that disappears apocenosis, inducing diuresis for treating strangurtia etc., mainly lives
Property ingredient be flavone compound and volatile oil.Fructus Forsythiae is one of China's clinic conventional Chinese medicine, and forsythia mainly contains terpene
The compounds such as class, lignanoids, flavonoids have the effects that antibacterial anti-inflammatory, antiviral, antiendotoxin.Rhizoma anemarrhenae is known for Liliaceae
The dry rhizome of female platymiscium rhizoma anemarrhenae has the effect of clearing heat-fire, nourishing Yin and moistening dryness, clinically can be used for viral disease
Treatment.
It retrieves about ganoderma lucidum in the application in treatment hand-foot-and-mouth disease and enterovirns type 71, discovery related patents have: in
State's number of patent application is 201110162657.4 ganoderic acid Y answering in preparation treatment or prevention enterovirns type 71 infection medicine
With and number of patent application 201110436439.5 ganoderic acid Lanosta-7,9 (11) 24-trien-3-one15,26-
Application of the dihydroxy in preparation prevention and/or treatment enterovirns type 71 infection medicine.The anti-enterovirns type 71 of ganoderma lucidum
Patent is concentrated mainly on ganoderic acid, there is not yet the research of ganoderma lucidum polysaccharide compound prevention and treatment hand-foot-and-mouth disease and anti-enterovirns type 71
And patent.
Summary of the invention
Through inventor the study found that being prepared with ganoderma lucidum polysaccharide compounding Chinese Medicinal Houttuynia Cordata Thunb, Fructus Forsythiae and anemarrhena asphodefoides extract
Compound, can significantly play the role of preventing and treating hand-foot-and-mouth disease and anti-enterovirns type 71.The purpose of the present invention is to provide one
The ganoderma lucidum polysaccharide compound and preparation method thereof of kind prevention and treatment hand-foot-and-mouth disease.The compound, which can be further prepared into, has prevention and treatment brothers
The drug or health care product of stomatosis and enterovirns type 71 effect, it is matched with clinical treatment drug, significant prevention and treatment hand can be played
The function of sufficient stomatosis, and the toxic side effect of drug can be reduced.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease, is made of: ganoderma lucidum polysaccharide 20-30 following parts by weight of component
Part, cordate houttuynia, Fructus Forsythiae, 3-5 parts of rhizoma anemarrhenae compound Chinese medicine extract, 5-10 parts of sucrose, 0.03-0.05 parts of Aspartame, essence
0.03-0.08 parts, 200-300 parts of pure water.
The ganoderma lucidum polysaccharide the preparation method comprises the following steps:
(1) ganoderma lucidum fruitbody cleans 2-3 times, pulverizes after 55-60 °C of drying, crosses 80-100 mesh, spare；
(2) ganoderma powder that will be prepared in step (1) is by weight that 95% ethyl alcohol, 65- is added in 1:3-4 with volume ratio g/mL
80 °C of reflux 1-2h remove liposoluble substance, and 3000-4500 r/min is centrifuged 10-15 min, collect filter residue, and 50-60 DEG C dries,
80-100 mesh was pulverized, it is spare；
(3) pure water is added in the ganoderma powder for taking step (2) to obtain, wherein ganoderma powder weight and pure water volume ratio
G/mL is 1:20-35, and 2-4 h is extracted under 75-85 °C, and 3000-4500 r/min is centrifuged 15-25 min, takes supernatant spare, will
The residue of centrifugation carries out above-mentioned steps and extracts 2-3 times, merges supernatant；
(4) step (3) supernatant is subjected to rotary evaporation in the case where 55-65 °C and revolving speed is 50-80r/min, by supernatant
Concentration is original 1/10-1/5；
(5) 3-4 times of 95% ethyl alcohol is added by volume, is placed in alcohol in 0-4 °C for the supernatant that step (4) is concentrated
Heavy 5-10 h, 3000 ~ 4500 r/min are centrifuged 15-25 min and crushed 50-100 mesh after sediment freeze-drying, obtain powder
End is spare；
(6) pure water is added to the powder prepared obtained by step (5) to be redissolved, powder weight and pure water volume ratio g/
ML is 1:5-8, with one or more of removing protein 3-5 times in Sevag method, polyamide adduct, trichloroacetic acid method, enzyme process, obtains ganoderma lucidum
Thick many candies solution；
(7) the ganoderma lucidum crude polysaccharide solution obtained to step (6) is that 1:15-20 uses AB-8 by volume and weight ratio L/g
Resin, 50-80 r/min oscillation 3-4 h decolourizes under the conditions of 20-25 °C, obtained polysaccharide solution；
(8) polysaccharide solution for obtaining step (7) is packed into bag filter, 10000 or more molecular cut off, distilled water dialysis
16-20 h, every 4 h change primary distilled water；
(9) polysaccharide solution for taking step (8) to obtain is eluted through DEAD Sephadex A-25 ion exchange column, elution
Obtained component through Sephadex G-200 sephadex column, is collected single eluting peak solution, after freeze-drying, be crushed again
50-100 mesh obtains ganoderma lucidum holosaccharide powder, spare.
