CN106539944A - A kind of effective ingredient of calyx from generation to generation and preparation method and the application in slimming medicine is prepared - Google Patents
A kind of effective ingredient of calyx from generation to generation and preparation method and the application in slimming medicine is prepared Download PDFInfo
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- CN106539944A CN106539944A CN201610998025.4A CN201610998025A CN106539944A CN 106539944 A CN106539944 A CN 106539944A CN 201610998025 A CN201610998025 A CN 201610998025A CN 106539944 A CN106539944 A CN 106539944A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
- A61K36/752—Citrus, e.g. lime, orange or lemon
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- 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
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention belongs to the field of Chinese medicines, discloses a kind of effective ingredient of calyx from generation to generation and preparation method with the application in slimming medicine is prepared.Methods described is that (1) calyx will dry, pulverize and sieve from generation to generation, obtains calyx coarse powder from generation to generation；(2) using ethanol solution heating and refluxing extraction calyx coarse powder from generation to generation, then extracting solution is centrifuged, and by supernatant concentration, is dried, obtains calyx alcohol extracts from generation to generation；(3) water dissolution will be distilled by calyx alcohol extracts extractum from generation to generation, as sample solution；After by macroporous resin pretreatment, sample solution is added in macroporous resin, eluting is carried out using water, again using the ethanol solution gradient elution of variable concentrations, each component corresponding to different concentration ethanol eluant solution is collected, concentrating under reduced pressure is carried out respectively, it is dried to constant weight, obtains calyx effective ingredient from generation to generation.The effective ingredient of the present invention has significant anti-obesity activity, and using dosage is low, and the toxic and side effects under same dosage are relatively low.
Description
Technical field
The invention belongs to the field of Chinese medicines, and in particular to a kind of calyx effective ingredient and preparation method thereof from generation to generation with subtract in preparation
Application in fertile medicine.
Background technology
Metabolism syndrome is the general name of various Developmental and Metabolic Disorder disease groups, including fat, insulin resistant, and dyslipidemia is high
Insulinemia, hypertension etc..In these Developmental and Metabolic Disorder, obesity is stood in the breach, and becomes the primary embodiment of other Developmental and Metabolic Disorder,
The health of the mankind has been endangered in the world.Various diseases such as type Ⅱdiabetes mellitus, hypertension, cardiovascular and cerebrovascular disease, tumor
The generation of disease, development all have close relationship with obesity.The global health for becoming harm human health fat at present is asked
Topic, it is closely related with the generation of various diseases such as type Ⅱdiabetes mellitus, hypertension, cardiovascular and cerebrovascular disease, tumor, development.
3T3-L1 PECTORAL LIMB SKELETONs are isolated from the Swiss 3T3 cells of mice embryonic (17～19d), specifically can be lured
Lead the adipose cell for being divided into maturation.Due to drawing materials for mice Preadipocyte In Vitro it is more convenient, and with regard to body fat before mice
The culture studies technology of fat cell is more ripe so that 3T3-L1 PECTORAL LIMB SKELETONs have become research lipid metabolism generally acknowledged in the world
Cell model.Increasing research finds that Chinese medicine can pass through to suppress the propagation of 3T3-L1 PECTORAL LIMB SKELETONs to play prevention or subtract
Effect of fertilizer.Such as in Hua Qiang etc. using the apigenin of mtt assay and oil red O staining method research variable concentrations to PECTORAL LIMB SKELETON
Propagation and the impact broken up, test result indicate that, apigenin can significantly suppress the propagation of 3T3-L1 PECTORAL LIMB SKELETONs with
And differentiation, and be in dose dependent, point out apigenin there be prevention or treat fat effect.
