CN107594533A - A kind of pinctada fucata glycosaminoglycan chewable tablets and preparation method thereof - Google Patents
A kind of pinctada fucata glycosaminoglycan chewable tablets and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of pinctada fucata glycosaminoglycan chewable tablets and preparation method thereof, include following components in percentage by weight：Pinctada fucata glycosaminoglycan 15% 25%, hydroxypropyl methyl cellulose 3.5% 4.5%, mannitol 20% 25%, microcrystalline cellulose 40% 50%, magnesium stearate 1% 2%, surplus is lactose.Pinctada fucata glycosaminoglycan chewable tablets piece type prepared by the present invention is that biplane is circular, and shape is complete, and surface is smooth, and color and luster is white, uniformity, without sense of eking out a living, has good organoleptic quality, has no toxic side effect, and helps to maintain the normal immunologic function of body.The raw material sources of the inventive method are extensive, and cost is cheap, and technical matters is scientific and reasonable, preparation method is simple and easy, it is adapted to industrialized production, technological innovation is provided for the high-valued application of pinctada fucata shellfish meat, the development to promotion pearl aquaculture and processing industry is significant.
Description
Technical field
The present invention relates to health food processing technique field, more particularly to a kind of pinctada fucata glycosaminoglycan chewable tablets and
Its preparation method.
Background technology
Pinctada fucata Pinctadafucata martensii (Dunker), also known as pteria martensii, are south Chinas
The principal item of sea water pearls cultivation, is distributed in Guangdong, Guangxi and Hainan coastal area.
In addition to rich in good protein and several mineral materials, central glycosaminoglycan also particularly induces one pinctada fucata
Concern.Glycosaminoglycan (Glycosaminoglycans, GAGs), also referred to as glycosaminoglycan or acid mucopolysaccharide, it is by the two of repetition
The long chain polysaccharides that sugared unit is formed, are present in the tissue of most marine animals, and structure and property and known terrestrial plant
Polysaccharide is different, has been reported to have a variety of effects such as enhancing immunologic function, anticoagulation, hypoglycemic, antitumor.According to the literature,
Glycosaminoglycan can stimulate maturation, differentiation and the breeding of the various immunocompetent cells (BMDC, bone-marrow-derived lymphocyte) of body,
Increase the generation of cell factor such as macrophage non-specific cell poison, induction IL-1, IL-2, TNF-α, IFN-γ, IL-8 and
The expression of cytokine receptor, antigen submission, antibody is promoted to be formed, classical pathway and change approach of complement activation system etc. are made
With.Pinctada fucata glycosaminoglycan is a kind of bioactive polysaccharide with immunoloregulation function of natural origin, mainly by D-
The monose such as Glucosamine, D- aminoglucoses uronic acid, glucose, mannose, galactolipin, xylose, fucose are formed.
After pearl is adopted in pearl agriculture, annual remaining processing byproduct pearl oyster meat about 3500-4000 tons, are mostly fresh or thick
Eaten after processing, sold with fresh goods based on, cheap (about 5-10 members/kg), in Guangdong Leizhou City and Xuwen County, Beihai Fisheries Base Guangxi Province
The ground such as city, Hainan cause the huge waste of resource, and deep processing rate is relatively low, and preservation term is short, easily putrid and deteriorated, cause pearl oyster meat
Utilization are restricted.
As 103039792A patent applications disclose " it is a kind of be used to strengthening immune glycosaminoglycan from Pinctada martensii soft capsule and its
Preparation method ", its technique are by nacre whole viscera homogenate enzymolysis, centrifugation alcohol precipitation, dry the glycosaminoglycan of washing, and are added auxiliary
Soft capsule is made in material.But the defects of above method, is：Pearl oyster meat influences the effect subsequently digested without sofening treatment,
Experiments verify that obtained thick glycosaminoglycan purity is only 30%-45%, the too low Immune-enhancing effect effect that can influence that product are made of purity
Fruit.
Chewable tablets refers to the tablet that clothes make piece swallow after dissolving is chewed or suck in oral cavity, convenient to take, can swallow, chew
Containing suck or it is water-dispersible after take.Tablet surface area after chewing increases, and can promote the dissolving and absorption of medicine in vivo, even if
It is also ensured that take on time under exsiccosis, it is especially suitable for old man, child, the crowd for swallowing difficult and gastrointestinal function difference,
The burden of intestines and stomach can be reduced.
The content of the invention
First purpose of the present invention provides a kind of pinctada fucata glycosaminoglycan chewable tablets, can help to maintain normally to exempt from
Epidemic disease function, and solve the problems, such as that pinctada fucata shellfish meat higher value application rate is relatively low.
Second object of the present invention is to provide the preparation method of above-mentioned pinctada fucata glycosaminoglycan chewable tablets.