The essence includes one of vanilla, butter essence, flavoring apple essence, grape essence, flavoring banana essence or several
Kind.
The cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae compound Chinese medicine extract the preparation method comprises the following steps:
(1) cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae are respectively washed 2-3 times, are pulverized after 55-60 °C of drying, are crossed 80-100 mesh,
It is spare；
(2) Heartleaf Houttuynia Herb for taking step (1) to prepare is that 1:3-5 addition mass concentration is with volume ratio g/mL by weight
The ethanol solution of 70%-80%, microwave-ultrasonic cooperate with assisted extraction 10-20min, filtering；Filter residue is repeated to mention by above-mentioned steps
It takes 2-3 times, merging filtrate, 10-20 min is centrifuged under the conditions of 3000-4000 r/min, supernatant is taken, in 45-55 °C of condition
Under be concentrated into medicinal extract shape, crushed 50-80 mesh after freeze-drying, obtain houttuynia extract；
(3) Weeping Forsythia Capsule for taking step (1) to prepare is that 1:3-4 addition mass concentration is with volume ratio g/mL by weight
The ethanol solution of 70%-80%, microwave-ultrasonic cooperate with assisted extraction 10-15min, filtering；Filter residue is repeated to mention by above-mentioned steps
It takes 2-3 times, merging filtrate, 10-20 min is centrifuged under the conditions of 3000-4000 r/min, supernatant is taken, in 45-55 °C of condition
Under be concentrated into medicinal extract shape, crushed 50-80 mesh after freeze-drying, obtain forsythia suspense extraction；
(4) Common Anemarrhena Rhizome for taking step (1) to prepare is that 1:4-6 addition mass concentration is with volume ratio g/mL by weight
The ethanol solution of 70%-80%, microwave-ultrasonic cooperate with assisted extraction 15-20 min, filtering；Filter residue is repeated by above-mentioned steps
It extracts 2-3 times, merging filtrate, 10-20 min is centrifuged under the conditions of 3000-4000 r/min, supernatant is taken, in 45-55 °C of item
It is concentrated into medicinal extract shape under part, crushed 50-80 mesh after freeze-drying, obtain anemarrhena asphodefoides extract；
(5) step (2) houttuynia extract, step (3) forsythia suspense extraction, step (4) anemarrhena asphodefoides extract powder are taken, according to
Weight ratio 2:1:1, which is mixed, is made Chinese medicinal compound extract.
A kind of preparation method of ganoderma lucidum polysaccharide compound that treating hand-foot-and-mouth disease includes the following:
(1) according to formula, ganoderma lucidum holosaccharide is taken, is mixed with cordate houttuynia, Fructus Forsythiae and rhizoma anemarrhenae Chinese medicinal compound extract, then plus
Enter sucrose, essence, Aspartame, pure water, the original solution of composite oral liquid is made in stirring 20-30 min；Adjust solution ph
For 6.9-7.5；
(2) the solution left standstill 2-4 h for obtaining step (1), takes supernatant through hollow-fibre membrane ultrafiltration apparatus ultrafiltration, obtains mouth
Take liquid；
(3) the deployed oral solution of step (2) is used into filling, locking closure while hot, through water dip sterilization and after the assay was approved
For product.