(Citrus aurantium L.var.amara Engl) is a change of Rutaceae citrus plant Citrus aurantium Linn. from generation to generation
Kind, CitrusaurantiumL.Var.amara Engl. is the dry product of its alabastrum, with good pharmacological action, is usually used in treating depression of QI, abdominal distention, food stagnation not
Change, nausea and vomiting etc..At present both at home and abroad to the research of CitrusaurantiumL.Var.amara Engl. mainly based on its volatile oil compound, and to CitrusaurantiumL.Var.amara Engl. its
The research report of its effective ingredient does not almost have.The natural health-care products that CitrusaurantiumL.Var.amara Engl. is worth as medicine-food two-purpose, its effective ingredient
The research of the aspect such as extraction purification and pharmacologically active need further further investigated, strengthens the biological utilisation of CitrusaurantiumL.Var.amara Engl. comprehensively,
For developing new drug, clinical application is instructed to have great importance.
The content of the invention
In order to overcome the deficiencies in the prior art and shortcoming, the primary and foremost purpose of the present invention is that a kind of calyx from generation to generation of offer is effective
The preparation method of composition.
Another object of the present invention is to provide the calyx effective ingredient from generation to generation that above-mentioned preparation method is prepared.
It is still another object of the present invention to provide application of the above-mentioned effective ingredient of calyx from generation to generation in slimming medicine is prepared.
The purpose of the present invention is realized by following proposal：
A kind of preparation method of the effective ingredient of calyx from generation to generation, comprises the steps of：
(1) calyx (Citrus aurantium L.var.amara Engl) will dry, pulverize and sieve from generation to generation, obtain from generation to generation
Calyx coarse powder；
(2) using ethanol solution heating and refluxing extraction calyx coarse powder from generation to generation, then extracting solution is centrifuged, and supernatant is dense
Contracting, is dried, obtains calyx alcohol extracts from generation to generation；
(3) water dissolution will be distilled by calyx alcohol extracts from generation to generation, as sample solution；After macroporous resin pretreatment, will
During sample solution adds macroporous resin, eluting is carried out using water, then using the ethanol solution gradient elution of variable concentrations, collect different
Each component corresponding to concentration ethanol eluant solution, carries out concentrating under reduced pressure respectively, is dried to constant weight, obtains calyx from generation to generation effective
Composition.
When the ethanol solution of variable concentrations be ethanol water that volume fraction is 30%, volume fraction be 50% ethanol water
Solution and volume fraction are 70% ethanol water, and the effective ingredient of calyx from generation to generation is designated as CAV-30%, CAV- respectively
50%th, CAV-70%.
Gradient elution described in step (3) to be referred to and carry out eluting successively using the ascending ethanol solution of concentration.It is described
The ethanol solution of variable concentrations is ethanol water that volume fraction is 30%, volume fraction is 50% ethanol water and volume
Fraction is 70% ethanol water.
Baking temperature described in step (1) is 40～60 DEG C；The drying time is 18～24h；Described pulverizes and sieves
For 40～60 mesh sieves.
The volume fraction of ethanol solution described in step (2) is 70%～90%.Step (2) ethanol solution with from generation to generation
Volume mass ratio (ratio of water to material) of calyx coarse powder is (10～30) mL:1g.
The number of times of heating and refluxing extraction described in step (2) is 2～5 times, and the temperature extracted every time is 80～100 DEG C；Often
The time of secondary extraction is 1～3h.
The rotating speed being centrifuged described in step (2) is 3000～5000r/min；The time of the centrifugation is 8～12min.Step
Suddenly concentration described in (2) is concentrating under reduced pressure；The temperature of the concentrating under reduced pressure is 40～60 DEG C；The temperature of the drying is 40～60
DEG C, the dry time is 18～24h.
The method of the pretreatment of step (3) macroporous adsorbent resin is：Macroporous adsorbent resin is first used distilled water immersion
12～24h, then first soaks 0.5～3h with the hydrochloric acid solution that mass fraction is 3～5%, and distillation is washed to neutrality, then uses quality
Fraction is that 3～5% sodium hydroxide solution soaks 0.5～3h, and distillation is washed to neutrality, obtains the macroporous absorption through pretreatment
Resin；
Macroporous adsorbent resin described in step (3) is selected as AB-8 resins.
