CN105287709B - A kind of Hickory Leaves extractive total flavone as well as preparation method and application thereof with anti-trioxypurine effect - Google Patents
A kind of Hickory Leaves extractive total flavone as well as preparation method and application thereof with anti-trioxypurine effect Download PDFInfo
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Abstract
The invention discloses a kind of Hickory Leaves extractive total flavone as well as preparation method and application thereof with anti-trioxypurine effect, the Hickory Leaves extractive total flavone contain pinostrobin, pinostrobin chalcone, Chrysin and the bright gross mass percentage of red bean bandit 50% or more.The present invention is to rely on Hickory Leaves, prepares the general flavone in Hickory Leaves, develops its effect, expands the new medicine resource of Hickory Leaves.The Hickory Leaves extractive total flavone can significantly reduce the serum uric acid level of rat model, can be used for preventing and treating drug and health product that hyperuricemia is major function.
Description
Technical field
The present invention relates to pharmaceutical technology field, in particular to a kind of Hickory Leaves extract with anti-trioxypurine effect is always yellow
Ketone as well as preparation method and application thereof.
Background technique
Uric acid is the metabolite of purine in endogenous purine and diet.In the purine metabolism of human body, ATP etc. passes through one
The metabolic response of series, produces hypoxanthine and xanthine, and the two generates uric acid under xanthine oxidase effect.When purine generation
When thanking to disorder, serum uric acid level is excessively high, leads to the generation of hyperuricemia (Hyperuricemia), then uric acid is caused to be saltoutd
Joint is crystallized and be deposited in out, gout is caused to be broken out, and is often involved kidney and is caused arteriosclerotic kidney and uric acid kidney stones, and
And it may cause Metabolic syndrome and seek peace relevant cardiovascular disease.In recent years, the disease incidence of hyperuricemia sharply increases, and has surpassed
10% is crossed, one of big chronic disease of 21 century 20 is classified as by the United Nations.
Prevention and treatment hyperuricemia class drug is broadly divided into uric acid and generates depressant (such as Allopurinol) and uricosuric excretion at present
Two kinds of medicine (such as Benzbromarone), although these drugs have apparent anti-trioxypurine to act on, side effect is also larger, such as fash, hair
Heat, hepatic renal dysfunction etc., thus the range limited its application.Therefore, developmental research drug effect is clear and the novel of safety resists
Antihyperuricemic disease drug is very necessary.According to the literature, inhibit in natural plant crude drugs source containing a large amount of xanthine oxidases
The ingredient of agent and uricosuric excretion, and small toxicity, it is highly-safe, there is preferable research and development prospect.
Chinese walnut (Carya cathayensis Sarg.) is Juglandaceae hickory plant, Hickory Leaves frequently as
Folk tradition Chinese herbal medicine, for the purposes such as clearing heat and detoxicating, existing research shows that Hickory Leaves extract contains a large amount of anti-oxidant work
Property substance, predominantly Flavonoid substances, have apparent anti-aging, antitumor, anti-angiogenesis and other effects.Currently, there is not yet
To the relevant report using Hickory Leaves extractive total flavone treatment hyperuricemia.Inventors discovered through research that hickory nut
Leaf extract pinostrobin can reduce rat model serum uric acid level, can be used for preventing and treating the drug that hyperuricemia is major function
And health food.
Summary of the invention
The purpose of the present invention is to provide a kind of Hickory Leaves extractive total flavones with anti-trioxypurine effect, provide simultaneously
Preparation method and the usage.
The technical solution adopted by the present invention to solve the technical problems is:
A kind of Hickory Leaves extractive total flavone with anti-trioxypurine effect, contains pinostrobin, pinostrobin chalcone, white poplar
Plain and bright red bean bandit gross mass percentage is 50% or more.
A kind of preparation method of the Hickory Leaves extractive total flavone with anti-trioxypurine effect, comprising the following steps:
(1) 35-65 DEG C of oven drying of Hickory Leaves is taken, 40-80 mesh is crushed to, is put into round-bottomed flask, volume hundred is added
Dividing concentration is 70-95% ethanol water, is extracted 20-100 minutes in 80-110 DEG C of normal pressure, repeats to extract 1-3 times;Then with filter
Paper filtering, merging filtrate;
(2) it is 45-65% that filtrate is concentrated under reduced pressure into filtrate concentration of alcohol under 300 ± 100pa vacuum degree, in centrifuging and taking
This supernatant is crossed SD200 large pore resin absorption column (filling SD200 aromatic series macroporous absorbent resin) by clear liquid, and loading speed is
1BV/h-8BV/h, loading volume 4-8BV;Under 20-35 DEG C, normal pressure, with 1BV/h-8BV/h flow velocity, with distillation water elution tree
Rouge, elution volume 5-10BV;It is 10- with concentration expressed in percentage by volume with 1BV/h-8BV/h flow velocity under 20-35 DEG C, normal pressure
30% ethanol aqueous wash deresination, elution volume 8-12BV;Under 20-35 DEG C, normal pressure, with 1BV/h-8BV/h flow velocity,
The ethanol aqueous wash deresination for being 40-90% with concentration expressed in percentage by volume, elution volume 8-12BV;Collect 40-90% ethyl alcohol
Eluent, concentration recycle ethyl alcohol, are dried under reduced pressure, obtain Hickory Leaves extractive total flavone, wherein pinostrobin, pinostrobin Cha Er
Ketone, Chrysin and the bright total content of red bean bandit are 50% or more.
