CN107595867A - A kind of method for preparing naringenin, hesperetin and its single glucoside mixture - Google Patents
A kind of method for preparing naringenin, hesperetin and its single glucoside mixture Download PDFInfo
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- CN107595867A CN107595867A CN201710735464.0A CN201710735464A CN107595867A CN 107595867 A CN107595867 A CN 107595867A CN 201710735464 A CN201710735464 A CN 201710735464A CN 107595867 A CN107595867 A CN 107595867A
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- hesperetin
- naringenin
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Abstract
The invention provides it is a kind of mainly contain naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides pharmaceutical composition, and it is raw material to utilize the medicinal material containing aurantiin, rue aurantiin, aurantiamarin, neohesperidin, its extract is hydrolyzed or extracted with hydrolyzing while can obtain this plant extracts.This product can be used for the development of food, health food and new Chinese medicine to produce.
Description
Technical field
The invention provides a kind of brand-new Chinese medicine (or plant) extract and preparation method thereof, and in particular to a kind of main
Containing naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides brand-new plant extracts and preparation method thereof, category
In Chinese medicine or researches on natural drugs field.
Background technology
Naringenin monoglycosides (Prunin), also known as eriodictyol-7- O -glucoside (Naringenin-7-O-
Glucoside), it is that rue aurantiin (Narirutin) or aurantiin (Naringin) hydrolysis are fallen a molecule rhamnose and obtained
Flavanone monoglycosides, white solid, 225 DEG C of fusing point, molecular formula is C21H22O10, molecular weight 434.39.Document shows shaddock ped
Plain monoglycosides have antibacterial, anti-oxidant, antiallergy, antiviral, reduction cholesterol, hypoglycemic, suppression platelet aggregation, promotion bone
Theca cell propagation, suppress the effect such as aldose reductase.
Hesperetin monoglycosides, also known as hesperetin -7-O- glucosides (hesperetin-7-O-glucoside), are orange peels
The flavanone monose that a molecule rhamnose obtains is fallen in glycosides (Hesperidin) or neohesperidin (Neohesperidin) hydrolysis
Glycosides, white solid, 206-207 DEG C of fusing point, molecular formula are C22H24O11, molecular weight 464.42.Hesperetin monoglycosides are thin to lung cancer
Born of the same parents, breast cancer, prostate cancer, melanoma, which have, suppresses growth.There is improvement for the osteoporosis of ovariectomized female rats
Effect, hesperetin monoglycosides can suppress the growth of helicobacter pylori in addition.
Naringenin (naringenin) is the aglycon of rue aurantiin (Narirutin) or aurantiin (Naringin), category
In flavanone, fusing point is 251 DEG C, and molecular formula is C15H12O5, molecular weight 272.25.Naringenin has anti-atherogenic hard
The bioactivity such as change, hypotensive, hypoglycemic, regulation blood fat, fat-reducing, suppression alpha-glucosidase activity, anti-oxidant, anti-inflammatory.Shaddock
Pi Su has antitumaous effect, energy to lung carcinoma cell, colon cancer cell, breast cancer cell, prostate gland cancer cell, melanoma cells
Enough prevent prostate gland cancer cell to be mutated, prevent or treat boar prostatoplasia diseases.
Hesperetin (hesperetin) is the glycosides of aurantiamarin (Hesperidin) or neohesperidin (Neohesperidin)
Member, belong to flavanone, fusing point is 227.5 DEG C, and molecular formula is C16H14O6, molecular weight 302.28.Hesperetin has to heart
There is protective effect, there is diastole effect to rat aortic smooth muscle, cardiac muscle cell apoptosis can be suppressed, it is anti-to reduce oxidative stress
, CBF should be increased.Hesperetin has Antianxiety Activity, and its effect is better than aurantiamarin.Hesperetin is by suppressing arachidonic acid
The thrombocytin secretion of induction, suppresses platelet aggregation.Hesperetin also has anti-oxidant, anti-allergic effects.
Naringenin, naringenin monoglycosides, hesperetin, the structural formula of hesperetin monoglycosides are as follows.