A kind of calyx effective ingredient from generation to generation is prepared by above-mentioned preparation method.
Application of the effective ingredient of calyx from generation to generation in slimming medicine is prepared.The effective ingredient of calyx from generation to generation can conduct
New slimming medicine, is applied to the treatment of obesity.
A kind of slimming medicine, containing in the above-mentioned effective ingredient of calyx from generation to generation (CAV-30%, CAV-50%, CAV-70%)
It is at least one；
The principle of the present invention：The effective ingredient of natural botanical source can pass through to suppress the propagation of PECTORAL LIMB SKELETON to play fat-reducing
Effect.Present invention discover that there is significant anti-obesity activity with confirmation active ingredient of natural plant, and using dosage is low, same
Toxic and side effects under dosage are relatively low.
The present invention is had the following advantages relative to prior art and effect：
(1) present invention firstly discovers that calyx effective ingredient (CAV-30%, CAV-50%, CAV-70%) in vitro can from generation to generation
Significantly inhibit the propagation of 3T3-L1 PECTORAL LIMB SKELETONs.
(2) present invention firstly discovers that calyx effective ingredient (CAV-70%) can significantly inhibit fat before 3T3-L1 in vitro from generation to generation
The differentiation of fat cell.
Description of the drawings
Fig. 1 is calyx from generation to generation effective ingredient CAV-30%, CAV-50% and CAV-70% couple that embodiment 4 is prepared
3T3-L1 PECTORAL LIMB SKELETON inhibited proliferation figures；
Fig. 2 is that the effective ingredient of the calyx from generation to generation CAV-70% that embodiment 4 is prepared breaks up to 3T3-L1 PECTORAL LIMB SKELETONs
The action diagram of early stage；
Fig. 3 is that the effective ingredient of the calyx from generation to generation CAV-70% that embodiment 4 is prepared breaks up to 3T3-L1 PECTORAL LIMB SKELETONs
The action diagram of overall process.
Specific embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited
In this.
In embodiment, mice 3T3-L1 PECTORAL LIMB SKELETONs are purchased from Shanghai life science institute of Chinese Academy of Sciences cellular resources
The heart；Calyx is purchased from the peaceful medicinal material market in Guangzhou from generation to generation.
Embodiment 1 prepares calyx alcohol extracts CAV-I from generation to generation from calyx from generation to generation
(1) calyx (Citrus aurantium L.var.amara Engl) will be placed in electric drying oven with forced convection from generation to generation
50 DEG C are dried 24h, after being crushed with pulverizer, cross 60 mesh sieves, obtain calyx coarse powder from generation to generation standby；
(2), in conical flask, the volume fraction for adding 3000mL is 80% ethanol solution to weigh calyx coarse powder 100g from generation to generation,
Fully shaking shakes up, 90 DEG C of water-bath heating and refluxing extraction 3 times, heats 3h every time；After heating terminates, united extraction liquid,
10min is centrifuged under 4000r/min, supernatant liquid is taken, by supernatant Rotary Evaporators at 50 DEG C concentrating under reduced pressure, 45 DEG C are true
Empty drying baker is dried 20h, obtains calyx alcohol extracts CAV-I from generation to generation.
Embodiment 2 prepares calyx alcohol extracts CAV-I from generation to generation from calyx from generation to generation
(1) calyx (Citrus aurantium L.var.amara Engl) will be placed in electric drying oven with forced convection from generation to generation
50 DEG C are dried 24h, after being crushed with pulverizer, cross 60 mesh sieves, obtain calyx coarse powder from generation to generation standby；
(2), in conical flask, the volume fraction for adding 2000mL is 90% ethanol solution to weigh calyx coarse powder 100g from generation to generation,
Fully shaking shakes up, 100 DEG C of water-bath heating and refluxing extraction 4 times, heats 2h every time；After heating terminates, united extraction liquid,
8min is centrifuged under 5000r/min, supernatant liquid is taken, by supernatant Rotary Evaporators at 40 DEG C concentrating under reduced pressure, 50 DEG C are true
Empty drying baker is dried 18h, obtains final product calyx alcohol extracts CAV-I from generation to generation.
