CN110251462A - A kind of oral solution with blood pressure lowering, anti thrombotic action - Google Patents
A kind of oral solution with blood pressure lowering, anti thrombotic action Download PDFInfo
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Abstract
The present invention provides a kind of oral solution with blood pressure lowering, anti thrombotic action, belongs to biologic product technology field, the preparation method of the oral solution includes: the extraction of laminarin；Fucoidin isolates and purifies；Free-radical oxidation degradation fucoidin；Dissolution；Coarse filtration；Flavor mixing；Sterilization；Filling, sealing；Wherein, choline chloride is added in fucoidin Oxidative Degradation Process.Oral solution molecular weight provided by the invention is small, easily absorbs, and sulfate radical content is high, and bioactivity is high, and further improves the anti thrombotic action of fucoidin from the approach for improving hemorheology.
Description
Technical field
The invention belongs to biologic product technology fields, and in particular to a kind of oral solution with blood pressure lowering, anti thrombotic action.
Background technique
1913, Kylin extracted a kind of polysaccharide containing L-fucose from Laminaria digitata, was named as Fucoidan, in
Literary fame is known as fucoidin, fucoidin or fucoidin.Hereafter, and successively report in the molecule of fucoidin and contain sulfuric acid
Ester, galactolipin, xylose, uronic acid etc., fucoidin are highly heterogeneous polysaccharide.Studies have shown that fucoidin has many
Biological activity has the function of anticoagulation, antiviral, antithrombotic, blood pressure lowering and antitumor etc., especially its preferable anticoagulation
Activity increasingly causes people widely to pay close attention to, and has biggish market potential in terms of anticoagulation medicine.
The laminaria culture amount of China at home and abroad maintains the leading position, and fucosan content reaches 3.6%, is that China is brown
The highest a kind of brown alga of fucose content in algae resource can be used as the raw material for developing and utilizing fucoidan from Laminaria japonica, processing
Kelp in sodium alginate, iodine and mannitol rich in, and process and generate a large amount of fucoidins therewith, through frequently as useless
Gurry abandons, and causes the very big wasting of resources.And at present about fucoidan especially low molecule quality kelp rock
The research of the preparation of algae glycan sulfuric ester and application is also relatively fewer.
The prior art such as Authorization Notice No. is the Chinese invention patent of 104256252 B of CN, which is related to a kind of auxiliary
Antihypertensive healthcare product, provides a kind of step by step arithmetic and bioconversion coupling prepares the side of aided blood pressure-lowering health care kelp product
Method belongs to marine food intensive processing field.This method using high-quality kelp as raw material, using circulating ultrasonic substep extracting technology with
Bioconversion is coupled technology of preparing, and preparation step is successively are as follows: pretreatment of raw material, alcohol extracting mannitol, acid mention alcohol precipitation algin and
Fucoidin, bioconversion, ultrafiltration obtain blood pressure lowering physiological activity component and carry out scientific compatibility.The inventive method is directed to me
The large marine product kelp of state's tradition carries out intensive processing and comprehensive utilization, obtains having the mannitol of antihypertensive function, brown alga
Five glue, fucoidin, kelp dietary fiber and kelp blood pressure peptide products.Scientific compatibility, exploitation are carried out to this five products
Series has the novel kelp blood pressure functional article of significant auxiliary function for lowering blood pressure.Product ACE inhibitory activity IC50 is
0.68~1.15mg/g significantly reduces SHR rat blood pressure (- 12.4~-17.5) mmHg.
Summary of the invention
The purpose of the present invention is to provide a kind of with blood pressure lowering, the oral solution of anti thrombotic action and preparation method thereof, should
Method can shorten the covalent bond distance of sulfate group, increase covalent bond strength, reduce falling off for sulfate radical, brown alga in the oral solution
Xanthan molecule amount is low, easily absorbs, and can reduce the formation of saturated fatty acid in erythrocyte membrane, improves Surface of Erythrocytes receptor saliva
Acid content reduces blood viscosity, has preferable antithrombotic effectiveness.