Technical scheme is used by solving first purpose of the invention：
A kind of pinctada fucata glycosaminoglycan chewable tablets, by weight percentage including following components：
Technical scheme is used by solving second purpose of the invention：
A kind of preparation method of pinctada fucata glycosaminoglycan chewable tablets, comprises the following steps：
1) take pinctada fucata meat tissue to add flooding after being homogenized, adjust pH, add neutral proteinase and trypsase is answered
Synthase solution, centrifuging and taking supernatant after enzyme deactivation, after alcohol precipitation filters and washs precipitation, obtain thick glycosaminoglycan；
2) the thick glycosaminoglycan of step 1) is taken, concentration of being dialysed after ion-exchange chromatography, is freeze-dried to obtain Hepu pearl oyster
Shellfish glycosaminoglycan；
3) pinctada fucata glycosaminoglycan and microcrystalline cellulose, mannitol, lactose are mixed evenly in proportion, then sprayed
Spill 10% Gonak softwood, be granulated, dry, addition magnesium stearate tabletting after whole grain, after sterilization packaging
Obtain pinctada fucata glycosaminoglycan chewable tablets finished product.
Preferably, the solid-liquid ratio that flooding is added in step 1) is 1:2, the 45-55 DEG C of thermostatted water under magnetic stirring apparatus effect
4-6h is bathed, with the pH to 7.0-7.6 of 0.1mol/L sodium hydrate regulator solution.
Preferably, in step 1) complex enzyme hydrolysis be with every gram of pinctada fucata shellfish meat add 10-15mg neutral proteinases and
The mass ratio of trypsase combined hydrolysis, wherein neutral proteinase and trypsase is 7:8, the 45- under magnetic stirring apparatus effect
55 DEG C of water-bath 4h, after terminating in 100 DEG C of water-baths inactivated proteases 10min, be cooled to room temperature, 8000-12000rpm centrifugations 15-
20min, supernatant is collected, obtains pinctada fucata glycosaminoglycan mother liquor.
Preferably, it is to take supernatant to add absolute ethyl alcohol to final concentration 75% that alcohol precipitation, which filters and washs precipitation, in step 1), 4 DEG C
Lower precipitation 8-12h, then 15-20min is centrifuged with 10000-12000rpm, precipitation is alternately washed 4-5 times with absolute ethyl alcohol-acetone,
Thick glycosaminoglycan is obtained in 0.030~0.035kPa, -50~-55 DEG C of vacuum freezedrying 12-36h.
Preferably, it is that thick glycosaminoglycan is entered using DEAE-52 anion-exchange columns that step 2) intermediate ion, which exchanges column chromatography,
Row purifying, applied sample amount 10mL, flow velocity 1mL/min, often pipe collect 10mL, mobile phase be respectively pure water, 0.5mol/mLNaAc,
1.5mol/mLNaCl, dialysed 24h with molecular cut off 8KD-14KD bag filter, in 0.030~0.035kPa, -50~-55
DEG C vacuum freezedrying 12-36h obtains pinctada fucata glycosaminoglycan.
Preferably, softwood processed in step 3), to be granulated be that the raw material after mixing in proportion is crossed into 100 mesh sieves, to mixed material
The 10% hydroxypropyl methyl cellulose aqueous solution is slowly sprayed by 4.5mL/10g, while is stirred continuously uniformly, softwood, mistake is made
16 mesh sieves, make softwood become particle.
Preferably, drying is that manufactured wet granular is dried into moisture under the conditions of 60-70 DEG C to be less than in step 3)
Less than 5%.
Preferably, whole grain tabletting is to crush granular solids raw material after drying in step 3), crosses 80 mesh sieves, adds quality
Fraction 1%-2% magnesium stearate, with single-punch tablet press tabletting.
Preferably, sterilization packaging is to carry out ultraviolet-sterilization using ultraviolet disinfecting machine in step 3), hinders wet bag with food-grade
Pack is packed by 20 specifications per pouch, then pouch-packaged is packaged into big packaging according to certain amount.
The beneficial effects of the invention are as follows：
1) before enzymolysis processing is carried out, hot water extraction first is carried out to raw material, enzymolysis time, saving enzyme use are saved to reach
Measure, the more preferable purpose for removing foreign protein in finished product, the hot water extraction before enzymolysis expands volume of material, makes protein in raw material
Prototype structure part changes, is denatured.
2) with biological enzymolysis process technology processing pinctada fucata shellfish meat, semifinished product extraction step is simplified, and add
Anion exchange chromatography purifies glycosaminoglycan, makes to be made product purity up to more than 85%, ensure that finished product maintains body normal
The effect of immunologic function, it is that one kind can keep the original bioactivity of glycosaminoglycan, and can improves glycosaminoglycan and carried to greatest extent
Take the new technology of rate.In addition, the residue in extraction process can prepare corresponding polypeptide products etc. according to conventional technological invention.