Preferably, 70-95% ethanol water dosage is that every g Hickory Leaves crushed material adds 10mL in step (1).
Preferably, parameter of noncentricity in step (2) are as follows: 2000-4000 rpms, be centrifuged 20 minutes.With anti-trioxypurine
The Hickory Leaves extractive total flavone of effect is preparing the purposes in antihyperuricemic disease drug.
Above-mentioned Hickory Leaves extractive total flavone can with the auxiliary material approved on medicament or food, using Modern preparations skill
Capsule, tablet, powder, oral solution, soft capsule, pill, gel based on Hickory Leaves extractive total flavone is made in art
The various dosage forms such as agent, sustained release preparation.
The beneficial effects of the present invention are: the present invention is to rely on Hickory Leaves, the general flavone in Hickory Leaves is prepared, is developed
Its effect expands the new medicine resource of Hickory Leaves.The experiment of inventor shows: the Hickory Leaves extractive total flavone can be shown
The serum uric acid level for reducing rat model is write, can be used for preventing and treating drug and health product that hyperuricemia is major function.
Specific embodiment
Below by specific embodiment, technical scheme of the present invention will be further explained in detail.
In the present invention, if not refering in particular to, used raw material and equipment etc. are commercially available or commonly used in the art.
Method in following embodiments is unless otherwise instructed the conventional method of this field.
Embodiment
A kind of preparation method of the Hickory Leaves extractive total flavone with anti-trioxypurine effect, comprising the following steps: (1) take
35-65 DEG C of oven drying of Hickory Leaves, is crushed to 40-80 mesh, is put into round-bottomed flask, and addition concentration expressed in percentage by volume is 70-
95% ethanol water (every g Hickory Leaves crushed material adds 10mL) extracts 20-100 minutes in 80-110 DEG C of normal pressure, repeats to extract
1-3 times;Then it is filtered with filter paper, merging filtrate;
(2) filtrate is concentrated under reduced pressure under 300 ± 100pa vacuum degree, makes concentration of alcohol 45-65% in filtrate, centrifugation
(2000-4000 rpms, be centrifuged 20 minutes) takes supernatant, this supernatant is crossed SD200 large pore resin absorption column, loading speed
Degree is 1BV/h-8BV/h, loading volume 4-8BV;Under 20-35 DEG C, normal pressure, with 1BV/h-8BV/h flow velocity, washed with distillation
Deresination, elution volume 5-10BV;Under 20-35 DEG C, normal pressure, with 1BV/h-8BV/h flow velocity, it is with concentration expressed in percentage by volume
The ethanol aqueous wash deresination of 10-30%, elution volume 8-12BV;Under 20-35 DEG C, normal pressure, flowed with 1BV/h-8BV/h
Speed, the ethanol aqueous wash deresination for being 40-90% with concentration expressed in percentage by volume, elution volume 8-12BV;Collect 40-90% second
Alcohol eluen, concentration recycle ethyl alcohol, are dried under reduced pressure, obtain Hickory Leaves extractive total flavone, wherein pinostrobin, pinostrobin are looked into
Your ketone, Chrysin and the bright total content of red bean bandit are 50% or more.
Specific embodiment 1
After Hickory Leaves are dry, it is mechanically pulverized, weighs Hickory Leaves powder 200g, add the volume basis of 10 times of amounts (g:mL)
Than the ethanol solution for 95%, extract 2 times, each 30min, merging filtrate is concentrated under reduced pressure under 300 ± 100pa vacuum degree, makes
Concentration of alcohol is 55% in filtrate, 3000 rpms, is centrifuged 20 minutes, takes supernatant.Weigh SD200 aromatic series macroporous absorption tree
Rouge (Zhejiang Province Zhengguan Industry Co., Ltd) 4.0g is put into chromatographic column (15mm × 350mm i.d), with the flow velocity of 2BV/h
Carry out loading, loading volume 6.5BV.10BV water elution is first used, then is washed with the ethanol solution that 10BV percent by volume is 20%
It is de-, it is finally eluted with the ethanol solution that 10BV percent by volume is 70%, elution speed 4BV/h collects 70% ethanol elution
Liquid, decompression rotary evaporated to dryness is dry, obtains Hickory Leaves extractive total flavone, wherein pinostrobin, pinostrobin chalcone, Chrysin
And the bright total content of red bean bandit is 55.3% or more.