The technical solution that the present invention is taken to achieve the above object are as follows:
A kind of oral solution with blood pressure lowering, anti thrombotic action, the preparation method of oral solution are provided, comprising:
The extraction of S1, laminarin；
S2, fucoidin isolate and purify；
S3, free-radical oxidation degradation fucoidin；
S4, dissolution；
S5, coarse filtration；
S6, flavor mixing；
S7, sterilization；
S8, filling, sealing；
Wherein, choline chloride is added in step S3 fucoidin Oxidative Degradation Process.Free radical cracking algal polysaccharide
Production method is relatively easy, and product quality is easily controllable, is advantageously implemented industrialized production, but excessively violent reaction condition
The structure of sugar chain can be destroyed, engaging sulphate degree also will increase, and when fucoidin oxidative degradation adds choline chloride, sulfate group
Positively charged choline is adsorbed, the O on ester group shortens the covalent bond of sulfate group from the hydroxyl electron donating group electrophilic of choline
It is long, covalent bond strength is increased, to reduce falling off for sulfate radical, sulfate radical content is improved, improves low molecule fucoidin
Bioactivity.
In some embodiments, the method for step S3 free-radical oxidation degradation fucoidin includes:
A, 85-100mg/mL fucoidin solution is prepared, copper acetate is added to 0.038-0.042mol/L；
B, choline chloride is added to 0.032-0.036mol/L, stands 0.8-1.2h；
C, at 59-61 DEG C, quality volume fraction is added as the H of 4.4-4.6% with constant flow rate2O2Solution reacts 5-6h；
D, 0.22 μm of membrane filtration of reaction solution,
E, filtrate is taken, is eluted with chelating resin；
F, concentrated freeze-dried after revolving, obtain low-molecular-weight algal carbohydrate gum；
Wherein, the step a- step c whole process keeps pH in 7.2-7.8.Free-radical oxidation drop provided by the invention
The obtained molecular weight of product of solution condition is lower, while sulfate radical content is higher, therefore bioactivity with higher and biology
Availability.
In some embodiments, step S1 uses ultrasonic extraction method laminarin.Using ultrasonic extraction method kelp
Polysaccharide yield and bioactivity are higher, while reducing the precipitation of the impurity such as pigment, simplify extraction purification process.
In some embodiments, the method for ultrasonic wave extraction laminarin are as follows: according to 1:70-75 (g/mL) ratio to
Dry Kelp Powder is added in distilled water, ultrasonication 55-65min at 62-68 DEG C.Kelp has very strong swellability, solid-to-liquid ratio
Be swollen insufficient when too high, recovery rate is low, more than appropriate swellbility solid-to-liquid ratio when, swelling excessively also results in recovery rate decline；
Ultrasonic wave is by mechanical shear stress come assisted extraction, and ultrasonic time is too short, and mechanical shearing effect is insufficient, and recovery rate is not
Height, ultrasonic time is too long, can make macromolecular polysaccharide chain rupture, also reduce recovery rate；Temperature increases the surface that can make liquid medium
Tension and viscosity reduce, and accelerate the diffusion of solution, promote intracellular polysaccharide to external diffusion, but when the temperature is excessively high, polysaccharide structures
Part can be destroyed.
In some embodiments, step S2 is isolated and purified using ethyl alcohol reprecipitation method.Ethanol precipitation method behaviour
It is easy to control to make easy and condition, is suitble to isolate and purify on a large scale.
In some embodiments, the method that ethyl alcohol reprecipitation isolates and purifies fucoidin are as follows: after ultrasonication, mistake
Filter concentrates the filtrate to 18-25% original volume, and it is 28-32% that 90-95% ethyl alcohol to concentration, which is added,；Filtering, then filtrate is concentrated
To 1/5 volume, ethyl alcohol to the concentration that 90-95% is added is 59-61%, stands 11-13h；Collect precipitating, freeze-drying.This hair
Filtrate cycles of concentration, ethyl alcohol dosage and the ethyl alcohol mass fraction of bright offer are that best alcohol analyses condition, and fucoidin yield is higher.
In some embodiments, ungrease treatment is carried out to kelp before step S1 extracts laminarin.Polysaccharide is polarity
Strong macromolecular compound, carrying out degreasing to raw material can make aqueous solution be easier to go deep into inside raw material, be conducive to the molten of polysaccharide
Out.
In some embodiments, the method for step S6 flavor mixing are as follows: be added in filtered fucoidin solution
The beta-cyclodextrin of 0.13-0.17% (w/w), the xylitol of 13-17% (w/w), the citric acid of 0.13-0.17% (w/w) and suitable
The essence of amount, stirs evenly.Flavor modulation technique provided by the invention, from the face of the sense of taste, smell, vision etc. adjustment product
Color, fragrance make product sour and sweet palatability, pure in mouth feel.
