CN102550920B - Bifidus factor oral liquid and preparation method thereof - Google Patents
Bifidus factor oral liquid and preparation method thereof Download PDFInfo
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- CN102550920B CN102550920B CN 201210036233 CN201210036233A CN102550920B CN 102550920 B CN102550920 B CN 102550920B CN 201210036233 CN201210036233 CN 201210036233 CN 201210036233 A CN201210036233 A CN 201210036233A CN 102550920 B CN102550920 B CN 102550920B
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Abstract
The invention discloses a bifidus factor oral liquid. The bifidus factor oral liquid is formed by mixing Coriolus versicolor, complex amino acid powder, xylo-oligosaccharide, taurine, zinc gluconate, aspartame and potassium sorbate in a certain mass ratio. A preparation method for the bifidus factor oral liquid comprises the following steps of: crushing the Coriolus versicolor, degreasing, performing enzymolysis by using cellulose, treating by using microwaves, and performing ethanol precipitation to obtain Coriolus versicolor polysaccharide; and performing chromatography purification on the Coriolus versicolor polysaccharide and the xylo-oligosaccharide by using a diethylaminoethyl (DEAE)-cellulose column, homogenizing the obtained product and other raw materials and excipients under high pressure, performing thin mixing, sterilizing and packaging. The xylo-oligosaccharide is compatible with other raw materials and excipients, so that the bifidus factor oral liquid has a good health-care function of strengthening the immunity, and can also adjust intestinal floras, balance the nutrition of organisms, and reduce blood pressure, serum, cholesterol and blood sugar; and due to the scientific rationality of the production process, the invention has the advantages that: the bifidus factor oral liquid is easy to absorb, is high in stability, is suitable for production, and the like.
Description
Technical field
The present invention relates to a kind of health care oral liquid and preparation method thereof, what be specifically related to is a kind of Bifidus factor oral liquid and preparation method thereof.
Background technology
Oilgosaccharkdes is to make Bifidobacterium in the enteron aisle internal breeding, promote the material of bifidobacterium growth breeding, mainly comprise the extract of oligosaccharides, polysaccharose substance, short-chain fat acids material, protein hydrolysate and some protein substances, natural plants and Chinese herbal medicine.
The majority of suitability for industrialized production is oligosaccharides at present.In all functions compound sugar, xylo-oligosaccharide is a kind of that function is the strongest, effective dose is minimum, so xylo-oligosaccharide can be widely used in food, the health products.Function is strong, effective dose is little because xylo-oligosaccharide has (gram every days 0.7-1.4), the characteristic property of good stability, greatly reduce production cost at aspects such as raw materials for production, operations, improve value-added content of product, met modern production enterprise, consumer's needs.
Oral liquid is as a kind of novel form, absorb the process characteristic of traditional Chinese medicine, further refining, concentrated, embedding of decoction, sterilization get, have taking dose little, absorb the advantages such as very fast, steady quality, easy to carry and use, easy preservation, especially suitable suitability for industrialized production.But because complicated component in the oral liquid, performance, extraction process are different, cause on the market many oral liquids to exist to be long placed in and muddiness even deposited phenomenon occur, this is because wherein contain the inabundant mixing of a large amount of impurity and each active principle, this has not only affected the stability of product, also greatly reduces the absorptivity of human body oral disposition liquid.
Summary of the invention
The purpose of this invention is to provide a kind of is the oral liquid that main supplementary material is made by rainbow conk, composite aminophenol powder, xylo-oligosaccharide, taurine, zinc gluconate, Aspartame, potassium sorbate, purified water, make things convenient for human body to take, human body is had the regulating intestinal canal flora, promote Bifidobacterium Bifidum propagation; Strengthen immunity; The health care of balanced body nutrition etc.
Because the xylo-oligosaccharide and the coriolan that extract through each technique all are mixed with the impurity such as albumen, pigment and small-molecule substance, this not only affects product purity, stability, has also greatly restricted the performance of they effects.