Document shows, the plant containing Pu Luning up to the present found or Chinese medicine have loosestrife (content is
0.003%), clinopodium grass, calamint, eucommia Bark male flower, the Flowers of Helichrysum bracteatum, MurrayatetrameraC.C.Huang；Contain hesperetin list
The plant of glucosides or Chinese medicine have the dried immature fruit of citron orange, Artemisia anomala, yellow chrysanthemum flower, peppermint, spearmint, ching-chieh, Lindley Butterflybush Herb, toddalia,
Bidensfrondosa、MurrayatetrameraC.C.Huang；Plant or Chinese medicine containing naringenin have the dried immature fruit of citron orange, and (content is
0-0.2%), Fructus Aurantii (content 0-0.05%), dendrobium candidum (content 0.02%-0.1%), the radix paeoniae rubrathe, (content is vine tea
0.01%), shaddock ped, the root of Chinese clematis, Chinese lobelia, typha angustifolia, clinopodium polycephalum, citrus medic L., FTUCTUS CITRI IMMATURI；Plant or Chinese medicine containing hesperetin
There is a dried immature fruit of citron orange (content 0-0.1%), (content is for yellow meal flower (content 0.013%), vine tea (content 0.01%), Chinese lobelia
0.008%), mountain breeze (content 0.003%), motherwort (content 0.002%), water chestnut skin, citrus chachiensis hortorum, FTUCTUS CITRI IMMATURI, flying dragon
Slap blood, Artemisia spharocephala.There are FTUCTUS CITRI IMMATURI, vine tea, Chinese lobelia containing hesperetin, the plant of naringenin or Chinese medicine simultaneously；Simultaneously containing orange
Pi Su, the plant of hesperetin monoglycosides or Chinese medicine have toddalia；Simultaneously containing Pu Luning, hesperetin monoglycosides plant or in
Medicine has MurrayatetrameraC.C.Huang.Simultaneously containing hesperetin monoglycosides, naringenin, the plant of hesperetin or Chinese medicine
There is the dried immature fruit of citron orange.Simultaneously containing naringenin, naringenin monoglycosides, hesperetin, the plant of four kinds of hesperetin monoglycosides or Chinese medicine there is not yet
Report.
Although in above-mentioned plant or Chinese medicine containing naringenin, naringenin monoglycosides, hesperetin, a kind of hesperetin monoglycosides
Or 2 kinds or 3 kinds, but their content is very low, it is difficult to obtain the extract using these compositions as main component, for researching and developing or
Produce corresponding food, health food and new Chinese medicine.
Up to now without the plant found simultaneously containing naringenin monoglycosides, hesperetin monoglycosides, naringenin and hesperetin
Thing, more without document report simultaneously containing naringenin monoglycosides, hesperetin monoglycosides, naringenin and hesperetin two kinds or three
Kind or four kinds of plant extracts and preparation method thereof.
The content of the invention
This 4 kinds of native compounds of naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin have diversified
Bioactivity.According to the multicomponent Mutiple Targets theory of traditional Chinese medicine, if we are it may be speculated that can obtain or prepare with shaddock ped
Two or three or four kinds in this 4 kinds of native compounds of plain monoglycosides, hesperetin monoglycosides, naringenin, hesperetin is main
If the Chinese medical extract or active component of composition, this extract may have the life more stronger than any one single compound
Thing activity, while it is also possible to show new bioactivity.
Exist in the Nature containing naringenin or naringenin monoglycosides or the plant of hesperetin or hesperetin monoglycosides, but its
Species is few and content is also very low.Therefore be difficult obtain with one or both of these four compositions or three kinds or four kinds for mainly into
The extract or active component divided.
Fortunately in nature exist largely containing rue aurantiin, aurantiin, aurantiamarin, neohesperidin plant or
Chinese medicine.By hydrolyzing these compositions, we can be obtained with naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin
In two or three or four kinds be main component extract or active component.
Based on this thinking, inventor completes this hair on the basis of detailed consulting literatures by substantial amounts of experimental study
It is bright.