3) pinctada fucata glycosaminoglycan chewable tablets piece type prepared by the present invention is circular for biplane, and shape is complete, surface
Smooth, color and luster is white, uniformity, without sense of eking out a living, has good organoleptic quality, has no toxic side effect, contribute to maintenance machine
The normal immunologic function of body, is made chewing sheet form, instant edible.Preparation method is simple and easy, is adapted to industrialized production, to close
The high-valued application of Pu nacre shellfish meat provides technological innovation, has important meaning to the development for promoting pearl aquaculture and processing industry
Justice, there are preferable market prospects.
4) pinctada fucata glycosaminoglycan chewable tablets of the invention has obvious health-care efficacy, action temperature and and nontoxic secondary work
With, contribute to maintain the normal immunologic function of human organism, there are wide market prospects, advantageously account for pinctada fucata high level
Change the problem of utilizing, improve the added value of low value pearl oyster meat, avoid the wasting of resources, while solve environment caused by shellfish meat discarding
Pollution problem, promote the sustainable and healthy development of our province pearl culture and secondary industry, have great importance to environment and resource.
Brief description of the drawings
Fig. 1 is the block diagram of protective effect of the pinctada fucata glycosaminoglycan chewable tablets of the present invention to spleen；
Fig. 2 is the block diagram of the effect of pinctada fucata glycosaminoglycan chewable tablets Number of Peripheral Blood Leucocyte of the present invention；
Fig. 3 is the block diagram of effect of the pinctada fucata glycosaminoglycan chewable tablets of the present invention to bone marrow cell；
Fig. 4 is the column of effect of the pinctada fucata glycosaminoglycan chewable tablets of the present invention to spleen lymphocyte proliferation ability
Figure；
Fig. 5 is the block diagram of effect of the pinctada fucata glycosaminoglycan chewable tablets of the present invention to spleen NK cytoactives；
Fig. 6 is the column of effect of the pinctada fucata glycosaminoglycan chewable tablets of the present invention to peritoneal macrophage phagocytic activity
Figure；
Fig. 7 is the block diagram of the pinctada fucata glycosaminoglycan chewable tablets of the present invention effect horizontal to serum IL-2.
Embodiment
With drawings and examples, the invention will be further described below, so that those skilled in the art can preferably manage
The solution present invention can be simultaneously practiced, but illustrated embodiment is not as a limitation of the invention.
Embodiment 1
A kind of pinctada fucata glycosaminoglycan chewable tablets for helping to maintain normal immunological function, the chewable tablets piece weight are
201.3mg ± 2.9mg, comprising pinctada fucata glycosaminoglycan 25%, hydroxypropyl methyl cellulose 4.5%, mannitol 20%, micro-
Crystalline cellulose 40%, magnesium stearate 1.5%, surplus are lactose.
A kind of preparation method of pinctada fucata glycosaminoglycan chewable tablets, the preparation method comprise the following steps：
1) pinctada fucata meat tissue 2590g is taken, using solid-liquid ratio as 1 after homogenate:2 add flooding, make in magnetic stirring apparatus
With lower 55 DEG C of water bath with thermostatic control 4h, with the pH to 7.2 of 0.1mol/L sodium hydrate regulator solution, with every gram of pinctada fucata shellfish meat
(mass ratio of neutral proteinase and trypsase is 7 for addition 12mg neutral proteinases and trypsase combined hydrolysis:8), in magnetic
Power agitator acts on lower 55 DEG C of water-bath 4h.After end in 100 DEG C of water-baths inactivated proteases 10min, be cooled to room temperature,
12000rpm centrifuges 15min, collects supernatant, obtains pinctada fucata glycosaminoglycan mother liquor, take supernatant to add absolute ethyl alcohol extremely
Final concentration 75%, 8-12h is precipitated at 4 DEG C, then 15min, precipitation absolute ethyl alcohol-acetone alternately washing 4 are centrifuged with 12000rpm
Secondary, in 0.034kPa, -51 DEG C of vacuum freezedrying 24h obtain thick glycosaminoglycan 32.12g, and crude product yield is 1.24%；
2) the thick glycosaminoglycan of step 1) is taken, thick glycosaminoglycan is purified using DEAE-52 anion-exchange columns, on
Sample amount 10mL, flow velocity 1mL/min, often pipe collection 10mL, mobile phase is respectively pure water, 0.5mol/mLNaAc, 1.5mol/
MLNaCl, with molecular cut off 14KD bag filter dialysis 24h, closed in 0.032kPa, -51 DEG C of vacuum freezedrying 30h
Pu glycosaminoglycan from Pinctada martensii 12.46g, highly finished product yield are 0.481%；
3) pinctada fucata glycosaminoglycan and microcrystalline cellulose, mannitol, lactose are mixed evenly according to the above ratio
100 mesh sieves are crossed afterwards, and the hydroxypropyl methyl cellulose for slowly spraying mass concentration 10% by 4.5mL/10g to mixed material is water-soluble
Liquid, while be stirred continuously uniformly, neither too hard, nor too soft softwood is made, crosses 16 mesh sieves, softwood is become particle, by manufactured wet granular
Spread in enamel tray, moisture is dried under the conditions of 65 DEG C less than less than 5%, is crushed with pulverizer, 80 mesh sieves is crossed, adds
Enter the magnesium stearate of mass fraction 1.5%, with single-punch tablet press tabletting, carried out using TR-ZW-1001 ultraviolet disinfecting machines ultraviolet
Sterilization, hinder wet packaging bag with food-grade and packed by 20 specifications per pouch, then pouch-packaged is packaged into according to certain amount
Big packaging, is made pinctada fucata glycosaminoglycan chewable tablets 220 altogether, and piece weight is 201.3mg ± 2.9mg, every pearl containing Hepu
Female shellfish glycosaminoglycan 0.05g.