In some embodiments, biotin is grafted on fucoidin.The fucoidin of low molecule quality has certain
Antithrombotic acitivity, hemorrhage risk is relatively low, and the fucoidin of low molecule quality mainly passes through anticoagulation, D-dimer
Anti thrombotic action is played with fibrinolytic function.The fucoidin of low molecule quality has certain antithrombotic acitivity, hemorrhage risk
Relatively low, the fucoidin of low molecule quality mainly passes through anticoagulation, D-dimer and fibrinolytic function and plays antithrombotic
Effect.Have a major reason in thrombosis factor precisely due to hemorheology change, cause blood viscousness, lectin from hemolymph
Slowly, a possibility that increasing impaired vascular endothelium and embolism formation.After being grafted biotin on fucoidin, red blood cell can be improved
Interior SOD is horizontal, improves the oxidation characteristic of Surface of Erythrocytes, the injury for fighting and blocking oxygen radical to red blood cell, reduces thin
The formation of saturated fatty acid in after birth, improve cell membrane mobility, reduce viscosity, while can be improved Surface of Erythrocytes by
Body sialic acid content reduces the aggregate index of red blood cell, in short, grafting biotin after fucoidin can make whole blood viscosity,
Plasma Viscosity, hematocrit, erythrocyte sedimentation rate (ESR) and ESR equation K value significantly reduce, and improve the deformability of haemocyte,
To be obviously improved hemorheology, antithrombotic effectiveness is improved.
In some embodiments, oral solution also has the function of antiviral and antitumor and improves immunity.The present invention
The fucoidin of offer is oral to be extracted from kelp, and by degradation, modification has molecular weight small, easy the characteristics of absorbing, and disease-resistant
Bioactivity that is malicious, antitumor and improving immunity is higher.
The invention has the benefit that
1) present invention is improved by the method to free-radical oxidation degradation fucoidin, shortens the covalent of sulfate group
Bond distance increases covalent bond strength, reduces falling off for sulfate radical, improves sulfate radical content, improves the biology of low molecule fucoidin
Activity；
2) present invention is by optimizing ultrasonication condition, alcohol precipitation condition, improving more to kelp ungrease treatment
The recovery rate of sugar and the yield of laminaran；
3) present invention improves the oxidation characteristic of Surface of Erythrocytes, subtracts by being grafted polypeptide to low-molecular-weight algal carbohydrate gum
The formation of saturated fatty acid in few cell membrane improves the mobility of cell membrane, reduces viscosity, while can be improved erythrocyte membrane table
Face receptor sialic acid content reduces the aggregate index of red blood cell, improves antithrombotic effectiveness.
Detailed description of the invention
Fig. 1 is the schematic diagram of test example 1 of the invention；
Fig. 2 is the schematic diagram of test example 2 of the invention；
Fig. 3 is the schematic diagram of test example 3 of the invention；
Fig. 4 is the schematic diagram of test example 4 of the invention；
Fig. 5 is Surface of Erythrocytes sialic acid content of the invention, the schematic diagram of SOD, MDA level；
Fig. 6 is 150s of the invention-1And 10s-1Whole blood viscosity under shear rate, the schematic diagram of plasma viscosity；
Fig. 7 be hematocrit of the invention, erythrocyte sedimentation rate (ESR), ESR equation K value schematic diagram.
Specific embodiment
Present invention is further described in detail with reference to embodiments:
Embodiment 1:
A kind of preparation method of fucoidin, comprising:
Kelp dry powder 5g is weighed, after distilled water 312.5mL is added, ultrasonic echography handles 60min at 65 DEG C, and 1/ is added afterwards
The sevage reagent of 3 volumes, after magnetic stirrer 30min, 1000rpm is centrifuged 15min, collects supernatant, is repeated 5 times, and uses
Combined closure system is the dialysis of 3500 films, is concentrated under reduced pressure into 1/5 volume, and it is 30% that ethyl alcohol to the concentration that mass fraction is 95%, which is added, quiet
It sets, filters, then concentrate the filtrate to 1/5 volume.It is 60% that 95% ethyl alcohol to concentration, which is added, stands 12h, collects precipitating, freezing
It dries to constant weight.