Rue aurantiin, aurantiin, aurantiamarin, neohesperidin are widely present in rutaceae.Parts of generic medicinal plants trifoliate orange
Content is higher in reality, Fructus Aurantii, Exocarpium Citri Rubrum, Exocarpium Citri Grandis, rascal and dried orange peel.
In order to found from the dried immature fruit of citron orange, Fructus Aurantii, Exocarpium Citri Rubrum, Exocarpium Citri Grandis, rascal and dried orange peel above-mentioned four kinds of compositions i.e. rue aurantiin,
The higher medicinal material of aurantiin, aurantiamarin, neohesperidin content, the present invention have carried out assay to above-mentioned medicinal material first.Measure
As a result show, the medicinal material content difference of separate sources is larger.By taking the preferable medicinal material of quality of medicinal material as an example, rue aurantiin in dried orange peel
Content be about 1%, the content of aurantiamarin is typically 6% or so.The content of rue aurantiin is about 2% in rascal, new orange peel
The content of glycosides is about 1%, and aurantiamarin is typically 10% or so.Aurantiamarin and neohesperidin account for 1% in Exocarpium Citri Rubrum, and aurantiin contains
Amount is typically 5% or so.The content of aurantiin is about 4% in Exocarpium Citri Grandis, and aurantiamarin and neohesperidin account for 1%.Rue in Fructus Aurantii
Fragrant naringin content is about 2%, and naringin content general 6% or so, content of hesperidin is generally greater than 2%, and neohesperidin is about
4%.Rue naringin content is about 3% in the dried immature fruit of citron orange, and naringin content is typically 11% or so, and content of hesperidin is about 2%, newly
Aurantiamarin is generally greater than 11%.
It is an object of the invention to provide it is a kind of with naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin two
Kind or extract or active component that three kinds or four kinds are main component, and this extract or active component are to utilize to contain rue
What fragrant aurantiin or aurantiin or the medicinal material of aurantiamarin or neohesperidin were raw material to prepare.Specifically to contain rue aurantiin
Or aurantiin or the medicinal material of aurantiamarin or neohesperidin are raw material, by acid adding or alkali carries are added to take, it is simultaneously complete in extraction process
Into hydrolysis, naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin mixture are obtained, or first with above-mentioned medicinal material system
Standby extract, then extract is hydrolyzed and obtains naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin mixing
Thing.Result of study shows that first method is due to second method.
Concrete technical scheme of the present invention is as follows：
Method one
(1) medicinal material or plant containing rue aurantiin and/or aurantiin and aurantiamarin and/or neohesperidin simultaneously are taken,
Crush
(2) get it filled material or plant powder adds suitable quantity of water or organic solvent, adds a certain amount of acid, heating extraction, carries
Liquid is taken to filter while hot
(3) recycling design, obtain containing naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides plant extracts
(4) plant extracts for taking step (3) to obtain is extracted with ethyl acetate, combining extraction liquid, reclaims ethyl acetate, obtains
Naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides the content plant extracts high compared with (3).
(5) take step (3) or the plant extracts of (4) acquisition to carry out silica gel column chromatography, collect containing naringenin, naringenin
Monoglycosides, hesperetin, the component of hesperetin monoglycosides, merge, obtain naringenin, naringenin monoglycosides, hesperetin, hesperetin monose
The higher plant extracts of glycosides content.
Method two
(1) medicinal material or plant containing rue aurantiin and/or aurantiin and aurantiamarin and/or neohesperidin simultaneously are taken,
Crush
(2) get it filled material or plant powder adds suitable quantity of water or organic solvent is extracted, and extract water or organic solvent are molten
Solution, adds a certain amount of acid, and heating carries out sour water solution, recycling design, obtained containing naringenin, naringenin monoglycosides, hesperetin, orange
The plant extracts of skin element monoglycosides
(3) plant extracts for taking step (2) to obtain, is extracted with ethyl acetate, obtains naringenin, naringenin
Monoglycosides, hesperetin, hesperetin monoglycosides the content plant extracts high compared with (2).
(4) take step (2) or the plant extracts of (3) acquisition to carry out silica gel column chromatography, collect containing naringenin, naringenin
Monoglycosides, hesperetin, the component of hesperetin monoglycosides, merge, recycling design, obtain naringenin, naringenin monoglycosides, hesperetin,
The higher plant extracts of hesperetin monoglycosides content.