According to the regulation of GB 16740-2014 national food safety standard health foods, gained final product quality index such as table 1
It is shown, meet the requirements.
The pinctada fucata glycosaminoglycan chewable tablets quality index of table 1 detects
In order to verify, regulation of the obtained pinctada fucata glycosaminoglycan chewable tablets to immunologic function is made in above-described embodiment
With and its security, by building immunological function repression mouse model and oral acute toxicity rat model, observe above-mentioned implementation
Influence and Acute oral tox-hty test of the product to immune organ, immunocyte and cell factor etc. are made in example.
According to glycosaminoglycan active evaluation test result early stage and adult-mouse dose reduction formula, pinctada fucata sugar
The mouse effective dose of amine glycan is 0.25mg/d, is adult effective dose 100mg/d according to formula scales, equivalent to two panels nozzle
Chew the GAG content of piece.
From obtained pinctada fucata glycosaminoglycan chewable tablets in embodiment 1, built using normal experiment method immune
Function inhibitio Balb/c inbred mouses model (SPF levels, male and female half and half, body weight 16-18g), 60 Balb/c mouse are random
It is divided into 6 test groups, glycosaminoglycan chewable tablets low dosage (0.125mg/d), middle dosage (0.25mg/d), high dose is set respectively
(0.5mg/d) three groups, using lentinan film (0.25mg/d) as positive controls, gavage given the test agent totally 21 days, see
Examine and influence of the product to immune organ, immunocyte and cell factor etc. is made in example 1.
Test data result is as follows：
1) protective effect of the pinctada fucata glycosaminoglycan chewable tablets to spleen
As shown in figure 1, compared with Normal group, endoxan causes major injury to mouse spleen, causes spleen etc.
The weight saving of immune organ, illustrate model construction success (P<0.01).Compared with model group, lentinan film, Hepu pearl oyster
Shellfish glycosaminoglycan chewable tablets middle dose group (0.25mg/d) and high dose group (0.5mg/d) have improvement to make mouse spleen damage
With (P<0.01), illustrate that lentinan film and pinctada fucata glycosaminoglycan chewable tablets are equal to spleen atrophy caused by endoxan
With obvious improvement.Glycosaminoglycan chewable tablets low dose group (0.125mg/d), middle dose group (0.25mg/d), high agent
Dose-effect relationship (P is presented in amount group (0.5mg/d)<0.05), but compared with normal group, each group does not recover to Normal group level
(P<0.05)。
Fig. 1 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
2) effect of pinctada fucata glycosaminoglycan chewable tablets Number of Peripheral Blood Leucocyte
As shown in Fig. 2 compared with normal group, endoxan seriously reduces mouse peripheral blood leucocyte level, illustrates mould
Type successfully constructs (P<0.01).Compared with model group, lentinan film, pinctada fucata glycosaminoglycan chewable tablets low dose group
(0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d) are under mouse peripheral blood leucocyte level
Drop has improvement result (P<0.05), illustrate that it can make peripheral white blood cells are horizontal to recover, show lentinan film, pinctada fucata
Reduction of the glycosaminoglycan chewable tablets for leucocyte plays a protective role.Pinctada fucata glycosaminoglycan chewable tablets low dose group
Dose-effect relationship (P is presented in (0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d)<0.05), but with it is normal
Group is compared, and each group does not recover to Normal group level (P<0.01).