Embodiment 3 prepares calyx alcohol extracts CAV-I from generation to generation from calyx from generation to generation
(1) calyx (Citrus aurantium L.var.amara Engl) will be placed in electric drying oven with forced convection from generation to generation
45 DEG C are dried 18h, after being crushed with pulverizer, cross 50 mesh sieves, obtain calyx coarse powder from generation to generation standby；
(2), in conical flask, the volume fraction for adding 1500mL is 70% ethanol solution to weigh calyx coarse powder 100g from generation to generation,
Fully shaking shakes up, 90 DEG C of water-bath heating and refluxing extraction 3 times, heats 3h every time；After heating terminates, united extraction liquid,
10min is centrifuged under 4000r/min, supernatant liquid is taken, by supernatant Rotary Evaporators at 60 DEG C concentrating under reduced pressure, 55 DEG C are true
Empty drying baker is dried 16h, obtains final product calyx alcohol extracts CAV-I from generation to generation.
Embodiment 4 prepares calyx effective ingredient CAV-30%, from generation to generation calyx effective ingredient from generation to generation from calyx from generation to generation
CAV-50% and from generation to generation calyx effective ingredient CAV-70%
The alcohol extracts of calyx from generation to generation prepared in embodiment 1 are distilled after water dissolution, 4000r/min at a high speed from
Heart 10min, takes supernatant as sample solution；Sample solution is slowly added into after above the macroporous resin of pretreatment, is first used
Distillation 5 column volumes of water elution, wash away water-solubility impurity；Then volume fraction is used to be 30% ethanol solution, volume fraction successively
It is that 50% ethanol solution and volume fraction distinguish 5 column volumes of eluting for 70% ethanol solution, collects each component, 50 DEG C of decompressions
Concentration and recovery ethanol, 50 DEG C are dried under vacuum to constant weight, respectively obtain 30% elution fraction, 50% elution fraction and 70% eluting portion
Point i.e. calyx effective ingredient CAV-30%, from generation to generation calyx effective ingredient CAV-50%, from generation to generation calyx effective ingredient CAV- from generation to generation
70%.
The method of the pretreatment of described macroporous adsorbent resin is：Macroporous adsorbent resin is first used distilled water immersion 16h, so
Afterwards first with hydrochloric acid solution that mass fraction is 4% immersion 2h, distillation is washed to neutrality, then with the hydroxide that mass fraction is 4%
Sodium solution soaks 2h, and distillation is washed to neutrality, obtains the macroporous adsorbent resin through pretreatment；Described macroporous adsorbent resin choosing
With for AB-8 resins.
Embodiment 5 prepares calyx effective ingredient CAV-30%, from generation to generation calyx effective ingredient from generation to generation from calyx from generation to generation
CAV-50% and from generation to generation calyx effective ingredient CAV-70%
The alcohol extracts of calyx from generation to generation prepared in embodiment 1 are distilled after water dissolution, 3000r/min at a high speed from
Heart 12min, takes supernatant as sample solution；After sample solution is slowly added into above the macroporous resin of pretreatment, first with distillation
4 column volumes of water elution, wash away water-solubility impurity, are then that 30% ethanol solution, volume fraction are with volume fraction successively
50% ethanol solution and volume fraction are that 70% ethanol solution distinguishes 5 column volumes of eluting, collect each component, and 45 DEG C of decompressions are dense
Retract and receive ethanol, 45 DEG C are dried under vacuum to constant weight, respectively obtain 30% elution fraction, 50% elution fraction and 70% elution fraction
That is calyx effective ingredient CAV-30%, from generation to generation calyx effective ingredient CAV-50%, from generation to generation calyx effective ingredient CAV-70% from generation to generation.
