CN106728187A - A kind of extract of calyx from generation to generation and preparation method and the application in slimming medicine is prepared - Google Patents
A kind of extract of calyx from generation to generation and preparation method and the application in slimming medicine is prepared Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
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- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
The invention belongs to the field of Chinese medicines, a kind of extract of calyx from generation to generation and preparation method are disclosed with the application in slimming medicine is prepared.Methods described is, using ethanol solution heating and refluxing extraction calyx meal from generation to generation, then extract solution to be centrifuged, and supernatant concentration is dried to constant weight, obtains calyx alcohol extracts from generation to generation；Then by alcohol extracts distillation water dissolves, petroleum ether extraction is added, removes petroleum ether phase；Add chloroform to be extracted in the solution after removing petroleum ether phase, repeat extracted several times, chloroform phase is collected respectively and the solution after chloroform phase is taken out, chloroform is mutually concentrated and is evaporated, obtain chloroform phase extract；(4) to adding ethyl acetate to extract in the solution taken out after chloroform phase, combined ethyl acetate phase, concentration is evaporated, and obtains ethyl acetate phase extract.The extract has significant anti-obesity activity, and dosage is low, and the toxic and side effect under same dosage is relatively low.
Description
Technical field
The invention belongs to the field of Chinese medicines, and in particular to a kind of calyx extract and preparation method thereof from generation to generation is lost weight with preparation
Application in medicine.
Background technology
Metabolic syndrome is the general name of various metabolic disorder diseases group, including fat, insulin resistance, and dyslipidemia is high
Insulinemia, hypertension etc..In these metabolic disorders, obesity is stood in the breach, the primary embodiment as other metabolic disorders,
The health of the mankind has been endangered in the world.Various diseases such as type II diabetes, hypertension, cardiovascular and cerebrovascular disease, tumour
The generation of disease, development all have close relationship with obesity.The global health for having turned into harm human health fat at present is asked
Topic, its generation with various diseases such as type II diabetes, hypertension, cardiovascular and cerebrovascular disease, tumour, development are closely related.
3T3-L1 PECTORAL LIMB SKELETONs are isolated from the Swiss 3T3 cells of mice embryonic (17~19d), can specifically be lured
Lead the fat cell for being divided into maturation.Because the materials of mouse Preadipocyte In Vitro are more convenient, and on body fat before mouse
The culture studies technology of fat cell is more ripe so that 3T3-L1 PECTORAL LIMB SKELETONs have turned into research fat metabolism generally acknowledged in the world
Cell model.Increasing research finds that Chinese medicine can be by suppressing the propagation performance prevention of 3T3-L1 PECTORAL LIMB SKELETONs or subtracting
Effect of fertilizer.Such as the apiolin of various concentrations is studied to PECTORAL LIMB SKELETON using mtt assay and oil red O staining method in magnificent strong grade
Propagation and the influence of differentiation, test result indicate that, apiolin can significantly suppress the propagation of 3T3-L1 PECTORAL LIMB SKELETONs with
And differentiation, and in dose dependent, point out apiolin to have prevention or the fat effect for the treatment of.
(Citrus aurantium L.var.amara Engl) is a change of Rutaceae citrus plant bitter orange from generation to generation
Kind, bitter orange flower is the dry product of its bud, with good pharmacological action, is usually used in treatment depression of QI, abdominal distention, having indigestion not
Change, n and V etc..It is main to the research of bitter orange flower based on its volatile oil compound both at home and abroad at present, and to bitter orange flower its
The research report of its active ingredient does not almost have.The natural health-care products that bitter orange flower is worth as medicine-food two-purpose, its active ingredient
The research of the aspect such as extraction purification and pharmacological activity need further further investigated, and the biological utilisation of bitter orange flower is strengthened 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, primary and foremost purpose of the invention is to provide a kind of extraction of calyx from generation to generation
The preparation method of thing.