Fig. 2 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
3) effect of the pinctada fucata glycosaminoglycan chewable tablets to bone marrow cell
As shown in figure 3, compared with normal group, endoxan causes major injury to bone marrow cells in mice, illustrates model structure
Build up work((P<0.01).Compared with model group, lentinan film, pinctada fucata glycosaminoglycan chewable tablets low dose group
(0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d) have improvement to make bone marrow cells in mice damage
With (P<0.05).Pinctada fucata glycosaminoglycan chewable tablets low dose group (0.125mg/d), middle dose group (0.25mg/d), height
Dose-effect relationship (P is presented in dosage group (0.5mg/d)<0.05), and compared with normal group, middle dose group (0.25mg/d) and high dose
Group (0.5mg/d) has returned to normal group level (P>0.05).
Fig. 3 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
4) effect of the pinctada fucata glycosaminoglycan chewable tablets to spleen lymphocyte proliferation ability
As shown in figure 4, compared with normal group, endoxan causes major injury to mice spleen lymphocytes proliferation ability,
Illustrate model construction success (P<0.01).Compared with model group, lentinan film, low dose of pinctada fucata glycosaminoglycan chewable tablets
Amount group (0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d) are to mice spleen lymphocytes proliferation energy
The reduction of power has improvement result (P<0.05).Pinctada fucata glycosaminoglycan chewable tablets low dose group (0.125mg/d), middle dosage
Dose-effect relationship (P is presented in group (0.25mg/d), high dose group (0.5mg/d)<0.05), but compared with normal group, each group is not extensive
Normal group level (P is arrived again<0.01).
Fig. 4 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
5) effect of the pinctada fucata glycosaminoglycan chewable tablets to spleen NK cell killing activities
As shown in figure 5, compared with normal group, endoxan causes major injury to mice spleen NK cell killing activities, says
Bright model construction success (P<0.01).Compared with model group, lentinan film, pinctada fucata glycosaminoglycan chewable tablets low dosage
Group (0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d) are to mice spleen NK cell killing activities
Reduction has improvement result (P<0.01).Pinctada fucata glycosaminoglycan chewable tablets low dose group (0.125mg/d), middle dose group
Dose-effect relationship (P is presented in (0.25mg/d), high dose group (0.5mg/d) group<0.05), and compared with normal group, high dose group
(0.5mg/d) has returned to normal group level (P>0.05).
Fig. 5 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
6) effect of the pinctada fucata glycosaminoglycan chewable tablets to peritoneal macrophage phagocytic activity
As shown in fig. 6, compared with normal group, endoxan causes seriously to damage to Turnover of Mouse Peritoneal Macrophages phagocytic activity
Wound, illustrate model construction success (P<0.01).Compared with model group, lentinan film, pinctada fucata glycosaminoglycan chewable tablets
Low dose group (0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d) are to Turnover of Mouse Peritoneal Macrophages
The reduction of phagocytic activity has improvement result (P<0.05).Pinctada fucata glycosaminoglycan chewable tablets low dose group (0.125mg/d),
Dose-effect relationship (P is presented in middle dose group (0.25mg/d), high dose group (0.5mg/d)<0.05), but compared with normal group, each group
Normal group level (P is not recovered to<0.05).
Fig. 6 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
7) the pinctada fucata glycosaminoglycan chewable tablets effect horizontal to serum IL-2
As shown in fig. 7, compared with normal group, endoxan causes major injury to mice serum IL-2 levels, illustrates mould
Type successfully constructs (P<0.01).Compared with model group, lentinan film, pinctada fucata glycosaminoglycan chewable tablets middle dose group
To mice serum IL-2, horizontal reduction has improvement result (P for (0.25mg/d), high dose group (0.5mg/d)<0.01).Hepu
Glycosaminoglycan from Pinctada martensii chewable tablets low dose group (0.125mg/d), middle dose group (0.25mg/d), high dose group (0.5mg/d)
Dose-effect relationship (P is presented<0.05), but compared with normal group, each group does not recover to Normal group level (P<0.01).
Fig. 7 is noted：*P<0.05and**P<0.01vs. normal groups,#P<0.05and##P<0.01vs. model groups (n=10)
8) conclusion
After gavage given the test agent three weeks, pinctada fucata glycosaminoglycan chewable tablets is to being immunized suppression caused by endoxan
The positive correlation effect of dose dependent is presented in system, and middle dosage (0.25mg/d) is that effective dose (is people according to body surface area conversion
Effective dose be 100mg/d), be embodied in：Accelerate the extensive of index and spleen index, peripheral white blood cell and bone marrow cell
It is multiple, strengthen spleen lymphocyte proliferation and conversion capability, spleen NK cell killing activities and peritoneal macrophage phagocytic activity, raise blood
Clear IL-2 is horizontal, while can also preferably improve the quality of life of mouse.Compared with lentinan film, pinctada fucata osamine gathers
The immunoregulation effect of sugar chawing tablet is suitable, works gentle, has improvement result to multiple indexs, be applicable to sub-health population
The raising of immunity of organisms.