The method of the pretreatment of described macroporous adsorbent resin is：Macroporous adsorbent resin is first used distilled water immersion 12h, so
Afterwards first with hydrochloric acid solution that mass fraction is 5% immersion 0.5h, distillation is washed to neutrality, then with the hydrogen-oxygen that mass fraction is 5%
Change sodium solution immersion 0.5h, distillation is washed to neutrality, obtains the macroporous adsorbent resin through pretreatment；Described macroporous absorption tree
Fat is selected as AB-8 resins.
Embodiment 6 prepares calyx effective ingredient CAV-30%, from generation to generation calyx effective ingredient from generation to generation from calyx from generation to generation
CAV-50% and from generation to generation calyx effective ingredient CAV-70%
The alcohol extracts of calyx from generation to generation prepared in embodiment 1 are distilled after water dissolution, 5000r/min at a high speed from
Heart 8min, takes supernatant as sample solution；After sample solution is slowly added into above macroporous resin, first with distillation water elution 6
Column volume, washes away water-solubility impurity, then uses volume fraction to be 50% ethanol solution for 30% ethanol solution, volume fraction successively
It is that 70% ethanol solution distinguishes 6 column volumes of eluting with volume fraction, collects each component, 55 DEG C of concentrating under reduced pressure reclaims ethanol,
55 DEG C are dried under vacuum to constant weight, respectively obtain 30% elution fraction, 50% elution fraction and 70% elution fraction i.e. calyx from generation to generation
Effective ingredient CAV-30%, from generation to generation calyx effective ingredient CAV-50%, from generation to generation calyx effective ingredient CAV-70%.
The method of the pretreatment of described macroporous adsorbent resin is：Macroporous adsorbent resin is first used distilled water immersion 24h, so
Afterwards first with hydrochloric acid solution that mass fraction is 3% immersion 3h, distillation is washed to neutrality, then with the hydroxide that mass fraction is 3%
Sodium solution soaks 3h, and distillation is washed to neutrality, obtains the macroporous adsorbent resin through pretreatment；The macroporous adsorbent resin is selected
For AB-8 resins.
7 mtt assay of embodiment detects calyx effective ingredient CAV-30%, from generation to generation calyx effective ingredient CAV-50%, generation from generation to generation
The impact that seville orange flower calyx effective ingredient CAV-70% breeds to 3T3-L1 PECTORAL LIMB SKELETONs
The mice 3T3-L1 PECTORAL LIMB SKELETONs of growth logarithmic (log) phase are taken respectively, and 1000rpm centrifugation 5min, abandoning supernatant are used
It is 5 × 10 containing the DMEM culture medium adjustment cell number that mass fraction is 10% hyclone4/ mL, 100 μ L are inoculated in the culture of 96 holes
In plate, 5%CO is placed in2In incubator, after 37 DEG C of culture 24h, abandoning supernatant, each group are separately added into (real containing CAV-30%
Apply example 4 to be obtained), CAV-50% (embodiment 4 be obtained), CAV-70% (embodiment 4 is obtained) DMEM culture medium (CAV-30%,
The final concentration of CAV-50%, CAV-70% is the 50 μ g/mL of μ g/mL, 100,200,400 and 800 respectively).In 37 DEG C, 5%CO2
24h is cultivated in incubator, abandoning supernatant, PBS add DMEM basal mediums of the 200 μ L without serum twice, per hole
With 20 μ L tetrazolium bromides (MTT), continue incubation 4h in being put into incubator, abandon supernatant, 150 μ L dimethyl sulfoxide are added per hole
(DMSO), avoid light place uniformly shakes 10min in micro oscillator, measures the OD values under 490nm wavelength with microplate reader, calculates
Cell proliferation inhibition rate；
Fig. 1 is calyx from generation to generation effective ingredient CAV-30%, CAV-50% and CAV-70% couple that embodiment 4 is prepared
3T3-L1 PECTORAL LIMB SKELETON inhibited proliferation figures.Calyx effective ingredient CAV-30% (Fig. 1), from generation to generation calyx effective ingredient from generation to generation
CAV-50% (Fig. 1), from generation to generation calyx effective ingredient CAV-70% (Fig. 1) in the range of 50～800 μ g/mL to mice 3T3-L1 before
Adipose cell shows significant inhibited proliferation, and CAV-VI-70% inhibitions are most notable, therefore selects CAV-
70% carries out next step experiment.