For addressing the above problem, another object of the present invention also is: the preparation technology of this Bifidus factor oral liquid is provided, and, stability height high with the active ingredient purity of guaranteeing product is easy to absorption of human body.
The present invention is achieved by the following technical solutions:
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 10-20g, composite aminophenol powder 60-80g, xylo-oligosaccharide 45-55g, taurine 7.5-8.5g, zinc gluconate 5-8g, Aspartame 4-6g, potassium sorbate 1-3g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with the 80-95% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 3-5g, in 55 ℃ of enzymolysis 15-30min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 10-30min in the microwave of 300-500w;
E, alcohol precipitation: filter to get filtrate, filtrate is with 50-70% ethanol precipitation, and the sediment vacuum drying gets coriolan a.
Two, the edulcoration purification of xylo-oligosaccharide and coriolan
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution.
Three, the supplementary material homogeneous is rare joins
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 50-80Mpa 15-30min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
Wherein, composite aminophenol powder comprises: L-Leu, ILE, L lysine HCL, L-Phe, L-threonine, Valine, L-Trp, L-Methionine, and
Each amino acid whose mass ratio is L-Leu: ILE: L lysine HCL: L-Phe: L-threonine: Valine: L-Trp: L-Methionine=18:6:25:5:4:7:5:18.
Wherein, Aspartame is the flavouring of oral liquid, and potassium sorbate is the anticorrisive agent of oral liquid, and purified water is diluent.
This product has following health care:
1, regulating intestinal canal flora promotes Bifidobacterium Bifidum propagation;
2, improve immunity of organism, the nutrition of balance body;
3, reduce blood pressure, serum cholesterol and blood sugar;
4, improve endocrinosity;
5, low in calories;
6, generate nutriment.
The present invention is further described below in conjunction with specific embodiment.
The specific embodiment
The present invention is described further below in conjunction with specific embodiment, to help understanding content of the present invention.
Embodiment 1:
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 90% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 5g, in 55 ℃ of enzymolysis 20min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 15min in the microwave of 400w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 60% ethanol, and the sediment vacuum drying gets coriolan a.
Two, the edulcoration purification of xylo-oligosaccharide and coriolan
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution.
Three, the supplementary material homogeneous is rare joins:
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 60Mpa 20min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
Wherein, composite aminophenol powder comprises: L-Leu, ILE, L lysine HCL, L-Phe, L-threonine, Valine, L-Trp, L-Methionine, and each amino acid whose mass ratio is L-Leu: ILE: L lysine HCL: L-Phe: L-threonine: Valine: L-Trp: L-Methionine=18:6:25:5:4:7:5:18.
Wherein, Aspartame is the flavouring of oral liquid, and potassium sorbate is the anticorrisive agent of oral liquid, and purified water is diluent.
Embodiment 2
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 15g, composite aminophenol powder 70g, xylo-oligosaccharide 55g, taurine 8.5g, zinc gluconate 7g, Aspartame 6g, potassium sorbate 3g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan:
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 85% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 4g, in 55 ℃ of enzymolysis 30min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 20min in the microwave of 350w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 70% ethanol, and the sediment vacuum drying gets coriolan a.
Two, the edulcoration purification of xylo-oligosaccharide and coriolan:
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution.
Three, the supplementary material homogeneous is rare joins:
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 70Mpa 15min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid pH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
Wherein, composite aminophenol powder comprises: L-Leu, ILE, L lysine HCL, L-Phe, L-threonine, Valine, L-Trp, L-Methionine, and each amino acid whose mass ratio is L-Leu: ILE: L lysine HCL: L-Phe: L-threonine: Valine: L-Trp: L-Methionine=18:6:25:5:4:7:5:18.
Wherein, Aspartame is the flavouring of oral liquid, and potassium sorbate is the anticorrisive agent of oral liquid, and purified water is diluent.