Acute oral tox-hty test refers to the experiment contaminated once or repeatedly in 24h, and the main half for determining tested material causes
Dead amount or concentration, and the poisoning manifestations of viewing test animal.In the experiment, pinctada fucata glycosaminoglycan chewable tablets group, solvent
Each 20 SD rats of control group are giving tested material process and in the 14d observation periods, rat expression behaviour, activity situation, breathing, appearance
Gesture etc. is showed no exception, no rats death.After zootomy, heart, liver, spleen, stomach, intestines, kidney and the life of rat are visually observed
The internal organs such as gland profile, color, size and ratio are grown, is showed no any exception, lesion does not occur for Rats Organs and Tissues.Rat weight is just
Often increase, the rat weight of pinctada fucata glycosaminoglycan chewable tablets group is more equal in each minute point and solvent control group
There was no significant difference (P>0.05), as shown in table 2.Therefore, under the experimental condition, pinctada fucata glycosaminoglycan chewable tablets pair
SD rat orals maximum tolerated dose (MTD) is higher than 15g/kg, and toxicity grading is nontoxic.
The SD rat peroral acute toxicities of table 2 test body weight result
Embodiment 2
A kind of pinctada fucata glycosaminoglycan chewable tablets for helping to maintain normal immunological function, the chewable tablets piece weight are
200.8mg ± 3.4mg, comprising pinctada fucata glycosaminoglycan 20%, hydroxypropyl methyl cellulose 4.0%, mannitol 20%, micro-
Crystalline cellulose 45%, magnesium stearate 2%, lactose 9%.
A kind of preparation method of pinctada fucata glycosaminoglycan chewable tablets, the preparation method comprise the following steps：
1) pinctada fucata meat tissue 1890g is taken, using solid-liquid ratio as 1 after homogenate:2 add flooding, make in magnetic stirring apparatus
With lower 55 DEG C of water bath with thermostatic control 5h, with the pH to 7.6 of 0.1mol/L sodium hydrate regulator solution, with every gram of pinctada fucata shellfish meat
(mass ratio of neutral proteinase and trypsase is 7 for addition 11mg neutral proteinases and trypsase combined hydrolysis:8), in magnetic
Power agitator acts on lower 55 DEG C of water-bath 4h.After end in 100 DEG C of water-baths inactivated proteases 10min, be cooled to room temperature,
12000rpm centrifuges 20min, collects supernatant, obtains pinctada fucata glycosaminoglycan mother liquor, take supernatant to add absolute ethyl alcohol extremely
Final concentration 75%, 8-12h is precipitated at 4 DEG C, then 15min, precipitation absolute ethyl alcohol-acetone alternately washing 4 are centrifuged with 12000rpm
Secondary, in 0.032kPa, -54 DEG C of vacuum freezedrying 18h obtain thick glycosaminoglycan 19.09g, and crude product yield is 1.01%；
2) the thick glycosaminoglycan of step 1) is taken, thick glycosaminoglycan is purified using DEAE-52 anion-exchange columns, on
Sample amount 10mL, flow velocity 1mL/min, often pipe collection 10mL, mobile phase is respectively pure water, 0.5mol/mLNaAc, 1.5mol/
MLNaCl, with molecular cut off 8KD bag filter dialysis 24h, obtained after concentration in 0.032kPa, -54 DEG C of vacuum freezedrying 18h
To pinctada fucata glycosaminoglycan 10.04g, highly finished product yield is 0.531%；
3) pinctada fucata glycosaminoglycan and microcrystalline cellulose, mannitol, lactose are mixed evenly according to the above ratio
100 mesh sieves are crossed afterwards, and the hydroxypropyl methyl cellulose for slowly spraying mass concentration 10% by 4.0mL/10g to mixed material is water-soluble
Liquid, while be stirred continuously uniformly, neither too hard, nor too soft softwood is made, crosses 16 mesh sieves, softwood is become particle, by manufactured wet granular
Spread in enamel tray, moisture is dried under the conditions of 60 DEG C less than less than 5%, is crushed with pulverizer, 80 mesh sieves is crossed, adds
Enter the magnesium stearate of mass fraction 2%, with single-punch tablet press tabletting, ultraviolet kill is carried out using TR-ZW-1001 ultraviolet disinfecting machines
Bacterium, hinder wet packaging bag with food-grade and packed by 20 specifications per pouch, then pouch-packaged is packaged into greatly according to certain amount
Packaging, is made pinctada fucata glycosaminoglycan chewable tablets 210 altogether, and piece weight is 200.8mg ± 3.4mg, every pearl oyster containing Hepu
Shellfish glycosaminoglycan 0.04g.