Another object of the present invention is to provide the calyx extract 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 extract 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 extract of calyx from generation to generation, comprises the following steps：
(1) sieving will be dry, pulverize by calyx (Citrus aurantium L.var.amara Engl) from generation to generation, will be obtained from generation to generation
Calyx meal；
(2) ethanol solution heating and refluxing extraction calyx meal from generation to generation is used, then extract solution is centrifuged, and supernatant is dense
Contracting, dries to constant weight, obtains calyx alcohol extracts (being designated as CAV-I) from generation to generation；
(3) water dissolves will be distilled by calyx alcohol extracts from generation to generation, and will add petroleum ether to be extracted, repeat extracted several times,
Removal petroleum ether phase；Add chloroform to be extracted in the solution after removing petroleum ether phase, repeat extracted several times, chlorine is collected respectively
Imitative solution mutually and after taking-up chloroform phase, chloroform is mutually concentrated and is evaporated, and obtains chloroform phase extract (being designated as CAV-II)；
(4) to adding ethyl acetate to be extracted in the solution taken out after chloroform phase, extracted several times are repeated, merges acetic acid second
Ester phase, concentration is evaporated, and obtains ethyl acetate phase extract (being designated as CAV-III).The extract of calyx from generation to generation is calyx from generation to generation
Ethanol crude extract, chloroform phase extract and/or ethyl acetate phase extract.
Dry temperature described in step (1) is 40~60 DEG C；The drying time is 18~24h；It is described to pulverize and sieve
It is 40~60 mesh sieves.
The volume fraction of ethanol solution described in step (2) is 70~90%.
Ethanol solution described in step (2) is (10~30) mL with the volume mass ratio of calyx meal from generation to generation:1g.
The temperature of heating and refluxing extraction described in step (2) is 80~100 DEG C；The heating and refluxing extraction number of times is 2~5
Secondary, the time extracted every time is 1~3h.
The rotating speed being centrifuged described in step (2) is 3000~5000r/min；The time of the centrifugation is 8~12min.
Dry temperature described in step (2) is 40~60 DEG C.
Step (3) the calyx alcohol extracts from generation to generation and the mass volume ratio of distilled water are 1g：(10~20) mL.
The mode of the concentration described in step (2), (3) and (4) is concentrated under reduced pressure；
The time extracted described in step (3) is all 30~60min；The time extracted described in step (4) for 30~
60min；
Extraction times described in step (3) are all 3~5 times；Extraction times described in step (4) are all 3~5 times.
A kind of calyx extract (CAV-I, CAV-II, CAV-III) from generation to generation, is prepared by above-mentioned preparation method；
Application of the described extract of the calyx from generation to generation (CAV-I, CAV-II, CAV-III) in slimming medicine is prepared；
The described extract of calyx from generation to generation (CAV-I, CAV-II, CAV-III) can be applied to as new slimming medicine
The treatment of obesity.
A kind of slimming medicine, contains at least in the above-mentioned extract of calyx from generation to generation (CAV-I, CAV-II, CAV-III)
Kind；
Principle of the invention：The active ingredient of natural botanical source can play fat-reducing by suppressing the propagation of PECTORAL LIMB SKELETON
Effect.Present invention discover that there is significant anti-obesity activity with confirmation active ingredient of natural plant, and dosage is low, same
Toxic and side effect under dosage is relatively low.
The present invention has the following advantages and effect relative to prior art：
(1) present invention firstly discovers that calyx extract (CAV-I, CAV-II, CAV-III) can be significantly inhibited in vitro from generation to generation
The propagation of 3T3-L1 PECTORAL LIMB SKELETONs.
(2) present invention firstly discovers that fat is thin before calyx extract (CAV-II) can significantly inhibit 3T3-L1 in vitro from generation to generation
The differentiation of born of the same parents.
Brief description of the drawings
Fig. 1 be embodiment 1 prepare from generation to generation calyx alcohol extracts CAV-I to 3T3-L1 PECTORAL LIMB SKELETON Proliferation Abilities
Action diagram；
Fig. 2 is the extract of the calyx from generation to generation CAV-II for preparing of embodiment 4 to 3T3-L1 PECTORAL LIMB SKELETON Proliferation Abilities
Action diagram；
Fig. 3 is the extract of the calyx from generation to generation CAV-III for preparing of embodiment 4 to 3T3-L1 PECTORAL LIMB SKELETON Proliferation Abilities
Action diagram；
Fig. 4 is that the extract of the calyx from generation to generation CAV-II that embodiment 4 is prepared breaks up early stage to 3T3-L1 PECTORAL LIMB SKELETONs
Action diagram；
Fig. 5 is that the extract of the calyx from generation to generation CAV-II that embodiment 4 is prepared breaks up full mistake to 3T3-L1 PECTORAL LIMB SKELETONs
The action diagram of journey.