The present invention has the advantages that ganoderma lucidum holosaccharide and cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae compound Chinese medicine extract, through pharmacodynamics
Experiment, has the effect of very strong anti-enterovirns type 71.There is not yet the anti-enterovirns type 71 of ganoderma lucidum polysaccharide compound and anti-
Control the correlative study of hand-foot-and-mouth disease.The technological operation is easy, and cost price is low, securely and reliably, is suitble to industrial scale production.
Detailed description of the invention
Anti- enterovirns type 71 test result outside Fig. 1 ganoderma lucidum polysaccharide composite body.
Specific embodiment
Embodiment 1
A kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease, is made of following parts by weight of component: 20 parts of ganoderma lucidum polysaccharide, fish
Raw meat grass, Fructus Forsythiae, 3 parts of rhizoma anemarrhenae compound Chinese medicine extract, 5 parts of sucrose, 0.03 part of Aspartame, 0.03 part of essence, pure water 200
Part.
The ganoderma lucidum polysaccharide the preparation method comprises the following steps:
(1) ganoderma lucidum fruitbody cleans 2 times, pulverizes after 55 °C of drying, crosses 80 meshes, spare；
(2) ganoderma powder that will be prepared in step (1) is that 1:3 is added 95% ethyl alcohol with volume ratio g/mL by weight, 65 °C
The 1h that flows back removes liposoluble substance, and 3000 r/min are centrifuged 10min, collects filter residue, and 50 DEG C of drying pulverized 80 meshes,
It is spare；
(3) pure water is added in the ganoderma powder for taking step (2) to obtain, wherein ganoderma powder weight and pure water volume ratio
G/mL is 1:20, and 2 h are extracted under 75 °C, and 3000 r/min are centrifuged 15 min, take supernatant spare, the residue of centrifugation is carried out
It states step to extract 2 times, merges supernatant；
(4) step (3) supernatant is subjected to rotary evaporation in the case where 55 °C and revolving speed is 50r/min, is by supernatant concentration
Originally 1/10；
(5) 3 times of 95% ethyl alcohol is added by volume, is placed in alcohol precipitation in 0 °C for the supernatant that step (4) is concentrated
5h, 3000r/min are centrifuged 15min and crushed 50 meshes after sediment freeze-drying, it is spare to obtain powder；
(6) pure water is added to the powder prepared obtained by step (5) to be redissolved, powder weight and pure water volume ratio g/
ML is 1:5, with Sevag method removing protein 3 times, obtains ganoderma lucidum crude polysaccharide solution；
(7) the ganoderma lucidum crude polysaccharide solution obtained to step (6) uses AB-8 tree with weight ratio L/g by volume for 1:15
Rouge, 50 r/min vibrate 3 h and decolourize under the conditions of 20 °C, obtained polysaccharide solution；
(8) polysaccharide solution for obtaining step (7) is packed into bag filter, 10000 or more molecular cut off, distilled water dialysis 16
H, every 4 h change primary distilled water；
(9) polysaccharide solution for taking step (8) to obtain is eluted through DEAD Sephadex A-25 ion exchange column, elution
Obtained component through Sephadex G-200 sephadex column, is collected single eluting peak solution, after freeze-drying, be crushed again
50 meshes obtain ganoderma lucidum holosaccharide powder, spare.
The essence is vanilla.
The cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae compound Chinese medicine extract the preparation method comprises the following steps:
(1) cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae are respectively washed 2 times, are pulverized after 45 °C of drying, and 80 meshes are crossed, spare；
(2) it is 70% that the Heartleaf Houttuynia Herb for taking step (1) to prepare is 1:3 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 10min, filtering；Filter residue is repeated to extract 2 times by above-mentioned steps, is merged
Filtrate is centrifuged 10 min under the conditions of 3000 r/min, takes supernatant, and medicinal extract shape is concentrated under the conditions of 50 °C, freeze-drying
After crushed 50 meshes, obtain houttuynia extract；
(3) it is 70% that the Weeping Forsythia Capsule for taking step (1) to prepare is 1:3 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 10min, filtering；Filter residue is repeated to extract 2 times by above-mentioned steps, merges filter
Liquid is centrifuged 10 min under the conditions of 3000 r/min, takes supernatant, medicinal extract shape is concentrated under the conditions of 50 °C, after freeze-drying
It crushed 50 meshes, obtain forsythia suspense extraction；
(4) it is 70% that the Common Anemarrhena Rhizome for taking step (1) to prepare is 1:4 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with 15 min of assisted extraction, filtering；Filter residue is repeated to extract 2 times by above-mentioned steps, is merged
Filtrate is centrifuged 10min under the conditions of 3000 r/min, takes supernatant, medicinal extract shape is concentrated under the conditions of 50 °C, after freeze-drying
It crushed 50 meshes, obtain anemarrhena asphodefoides extract；
(5) step (2) houttuynia extract, step (3) forsythia suspense extraction, step (4) anemarrhena asphodefoides extract powder are taken, according to
Weight ratio 2:1:1, which is mixed, is made compound Chinese medicine extract.
A kind of preparation method of ganoderma lucidum polysaccharide compound that treating hand-foot-and-mouth disease includes the following:
(1) according to formula, ganoderma lucidum holosaccharide is taken, is mixed with cordate houttuynia, Fructus Forsythiae and rhizoma anemarrhenae compound Chinese medicine extract, then plus
Enter sucrose, essence, Aspartame, pure water, the original solution of composite oral liquid is made in stirring 20min；Adjusting solution ph is
6.9；
(2) the solution left standstill 2h for obtaining step (1) takes supernatant through hollow-fibre membrane ultrafiltration apparatus ultrafiltration, obtains oral
Liquid；
(3) the deployed oral solution of step (2) is used into filling, locking closure while hot, through water dip sterilization and after the assay was approved
For product.
Embodiment 2
A kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease, is made of following parts by weight of component: 30 parts of ganoderma lucidum polysaccharide, fish
Raw meat grass, Fructus Forsythiae, 4 parts of rhizoma anemarrhenae compound Chinese medicine extract, 10 parts of sucrose, 0.05 part of Aspartame, 0.08 part of essence, pure water 300
Part.
The ganoderma lucidum polysaccharide the preparation method comprises the following steps:
(1) ganoderma lucidum fruitbody cleans 3 times, pulverizes, sieves with 100 mesh sieve after 60 °C of drying, spare；
(2) ganoderma powder that will be prepared in step (1) is that 1:4 is added 95% ethyl alcohol with volume ratio g/mL by weight, 80 °
C reflux 2h removes liposoluble substance, and 4500 r/min are centrifuged 15 min, collects filter residue, and 60 DEG C of drying pulverized 100
Mesh, it is spare；
(3) pure water is added in the ganoderma powder for taking step (2) to obtain, wherein ganoderma powder weight and pure water volume ratio
G/mL is 1:35, and 4 h are extracted under 85 °C, and 4500 r/min are centrifuged 25 min, take supernatant spare, and the residue of centrifugation is carried out
Above-mentioned steps are extracted 3 times, and supernatant is merged；
(4) step (3) supernatant is subjected to rotary evaporation in the case where 65 °C and revolving speed is 80r/min, is by supernatant concentration
Originally 1/5；
(5) 4 times of 95% ethyl alcohol is added by volume, is placed in alcohol precipitation 10 in 4 °C for the supernatant that step (4) is concentrated
H, 4500 r/min are centrifuged 25 min, and after sediment freeze-drying, crushing is sieved with 100 mesh sieve, and it is spare to obtain powder；
(6) pure water is added to the powder prepared obtained by step (5) to be redissolved, powder weight and pure water volume ratio g/
ML is 1:8, with polyamide adduct removing protein 5 times, obtains ganoderma lucidum crude polysaccharide solution；
(7) the ganoderma lucidum crude polysaccharide solution obtained to step (6) uses AB-8 tree with weight ratio L/g by volume for 1:20
Rouge, 80 r/min vibrate 4 h and decolourize under the conditions of 25 °C, obtained polysaccharide solution；
(8) polysaccharide solution for obtaining step (7) is packed into bag filter, 10000 or more molecular cut off, distilled water dialysis 20
H, every 4 h change primary distilled water；
(9) polysaccharide solution for taking step (8) to obtain is eluted through DEAD Sephadex A-25 ion exchange column, elution
Obtained component through Sephadex G-200 sephadex column, is collected single eluting peak solution, after freeze-drying, be crushed again
100 meshes obtain ganoderma lucidum holosaccharide powder, spare.