Embodiment 3
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 10g, composite aminophenol powder 75g, xylo-oligosaccharide 55g, taurine 8g, zinc gluconate 7g, Aspartame 6g, potassium sorbate 3g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 95% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 5g, in 55 ℃ of enzymolysis 25min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 10min in the microwave of 500w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 70% ethanol, and the sediment vacuum drying gets coriolan a.
Two, the edulcoration purification of xylo-oligosaccharide and coriolan
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution.
Three, the supplementary material homogeneous is rare joins
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 80Mpa 15min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
Wherein, composite aminophenol powder comprises: L-Leu, ILE, L lysine HCL, L-Phe, L-threonine, Valine, L-Trp, L-Methionine, and each amino acid whose mass ratio is L-Leu: ILE: L lysine HCL: L-Phe: L-threonine: Valine: L-Trp: L-Methionine=18:6:25:5:4:7:5:18.
Wherein, Aspartame is the flavouring of oral liquid, and potassium sorbate is the anticorrisive agent of oral liquid, and purified water is diluent.
Embodiment 4
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Among the Bifidus factor oral liquid preparation technology of the present invention, only coriolan is carried out DEAE-cellulose chromatography purifying, concrete operations are as follows: coriolan a is dissolved in the 20ml sodium phosphate buffer, upper DEAE-cellulose column, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution;
All the other are with embodiment 1.
Embodiment 5
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Among the Bifidus factor oral liquid preparation technology of the present invention, coriolan and xylo-oligosaccharide are not carried out DEAE-cellulose chromatography purifying;
All the other are with embodiment 1.
Embodiment 6
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Among the Bifidus factor oral liquid preparation technology of the present invention, oral disposition liquid does not carry out high-pressure homogeneously, only it is carried out that supplementary material is rare joins, and concrete operations are as follows:
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
All the other are with embodiment 1.
Comparative Examples 1
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 90% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 5g, in 55 ℃ of enzymolysis 20min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 15min in the microwave of 400w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 60% ethanol, and the sediment vacuum drying gets coriolan a.
Two, supplementary material is rare joins
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and coriolan a stirring and dissolving;
B, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
All the other are with embodiment 1.
Comparative Examples 2
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 90% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 5g, in 55 ℃ of enzymolysis 20min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 30min in the microwave of 200w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 60% ethanol, and the sediment vacuum drying gets coriolan a.
Two, the edulcoration purification of xylo-oligosaccharide and coriolan
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution.
Three, the supplementary material homogeneous is rare joins:
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 40Mpa 30min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
All the other are with embodiment 1.
Comparative Examples 3
Bifidus factor oral liquid, made by the supplementary material of following weight proportioning:
Rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
Below make altogether 1000ml.
Bifidus factor oral liquid preparation technology flow process of the present invention is as follows:
One, the extraction of coriolan
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 90% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 5g, in 55 ℃ of enzymolysis 20min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 15min in the microwave of 600w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 60% ethanol, and the sediment vacuum drying gets coriolan a.
Two, the edulcoration purification of xylo-oligosaccharide and coriolan
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution.
Three, the supplementary material homogeneous is rare joins
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to about 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 90Mpa 30min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid PH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
All the other are with embodiment 1.
Comparative Examples 4
Select the supplementary material of following weight proportioning: red date 60g, ginger 50g, xylo-oligosaccharide 40g, oral glucose 100g;
The process using publication number is the embodiment 1 in the CN101396115A patent, and step is as follows: the preparation of jujube extract: select fresh clean red date 60g, the clear water flushing drains away the water,
Enter cooker, add the 120g 100 ℃ of boilings 15 minutes of purifying waste water, be cooled to making beating below 50 ℃, entered the hold-up tank lixiviate 30 minutes, plate-frame filtering, supernatant add 80g and purify waste water and stir stored for future use.
The preparation of ginger extract: select fresh clean ginger 50g, the clear water flushing drains away the water, section, add the 100g 100 ℃ of boilings 5 minutes of purifying waste water, be cooled to making beating below 50 ℃, entered the hold-up tank lixiviate 30 minutes, plate-frame filtering, supernatant adding 60g purifies waste water and stirs stored for future use.
