CN102125203B - Glycosaminoglycan ultrafine particle-containing health food and production method thereof - Google Patents

Glycosaminoglycan ultrafine particle-containing health food and production method thereof Download PDF

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CN102125203B
CN102125203B CN 201010602614 CN201010602614A CN102125203B CN 102125203 B CN102125203 B CN 102125203B CN 201010602614 CN201010602614 CN 201010602614 CN 201010602614 A CN201010602614 A CN 201010602614A CN 102125203 B CN102125203 B CN 102125203B
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glycosaminoglycan
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许梓荣
许英蕾
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PUAI BIOENGINEERING CO Ltd NINGBO
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Abstract

The invention relates to health food and a production method thereof, and aims to provide glycosaminoglycan ultrafine particle-containing health food and a production method thereof. The health food comprises a food-grade auxiliary material, and glycosaminoglycan ultrafine particles with the particle size of 5 to 500nm. The method comprises the following steps of: adding glycosaminoglycan into dilute acid solution to prepare acid solution of polysaccharide; adding an emulsifier to form fine particle emulsion; preparing glycosaminoglycan ultrafine particle suspension with high dispersibility by utilizing a physical grinding method; and adding the food-grade auxiliary material, and adjusting the pH value of the solution to obtain glycosaminoglycan ultrafine particle-containing health oral liquid with stable properties. The glycosaminoglycan is prepared into the glycosaminoglycan ultrafine particles and applied to the health food, so that the bioavailability of the glycosaminoglycan in vivo is dramatically improved; the ultrafine particles have uniform particle size distribution and stable properties and do not have potential safety hazard; and the product is applied to patients suffering from tumors and having low immunity and normal people.