The essence includes vanilla, butter essence, flavoring apple essence, and each weight ratio is 1:1:1.
The cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae compound Chinese medicine extract the preparation method comprises the following steps:
(1) cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae are respectively washed 3 times, are pulverized, are sieved with 100 mesh sieve after 50 °C of drying, spare；
(2) Heartleaf Houttuynia Herb for taking step (1) to prepare is that 1:5 addition mass concentration is with volume ratio g/mL by weight
80% ethanol solution, microwave-ultrasonic cooperate with assisted extraction 20min, filtering；Filter residue is repeated to extract 3 times by above-mentioned steps,
Merging filtrate is centrifuged 20 min under the conditions of 4000 r/min, takes supernatant, and medicinal extract shape is concentrated under the conditions of 55 °C, freezing
Crushed after being dried crosses 80 meshes, obtains houttuynia extract；
(3) it is 80% that the Weeping Forsythia Capsule for taking step (1) to prepare is 1:4 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 15min, filtering；Filter residue is repeated to extract 3 times by above-mentioned steps, is merged
Filtrate is centrifuged 20 min under the conditions of 4000 r/min, takes supernatant, and medicinal extract shape is concentrated under the conditions of 55 °C, freeze-drying
After crushed 80 meshes, obtain forsythia suspense extraction；
(4) it is 80% that the Common Anemarrhena Rhizome for taking step (1) to prepare is 1:6 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with 20 min of assisted extraction, filtering；Filter residue is repeated to extract 3 times by above-mentioned steps, is closed
And filtrate, it is centrifuged 20 min under the conditions of 4000 r/min, takes supernatant, medicinal extract shape is concentrated under the conditions of 55 °C, freezing is dry
It crushed 80 meshes after dry, obtain anemarrhena asphodefoides extract；
(5) step (2) houttuynia extract, step (3) forsythia suspense extraction, step (4) anemarrhena asphodefoides extract powder are taken, according to
Weight ratio 2:1:1, which is mixed, is made compound Chinese medicine extract.
A kind of preparation method of ganoderma lucidum polysaccharide compound that treating hand-foot-and-mouth disease includes the following:
(1) according to formula, ganoderma lucidum holosaccharide is taken, is mixed with cordate houttuynia, Fructus Forsythiae and rhizoma anemarrhenae Chinese medicinal compound extract, then plus
Enter sucrose, essence, Aspartame, pure water, the original solution of composite oral liquid is made in 30 min of stirring；Adjusting solution ph is
7.5；
(2) 4 h of solution left standstill for obtaining step (1), takes supernatant through hollow-fibre membrane ultrafiltration apparatus ultrafiltration, obtains oral
Liquid；
(3) the deployed oral solution of step (2) is used into filling, locking closure while hot, through water dip sterilization and after the assay was approved
For product.
Embodiment 3
A kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease, is made of following parts by weight of component: 25 parts of ganoderma lucidum polysaccharide, fish
Raw meat grass, Fructus Forsythiae, 5 parts of rhizoma anemarrhenae compound Chinese medicine extract, 7 parts of sucrose, 0.04 part of Aspartame, 0.05 part of essence, pure water 250
Part.