Specific embodiment 2
It after Hickory Leaves are dry, are mechanically pulverized, weigh Hickory Leaves powder 400g, adding 10 times of amount percents by volume is 95%
Ethanol solution, extract 2 times, each 30min, merging filtrate is concentrated under reduced pressure under 300 ± 100pa vacuum degree, makes second in filtrate
Determining alcohol is 55%, 3000 rpms, is centrifuged 20 minutes, takes supernatant.SD200 macroporous absorbent resin 200.0g is weighed, is put into
In chromatographic column (15mm × 350mm i.d), loading, loading volume 6.5BV are carried out with the flow velocity of 2BV/h.First washed with 10BV
It is de-, then the ethanol solution elution for being 20% with 10BV percent by volume, it is finally molten for 70% ethyl alcohol with 10BV percent by volume
Liquid elution, elution speed 4BV/h collect 70% ethanol eluate, and decompression rotary evaporated to dryness is dry, obtain Hickory Leaves and extract
Object general flavone, wherein the bright total content of pinostrobin, pinostrobin chalcone, Chrysin, red bean bandit is 52.2% or more.
The bright total content measuring method of pinostrobin, pinostrobin chalcone, Chrysin, red bean bandit is referring to the efficient liquid of the such as Shen Yong
Phase chromatography measures 5 flavone aglycone content [J] Pharmaceutical Analysis magazines in Hickory Leaves, 05 phase in 2013.
Effect test
Hickory Leaves extractive total flavone finds the hyperuricemia model rat prepared to fat emulsion through long term administration
Serum uric acid level have reduce effect significantly.
1 experimental material and instrument
1.1 materials and instrument
1.1 animal SD rats, male, weight (200-500g), 60.Quality certification number: scxk (Zhejiang) 20080033.
1.2 drugs are prepared
1.2.1 test drug
Hickory Leaves extractive total flavone, being configured to concentration with 0.3%CMC-Na is 200mg/kg and 100.0mg/kg.
1.2.2 positive drug allopurinol tablet, specification 100mg × 200 piece, Jiangsu side force pharmaceutical factory Co., Ltd, 10mg/
Kg and benzbromarone tablets, specification 50mg × 10 piece, the German pharmaceutical factory He Man great, 5mg/kg.
1.3 instrument
Electronic balance CP114 (Shanghai Ao Haosi Instrument Ltd.);Electric jacket (the limited public affairs of the bold and unconstrained laboratory apparatus of Nantong benefit
Department);GG17 Rotary Evaporators (Shensheng Science & Tech. Co., Ltd., Shanghai);SZ-93A dual pure water distiller (Shanghai Asia Rong Sheng automatically
Change instrument plant);Refrigerator (Qingdao HaiEr Co., Ltd);Dionex high performance liquid chromatography (Dai An company, the U.S.);TBA-
AOFR (ACCUTE) automatic clinical chemistry analyzer (Japanese wide medical Co., Ltd., Toshiba three).
2 experimental methods
2.1 administration
60 rats are randomly divided into normal group according to weight, model control group, positive controls, Hickory Leaves extract
General flavone high dose group (200mg/kg), Hickory Leaves extractive total flavone low dose group (100mg/kg), normal group give it is pure
Change water, model group gives fat emulsion.Tap water is given in gastric infusion, model group and normal stock stomach-filling, and administration group gives correspondence
Drug, administration capacity are 1mL/100g weight.
2.2 observation index
Successive administration 8 weeks, take hematometry rat blood serum uric acid (UA) horizontal every 2 weeks.
2.3 statistics and analysis
Measurement data data result withIt indicates, group difference uses t-test.
3 experimental results
Normal group, model group and each administration group serum uric acid situation of change during 3.1 administrations
Compared with normal group, model group rats serum uric acid is significantly raised (P < 0.01);Compared with model group, it is administered 4 weeks
The high low dose group of Hickory Leaves extractive total flavone can be substantially reduced model group rats UA level (P < 0.05), and anti-trioxypurine effect
It is steady to continue.It the results are shown in Table 1.
The variation (X ± S, n=10) of 1 successive administration rat UA level of table
Note: compared with normal group, P < 0.01 * P < 0.05, * *;Compared with model control group, #P < 0.05, ##P < 0.01.
Above-mentioned embodiment is only a preferred solution of the present invention, not the present invention is made in any form
Limitation, there are also other variations and modifications on the premise of not exceeding the technical scheme recorded in the claims.