In material-compound tank, add xylo-oligosaccharide 70 syrup 40g, jujube extract 200g, ginger extract 100g, oral glucose 100g adds the polishing 1000ml that purifies waste water, stirs, plate-frame filtering, canned, sterilization, check, warehouse-in.
Test example 1
The main component of Bifidus factor oral liquid is stachyose, is formed through water extraction by lamp.Stachyose has at double hyperplasia of the interior Bifidobacterium of the enteron aisle of making, and the regulating intestinal canal microecological balance reduces the effects such as blood endotoxin.Now adopt the high-performance liquid chromatogram determination content of stachyose, wherein stachyose reference substance (self-control, purity〉98 %).
Test method: chromatographic column is domestic Spherisorb NH2 post, mobile phase acetonitrile-water (70: 30), column temperature room temperature, flow velocity 1.2ml/min.Precision is measured the Bifidus factor oral liquid 5ml that embodiment 1-6 and Comparative Examples 1-3 make, and puts in the 100ml measuring bottle, and thin up is to scale, mixing, and the centrifugal 5min that takes a morsel, 10000r/min gets supernatant as sample solution.Accurate absorption reference substance solution and embodiment make solution 10～20 microlitres respectively, inject high performance liquid chromatograph, and external standard method is calculated, and get final product.Wherein content of stachyose is listed in table 1:
Table 1 content of stachyose relatively
| Embodiment | 1 | 2 | 3 | 4 | 5 | 6 | Comparative Examples 1 | Comparative Examples 2 | Comparative Examples 3 | Comparative Examples 4 |
| Content mg/ ml | 384.7 | 355.2 | 361.4 | 302.9 | 305.8 | 350.6 | 264.1 | 301.1 | 289.2 | 212.3 |
Through content of stachyose relatively, visible xylo-oligosaccharide is through embodiment 1-3 and the embodiment 6 of DEAE-cellulose chromatography purification process, and the gained active constituent content is apparently higher than other embodiment and Comparative Examples, and is wherein best with embodiment 1 effect.
Test example 2 accelerated tests
The Bifidus factor oral liquid that embodiment 1-6 and Comparative Examples 1-3 are made, place respectively 40 ℃ of temperature, preserved 6 months in the environment of relative humidity 75%, take a sample respectively once 3 months, 6 the end of month at duration of test, measure respectively its precipitation capacity and clarity, the results are shown in Table 2:
The comparison of table 2 precipitation capacity and clarity
The high-pressure homogeneous affinity that can strengthen between each component can improve simultaneously the stability of oral liquid, and granularity and density contrast between reducing between component prevent the oral liquid layering and precipitating, and make even tissue sticky, delicate mouthfeel.In accelerated test, clarification is without precipitation, and mouthfeel is better, and is wherein the most stable with embodiment 1 quality through high-pressure homogeneous embodiment 1-5.
Experimental example
Choose SPF level Kunming mouse, provided by Shandong University zoopery center.22 ℃ ± 2 ℃ of experimental situation temperature, relative humidity 50% ± 10%.
Dosage is selected and sample treatment: give respectively embodiment 1-6 and the made Bifidus factor oral liquid of Comparative Examples 1-3, and the concentration of tested material is 166.7ml/L, control group replaces tested material with isopyknic distilled water, gives continuously to survey every immune indexes behind the 30d.Select immediately 11 experiment mices for every group, and feed with the mouse special feed and to raise.