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, mouse 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 extract 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 dry 24h, after being crushed with pulverizer, cross 60 mesh sieves, obtain calyx meal from generation to generation standby；
(2) in conical flask, the volume fraction for adding 3000mL is 80% ethanol solution to weigh calyx meal 100g from generation to generation,
Fully shaking is shaken up, 90 DEG C of water-bath heating and refluxing extraction 3 times, and 3h is heated every time；After heating terminates, merge extract solution,
10min is centrifuged under 4000r/min, supernatant liquid is taken, supernatant Rotary Evaporators are concentrated under reduced pressure at 50 DEG C, vacuum is done
The 50 DEG C of dryings of dry case obtain final product calyx alcohol extracts from generation to generation and are designated as CAV-I i.e. calyx extract CAV-I from generation to generation to constant weight.
Embodiment 2 prepares calyx extract 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 dry 24h, after being crushed with pulverizer, cross 60 mesh sieves, obtain calyx meal from generation to generation standby；
(2) in conical flask, the volume fraction for adding 2000mL is 90% ethanol solution to weigh calyx meal 100g from generation to generation,
Fully shaking is shaken up, 100 DEG C of water-bath heating and refluxing extraction 4 times, and 2h is heated every time；After heating terminates, merge extract solution,
8min is centrifuged under 5000r/min, supernatant liquid is taken, supernatant Rotary Evaporators are concentrated under reduced pressure at 40 DEG C, vacuum is done
The 45 DEG C of dryings of dry case obtain final product calyx alcohol extracts from generation to generation and are designated as CAV-I i.e. calyx extract CAV-I from generation to generation to constant weight.
Embodiment 3 prepares calyx extract 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 dry 18h, after being crushed with pulverizer, cross 50 mesh sieves, obtain calyx meal from generation to generation standby；
(2) in conical flask, the volume fraction for adding 1500mL is 70% ethanol solution to weigh calyx meal 100g from generation to generation,
Fully shaking is shaken up, 90 DEG C of water-bath heating and refluxing extraction 3 times, and 3h is heated every time；After heating terminates, merge extract solution,
10min is centrifuged under 4000r/min, supernatant liquid is taken, supernatant Rotary Evaporators are concentrated under reduced pressure at 60 DEG C, vacuum is done
The 45 DEG C of dryings of dry case obtain final product calyx alcohol extracts from generation to generation and are designated as CAV-I i.e. calyx extract CAV-I from generation to generation to constant weight.
Embodiment 4 prepares calyx extract CAV-III and CAV-II from generation to generation from calyx from generation to generation
(1) (calyx ethanol is thick from generation to generation for the alcohol extracts of the calyx from generation to generation distillation water dissolves that will be prepared in embodiment 1
Extract quality (g) is 1 with the mass volume ratio of distilled water volume (mL):10), add the petroleum ether isometric with distilled water molten
Agent, fully shaking is mixed, and stands 40min, after two-phase solvent is layered completely, suctions out the petroleum ether phase on upper strata, repeats extraction three
It is secondary, petroleum ether layer is discarded, obtain aqueous phase solution；
(2) the isometric chloroform solvent of liquid compatible with water, fully shaking are added in the aqueous phase solution after petroleum ether extraction
Mix, stand 40min, after two-phase solvent is layered completely, collects chloroform phase and take out the solution after chloroform phase, repeat extraction three
It is secondary, chloroform phase is merged respectively and the solution after chloroform phase is taken out, chloroform is concentrated under reduced pressure in 60 DEG C, and 60 DEG C are dried under vacuum to perseverance
Weight, obtains chloroform phase extract and is designated as CAV-II i.e. calyx extract CAV-II from generation to generation；
(3) isometric ethyl acetate solvent is added in the aqueous solution after removing chloroform phase, fully shaking is mixed, and is stood
40min, after two-phase solvent is layered completely, suctions out the ethyl acetate phase on upper strata, repeats extraction three times, merges gained ethyl acetate
Phase extract, 60 DEG C are concentrated under reduced pressure, and are dried under vacuum to constant weight, obtain ethyl acetate phase extract and are designated as CAV-III i.e. bitter orange flower
Calyx extract CAV-III.