Described organic solvent is the methanol or ethanol that concentration of volume percent is 40%-100%.Described acid is hydrochloric acid
Or sulfuric acid or phosphoric acid or acetic acid, concentration 0.6mol/L-2mol/L.Described heating-up temperature is 60 DEG C -100 DEG C.Described carries
It is 1-3h to take the time.The extraction is 1-2 times with the dosage of ethyl acetate, and extraction times are 3-5 times.The medicinal material be the dried immature fruit of citron orange,
Fructus Aurantii, rascal, dried orange peel, Exocarpium Citri Rubrum.The ethanol of organic solvent 80%.Preferred acid is hydrochloric acid, concentration of hydrochloric acid 1.5mol/L.It is preferred that
Heating extraction and hydrolysis temperature are 75 DEG C, time 2.5h.
The present invention there is provided first it is a kind of available for food especially health food and new Chinese medicine research and development with production
New pharmaceutical composition, extract or active component.
Secondly, it is raw material system to have invented using containing rue aurantiin or the medicinal material of aurantiin or aurantiamarin or neohesperidin
The method of standby naringenin, hesperetin and its single glucoside mixture, has especially invented the side of extraction and hydrolysis while progress
Method, the method reduce intermediate link, save organic solvent usage amount relative to traditional method first extracted and hydrolyzed afterwards,
Low with cost, yield is high, takes short, and power consumption is small, the advantages of suitable industrialized mass production.
Applicant is found surprisingly that, any two kinds, three kinds of naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin
Or four kinds of combination has more preferable application effect relative to monomer.Especially in antianaphylactic purposes.
Embodiment
With reference to embodiment, the present invention will be further described.
Embodiment 1
Dried immature fruit of citron orange 10g is taken, is crushed, sieving, is dissolved in the ethanol of 200mL 85%, adding hydrochloric acid makes its concentration reach 1.6mol/L,
Stirred in the case where temperature is 75 DEG C of water bath conditions, hydrolyzed in extraction, react 2.5h；Ethanol is reclaimed, obtains brown solid 4.8g.It is logical
HPLC measure is crossed, the content of four compounds is respectively naringenin monoglycosides 6.3% in this mixture, hesperetin monoglycosides
4.1%, naringenin 3.9%, hesperetin 2.4%.
Embodiment 2
Dried immature fruit of citron orange 10g is taken, is crushed, sieving, is dissolved in the ethanol of 200mL 85%, adding hydrochloric acid makes its concentration reach 1.6mol/L,
Stirred in the case where temperature is 75 DEG C of water bath conditions, hydrolyzed in extraction, react 2.5h；Ethanol is reclaimed, is extracted with ethyl acetate
(200mL × 4), ethyl acetate is recovered under reduced pressure and obtains yellow solid 3.49g, is determined by HPLC, four chemical combination in this mixture
The content of thing is respectively naringenin monoglycosides 13%, hesperetin monoglycosides 11%, naringenin 7%, hesperetin 5%.Pu Luning's
Conversion ratio 46.0%；The conversion ratio 39.0% of hesperetin monoglycosides；The conversion ratio 40.1% of naringenin；The conversion ratio of hesperetin
26.3%.
Embodiment 3
Dried immature fruit of citron orange 10g is taken, is crushed, sieving, is dissolved in the ethanol of 200mL 85%, adding hydrochloric acid makes its concentration reach 1.6mol/L,
Stirred in the case where temperature is 75 DEG C of water bath conditions, hydrolyzed in extraction, react 2.5h；Ethanol is reclaimed, is extracted with ethyl acetate
(200mL × 4), ethyl acetate is recovered under reduced pressure and obtains yellow solid 3.40g, take this mixture to think that mobile phase carries out silica gel column layer
Analysis, collect containing naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides component, merge, solvent is recovered under reduced pressure, obtains
Obtain 2.4 grams of content highest plant extracts.Determined by HPLC, the content of four compounds is respectively shaddock ped in this mixture
Plain monoglycosides 18.0%, hesperetin monoglycosides 15.4%, naringenin 10.2%, hesperetin 7.6%.