The ganoderma lucidum polysaccharide the preparation method comprises the following steps:
(1) ganoderma lucidum fruitbody cleans 3 times, pulverizes, sieves with 100 mesh sieve after 60 °C of drying, spare；
(2) ganoderma powder that will be prepared in step (1) is that 1:4 is added 95% ethyl alcohol with volume ratio g/mL by weight, 70 °C
The 2h that flows back removes liposoluble substance, and 4000 r/min are centrifuged 12 min, collects filter residue, and 55 DEG C of drying pulverized 80 meshes,
It is spare；
(3) pure water is added in the ganoderma powder for taking step (2) to obtain, wherein ganoderma powder weight and pure water volume ratio
G/mL is 1:30, and 3h is extracted under 80 °C, and 4000 r/min are centrifuged 20 min, take supernatant spare, the residue of centrifugation is carried out
It states step to extract 3 times, merges supernatant；
(4) step (3) supernatant is subjected to rotary evaporation in the case where 60 °C and revolving speed is 60r/min, is by supernatant concentration
Originally 1/5；
(5) 4 times of 95% ethyl alcohol is added by volume, is placed in alcohol precipitation 8 in 2 °C for the supernatant that step (4) is concentrated
H, 4000 r/min are centrifuged 20 min and crushed 80 meshes after sediment freeze-drying, it is spare to obtain powder；
(6) pure water is added to the powder prepared obtained by step (5) to be redissolved, powder weight and pure water volume ratio g/
ML is 1:7, with trichloroacetic acid method removing protein 4 times, obtains ganoderma lucidum crude polysaccharide solution；
(7) the ganoderma lucidum crude polysaccharide solution obtained to step (6) uses AB-8 tree with weight ratio L/g by volume for 1:18
Rouge, 70 r/min vibrate 4 h and decolourize under the conditions of 24 °C, obtained polysaccharide solution；
(8) polysaccharide solution for obtaining step (7) is packed into bag filter, 10000 or more molecular cut off, distilled water dialysis 20
H, every 4 h change primary distilled water；
(9) polysaccharide solution for taking step (8) to obtain is eluted through DEAD Sephadex A-25 ion exchange column, elution
Obtained component through Sephadex G-200 sephadex column, is collected single eluting peak solution, after freeze-drying, be crushed again
80 meshes obtain ganoderma lucidum holosaccharide powder, spare.
The essence is grape essence.
The cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae compound Chinese medicine extract the preparation method comprises the following steps:
(1) cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae are respectively washed 3 times, are pulverized after 55 °C of drying, and 90 meshes are crossed, spare；
(2) it is 70% that the Heartleaf Houttuynia Herb for taking step (1) to prepare is 1:4 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 15min, filtering；Filter residue is repeated to extract 3 times by above-mentioned steps, is merged
Filtrate is centrifuged 15 min under the conditions of 4000 r/min, takes supernatant, and medicinal extract shape is concentrated under the conditions of 45 °C, freeze-drying
After crushed 60 meshes, obtain houttuynia extract；
(3) it is 80% that the Weeping Forsythia Capsule for taking step (1) to prepare is 1:4 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 15min, filtering；Filter residue is repeated to extract 3 times by above-mentioned steps, merges filter
Liquid is centrifuged 15 min under the conditions of 4000 r/min, takes supernatant, medicinal extract shape is concentrated under the conditions of 45 °C, after freeze-drying
It crushed 70 meshes, obtain forsythia suspense extraction；
(4) it is 80% that the Common Anemarrhena Rhizome for taking step (1) to prepare is 1:5 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with 17 min of assisted extraction, filtering；Filter residue is repeated to extract 2 times by above-mentioned steps, is merged
Filtrate is centrifuged 15min under the conditions of 3000r/min, takes supernatant, medicinal extract shape is concentrated under the conditions of 45 °C, after freeze-drying
It crushed 60 meshes, obtain anemarrhena asphodefoides extract；
(5) step (2) houttuynia extract, step (3) forsythia suspense extraction, step (4) anemarrhena asphodefoides extract powder are taken, according to
Weight ratio 2:1:1, which is mixed, is made compound Chinese medicine extract.
A kind of preparation method of ganoderma lucidum polysaccharide compound that treating hand-foot-and-mouth disease includes the following:
(1) according to formula, ganoderma lucidum holosaccharide is taken, is mixed with cordate houttuynia, Fructus Forsythiae and rhizoma anemarrhenae compound Chinese medicine extract, then plus
Enter sucrose, essence, Aspartame, pure water, the original solution of composite oral liquid is made in 25 min of stirring；Adjusting solution ph is
7.2；
(2) 3 h of solution left standstill for obtaining step (1), takes supernatant through hollow-fibre membrane ultrafiltration apparatus ultrafiltration, obtains oral
Liquid；
(3) the deployed oral solution of step (2) is used into filling, locking closure while hot, through water dip sterilization and after the assay was approved
For product.