Embodiment 2:
A kind of preparation method of fucoidin, comprising:
80g kelp dry powder is taken, the ethyl alcohol that the mass fraction of 5 times of volumes is 95%, ultrasonic degreasing 25min, vacuum is added
Under the conditions of filtered, collect filter residue, be repeated 3 times, dry filter residue, smashed 80 meshes.
80 mesh kelp dry powder 5g are weighed, after distilled water 312.5mL is added, ultrasonic echography handles 60min at 65 DEG C, rear to add
Enter the sevage reagent of 1/3 volume, after magnetic stirrer 30min, 1000rpm is centrifuged 15min, collects supernatant, repeats 5
It is secondary, it is the dialysis of 3500 films with combined closure system, is concentrated under reduced pressure into 1/5 volume, ethyl alcohol to the concentration that mass fraction is 95%, which is added, is
30%, it stands, filtering, then concentrate the filtrate to 1/5 volume.It is 60% that 95% ethyl alcohol to concentration, which is added, stands 12h, and it is heavy to collect
It forms sediment, freeze-drying to constant weight.
It takes 4.5g fucoidin to be dissolved in distilled water, is configured to 100mg/mL solution, copper acetate is then added to Cu2+Concentration is
0.035mol/L is added choline chloride to 0.032mol/L, is stood 1h, water with 2mol/L NaOH solution tune pH to 7.5 or so
Bath is heated to solution temperature and reaches 60 DEG C, then with the speed of 12mL/h be added quality volume fraction I 4.5% H2O2Solution,
After reacting 5h stop that H is added2O2Solution, with 2mol/L NaOH solution tune pH to 7.5 or so.Reaction solution crosses the filter membrane of 0.22um,
Remove the Cu for forming precipitating2+.It adds Chelex100 chelating resin and removes remaining free Cu2+, it is with molecular cut off
The ultrafiltration apparatus of 1000Da removes most salt, then the bag filter dialysis desalting for being 500Da with molecular cut off, dense after revolving
Contracting freeze-drying, obtains low molecule fucoidin.
It weighs 2g low molecule fucoidin and is dissolved in formamide, stir 35min at 80 DEG C, biotin is dissolved in containing 1%1- (3-
Dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (EDC) formamide in, be added drop-wise in polysaccharide solution, react 5h after,
Reaction solution is poured out, with 85% ethanol precipitation, 95% ethanol washing precipitates 2 times, precipitating is dissolved in 200mL distilled water,
2000Da bag filter tap water dialyse 2d, distilled water dialyse 1d, it is concentrated freeze-dried, obtain biotin grafting fucoidin.
Embodiment 3:
A kind of preparation method with blood pressure lowering, the oral solution of anti thrombotic action, comprising:
Dissolution: algin obtained is dissolved in water, the alginate solution of 10mg/mL is obtained；
Coarse filtration: insoluble impurity is removed；
Flavor mixing: beta-cyclodextrin, 15% (w/w) of 0.15% (w/w) are added in filtered algin oligosaccharide solution
Xylitol, 0.15% (w/w) citric acid and suitable essence, stir evenly；
Sterilization: high pressure steam sterilization, 121 DEG C of sterilization 30min are used；
Filling, sealing: it is filling to use 20mL brown oral liquid glass bottle, it needs that empty Bottle and bottle cap is carried out to clean before filling to disappear
Poison.
Oral solution kelp fishy smell glue obtained is light, color be in dark brown or sepia, taste moderately sour and sweet, delicate mouthfeel,
There is no peculiar smell, appearance clear is visible by naked eyes impurity, allows to have a small amount of muddy or precipitating, sensory evaluation 98 to divide.
Comparative example 1:
Choline chloride is not added when fucoidin oxidative degradation, rest part and embodiment 2 are completely the same.
Comparative example 2:
The low molecule fucoidin after degradation is not modified, rest part and embodiment 2 are completely the same.
Test example 1:
The measurement of molecular weight
By the reaction solution after fucoidin oxidative degradation respectively with molecular cut off be 10kDa, 6kDa, 2.5kDa and
The ultrafiltration membrane of 300Da carries out ultrafiltration, obtains four component Fa (6k Da~10kDa), Fb (2.5kDa~6kDa), Fc (300Da
~2.5kDa) and Fd (being less than 300Da), it is concentrated freeze-dried respectively.