Embodiment 4
Fructus Aurantii 50g is taken, is crushed, sieving, is dissolved in the methanol of 1L 80%, adding sulfuric acid makes its concentration reach 1.2mol/L, in temperature
Spend to be stirred under 78 DEG C of water bath conditions, hydrolyzed in extraction, react 3h；Methanol is reclaimed, (400mL × 5) are extracted with ethyl acetate,
Ethyl acetate is recovered under reduced pressure and obtains yellow solid 15.04g, is determined by HPLC, the content of four compounds point in this mixture
Not Wei naringenin monoglycosides 11.9%, hesperetin monoglycosides 10.1%, naringenin 6.2%, hesperetin 5.4%.
Embodiment 5
Rascal 100g is taken, is crushed, sieving, is dissolved in the n-butanols of 3L 70%, adding phosphoric acid makes its concentration reach 1.8mol/L,
Stirred in the case where temperature is 70 DEG C of water bath conditions, hydrolyzed in extraction, react 2h；N-butanol is reclaimed, be extracted with ethyl acetate (1L ×
4) ethyl acetate, is recovered under reduced pressure and obtains yellow solid 24.91g, is determined by HPLC, the content of four compounds in this mixture
Respectively naringenin monoglycosides 2.4%, hesperetin monoglycosides 12.7%, naringenin 1.8%, hesperetin 8.7%.
Embodiment 6
Dried orange peel 500g is taken, is crushed, sieving, is dissolved in the ethanol of 12L 75%, adding hydrochloric acid makes its concentration reach 1.5mol/L,
Stirred in the case where temperature is 72 DEG C of water bath conditions, hydrolyzed in extraction, react 3h；Ethanol is reclaimed, be extracted with ethyl acetate (2L ×
5) ethyl acetate, is recovered under reduced pressure and obtains yellow solid 172.1g, is determined by HPLC, the content of four compounds in this mixture
Respectively naringenin monoglycosides 10.8%, hesperetin monoglycosides 10.1%, naringenin 8.6%, hesperetin 6.1%.
Embodiment 7
Exocarpium Citri Grandis 200g is taken, is crushed, sieving, is dissolved in the ethanol of 6L 90%, adding hydrochloric acid makes its concentration reach 1.0mol/L,
Hydrolyzed when stirring under temperature is 80 DEG C of water bath conditions is in extraction, react 3h；Ethanol is reclaimed, be extracted with ethyl acetate (800mL ×
5) ethyl acetate, is recovered under reduced pressure and obtains yellow solid 30.1g, is determined by HPLC, the content of four compounds in this mixture
Respectively naringenin monoglycosides 3.8%, hesperetin monoglycosides 2.4%, naringenin 4.3%, hesperetin 1.9%.
Embodiment 8
Exocarpium Citri Rubrum 200g is taken, is crushed, sieving, is dissolved in the ethanol of 6L 90%, adding hydrochloric acid makes its concentration reach 1.0mol/L,
Temperature is that stirring hydrolyzes in extraction under 80 DEG C of water bath conditions, reacts 3h；Ethanol is reclaimed, be extracted with ethyl acetate (800mL ×
5) ethyl acetate, is recovered under reduced pressure and obtains yellow solid 30.1g, is determined by HPLC, the content of four compounds in this mixture
Respectively naringenin monoglycosides 3.8%, hesperetin monoglycosides 2.4%, naringenin 4.3%, hesperetin 1.9%.
Influence of the test example 1 to the mast cell degranulation of monoclonal antibody antigen induction
1.1 experimental section
1.1.1 cell
Cell used in this experiment is RBL-2H3 cells, a subbreed in rat basophilic leukemia cell,
Buy from Shanghai Life Sciences Research Institute, Chinese Academy Of Sciences' cell resource center.