Claims (5)
1. a kind of Hickory Leaves extractive total flavone with anti-trioxypurine effect, it is characterised in that: looked into containing pinostrobin, pinostrobin
The gross mass percentage of your ketone, Chrysin and cardamomin is 50% or more;
The preparation of the Hickory Leaves extractive total flavone with anti-trioxypurine effect the following steps are included:
(1) 35-65 DEG C of oven drying of Hickory Leaves is taken, 40-80 mesh is crushed to, is put into round-bottomed flask, it is dense that volume basis is added
Degree is 70-95% ethanol water, is extracted 20-100 minutes in 80-110 DEG C of normal pressure, repeats to extract 1-3 times;Then with filter paper mistake
Filter, merging filtrate;
(2) it is 45-65%, centrifuging and taking supernatant that filtrate is concentrated under reduced pressure into concentration of alcohol in filtrate under 300 ± 100pa vacuum degree
This supernatant is crossed SD200 large pore resin absorption column by liquid, and loading speed is 1BV/h-8BV/h, loading volume 4-8BV;?
20-35 DEG C, under normal pressure, with 1BV/h-8BV/h flow velocity, with distillation water elution resin, elution volume 5-10BV;At 20-35 DEG C,
Under normal pressure, with 1BV/h-8BV/h flow velocity, the ethanol aqueous wash deresination for being 10-30% with concentration expressed in percentage by volume, elution volume
For 8-12BV;Under 20-35 DEG C, normal pressure, with 1BV/h-8BV/h flow velocity, the ethyl alcohol for being 40-90% with concentration expressed in percentage by volume is water-soluble
Liquid elutes resin, elution volume 8-12BV;40-90% ethanol eluate is collected, concentration recycles ethyl alcohol, is dried under reduced pressure, obtains
Hickory Leaves extractive total flavone, wherein the total content of pinostrobin, pinostrobin chalcone, Chrysin and cardamomin 50% with
On.
2. a kind of preparation method of the Hickory Leaves extractive total flavone with anti-trioxypurine effect, which is characterized in that including following
Step:
(1) 35-65 DEG C of oven drying of Hickory Leaves is taken, 40-80 mesh is crushed to, is put into round-bottomed flask, it is dense that volume basis is added
Degree is 70-95% ethanol water, is extracted 20-100 minutes in 80-110 DEG C of normal pressure, repeats to extract 1-3 times;Then with filter paper mistake
Filter, merging filtrate;
(2) it is 45-65%, centrifuging and taking supernatant that filtrate is concentrated under reduced pressure into concentration of alcohol in filtrate under 300 ± 100pa vacuum degree
This supernatant is crossed SD200 large pore resin absorption column by liquid, and loading speed is 1BV/h-8BV/h, loading volume 4-8BV;?
20-35 DEG C, under normal pressure, with 1BV/h-8BV/h flow velocity, with distillation water elution resin, elution volume 5-10BV;At 20-35 DEG C,
Under normal pressure, with 1BV/h-8BV/h flow velocity, the ethanol aqueous wash deresination for being 10-30% with concentration expressed in percentage by volume, elution volume
For 8-12BV;Under 20-35 DEG C, normal pressure, with 1BV/h-8BV/h flow velocity, the ethyl alcohol for being 40-90% with concentration expressed in percentage by volume is water-soluble
Liquid elutes resin, elution volume 8-12BV;40-90% ethanol eluate is collected, concentration recycles ethyl alcohol, is dried under reduced pressure, obtains
Hickory Leaves extractive total flavone, wherein the total content of pinostrobin, pinostrobin chalcone, Chrysin and cardamomin 50% with
On.
3. preparation method according to claim 2, it is characterised in that: 70-95% ethanol water dosage is in step (1)
Every g Hickory Leaves crushed material adds 10mL.
4. preparation method according to claim 2, it is characterised in that: parameter of noncentricity in step (2) are as follows: 2000-4000 turns
Per minute, it is centrifuged 20 minutes.
5. the Hickory Leaves extractive total flavone with anti-trioxypurine effect is in preparation hyperuricemia medicine as described in claim 1
Purposes in object.
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CN107252440A (en) * | 2017-06-05 | 2017-10-17 | 浙江中医药大学 | A kind of preparation method and purposes of the Hickory Leaves general flavone with hypoglycemic effect |
CN107519234A (en) * | 2017-08-28 | 2017-12-29 | 付林春 | A kind of preparation technology of Hickory Leaves extractive of general flavone |
CN111494569A (en) * | 2020-06-19 | 2020-08-07 | 陈正 | Preparation method of walnut leaf powder instant beverage for relieving gout |
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Inhibition of chrysin on xanthine oxidase activity and its inhibition mechanism;Suyun Lin等;《International Journal of Biological Macromolecules》;20150811;第274-282页 * |
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