Experimental example 1
The mouse spleen lymphocyte transformation experiment (mtt assay) that ConA induces: the aseptic spleen of getting, place the little plate that fills an amount of aseptic Hank liquid, make cell suspension, after filtering, wash 2 times 200 eye mesh screens each centrifugal 10min of 1000r/min with Hank liquid.Then cell is suspended in the complete culture solution of 1mlRPM640, the living cell counting number, adjusting cell concentration is 3000000/ml.Cell suspension minute two holes are added in 24 well culture plates, every hole 1ml, a hole adds 75 microlitre ConA liquid therein, 5%CO2 is put in contrast in another hole, after 38 ℃ of incubators are cultivated 68h, every hole sucks supernatant 0.7ml gently, adding 0.7ml does not contain the RPM640 nutrient solution of calf serum, and every hole adds 50 microlitre MTT simultaneously, continues to cultivate 4h.Every hole adds 1ml acid isopropyl alcohol, and the piping and druming mixing dissolves purple crystal fully, then liquid is moved in the cuvette, uses ultraviolet specrophotometer, measures its OD value (OD value) at 570nm wavelength place.Calculate the poor of test hole OD value, represent the Proliferation of lymphocytes ability with this.
The data obtained is measurement data, the results are shown in Table 3:
The impact of the mouse lymphocyte transformation experiment that table 3 Bifidus factor oral liquid is induced ConA
| Group | Number of animals (only) | Lymphopoiesis ability (OD value) |
| The water control group | 11 | 0.065±0.030 |
| Embodiment 1 | 11 | 0.114±0.024 |
| Embodiment 2 | 11 | 0.108±0.022 |
| Embodiment 3 | 11 | 0.111±0.025 |
| Embodiment 4 | 11 | 0.086±0.024 |
| Embodiment 5 | 11 | 0.079±0.031 |
| Embodiment 6 | 11 | 0.101±0.026 |
| Comparative Examples 1 | 11 | 0.074±0.019 |
| Comparative Examples 2 | 11 | 0.090±0.030 |
| Comparative Examples 3 | 11 | 0.089±0.021 |
| Comparative Examples 4 | 11 | 0.068±0.042 |
By as seen from Table 3: per os gives the Bifidus factor oral liquid 30 days that the different embodiment of mouse or Comparative Examples make, its lymphopoiesis ability and water control group have conspicuousness to improve, be that it has humidification to the mouse lymphocyte conversion capability, and embodiment 1-3 successful is better than Comparative Examples 4, and is wherein best with embodiment 1 effect again.
Experimental example 2
Antibody-producting cell is measured: get defiber sheep blood, with physiological saline washing 3 times, every mouse is through lumbar injection 2% hematocrit SRBC(2000r/min, 10min) 0.2ml, the mouse of SRBC immunity after 5 days put to death, get spleen, make cell suspension.After 10g/l agarose heating for dissolving, mix the packing small test tube with the double Hank liquid of equivalent, every pipe 0.5ml adds 10% hematocrit SRBC50 microlitre, splenocyte suspension 20 microlitres again in pipe, rapidly behind the mixing, be poured on the slide of brushing the agarose thin layer, after agar solidifies, the slide level buckled be placed on the horse, put into the CO2 incubator and hatch 1.5h, then complement (pressing the 1:8 dilution with the SA buffer solution) is added in the slide frame groove, behind the continuation incubation 1.5h, counting hemolysis plaque number.
The data obtained is measurement data, the results are shown in Table 4:
Table 4 Bifidus factor oral liquid is on the impact of mouse antibodies cellulation number
| Group | Number of animals (only) | Hemolysis plaque number (1000/ full spleen) |
| The water control group | 11 | 56.18±16.20 |
| Embodiment 1 | 11 | 76.02±20.34 |
| Embodiment 2 | 11 | 74.84±24.44 |
| Embodiment 3 | 11 | 75.78±21.38 |
| Embodiment 4 | 11 | 66.25±19.41 |
| Embodiment 5 | 11 | 64.03±20.15 |
| Embodiment 6 | 11 | 70.36±21.57 |
| Comparative Examples 1 | 11 | 62.74±18.36 |
| Comparative Examples 2 | 11 | 68.44±21.39 |
| Comparative Examples 3 | 11 | 63.59±16.83 |
| Comparative Examples 4 | 11 | 60.48±18.37 |
By as seen from Table 4, per os gave each embodiment of mouse or the prepared Bifidus factor oral liquid of Comparative Examples 30 days, the corresponding mouse group of embodiment 1-3 antibody-producting cell number and water control group and other groups are relatively, significant difference is all arranged, namely this oral liquid can strengthen mouse antibodies cellulation number, and embodiment 1 prepared Oilgosaccharkdes effect is optimum.