Description

A kind of health food and production method thereof that contains super fine particle of amino-polysaccharide as
Technical field
The present invention relates to a kind of health food and production method, particularly relate to a kind of health food and production method thereof that contains super fine particle of amino-polysaccharide as.
Background technology
Glycosaminoglycan is a kind of cellulosic natural polysaccharide that is similar to.It has tissue intersolubility, can not cause allergy and tissue rejection.Glycosaminoglycan degraded in vivo is to carry out under the effect of ferment, and is nontoxic, is convenient to remove from histoorgan, and final catabolite is a Glucosamine absorbed by the body.
Some studies show that glycosaminoglycan has and strengthen immunity, reducing blood lipid, protect health care such as liver.But immunocyte and complement system such as glycosaminoglycan activated macrophage, lymphocyte, NK cell also induce the various kinds of cell factor to produce, and improve body's immunity; Glycosaminoglycan can promote the drainage of body to lipid material in conjunction with the lipid material in the blood, thereby produces the health-care effect of reducing blood lipid; Glycosaminoglycan also can and suppress peroxidatic reaction of lipid and hepatic injury has the effect of protecting liver, lowering enzymes to chmice acute by Green Tea Extract damage.
But because the molecular proportion of glycosaminoglycan is bigger, be insoluble in water under neutrality or alkali condition, therefore the uptake in enteron aisle is few, and the bioavilability in the body is very low, is difficult to bring into play its distinctive biological effect.Some studies show that the macromolecular substances that will be difficult to absorb is built into micron or nano_scale particle can improve picked-up and the transport efficacy of enterocyte to it greatly, thereby improve the bioavilability in its body.The method for preparing the polysaccharide particulate has multiple, mainly comprise water oil two phase process, emulsifying technology, spray drying process etc., usually these preparation process complexity are not suitable for suitability for industrialized production, and since these methods with an organic solvent with surfactant, prepared particulate has the security doubt, is not suitable for the application in food and medicine.
Chinese invention patent " a kind of super fine particle of amino-polysaccharide as antineoplastic and preparation method thereof " (CN101002794) has disclosed a kind of super fine particle of amino-polysaccharide as antineoplastic and preparation method thereof.The glycosaminoglycan of this super fine particle of amino-polysaccharide as antineoplastic derives from organism, and viscosity average molecular weigh is 1 * 10 3~9 * 10 5, free amino group quantity accounts for total amino quantity 50%~100% in the molecule; Its preparation method may further comprise the steps: (1) adds the glycosaminoglycan of one or more molecular weight in 20~60 ℃ dilute acid soln, and configuration quality percentage is 0.1%~5% polysaccharide acid solution; (2) regulate polysaccharide acid solution pH value, make its pH value be 6-9, form the particulate emulsion; (3) the separate fine particle emulsion obtains particulate, and low temperature drying, obtains super fine particle of amino-polysaccharide as antineoplastic.The present invention makes ultrafine dust with glycosaminoglycan, has increased substantially its antitumor activity; The super fine particle of amino-polysaccharide as particle diameter of preparation method provided by the invention preparation is evenly distributed, easily separated and purifying and do not have potential safety hazard.
But, the examples of implementation that this patent documentation disclosed are directly to destroy various cancer cells or tumour cell life entity itself by super fine particle of amino-polysaccharide as under experimental condition, reach antineoplastic purpose, thereby be applied in the tumor treatment, do not relate to super fine particle of amino-polysaccharide as and strengthen immunity of organisms, regulate health cares such as gut flora and promotion digestion; Do not relate to specifically the mode which kind of is more conducive to practical application with yet and use super fine particle of amino-polysaccharide as.
Summary of the invention
The technical problem to be solved in the present invention is, overcomes deficiency of the prior art, and a kind of health food and production method thereof that contains super fine particle of amino-polysaccharide as is provided.
In order to solve the problems of the technologies described above, the present invention is achieved through the following technical solutions:
A kind of health food that contains super fine particle of amino-polysaccharide as is provided, comprises the food-grade auxiliary material; This health food comprises that also particle diameter is the super fine particle of amino-polysaccharide as of 5~500nm.
Among the present invention, the glycosaminoglycan that is used to prepare super fine particle of amino-polysaccharide as derives from organism, and its viscosity average molecular weigh is 1 * 10 3~9 * 10 5, free amino group quantity accounts for total amino quantity 50%~100% in the molecule.
Among the present invention, the concentration of the super fine particle of amino-polysaccharide as in the described health food is greater than 100ug/ml.
Among the present invention, described health food is an oral liquid formulation.
Among the present invention, described food-grade auxiliary material be colouring agent, flavor enhancement, stabilizing agent, antioxidant, anticorrisive agent, etc. ooze at least a in salting liquid or the phosphate buffer.
Further, the present invention also provides a kind of method of producing aforementioned health food, may further comprise the steps:
(1) in 20~60 ℃ dilute acid soln, adds glycosaminoglycan, be mixed with mass percent and be 0.05%~0.5% polysaccharide acid solution;
(2) press solution quality than the emulsifying agent that adds 0.01%~0.05%, form the particulate emulsion;
(3) adopt the physical crushing method to prepare super fine particle of amino-polysaccharide as suspension with good dispersion;
(4) add the food-grade auxiliary material, the pH value of regulator solution obtains the health oral liquid that contains super fine particle of amino-polysaccharide as of stable in properties between 4~6.
As a kind of improvement, the described diluted acid of step (1) is any one in acetate, hydrochloric acid, citric acid, the phosphoric acid, and the concentration of acid is less than 0.1M.
As a kind of improvement, the described emulsifying agent of step (2) is a food additives phosphate.
As a kind of improvement, the described physical crushing method of step (3) is any one or its combination in Ultrasonic Pulverization method or the liquid stream comminuting method.
Compared with prior art, the invention has the beneficial effects as follows:
(1) the present invention is prepared into glycosaminoglycan super fine particle of amino-polysaccharide as and is applied to health food, increased substantially its bioavilability in vivo, thereby body is produced obvious health care effect, can be used as the main active of health food and be widely used.
(2) the invention provides the method that a kind of production contains the health food of super fine particle of amino-polysaccharide as, the particle diameter of the super fine particle of amino-polysaccharide as in the health food that this method is produced is evenly distributed, and there is not potential safety hazard in stable in properties.
(3) can only to be applied to the antineoplaston institute of tumor patient different with the described super fine particle of amino-polysaccharide as of background technology, and as health food, super fine particle of amino-polysaccharide as is not only applicable to the tumor patient of hypoimmunity, and normal population also can be taken.
The specific embodiment
Further specify the present invention below in conjunction with instantiation.In following specific embodiment, unless description is arranged in addition, all solution % all are expressed as weight per volume.
Embodiment 1:
At 20 ℃, add a certain amount of glycosaminoglycan in 0.1% the acetum, be mixed with 0.2% polysaccharide acid solution; Add a certain amount of sodium phosphate trimer, make its concentration in solution reach 0.01%, form the particulate emulsion.Ultrasonic wave 800W/L disperseed 4 hours, obtained the transparency height, the super fine particle of amino-polysaccharide as suspension of favorable dispersibility; Get the 50ml suspension and add in the 50ml water, add 2g glucose then, the double edible caramel colorant of 0.1g with citric acid or sodium hydrogen phosphate regulator solution pH to 5, promptly gets the health drink that contains super fine particle of amino-polysaccharide as capable of direct drinking.
Embodiment 2:
At 60 ℃, add a certain amount of glycosaminoglycan in 0.2% the citric acid solution, be mixed with 0.5% polysaccharide acid solution.0.05% the ultra-low viscosity sodium alginate soln that slowly adds equal volume forms the particulate emulsion.Enter liquid stream pulverizer and be forced into 200MPa, every liter of circulation disperseed 4 hours, with citric acid or sodium hydrogen phosphate regulator solution pH to 4, obtained the transparency height, the super fine particle of amino-polysaccharide as suspension of favorable dispersibility.In 80ml water, add 20ml ultrafine dust suspension, 0.5g stevioside, 0.1g1% beta carotene dry powder, promptly get the health drink that contains super fine particle of amino-polysaccharide as capable of direct drinking.
Embodiment 3:
At 40 ℃, add a certain amount of glycosaminoglycan in 0.06% the phosphoric acid solution, be mixed with 0.05% polysaccharide acid solution, adding equal-volume concentration is 0.1% sodium pyrophosphate solution, forms the particulate emulsion.Every liter of liquid stream pulverizer pressurization 150MPa disperses that ultrasonic wave 600W disperseed 6 hours after 2 hours, with phosphoric acid or sodium hydrogen phosphate regulator solution pH to 6, obtains the transparency height, the super fine particle of amino-polysaccharide as suspension of favorable dispersibility; Get the 100ml suspension, add the 1g xylitol, 0.15 g eats caramel colorant, irradiation sterilization, and packing promptly gets the health drink that contains super fine particle of amino-polysaccharide as capable of direct drinking.(after adding the sodium pyrophosphate solution of equal volume 0.1%, the ultimate density of sodium pyrophosphate is 0.05% in the solution)
Embodiment 4:
Utilize fluorescein isothiocyanate (FITC) mark glycosaminoglycan, and the glycosaminoglycan of mark is prepared into ultrafine dust.Glycosaminoglycan that mark is crossed and super fine particle of amino-polysaccharide as are mixed with concentration respectively in nutrient solution be 0.25mg/ml, and with fluorescence microscope visible glycosaminoglycan mainly attach to cell surface at 37 ℃ after hatching 2 hours with the Caco-2 cell monolayer, and super fine particle of amino-polysaccharide as can be crossed over cell membrane and enters the cell interior (see figure 2).The Caco-2 cell monolayer is a kind of cell model of simulation gut epithelium tissue of generally acknowledging in the world at present, is mainly used in the absorption of assessment enteron aisle to medicine and nutrient.This shows that super fine particle of amino-polysaccharide as is easy to by intestinal absorption than glycosaminoglycan.
Embodiment 5:
24 of SD rats are divided into 3 groups at random, 8 every group, are respectively control group, test group I and test group II.The control group oral normal saline, the super fine particle of amino-polysaccharide as of the oral 80mg/kg b.w. of test group I, the super fine particle of amino-polysaccharide as of the oral 160mg/kg b.w. of test II group totally 21 days.After feeding experiment finishes, with the fasting 12 hours under the condition of freely drink water of all rats, barbital sodium (30mg/kg b.w.) intraperitoneal injection of anesthesia, femoral artery blood sampling execution.Get blood sample and spleen, measure in the serum immunoglobulin (Ig), complement and lysozyme level and carry out periphery blood lymphocyte conversioning test, spleen lymphocyte conversion test, result's (seeing Table 1-2) shows that oral super fine particle of amino-polysaccharide as can significantly improve the concentration of IgA in the rat blood serum, IgG, complement C 3With the lysozyme level, and has certain dose-effect relationship.In lymphocyte transformation test, at ConA A(ConA) when inducing or at lipopolysaccharides (LPS) when inducing, the SI (SI) of the test group I of oral super fine particle of amino-polysaccharide as and test group II is compared all with control group and is increased, and presents certain dose-effect relationship.This shows that oral super fine particle of amino-polysaccharide as can strengthen the immunologic function of rat; Get pH and short-chain fat acid content that cecal content carries out count of bacteria and bacterial enzyme activity analysis and measures content, result's (seeing Table 3-5) demonstration super fine particle of amino-polysaccharide as can be regulated the cecal content flora, reduce the activity of the sweet sour enzyme of β-glucose, improve the active of а-galactosidase, beta galactosidase and α-heteroside enzyme and can significantly improve the concentration of caecum propionic acid and butyric acid and the pH value of reduction cecal content; Get small intestine contents, mucous membrane of small intestine, measure the content digestive enzyme activity, the mucosal tissue enzymatic activity, and make histotomy and carry out the length and the crypts degree of depth that microscopy is measured intestinal villus, result's (seeing Table 6-8 and Fig. 2) shows that super fine particle of amino-polysaccharide as can improve the morphosis of mucous membrane of small intestine, significantly improve small intestine contents protease, lipase and diastatic activity, also can significantly improve the activity of duodenum, jejunum, ileal mucous membrane disaccharidase.Therefore oral super fine particle of amino-polysaccharide as also has the function of regulating gut flora and promoting digestion.
Table 1:SD rat peripheral blood and spleen lymphocyte conversion ratio
Statistical value is represented with mean+SD; Every group of 8 samples;
ConA: ConA A; LPS: lipopolysaccharides; SI: SI; * p<0.05.
The level of table 2:SD rat blood serum immunoglobulin (Ig), complement and lysozyme
Figure GDA0000331578192
*?p<0.05;**?p<0.01
Table 3:SD rat caecum flora count results
Figure GDA0000331578193
Annotate: the clump count of every gram cecal content is represented with its logarithm value; * p<0.05
The activity of bacterial enzyme in the table 4:SD rat cecal content
Figure GDA0000331578194
Enzyme unit alive (IU) definition: every gram cecal content per minute decomposes the micromole's number that discharges nitrophenol from substrate; * p<0.05
The variation of table 5:SD rat cecal content pH and chain rupture aliphatic acid (SCFA) content
Control group Test group I The test group II
pH 6.83±0.11 6.71±0.09* 6.45±0.05*
Acetate 56.41±8.37 62.40±10.36 64.40±7.06
Propionic acid 11.77±2.29 14.62±3.24* 19.52±5.22*
Butyric acid 8.39±5.17 11.97±3.68* 14.36±3.82*
Isobutyric acid 1.36±0.73 1.26±0.18 1.18±0.28
Valeric acid 1.95±0.63 2.09±0.27 2.20±0.27
Isovaleric acid 2.24±0.62 2.13±0.63 2.35±0.55
Total acid content 83.12±11.99 94.47±13.94 104.01±11.32
SCFA concentration is shown with SCFA micromole numerical table contained in every gram cecal content; * p<0.05
The table 6:SD rat small intestine height of naps and the crypts degree of depth
Figure GDA0000331578195
*p<0.05
Table 7:SD rat small intestine content digestive enzyme activity
Pancreatin (IU) Lipase (IU) Amylase (IU)
Control group 90.83±5.82 7.84±1.26 0.98±0.21
Test group I 106.51±12.59* 10.43±2.47* 1.30±0.44*
The test group II 113.85±14.66* 12.47±1.99* 1.43±0.30*
Unit (IU) definition of living of pancreatin enzyme: at pH8.0, under 37 ℃, the trypsase per minute that contains in every milligram of albumen makes absorbance change 0.003 and is the enzyme unit that lives;
Lipase activity unit (IU) definition: it is 1 enzyme unit that lives that every gram content albumen decomposes 1 micromole's triglycerides in per 30 minutes at 37 ℃;
Diastatic activity unit (IU) definition: every milligram of content albumen 37 ℃ with substrate-function 30 minutes, 10 milligrams of starch of hydrolysis are defined as 1 diastatic activity unit of force.
*p<0.05
The activity of table 8:SD rat small intestine mucous membrane enzyme
Figure GDA0000331578196
Enzyme unit alive (IU) definition: at pH6.0, the corresponding monose of every milligram of histone per minute hydrolysis 1 nanomole is defined as enzyme unit alive under 37 ℃ of conditions.
*p<0.05
At last, it is also to be noted that what more than enumerate only is some specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.