Experimental example: anti-enterovirns type 71 test outside ganoderma lucidum polysaccharide composite body
African green monkey kidney (Vero) cell is laid in 12 orifice plates, culture in 37 °C of incubators is put in, is paved with to cell monolayer
In 12 orifice plates when the 80%-90% in every hole, first with the ganoderma lucidum polysaccharide compound dissolved matter of the 5 mg/mL concentration of 100 μ L in every hole
2 h of pretreatment cell outwells complete DMEM culture medium, adds the virus liquid of enterovirns type 71 (moi=0.5) with normally
Method vero cells infection is respectively set normally to group, the drug-treated group of ganoderma lucidum polysaccharide compound and virus control group, 20 h
Observation ganoderma lucidum polysaccharide compound inhibits the effect of the cytopathic effect of enterovirns type 71 induction afterwards.It is found through test result,
Ganoderma lucidum polysaccharide compound processing group is added compared with virus control group, cytopathic effect significantly reduces, and illustrates that ganoderma lucidum polysaccharide is multiple
Close the cytopathic effect that object can effectively inhibit enterovirns type 71 to induce.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (3)
1. a kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease, it is characterised in that: be made of following parts by weight of component: ganoderma lucidum is more
It is 20-30 parts sugared, cordate houttuynia, Fructus Forsythiae, 3-5 parts of rhizoma anemarrhenae compound Chinese medicine extract, 5-10 parts of sucrose, 0.03-0.05 parts of Aspartame,
0.03-0.08 parts of essence, 200-300 parts of pure water；
The ganoderma lucidum polysaccharide the preparation method comprises the following steps:
(1) ganoderma lucidum fruitbody cleans 2-3 times, pulverizes after 55-60 °C of drying, crosses 80-100 mesh, spare；
(2) ganoderma powder that will be prepared in step (1) is that 1:3-4 is added 95% ethyl alcohol with volume ratio g/mL by weight, 65-80 °C
The 1-2h that flows back removes liposoluble substance, and 3000-4500 r/min is centrifuged 10-15 min, collects filter residue, 50-60 DEG C of drying, ultra micro
It crushed 80-100 mesh, it is spare；
(3) pure water is added in the ganoderma powder for taking step (2) to obtain, wherein ganoderma powder weight and pure water volume ratio g/mL
For 1:20-35,2-4 h is extracted under 75-85 °C, 3000-4500 r/min is centrifuged 15-25 min, takes supernatant spare, will be centrifuged
Residue carry out above-mentioned steps extract 2-3 time, merging supernatant；
(4) step (3) supernatant is subjected to rotary evaporation in the case where 55-65 °C and revolving speed is 50-80r/min, supernatant is concentrated
For original 1/10-1/5；
(5) 3-4 times of 95% ethyl alcohol is added by volume, is placed in alcohol precipitation 5- in 0-4 °C for the supernatant that step (4) is concentrated
10 h, 3000 ~ 4500 r/min are centrifuged 15-25 min and crushed 50-100 mesh after sediment freeze-drying, it is standby to obtain powder
With；
(6) pure water is added to the powder prepared obtained by step (5) to be redissolved, powder weight is with pure water volume ratio g/mL
It is slightly more to obtain ganoderma lucidum with one or more of removing protein 3-5 times in Sevag method, polyamide adduct, trichloroacetic acid method, enzyme process by 1:5-8
Sugar juice；
(7) the ganoderma lucidum crude polysaccharide solution obtained to step (6) is that 1:15-20 uses AB-8 resin by volume and weight ratio L/g,
50-80 r/min vibrates 3-4 h and decolourizes under the conditions of 20-25 °C, obtained polysaccharide solution；
(8) polysaccharide solution for obtaining step (7) is packed into bag filter, 10000 or more molecular cut off, distilled water dialysis 16-20
H, every 4 h change primary distilled water；
(9) polysaccharide solution for taking step (8) to obtain is eluted through DEAD Sephadex A-25 ion exchange column, is afforded
Component again through Sephadex G-200 sephadex column, collect single eluting peak solution, after freeze-drying, crushed 50-
100 meshes obtain ganoderma lucidum holosaccharide powder, spare；
The cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae compound Chinese medicine extract the preparation method comprises the following steps:
(1) cordate houttuynia, Fructus Forsythiae, rhizoma anemarrhenae are respectively washed 2-3 times, are pulverized after 45-55 °C of drying, and 80-100 mesh is crossed, spare；
(2) it is 70%- that the Heartleaf Houttuynia Herb for taking step (1) to prepare is 1:3-5 addition mass concentration with volume ratio g/mL by weight
80% ethanol solution, microwave-ultrasonic cooperate with assisted extraction 10-20min, filtering；Filter residue is repeated to extract 2- by above-mentioned steps
3 times, merging filtrate is centrifuged 10-20 min under the conditions of 3000-4000 r/min, takes supernatant, dense under the conditions of 45-55 °C
It is reduced to medicinal extract shape, 50-80 mesh is crushed after freeze-drying, obtains houttuynia extract；
(3) it is 70%-80% that the Weeping Forsythia Capsule for taking step (1) to prepare is 1:3-4 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 10-15min, filtering；Filter residue is repeated to extract 2-3 by above-mentioned steps
Secondary, merging filtrate is centrifuged 10-20 min under the conditions of 3000-4000 r/min, takes supernatant, be concentrated under the conditions of 45-55 °C
To medicinal extract shape, it crushed 50-80 mesh after freeze-drying, obtain forsythia suspense extraction；
(4) it is 70%-80% that the Common Anemarrhena Rhizome for taking step (1) to prepare is 1:4-6 addition mass concentration with volume ratio g/mL by weight
Ethanol solution, microwave-ultrasonic cooperate with assisted extraction 15-20 min, filtering；Filter residue is repeated to extract 2-3 by above-mentioned steps
Secondary, merging filtrate is centrifuged 10-20 min under the conditions of 3000-4000 r/min, takes supernatant, be concentrated under the conditions of 45-55 °C
To medicinal extract shape, it crushed 50-80 mesh after freeze-drying, obtain anemarrhena asphodefoides extract；
(5) step (2) houttuynia extract, step (3) forsythia suspense extraction, step (4) anemarrhena asphodefoides extract powder are taken, according to weight
It is mixed than 2:1:1 and compound Chinese medicine extract is made.
2. a kind of ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease according to claim 1, it is characterised in that: the essence
Including one or more of vanilla, butter essence, flavoring apple essence, grape essence, flavoring banana essence.
3. a kind of preparation method for the ganoderma lucidum polysaccharide compound for treating hand-foot-and-mouth disease as described in claim 1, it is characterised in that:
The method includes as follows:
(1) according to formula, ganoderma lucidum holosaccharide is taken, is mixed with cordate houttuynia, Fructus Forsythiae and rhizoma anemarrhenae Chinese medicinal compound extract, adds sugarcane
The original solution of composite oral liquid is made in sugar, essence, Aspartame, pure water, stirring 20-30 min；Adjusting solution ph is
6.9-7.5；
(2) the solution left standstill 2-4 h for obtaining step (1), takes supernatant through hollow-fibre membrane ultrafiltration apparatus ultrafiltration, obtains oral
Liquid；
(3) the deployed oral solution of step (2) is used into filling, locking closure while hot, sterilized and becomes product after the assay was approved.
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| CN102274234A (en) * | 2011-06-16 | 2011-12-14 | 武汉大学 | Application of ganoderic acid Y to preparation of medicament for treating or preventing enterovirus 71 infection |
| CN102526072A (en) * | 2011-12-23 | 2012-07-04 | 武汉大学 | Application of Lanosta-7,9(11),24-trien-3-one 15,26-dihydroxy-sterane triterpene in preparation of medicament for preventing and/or treating EV71 infection |
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| CN102274234A (en) * | 2011-06-16 | 2011-12-14 | 武汉大学 | Application of ganoderic acid Y to preparation of medicament for treating or preventing enterovirus 71 infection |
| CN102526072A (en) * | 2011-12-23 | 2012-07-04 | 武汉大学 | Application of Lanosta-7,9(11),24-trien-3-one 15,26-dihydroxy-sterane triterpene in preparation of medicament for preventing and/or treating EV71 infection |
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