Calyx effective ingredient CAV-70% breaks up the detection of 8 oil red O staining method of embodiment to 3T3-L1 PECTORAL LIMB SKELETONs from generation to generation
Impact
(1) this experiment is divided into two groups, is designated as I groups and II groups.Before taking the mice 3T3-L1 of growth logarithmic (log) phase respectively, fat is thin
Born of the same parents, 1000rpm centrifugation 5min, abandoning supernatant, with containing the DMEM culture medium adjustment cell that mass fraction is 10% hyclone
Number is 5 × 104/ mL, 100 μ L are inoculated in 96 well culture plates, are placed in 5%CO2In incubator, first with the training completely containing 10%FBS
Nutrient solution culture 2d, continues to be finished full nutrient solution culture 2d after cell is covered with, and makes cell reach contact inhibition, I groups and II components
10 μ g/mL insulins, the complete culture solution of 0.05mmol/L IBMX and 1 μm of ol/L Jia Ru not be contained, while adding CAV-70%
(final concentration is respectively 6.25,12.5,25,50 and 100 μ g/mL), while arranging matched group, every kind of process sets 5 repetitions.Culture
After 2d, I groups change the complete culture solution for comprising only 10 μ g/mL insulins, and II groups change the culture completely containing 10 μ g/mL insulins
Liquid, while adding CAV-70% (final concentration is respectively 6.25,12.5,25,50 and 100 μ g/mL), cultivates 2～4d, and I groups are used instead
Complete culture solution culture 4d, II groups are used instead containing CAV-70% (final concentration is respectively 6.25,12.5,25,50 and 100 μ g/mL)
Complete culture solution culture 4d, until differentiation terminates, 80% or so cell has been divided into the adipose cell of maturation；
(2) by induce differentiation into it is ripe after cell PBS carefully washing 3 times, the 4% poly first ferment solution with 100 μ L is solid
Determine 30min, then discard paraformaldehyde solution, washed with PBS
Dye 30min under O working solution room temperatures, after dyeing 30min, remove dye liquor, then plus 70% ethanol rapid cleanup cell reach decolouring
Purpose, plus PBS cell 3 times is subsequently adding 200 μ L isopropanols, determines light absorption value under 490nm；
As shown in Figures 2 and 3, Fig. 2 is the effective ingredient of calyx from generation to generation CAV-70% pair that embodiment 4 is prepared to test result
3T3-L1 PECTORAL LIMB SKELETONs break up the action diagram of early stage；Fig. 3 is the effective ingredient CAV- of calyx from generation to generation that embodiment 4 is prepared
70% pair of 3T3-L1 PECTORAL LIMB SKELETON breaks up the action diagram of overall process.I groups (Fig. 2) and II groups (Fig. 3) can significantly inhibit 3T3-
The differentiation of L1 PECTORAL LIMB SKELETONs, the Fat Accumulation in cell are gradually decreased, and fat drop diminishes, and II group inhibitions are than I group
Inhibition is more notable.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention not by above-described embodiment
Limit, other any spirit without departing from the present invention and the change, modification, replacement made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. a kind of preparation method of the effective ingredient of calyx from generation to generation, it is characterised in that：Comprise the steps of：
(1) calyx will dry, pulverize and sieve from generation to generation, obtain calyx coarse powder from generation to generation；
(2) using ethanol solution heating and refluxing extraction calyx coarse powder from generation to generation, then extracting solution is centrifuged, and by supernatant concentration,
It is dried, obtains calyx alcohol extracts from generation to generation；
(3) water dissolution will be distilled by calyx alcohol extracts extractum from generation to generation, as sample solution；After macroporous resin pretreatment, will
During sample solution adds macroporous resin, eluting is carried out using water, then using the ethanol solution gradient elution of variable concentrations, collect different
Each component corresponding to concentration ethanol eluant solution, carries out concentrating under reduced pressure respectively, is dried to constant weight, obtains calyx from generation to generation effective
Composition.