Embodiment 5 prepares calyx extract CAV-III and CAV-II from generation to generation from calyx from generation to generation
(1) (calyx ethanol is thick from generation to generation for the alcohol extracts of the calyx from generation to generation distillation water dissolves that will be prepared in embodiment 1
Extract quality (g) is 1 with the mass volume ratio of distilled water volume (mL):15), add the petroleum ether of 2 times of distilled water volumes molten
Agent, fully shaking is mixed, and stands 30min, after two-phase solvent is layered completely, suctions out the petroleum ether phase on upper strata, repeats extraction four
It is secondary, petroleum ether layer is discarded, obtain aqueous phase solution；
(2) 2 times of chloroform solvents of distilled water volume are added in the aqueous phase solution after petroleum ether extraction, fully shaking is mixed
It is even, 30min is stood, after two-phase solvent is layered completely, collects chloroform phase and take out the solution after chloroform phase, repeat extraction four
It is secondary, chloroform phase is merged respectively and the solution after chloroform phase is taken out, by chloroform phase, 50 DEG C are concentrated under reduced pressure, and are dried under vacuum to constant weight, obtain
It is calyx extract CAV-II from generation to generation to chloroform phase extract CAV-II；
(3) 2 times of ethyl acetate solvents of volume are added in the solution after removing chloroform phase, fully shaking is mixed, and is stood
30min, after two-phase solvent is layered completely, suctions out the ethyl acetate phase on upper strata, repeats extraction four times, merges gained ethyl acetate
Phase extract, 50 DEG C are concentrated under reduced pressure, and constant weight is dried under vacuum in 50 DEG C, are obtained ethyl acetate phase extract and are designated as CAV-III i.e.
Calyx extract CAV-III from generation to generation.
Embodiment 6 prepares calyx extract CAV-III and CAV-II from generation to generation from calyx from generation to generation
(1) (calyx ethanol is thick from generation to generation for the alcohol extracts of the calyx from generation to generation distillation water dissolves that will be prepared in embodiment 1
Extract quality (g) is 1 with the mass volume ratio of distilled water volume (mL):20), add the petroleum ether of 3 times of distilled water volumes molten
Agent, fully shaking is mixed, and stands 60min, after two-phase solvent is layered completely, suctions out the petroleum ether phase on upper strata, repeats extraction five
It is secondary, petroleum ether layer is discarded, obtain aqueous phase solution；
(2) 3 times of chloroform solvents of distilled water volume are added in the aqueous phase solution after petroleum ether extraction, fully shaking is mixed
It is even, 60min is stood, after two-phase solvent is layered completely, collects chloroform phase and take out the solution after chloroform phase, extraction 5 times is repeated,
Merge chloroform phase respectively and take out the solution after chloroform phase, chloroform is concentrated under reduced pressure in 55 DEG C, and 55 DEG C are dried under vacuum to constant weight,
Obtain chloroform phase extract CAV-II i.e. calyx extract CAV-II from generation to generation；
(3) 3 times of ethyl acetate solvents of volume are added in the solution after removing chloroform phase, fully shaking is mixed, and is stood
60min, after two-phase solvent is layered completely, suctions out the ethyl acetate phase on upper strata, repeats extraction five times, merges gained ethyl acetate
Phase extract, 55 DEG C are concentrated under reduced pressure, and are dried under vacuum to constant weight, obtain ethyl acetate phase extract and are designated as CAV-III.
The mtt assay of embodiment 7 detects that calyx extract CAV-I, CAV-II, CAV-III are to 3T3-L1 PECTORAL LIMB SKELETONs from generation to generation
The influence of propagation
The mouse 3T3-L1 PECTORAL LIMB SKELETONs of growth logarithmic phase are taken respectively, and 1000rpm centrifugation 5min, abandoning supernatant is used
It is 5 × 10 containing the DMEM culture mediums 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 is separately added into and contains CAV-I (embodiments
1 is obtained), CAV-II (embodiment 4 be obtained), DMEM culture mediums (the calyx extract from generation to generation of CAV-III (embodiment 4 is obtained)
The final concentration of CAV-I, CAV-II, CAV-III is respectively 50 μ g/mL, 100,200,400 and 800 μ g/mL).37 DEG C, 5%
CO224h, abandoning supernatant are cultivated in incubator, PBS adds DMEM basis cultures of the 200 μ L without serum twice, per hole
Base and 20 μ L tetrazolium bromides (MTT), continuation is incubated 4h in being put into incubator, abandons supernatant, and 150 μ L dimethyl sulfoxide (DMSO)s are added per hole
(DMSO), avoid light place uniformly shakes 10min in micro oscillator, and the OD values under 490nm wavelength are measured with ELIASA, calculates
Cell proliferation inhibition rate.