1.1.2 laboratory apparatus
The laboratory apparatus of table 1.1
1.1.3 experiment reagent and preparation
The experiment reagent of table 1.2
The experiment reagent of table 1.3 and its configuration
1.2 experimental method
1.2.1RBL-2H3 cell culture
Cryopreservation tube is taken out in liquid nitrogen container, and is transferred quickly in 37 DEG C of thermostat water baths, constantly shakes, makes it 1
Melting in minute turns into liquid.Cell is aseptically taken out afterwards, and centrifuge tube is transferred to after adding 9mL complete mediums
In, centrifuged 6 minutes under the conditions of 1000r, 25 DEG C.After the cell liquid supernatant discarding in centrifuge tube, after adding appropriate culture medium
It is inoculated in culture dish, puts 37 DEG C, cultivated in 5%CO2 incubators.After 24h, it is not adherent to remove to change fresh culture medium
Cell, passed on when cell is about paved with the area at ware bottom 80% or so.In passage, by culture dish from CO2 incubators
After taking-up, discarded culture medium is first drawn with liquid-transfering gun, the trypsase for adding 2mL afterwards is digested, and the time is about 1-2 points
Clock, when cell comes off from ware bottom, piping and druming auxiliary suitably can be carried out with liquid-transfering gun, add complete medium afterwards and terminate digestion,
Cell liquid is collected into centrifuge tube, after normal temperature centrifuges 5 minutes under the conditions of the 1000rpm, supernatant is abandoned and adds fresh culture and
Row passage, passage ratio are 1:2, passage in two days is once.
1.2.2MTT influence of the method detection medicine to RBL-2H3 cell-proliferation activities
From from incubator and the cell in exponential phase is picked out, is digested using pancreatin, is prepared
As cell suspension.Cell is counted with blood counting chamber, and adjusted cell number to 2 with DMEM complete mediums ×
104/mL.It is inoculated on 96 orifice plates, experiment sets blank group (being not added with cell), normal group (a refinement born of the same parents is not added with medicine), used
Medicine group (the μ of concentration gradient 3.125,6.25,12.5,25,50,75,100 of the various concentrations of DMED basal mediums dilution
Mol/L), per the μ L of hole 100, every group sets 6 multiple holes, when adding cell, is mixed after often adding one group, the sterile PBS of surrounding
Fluid-tight, it is placed in incubator and cultivates 24h, afterwards supernatant discarding, adds the required medicine of DMEM basal mediums and various concentrations,
Normal group cell is replaced with the DMSO of same concentrations.Continuing to take out cell after cultivating 20h, observation cell is in normal condition,
Lucifuge adds 15 μ LMTT per hole afterwards, and after being put into incubator culture 4h, lucifuge is put into centrifuge and centrifuged, and arrange parameter is to turn
Son 3,1500r, 5min, afterwards supernatant discarding.150 μ LDMSO are added per hole, 5min is shaken on low speed oscillator, fills crystallization
Divide dissolving, the OD values in each hole are detected under 570nm.
Calculation formula：
1.2.3 naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin monomer are to antigen induction RBL-2H3 cells
Discharge β-Hex influence
1.2.3.1 experiment packet
Cell is divided into normal group (N groups)：DNP-BSA (-), drug solution (-)；Model group (C groups)：DNP-BSA (+),
Drug solution (-)；Administration group (T groups)：DNP-BSA (+), drug solution (+)；Drug-negative control group (B groups)：DNP-BSA
(-), drug solution (+)；Naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin monomer are final concentration of：12.5μmol/
L
1.2.3.2 operating procedure
The cell in exponential phase taken out from incubator, after being digested, preparation turns into cell suspension,
Afterwards using blood counting chamber carry out cell count, afterwards with DMEM complete mediums by cell number adjust to 3.5 × 104/
mL.It is inoculated on 24 orifice plates, per the μ L of hole 400, when adding cell, is mixed after often adding one group.It is placed in incubator and cultivates
24h, afterwards in addition to normal group and drug-negative control group, model group and administration group are separately added into the anti-DNP IgE works of 10 μ L per hole
Make liquid, to final concentration of 0.125 μ g/mL.Drug concentration needed for dilution simultaneously, -4 DEG C stand-by.