According to the regulation of GB 16740-2014 national food safety standard health foods, gained final product quality index such as table 3
It is shown, meet the requirements.
The pinctada fucata glycosaminoglycan chewable tablets quality index of table 3 detects
Embodiment 3
A kind of pinctada fucata glycosaminoglycan chewable tablets for helping to maintain normal immunological function, the chewable tablets piece weight are
199.3mg ± 2.1mg, comprising pinctada fucata glycosaminoglycan 15%, hydroxypropyl methyl cellulose 4.5%, mannitol 25%, micro-
Crystalline cellulose 50%, magnesium stearate 1.5%, lactose 4%.
A kind of preparation method of pinctada fucata glycosaminoglycan chewable tablets, the preparation method comprise the following steps：
1) pinctada fucata meat tissue 990g is taken, using solid-liquid ratio as 1 after homogenate:2 add flooding, are acted in magnetic stirring apparatus
Lower 55 DEG C of water bath with thermostatic control 6h, with the pH to 7.2 of 0.1mol/L sodium hydrate regulator solution, added with every gram of pinctada fucata shellfish meat
Entering 15mg neutral proteinases and trypsase combined hydrolysis, (mass ratio of neutral proteinase and trypsase is 7:8), in magnetic force
Agitator acts on lower 55 DEG C of water-bath 4h.After end in 100 DEG C of water-baths inactivated proteases 10min, be cooled to room temperature, 12000rpm
15min is centrifuged, supernatant is collected, obtains pinctada fucata glycosaminoglycan mother liquor, take supernatant to add absolute ethyl alcohol to final concentration
75%, precipitate 8-12h at 4 DEG C, then 15min is centrifuged with 12000rpm, precipitation absolute ethyl alcohol-acetone alternately washing 5 times,
0.034kPa, -54 DEG C of vacuum freezedrying 12h obtain thick glycosaminoglycan 9.03g, and crude product yield is 0.912%；
2) the thick glycosaminoglycan of step 1) is taken, thick glycosaminoglycan is purified using DEAE-52 anion-exchange columns, on
Sample amount 10mL, flow velocity 1mL/min, often pipe collection 10mL, mobile phase is respectively pure water, 0.5mol/mLNaAc, 1.5mol/
MLNaCl, with molecular cut off 8KD bag filter dialysis 24h, Hepu is obtained in 0.032kPa, -52 DEG C of vacuum freezedrying 18h
Glycosaminoglycan from Pinctada martensii 4.92g, highly finished product yield are 0.497%；
3) pinctada fucata glycosaminoglycan and microcrystalline cellulose, mannitol, lactose are pressed into weight aforementioned proportion to mix
100 mesh sieves are crossed after even, the hydroxypropyl methyl cellulose for slowly spraying mass concentration 10% by 4.5mL/10g to mixed material is water-soluble
Liquid, while be stirred continuously uniformly, neither too hard, nor too soft softwood is made, crosses 16 mesh sieves, softwood is become particle, by manufactured wet granular
Spread in enamel tray, moisture is dried under the conditions of 70 DEG C less than less than 5%, is crushed with pulverizer, 80 mesh sieves is crossed, adds
Enter the magnesium stearate of mass fraction 1.5%, with single-punch tablet press tabletting, carried out using TR-ZW-1001 ultraviolet disinfecting machines ultraviolet
Sterilization, hinder wet packaging bag with food-grade and packed by 20 specifications per pouch, then pouch-packaged is packaged into according to certain amount
Big packaging, is made pinctada fucata glycosaminoglycan chewable tablets 120 altogether, and piece weight is 199.3mg ± 2.1mg, every pearl containing Hepu
Female shellfish glycosaminoglycan 0.03g.
According to the regulation of GB 16740-2014 national food safety standard health foods, gained final product quality index such as table 4
It is shown, meet the requirements.
The pinctada fucata glycosaminoglycan chewable tablets quality index of table 4 detects
It is not limiting the scope of the present invention with the above embodiment of the present invention, embodiments of the present invention are unlimited
In this, all this kind, according to the ordinary technical knowledge and customary means of this area, is not being departed from according to the above of the present invention
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification for the other diversified forms made to said structure of the present invention, replace or
Change, all should fall within the scope and spirit of the invention.