The measurement of total reducing sugar is carried out using phenol-sulfuric acid process；Molecular weight determination is carried out using high performance liquid chromatography.Efficiently
Liquid chromatogram is shown in Fig. 1.The determination condition of high performance liquid chromatography (HPLC) is as follows: chromatographic column is TSK-gel G2500PWxl,
Mobile phase be 0.2mol/L NaCl, elution speed 0.5mL/min, column temperature keep 40 DEG C, detected using Composition distribution, with
Dextran standard items make standard curve respectively using the logarithm of the retention time of standard items and molecular weight as transverse and longitudinal coordinate.It adopts
The weight average molecular weight (Mw) and number-average molecular weight (Mn) at sample elution peak are calculated with Agillent GPC software, calculate polydisperse system
Number D=Mw/Mn.
Graph of molecular weight distribution after the fucoidin oxidative degradation that high performance liquid chromatography measures is shown in Fig. 1.It is computed and obtains drop
Three components are mainly obtained after solution, the weight average molecular weight of each component is respectively 7.42kDa, 3.85kDa, 1.51kDa, polydisperse system
Number is respectively 1.024,1.048,1.006.
Test example 2:
The measurement of sulfate radical content:
According to the molecular weight of 13 components of gained of test example, after determining free-radical oxidation degradation, product is mainly attached most importance to respectively
Son amount is respectively three components of 7kDa, 4kDa, 1.5kDa, uses molecular cut off for 10kDa, 6kDa, 2.5kDa and 300Da
Ultrafiltration membrane and nanofiltration membrane catabolite is separated, quickly obtain F1 (6-10kDa), F2 (2.5-6kDa) and F3 and (be less than
2.5kDa) 3 parts, it is concentrated freeze-dried respectively, using the content of ion-chromatographic determination sulfate radical.
3 components to be measured are dried under reduced pressure 1h in phosphorus pentoxide in drier, 3mg is weighed and is put into ampoule bottle,
The trifluoroacetic acid dissolution of 1mL 2mol/L is added, with nitrogen tube sealing, rear 110 DEG C of hydrolysis 8h；Taking-up is cooled to room temperature, at 60 DEG C,
It with trifluoroacetic acid is dried with nitrogen, adds ultrapure water and sufficiently dissolves, with being dried with nitrogen, be repeated 3 times；Ultrapure water water is added to dissolve, it is fixed
Hold to 25mL.
Chromatographic condition are as follows: splitter is Dionex Ionpac AS11-HC (4mm × 250mm)；Guard column is Dionex
Ionpac AG11-HC(4mm×50mm)；Leacheate is 30mmol/L KOH, and suppressor is ASRS ULTRA II, inhibits electric current
238mA, flow velocity 1.0mL/min, sampling volume 25 μ L, program time 8min.Respectively with standard items SO4 2-Concentration and eluting peak
Peak area is that transverse and longitudinal coordinate makes standard curve, and each sample does three in parallel, takes the average value of sulfate radical peak area, substitutes into mark
The sulfate radical content of directrix curve calculating each component sample.The measurement result of sulfate radical content is shown in Fig. 2.
As seen from Figure 2, the F1 (6-10kDa), F2 (2.5-6kDa) and F3 (be less than 2.5kDa) of embodiment 2 three be not
It is above comparative example 1, especially F3 component with the sulfate radical content in the component of molecular weight ranges, this explanation, fucoidin oxygen
Choline chloride is added when changing degradation, sulfuric acid ester bond intensity can be increased, reduces falling off for sulfate radical, improves sulfate radical content.
Test example 3:
The measurement of blood pressure lowering ability:
Take 10 week old spontaneous hypertensive rats, weight 400-420g, systolic pressure > 180mmHg, after adaptable fed 1 week,
It is randomly divided into 2 groups, every group 20:
Control group: distilled water is given；
Administration group: 100mg/kg fucoidin is given；
By daily single, 21d is administered altogether, 2h measures the blood pressure of each group rat upon administration every 1d, and every rat is every
Secondary blood pressure replication 3 times, takes mean value.The measurement result of rat blood pressure is shown in Fig. 3.