After cultivating 12h in incubator, administration group adds each drug concentration, continues to cultivate 2.5h, abandons supernatant afterwards, uses
Tyrode ' s buffer buffer solutions wash cell twice, per the μ L of hole 500, add 200uLTyrode ' s buffer buffer solutions afterwards,
Wherein administration group adds 10 μ LDNP-BSA again with model group, to final concentration of 0.125 μ g/mL, blank group and negative control group
Add Tyrode ' the s buffer buffer solutions of equivalent.37 DEG C of incubation 1h.Ice bath 10min afterwards, stop exciting, in refrigerated centrifuge
Centrifuged in machine, parameter 14800r, 10min, 4 DEG C, collect each group supernatant afterwards, take 50 μ L to add into 96 orifice plates per hole, then
Add 50 μ L nitrite ions, 37 DEG C of incubation 1.5h.200uL terminate liquid stopped reactions are added, OD values are detected under 405nm.Experiment is public
Formula
Wherein：TOD is administration group OD values；BOD is drug-negative control group group OD values；COD is model group OD values；NOD is
Normal group OD values.
Claims (10)
- A kind of 1. pharmaceutical composition, it is characterised in that：Containing in naringenin monoglycosides, hesperetin monoglycosides, naringenin, hesperetin Any two kinds, three kinds or four kinds.
- 2. pharmaceutical composition as claimed in claim 1, it is characterised in that：Preparation method is as follows：(1) medicinal material or plant containing rue aurantiin and/or aurantiin and aurantiamarin and/or neohesperidin simultaneously are taken, is crushed(2) get it filled material or plant powder adds suitable quantity of water or organic solvent is extracted, and extract water or organic solvent dissolving, adds Enter a certain amount of acid, heating carries out sour water solution, recycling design, obtained containing naringenin, naringenin monoglycosides, hesperetin, hesperetin The plant extracts of monoglycosides(3) take step (2) obtain plant extracts, be extracted with ethyl acetate, obtain naringenin, naringenin monoglycosides, hesperetin, The hesperetin monoglycosides content plant extracts high compared with (2).(4) take step (2) or the plant extracts of (3) acquisition to carry out silica gel column chromatography, collect containing naringenin, naringenin monose Glycosides, hesperetin, the component of hesperetin monoglycosides, merge, recycling design, produce containing naringenin, naringenin monoglycosides, orange peel Element, the plant extracts of hesperetin monoglycosides；Or(1) medicinal material or plant containing rue aurantiin and/or aurantiin and aurantiamarin and/or neohesperidin simultaneously are taken, is crushed(2) get it filled material or plant powder adds suitable quantity of water or organic solvent, adds a certain amount of acid, heating extraction, extract solution Filter while hot(3) recycling design, obtain containing naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides plant extracts(4) plant extracts for taking step (3) to obtain is extracted with ethyl acetate, combining extraction liquid, reclaims ethyl acetate, obtains shaddock ped Element, naringenin monoglycosides, hesperetin, hesperetin monoglycosides the content plant extracts high compared with (3).(5) take step (3) or the plant extracts of (4) acquisition to carry out silica gel column chromatography, collect containing naringenin, naringenin monose Glycosides, hesperetin, the component of hesperetin monoglycosides, merge, obtain naringenin, naringenin monoglycosides, hesperetin, hesperetin monoglycosides and contain The higher plant extracts of amount.
- 3. want composition as claimed in claim 2, it is characterised in that：Described medicinal material or plant is the dried immature fruit of citron orange or Fructus Aurantii or tangerine Red or Exocarpium Citri Grandis or rascal or dried orange peel.
- 4. want composition as claimed in claim 2, it is characterised in that：Organic solvent is that concentration of volume percent is 40%- 100% methanol or ethanol.
- 5. as claimed in claim wants composition, it is characterised in that：Described acid is hydrochloric acid or sulfuric acid or phosphoric acid or acetic acid, dense Spend for 0.6mol/L-2mol/L.
- 6. as claimed in claim wants composition, it is characterised in that described heating-up temperature is 60 DEG C -100 DEG C.
- 7. as claimed in claim wants composition, it is characterised in that described extraction time is 1-3h.