Test result is as shown in Figures 1 to 3.Fig. 1 be embodiment 1 prepare from generation to generation calyx extract CAV-I to 3T3-L1
PECTORAL LIMB SKELETON inhibited proliferation figure；Fig. 2 be the extract of the calyx from generation to generation CAV-II for preparing of embodiment 4 to 3T3-L1 before
Lipocyte proliferation inhibitory action figure；Fig. 3 be the extract of the calyx from generation to generation CAV-III for preparing of embodiment 4 to 3T3-L1 before
Lipocyte proliferation inhibitory action figure.
CAV-I (Fig. 1) does not show significant propagation to mouse 3T3-L1 PECTORAL LIMB SKELETONs in 50~800 μ g/mL scopes
Inhibitory action.And from generation to generation calyx extract CAV-II (Fig. 2), calyx extract CAV-III (Fig. 3) is in 50~800 μ g/mL from generation to generation
In the range of significant inhibited proliferation is shown to mouse 3T3-L1 PECTORAL LIMB SKELETONs.Illustrate that calyx is by classifying screen from generation to generation
Choosing, the active component with effect of weight reducing is enriched with.Consider, further carried out from calyx extract CAV-II from generation to generation
Next step is tested.
The oil red O staining method of the embodiment 8 detection shadow that calyx extract CAV-II breaks up to 3T3-L1 PECTORAL LIMB SKELETONs from generation to generation
Ring
(1) this experiment is divided into two groups, is designated as I groups and II groups.Fat is thin before taking the mouse 3T3-L1 of growth logarithmic phase respectively
Born of the same parents, 1000rpm centrifugation 5min, abandoning supernatant adjusts cell with containing the DMEM culture mediums 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 cell is reached contact inhibition, I groups and II components
10 μ g/mL insulin, 0.05mmol/L IBMX and 1 μm of complete culture solution of ol/L Jia Ru not contained, calyx extract from generation to generation is added
CAV-II (final concentration is respectively 6.25,12.5,25,50 and 100 μ g/mL), while setting control group, every kind for the treatment of sets 5 weights
It is multiple.After culture 2d, I groups change the complete culture solution for comprising only 10 μ g/mL insulin, and II groups are changed and contain the complete of 10 μ g/mL insulin
Full nutrient solution, adds calyx extract CAV-II (final concentration is respectively 6.25,12.5,25,50 and 100 μ g/mL) from generation to generation, culture
2~4d, I group use complete culture solution culture 4d, II group instead and use containing calyx extract CAV-II from generation to generation that (final concentration is respectively instead
6.25th, 12.5,25,50 and 100 μ g/mL) complete culture solution culture 4d, until differentiation terminate, 80% or so cell is
It is divided into the fat cell of maturation；
(2) will induce differentiation into it is ripe after cell carefully washed with PBS 3 times, consolidated with the 4% poly first ferment solution of 100 μ L
Determine 30min, then discard paraformaldehyde solution, washed with PBS 3 times, add 100 μ L oil reds after drying at room temperature per hole
O working solutions dye 30min at room temperature, dye liquor are removed after dyeing 30min, then add 70% ethanol rapid cleanup cell to reach decolouring
Purpose, plus PBS cell 3 times is subsequently adding 200 μ L isopropanols, in determining light absorption value under 490nm；
As shown in Figures 4 and 5, Fig. 4 is the extract of the calyx from generation to generation CAV-II for preparing of embodiment 4 to 3T3- to test result
L1 PECTORAL LIMB SKELETONs break up the action diagram of early stage；Fig. 5 is the extract of the calyx from generation to generation CAV-II for preparing of embodiment 4 to 3T3-
L1 PECTORAL LIMB SKELETONs break up the action diagram of overall process.