As seen from Figure 3, embodiment 2 is apparently higher than comparative example 1 to the blood pressure lowering ability of spontaneous hypertensive rat, this
Illustrate, when fucoidin oxidative degradation adds choline chloride, can be improved sulfate radical content, to improve low molecule fucoidin
Hypotensive activity.
Test example 4:
The measurement of anti-thrombogenic capacity:
Acute blood is established in rat model: taking SD rat, is randomly divided into control group, model group, administration group, every group 20,
By the rat chloral hydrate anesthesia of model group and administration group, left common carotid artery is separated；It sprawls 3cm × 2cm film and always moves neck
Arteries and veins is isolated with surrounding tissue.There are 20 μ L FeCl with suction3The filter paper of 1cm × 1cm of solution wraps up left common carotid artery, removes paper afterwards
Piece sutures neck, instillation gentamicin sulphate after physiological saline cleaning.Administration group gastric infusion (algin oligosaccharide 100mg/
Kg), 1 time a day, continuous 5 days, control group, model group gave isometric(al) aquae destillata.After last dose 1h, with 10% hydration chlorine
Aldehyde anesthesia, dorsal position are fixed, clip 1cm thrombus section blood vessel, weighing.Blood vessel is splitted, removes in managing and weighs vascular wall again after thrombus
Weight.The calculation formula of wet weight of thrombus and thrombus thrombolysis rate is as follows:
The weight in wet base of one vascular wall of weight in wet base of wet weight of thrombus=thrombus and vascular wall
Thrombus thrombolysis rate=(one administration group wet weight of thrombus of model group wet weight of thrombus)/model group wet weight of thrombus × 100%
The measurement result of wet weight of thrombus and thrombus thrombolysis rate is shown in Fig. 4.
As seen from Figure 4, embodiment 2 to the thrombus thrombolysis rate of stasis syndrome rat model be apparently higher than comparative example 1,
Comparative example 2, this explanation, when fucoidin oxidative degradation, add choline chloride, can be improved sulfate radical content, and it is brown to improve low molecule
The antithrombotic acitivity of algae carbohydrate gum；After being grafted biotin on low molecule fucoidin, antithrombotic effectiveness can be improved.
Test example 5:
The detection of hemorheology index:
The preparation of erythrocyte membrane: by above-mentioned administration group rat, for 24 hours, eye socket takes blood to empty stomach after the last administration.With fresh heparin
Anticoagulation 4mL, 3000rpm centrifugation 5min, takes precipitating, and the isotonic PBS dilution of 5 times of volumes is added, is centrifuged 5min under 1500rpm,
Precipitating is taken, is repeated 3 times, red blood cell suspension is obtained, the Tris-HCl cooled solution of the 5mmol/L pH7.4 of 40 times of volumes is added,
A small amount of 0.1mmol/L PMSF is added and inhibits albumen, is ultrasonically treated 10min under 90Hz, 4h, 12000rpm centrifugation are stood at latter 4 DEG C
10min takes precipitating, and the Tris-HCl of 5mmol/L pH7.4 is added, and 12000rpm is centrifuged 10min, is repeated 3 times, obtains near-transparent
Film.It is suspended in PBS solution according to 1: 1 ratio.
Surface of Erythrocytes Biological characteristics: erythrocyte membrane is measured according to corresponding kit specification method respectively
Surface sialic acid content, Surface of Erythrocytes SOD, MDA are horizontal.Measurement result is shown in Fig. 5.
Hemorheology index detection: taking heparin anticoagulation 5mL takes whole blood 1mL to measure whole blood viscosity under different shear rates,
Blood is centrifuged 10min under the conditions of 3000rpm, blood plasma 1mL is taken to measure plasma viscosity.Micro-capillary tubes method measures red blood cell pressure
Product, wintrobe method measure erythrocyte sedimentation rate (ESR), calculate ESR equation K value according to the following formula:
K=hEsn/[-(1-H+lnH)]
Wherein, hEsnFor erythrocyte sedimentation rate (ESR), H is hematocrit.150s-1And 10s-1Whole blood viscosity under shear rate, blood
Slurry viscosity measurement result is shown in Fig. 6, hematocrit, erythrocyte sedimentation rate (ESR), ESR equation K value measurement result see Fig. 7.