2. the preparation method of calyx effective ingredient from generation to generation according to claim 1, it is characterised in that：Ladder described in step (3)
Degree eluting to be referred to and carry out eluting successively using the ascending ethanol solution of concentration.
3. the preparation method of calyx effective ingredient from generation to generation according to claim 1, it is characterised in that：The second of the variable concentrations
Alcoholic solution is ethanol water that volume fraction is 30%, volume fraction is 50% ethanol water and volume fraction is 70%
Ethanol water.
4. the preparation method of calyx effective ingredient from generation to generation according to claim 1, it is characterised in that：Second described in step (2)
The volume fraction of alcoholic solution is 70%～90%；
The number of times of heating and refluxing extraction described in step (2) is 2～5 times, and the temperature extracted every time is 80～100 DEG C；Carry every time
The time for taking is 1～3h.
5. the preparation method of calyx effective ingredient from generation to generation according to claim 1, it is characterised in that：Step (3) macropore
The method of the pretreatment of adsorbent resin is：Macroporous adsorbent resin is first used 12～24h of distilled water immersion, mass fraction is then first used
Hydrochloric acid solution for 3～5% soaks 0.5～3h, and distillation is washed to neutrality, then molten with the sodium hydroxide that mass fraction is 3～5%
Immersion steeps 0.5～3h, and distillation is washed to neutrality, obtains the macroporous adsorbent resin through pretreatment；
Macroporous adsorbent resin described in step (3) is selected as AB-8 resins.
6. the preparation method of calyx effective ingredient from generation to generation according to claim 1, it is characterised in that：Second described in step (2)
Alcoholic solution is (10～30) mL with the volume mass ratio of calyx coarse powder from generation to generation:1g；
The rotating speed being centrifuged described in step (2) is 3000～5000r/min；The time of the centrifugation is 8～12min.
7. the preparation method of calyx effective ingredient from generation to generation according to claim 1, it is characterised in that：It is dense described in step (2)
It is condensed to concentrating under reduced pressure；The temperature of the concentrating under reduced pressure is 40～60 DEG C；The temperature of the drying is 40～60 DEG C, the dry time
For 18～24h；Baking temperature described in step (1) is 40～60 DEG C；The drying time is 18～24h；Described crushing
Sieve as 40～60 mesh sieves.
8. calyx effective ingredient is prepared one kind by preparation method described in 1～7 any one of the claims from generation to generation.
9. application of the calyx effective ingredient in slimming medicine is prepared from generation to generation according to claim 8.
10. a kind of slimming medicine, containing at least one described in claim 8 from generation to generation in calyx effective ingredient.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108785397A (en) * | 2018-07-27 | 2018-11-13 | 华南理工大学 | A kind of vine of multiflower knotweed extract and preparation method and the application in preparing slimming medicine |
| CN116370536A (en) * | 2022-12-13 | 2023-07-04 | 南方医科大学 | Substituted flower flavone, preparation method thereof and application thereof in preparation of weight-losing medicine |
| CN116747262A (en) * | 2023-08-14 | 2023-09-15 | 云南英格生物技术有限公司 | Substituted flower active substance and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108785397A (en) * | 2018-07-27 | 2018-11-13 | 华南理工大学 | A kind of vine of multiflower knotweed extract and preparation method and the application in preparing slimming medicine |
| CN116370536A (en) * | 2022-12-13 | 2023-07-04 | 南方医科大学 | Substituted flower flavone, preparation method thereof and application thereof in preparation of weight-losing medicine |
| CN116747262A (en) * | 2023-08-14 | 2023-09-15 | 云南英格生物技术有限公司 | Substituted flower active substance and preparation method and application thereof |
| CN116747262B (en) * | 2023-08-14 | 2023-11-10 | 云南英格生物技术有限公司 | Substituted flower active substance and preparation method and application thereof |
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