I groups (Fig. 4) and II groups (Fig. 5) can significantly inhibit the differentiation of 3T3-L1 PECTORAL LIMB SKELETONs, the fat product in cell
Tired to gradually decrease, fat drop diminishes, and II groups inhibition is more notable than I group inhibitions.
Above-described embodiment is the present invention preferably implementation method, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from Spirit Essence of the 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 extract of calyx from generation to generation, it is characterised in that：Comprise the following steps：
(1) sieving will be dry, pulverize by calyx from generation to generation, will obtain calyx meal from generation to generation；
(2) ethanol solution heating and refluxing extraction calyx meal from generation to generation is used, then extract solution is centrifuged, and by supernatant concentration,
Dry to constant weight, obtain calyx alcohol extracts from generation to generation；
(3) water dissolves will be distilled by calyx alcohol extracts from generation to generation, and will add petroleum ether to be extracted, repeat extracted several times, removal
Petroleum ether phase；Add chloroform to be extracted in the solution after removing petroleum ether phase, repeat extracted several times, chloroform phase is collected respectively
Solution with after taking-up chloroform phase, chloroform is mutually concentrated and is evaporated, and obtains chloroform phase extract；
(4) to adding ethyl acetate to be extracted in the solution taken out after chloroform phase, extracted several times, combined ethyl acetate are repeated
Phase, concentration is evaporated, and obtains ethyl acetate phase extract；The extract of calyx from generation to generation is calyx ethanol crude extract, chlorine from generation to generation
Imitative phase extract and/or ethyl acetate phase extract.
2. the preparation method of calyx extract from generation to generation according to claim 1, it is characterised in that：Ethanol described in step (2)
The volume fraction of solution is 70~90%.
3. the preparation method of calyx extract from generation to generation according to claim 1, it is characterised in that：Heated described in step (2)
The temperature of refluxing extraction is 80~100 DEG C；The heating and refluxing extraction number of times is 2~5 times, and the time extracted every time is 1~3h.
4. the preparation method of calyx extract from generation to generation according to claim 1, it is characterised in that：Ethanol described in step (2)
Solution is (10~30) mL with the volume mass ratio of calyx meal from generation to generation:1g；
Step (3) the calyx alcohol extracts from generation to generation and the mass volume ratio of distilled water are 1g：(10~20) mL.
5. the preparation method of calyx extract from generation to generation according to claim 1, it is characterised in that：It is centrifuged described in step (2)
Rotating speed be 3000~5000r/min；The time of the centrifugation is 8~12min；
The time extracted described in step (3) is all 30~60min；The time extracted described in step (4) is 30~60min.
6. the preparation method of calyx extract from generation to generation according to claim 1, it is characterised in that：In step (2), (3) and (4)
The mode of described concentration is concentrated under reduced pressure, and the temperature of the concentration is 40~60 DEG C；
Extraction times described in step (3) are all 3~5 times；Extraction times described in step (4) are all 3~5 times.
7. the preparation method of calyx extract from generation to generation according to claim 1, it is characterised in that：Dried described in step (2)
Temperature be 40~60 DEG C；Dry temperature described in step (1) is 40~60 DEG C；The drying time is 18~24h；It is described
It is 40~60 mesh sieves to pulverize and sieve.
8. a kind of extract of calyx from generation to generation is prepared by the preparation method described in any one of claim 1~7.
9. application of the calyx extract in slimming medicine is prepared from generation to generation according to claim 8.
10. a kind of slimming medicine, contains at least one described in claim 8 from generation to generation in calyx extract.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102357142A (en) * | 2007-08-07 | 2012-02-22 | 北京北大维信生物科技有限公司 | Use of water extract or ethanol extract of Chinese herbal medicine seville orange flower in preparation of drug for losing weight and reducing blood fat or drug for inhibiting lipase activity |
-
2016
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102357142A (en) * | 2007-08-07 | 2012-02-22 | 北京北大维信生物科技有限公司 | Use of water extract or ethanol extract of Chinese herbal medicine seville orange flower in preparation of drug for losing weight and reducing blood fat or drug for inhibiting lipase activity |
Non-Patent Citations (2)
| Title |
|---|
| 杨丽: "代代花化学成分的研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑第03期》 * |
| 郝云芳: "代代花总黄酮对3T3-L1细胞增殖活性的影响", 《现代食品科技》 * |
Cited By (1)
| 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 |
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