As seen from Figure 5, the erythrocyte surface sialic acid content of embodiment 2 and Surface of Erythrocytes SOD level are bright
It is aobvious to be higher than comparative example 2；The hemorheology index of embodiment 2 is substantially change compared with model group, and comparative example 2 is with model
Compared to not substantially changeing, this illustrates group, can be by improving in red blood cell after being grafted biotin on low molecule fucoidin
SOD level and raising Surface of Erythrocytes receptor sialic acid content improve the mobility of cell membrane, reduce viscosity, reduce red thin
The aggregate index of born of the same parents further increases anti thrombotic action so as to improve hemorheology.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail
It repeats.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (10)
1. a kind of oral solution with blood pressure lowering, anti thrombotic action, it is characterised in that: the preparation method of the oral solution, comprising:
The extraction of S1, laminarin；
S2, fucoidin isolate and purify；
S3, free-radical oxidation degradation fucoidin；
S4, dissolution；
S5, coarse filtration；
S6, flavor mixing；
S7, sterilization；
S8, filling, sealing；
Wherein, choline chloride is added in the step S3 fucoidin Oxidative Degradation Process.
2. oral solution according to claim 1, it is characterised in that: the step S3 free-radical oxidation degradation fucoidin
Method includes:
A, 85-100mg/mL fucoidin solution is prepared, copper acetate is added to 0.038-0.042mol/L；
B, choline chloride is added to 0.032-0.036mol/L, stands 0.8-1.2h；
C, at 59-61 DEG C, quality volume fraction is added as the H of 4.4-4.6% with constant flow rate2O2Solution reacts 5-6h；
D, 0.22 μm of membrane filtration of reaction solution,
E, filtrate is taken, is eluted with chelating resin；
F, concentrated freeze-dried after revolving, obtain low-molecular-weight algal carbohydrate gum；
Wherein, the step a- step c whole process keeps pH in 7.2-7.8.
3. oral solution according to claim 1 or 2, it is characterised in that: the step S1 uses ultrasonic extraction method kelp
Polysaccharide.
4. oral solution according to claim 3, it is characterised in that: the method for the ultrasonic wave extraction laminarin are as follows: press
Dry Kelp Powder is added into distilled water according to the ratio of 1:70-75 (g/mL), ultrasonication 55-65min at 62-68 DEG C.
5. oral solution according to claim 1, it is characterised in that: the step S2 is separated using ethyl alcohol reprecipitation method
Purifying.
6. oral solution according to claim 5, it is characterised in that: the ethyl alcohol reprecipitation isolates and purifies the side of fucoidin
Method are as follows: after ultrasonication, filtering concentrates the filtrate to 18-25% original volume, and it is 28- that 90-95% ethyl alcohol to concentration, which is added,
32%；Filtering, then 1/5 volume is concentrated the filtrate to, ethyl alcohol to the concentration that 90-95% is added is 59-61%, stands 11-13h；
Collect precipitating, freeze-drying.
7. oral solution according to claim 1, it is characterised in that: the step S1 extract before laminarin to kelp into
Row ungrease treatment.
8. oral solution according to claim 1, it is characterised in that: the method for the step S6 flavor mixing are as follows: after filtering
Fucoidin solution in xylitol, the 0.13- of the beta-cyclodextrin of 0.13-0.17% (w/w), 13-17% (w/w) is added
The citric acid and suitable essence of 0.17% (w/w), stirs evenly.
9. oral solution according to claim 2, it is characterised in that: be grafted biotin on the fucoidin.
10. oral solution according to claim 1, it is characterised in that: the oral solution also have antiviral and antitumor and
Improve the effect of immunity.
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Cited By (2)
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| CN114014947A (en) * | 2022-01-10 | 2022-02-08 | 菏泽高新区优科生物科技有限公司 | Eutectic solvent for degrading algal polysaccharide and preparation method and application thereof |
| CN117510663A (en) * | 2023-11-07 | 2024-02-06 | 威海人生药业集团股份有限公司 | ACE (angiotensin converting enzyme) inhibiting low-molecular-weight fucoidan, and preparation method and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114014947A (en) * | 2022-01-10 | 2022-02-08 | 菏泽高新区优科生物科技有限公司 | Eutectic solvent for degrading algal polysaccharide and preparation method and application thereof |
| CN117510663A (en) * | 2023-11-07 | 2024-02-06 | 威海人生药业集团股份有限公司 | ACE (angiotensin converting enzyme) inhibiting low-molecular-weight fucoidan, and preparation method and application thereof |
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