Claims (1)

1. a production contains the method for the health food of super fine particle of amino-polysaccharide as, and this health food comprises the food-grade auxiliary material, and particle diameter is the super fine particle of amino-polysaccharide as of 5~500nm; It is characterized in that this method specifically comprises:
(1) in 20~60 ℃ dilute acid soln, adds glycosaminoglycan, be mixed with mass percent and be 0.05%~0.5% polysaccharide acid solution;
(2) press solution quality than the emulsifying agent that adds 0.01%~0.05%, form the particulate emulsion;
(3) adopt the physical crushing method to prepare super fine particle of amino-polysaccharide as suspension with good dispersion;
(4) add the food-grade auxiliary material, the pH value of regulator solution obtains the health oral liquid that contains super fine particle of amino-polysaccharide as of stable in properties between 4~6; The concentration of super fine particle of amino-polysaccharide as is greater than 100 μ g/ml in this oral liquid;
Described diluted acid is any one in acetate, hydrochloric acid, citric acid, the phosphoric acid, and the concentration of acid is less than 0.1M; Described glycosaminoglycan derives from organism, and its viscosity average molecular weigh is 1 * 10 3~9 * 10 5, free amino group quantity accounts for total amino quantity 50%~100% in the molecule; Described emulsifying agent is a food additives phosphate; Described physical crushing method is any one or its combination in Ultrasonic Pulverization method or the liquid stream comminuting method; Described food-grade auxiliary material be colouring agent, flavor enhancement, stabilizing agent, antioxidant, anticorrisive agent, etc. ooze at least a in salting liquid or the phosphate buffer.
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