Claims (10)
- A kind of 1. pinctada fucata glycosaminoglycan chewable tablets, it is characterised in that：Following components is included by weight percentage：
- 2. the preparation method of pinctada fucata glycosaminoglycan chewable tablets described in claim 1, it is characterised in that including following step Suddenly：1) take pinctada fucata meat tissue to add flooding after being homogenized, adjust pH, add neutral proteinase and trypsase complex enzyme Solution, centrifuging and taking supernatant after enzyme deactivation, after alcohol precipitation filters and washs precipitation, obtains thick glycosaminoglycan；2) the thick glycosaminoglycan of step 1) is taken, concentration of being dialysed after anion exchange chromatography, is freeze-dried to obtain pinctada fucata Glycosaminoglycan；3) pinctada fucata glycosaminoglycan and microcrystalline cellulose, mannitol, lactose are mixed evenly in proportion, then sprayed 10% Gonak softwood, it is granulated, dries, magnesium stearate tabletting is added after whole grain, obtained after sterilization packaging Pinctada fucata glycosaminoglycan chewable tablets finished product.
- 3. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that：Described The solid-liquid ratio for adding flooding in step 1) is 1:2, the 45-55 DEG C of water bath with thermostatic control 4-6h under magnetic stirring apparatus effect, use 0.1mol/ The pH of L sodium hydrate regulator solution to 7.0-7.6.
- 4. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that：Described Complex enzyme hydrolysis is to add 10-15mg neutral proteinases and trypsase combination water with every gram of pinctada fucata shellfish meat in step 1) Solution, the wherein mass ratio of neutral proteinase and trypsase are 7:8, the 45-55 DEG C of water-bath 4h under magnetic stirring apparatus effect, terminate The inactivated proteases 10min in 100 DEG C of water-baths afterwards, cooling, centrifuges 15-20min under the conditions of 8000-12000rpm, in collection Clear liquid, obtain pinctada fucata glycosaminoglycan mother liquor.
- 5. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that：Described It is to take supernatant to add absolute ethyl alcohol to final concentration 75% that alcohol precipitation, which filters and washs precipitation, in step 1), and 8-12h is precipitated at 4 DEG C, then 15-20min is centrifuged with 10000-12000rpm, precipitation absolute ethyl alcohol-acetone alternately washing 4-5 times, 0.030~ 0.035kPa, -50~-55 DEG C of vacuum freezedrying 12-36h obtain thick glycosaminoglycan.
- 6. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that：Described It is that thick glycosaminoglycan is purified using DEAE-52 anion-exchange columns that step 2) intermediate ion, which exchanges column chromatography, applied sample amount 10mL, flow velocity 1mL/min, often pipe collection 10mL, mobile phase is respectively pure water, 0.5mol/mLNaAc, 1.5mol/mLNaCl, is used Molecular cut off 8KD-14KD bag filter dialysis 24h, in 0.030~0.035kPa, -50~-55 DEG C of vacuum freezedrying 12- 36h obtains pinctada fucata glycosaminoglycan.
- 7. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that：Described Softwood processed in step 3), to be granulated be that the raw material after mixing in proportion is crossed into 100 mesh sieves, slow by 4.5mL/10g to mixed material The hydroxypropyl methyl cellulose aqueous solution of sprinkling 10%, while be stirred continuously uniformly, softwood is made, crosses 16 mesh sieves, becomes softwood Into particle.
- 8. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that described Drying is that manufactured wet granular is dried into moisture under the conditions of 60-70 DEG C less than 5% in step 3).
- 9. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that described Whole grain tabletting is to crush granular solids raw material after drying in step 3), crosses 80 mesh sieves, adds the hard of mass fraction 1%-2% Fatty acid magnesium, with single-punch tablet press tabletting.
- 10. the preparation method of pinctada fucata glycosaminoglycan chewable tablets according to claim 2, it is characterised in that described Step 3) in sterilization packaging be that ultraviolet-sterilization is carried out using ultraviolet disinfecting machine, hinder wet packaging bag by 20 per small with food-grade The specification packaging of bag, then pouch-packaged is packaged into big packaging according to certain amount.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110226713A (en) * | 2019-06-26 | 2019-09-13 | 甘肃犇旺旺生物科技有限公司 | A kind of ox bone cream chewable tablets and preparation method thereof |
| CN111011567A (en) * | 2019-11-28 | 2020-04-17 | 程红 | Tabletting candy with memory improving function and preparation method thereof |
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| CN101322551A (en) * | 2007-08-31 | 2008-12-17 | 辽源雷霓森药业有限公司 | Combination for increasing bone density and repairing articular cartilage |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101322551A (en) * | 2007-08-31 | 2008-12-17 | 辽源雷霓森药业有限公司 | Combination for increasing bone density and repairing articular cartilage |
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| 周小双等: "响应面法优化合浦珠母贝糖胺聚糖提取工艺", 《食品与发酵工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110226713A (en) * | 2019-06-26 | 2019-09-13 | 甘肃犇旺旺生物科技有限公司 | A kind of ox bone cream chewable tablets and preparation method thereof |
| CN111011567A (en) * | 2019-11-28 | 2020-04-17 | 程红 | Tabletting candy with memory improving function and preparation method thereof |
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Application publication date: 20180119 |