- 8. as claimed in claim wants composition, it is characterised in that extraction is 1-2 times with the dosage of ethyl acetate, extraction times For 3-5 times.
- 9. as claimed in claim wants composition, it is characterised in that medicinal material is the dried immature fruit of citron orange, and the ethanol of organic solvent 80%, acid is salt Acid, concentration of hydrochloric acid 1.5mol/L, heating extraction and hydrolysis temperature are 75 DEG C, time 2.5h.
- 10. the pharmaceutical composition described in claim 1 is preparing the purposes in treating Claritin.
Priority Applications (1)
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| CN113577158A (en) * | 2021-08-02 | 2021-11-02 | 杭州医学院 | Thoroughfare bitter orange effective component group for treating acute lung injury and preparation method and application thereof |
| CN113577088A (en) * | 2021-08-02 | 2021-11-02 | 杭州医学院 | Ququ fructus aurantii effective component group for treating viral pneumonia and preparation method and application thereof |
| CN115400138A (en) * | 2022-09-29 | 2022-11-29 | 河南科技大学 | Method for solubilizing hesperetin-7-O-glucoside and solubilizing compound thereof |
| CN115420815A (en) * | 2022-07-14 | 2022-12-02 | 神威药业集团有限公司 | Method for measuring contents of various active ingredients in powder for improving abnormal work and application thereof |
| CN115607538A (en) * | 2022-09-26 | 2023-01-17 | 浙江工商大学 | Application of hesperetin in preparation of medicine for inhibiting and/or treating wheat allergy |
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| CN116474025A (en) * | 2023-05-31 | 2023-07-25 | 中山大学附属第八医院（深圳福田） | Application of exocarpium Citri Grandis effective component in preparing medicine for preventing and treating cardiovascular diseases |
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| CN109517017A (en) * | 2018-12-05 | 2019-03-26 | 北京中医药大学 | Flavones reference extract and its preparation method and application |
| CN109932506A (en) * | 2019-03-27 | 2019-06-25 | 吉林大学 | Rue aurantiin is promoting and is maintaining the application in oral tolerance as adjuvant |
| CN113577158A (en) * | 2021-08-02 | 2021-11-02 | 杭州医学院 | Thoroughfare bitter orange effective component group for treating acute lung injury and preparation method and application thereof |
| CN113577088A (en) * | 2021-08-02 | 2021-11-02 | 杭州医学院 | Ququ fructus aurantii effective component group for treating viral pneumonia and preparation method and application thereof |
| CN113577088B (en) * | 2021-08-02 | 2022-08-26 | 杭州医学院 | Ququ fructus aurantii effective component group for treating viral pneumonia and preparation method and application thereof |
| CN115420815B (en) * | 2022-07-14 | 2024-03-26 | 神威药业集团有限公司 | Method for measuring content of various effective components in powder and application thereof |
| CN115420815A (en) * | 2022-07-14 | 2022-12-02 | 神威药业集团有限公司 | Method for measuring contents of various active ingredients in powder for improving abnormal work and application thereof |
| CN115607538A (en) * | 2022-09-26 | 2023-01-17 | 浙江工商大学 | Application of hesperetin in preparation of medicine for inhibiting and/or treating wheat allergy |
| CN115400138A (en) * | 2022-09-29 | 2022-11-29 | 河南科技大学 | Method for solubilizing hesperetin-7-O-glucoside and solubilizing compound thereof |
| CN115400138B (en) * | 2022-09-29 | 2023-09-19 | 河南科技大学 | Method for solubilising hesperetin-7-O-glucoside and solubilised complexes thereof |
| CN115645426B (en) * | 2022-10-27 | 2024-01-30 | 五邑大学 | Tyrosinase inhibitor and application thereof in preparation of drugs for treating melanoma |
| CN115645426A (en) * | 2022-10-27 | 2023-01-31 | 五邑大学 | Tyrosinase inhibitor and application thereof in preparation of drugs for treating melanoma |
| CN116474025A (en) * | 2023-05-31 | 2023-07-25 | 中山大学附属第八医院（深圳福田） | Application of exocarpium Citri Grandis effective component in preparing medicine for preventing and treating cardiovascular diseases |
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