Experimental example 3
The mensuration of mice serum hemolysin: get defiber sheep blood, with physiological saline washing 3 times, every mouse is through lumbar injection 2% hematocrit SRBC (2000r/min, 10min) 0.2ml, immunity was extractd eyeball of mouse and is got blood in centrifuge tube after 5 days, place about 1h, solidification blood and tube wall are peeled off, centrifugal, collect serum., with the serum doubling dilution different dilution factor serum are placed respectively in the Microhemagglutination breadboard with physiological saline, every hole 100 microlitres add 0.5%SRBC suspension 100 microlitres again, and mixing is put in the wet box, in 37 ℃ of incubation 3h, and record hemagglutination degree.The calculating antibody product.
The data obtained is measurement data, the results are shown in Table 5:
Table 5 Bifidus factor oral liquid is on the impact of mouse mice serum hemolysin level
| Group | Number of animals (only) | The antibody product |
| The water control group | 11 | 101.4±29.1 |
| Embodiment 1 | 11 | 134.9±30.8 |
| Embodiment 2 | 11 | 127.8±23.9 |
| Embodiment 3 | 11 | 132.5±21.4 |
| Embodiment 4 | 11 | 120.1±25.6 |
| Embodiment 5 | 11 | 116.3±22.3 |
| Embodiment 6 | 11 | 130.5±24.4 |
| Comparative Examples 1 | 11 | 108.0±28.2 |
| Comparative Examples 2 | 11 | 119.4±23.3 |
| Comparative Examples 3 | 11 | 112.6±26.5 |
| Comparative Examples 4 | 11 | 106.5±21.7 |
By as seen from Table 5, per os gave each embodiment of mouse or the prepared Bifidus factor oral liquid of Comparative Examples 30 days, the mouse antibodies product of embodiment 1-3 group is compared with other groups with the water control group, significant difference is all arranged, namely this oral liquid can improve the mice serum hemolysin level, and embodiment 1 best results.
Embodiment 4
NK cells in mice determination of activity (lactate dehydrogenase L DH determination method): 24h washes cultivations of going down to posterity of YAC-1 cell 3 times with Hank liquid with front before the test, is 400000/ml with RPM640 complete culture solution adjustment cell concentration.The mouse dislocation is put to death, the aseptic spleen of getting, the preparation splenocyte suspension is washed 3 times with Hank liquid, each centrifugal 10min of 1000rpm.Adding 1ml, to contain the RPM640 complete culture solution of 10% calf serum resuspended again, and with the blue dyeing counting of phenol (viable count should more than 95%), adjusting cell concentration be 20000000/ml, makes the effect target than being 50:1.
Get each 100 microlitre of target cell and splenocyte, add U-shaped 96 well culture plates, target cell Spontaneous release hole adds target cell and each 100 microlitre of nutrient solution, the maximum release aperture of target cell adds target cell and each 100 microlitre of 1%NP40, above-mentioned every three parallel holes of all establishing, 37 ℃, 5%CO2 incubator cultivation 4h, with 96 orifice plates with the centrifugal 5min of 1500r/min, every hole is drawn supernatant 100 microlitres and is changed in the ELISA Plate, add simultaneously LDH matrix liquid 100 microlitres, reaction 10min, then every hole adds the Hcl30 microlitre cessation reaction of 1mol/l, measures the OD value at the 490nm place with ELIASA.By formula calculate the NK cytoactive: NK cytoactive %=(reacting hole OD-Spontaneous release hole OD)/(maximum release aperture OD-Spontaneous release hole OD) * 100 the results are shown in Table 6:
Table 6 Bifidus factor oral liquid is on the impact of NK cells in mice activity
| Group | Number of animals (only) | NK cytoactive (%) |
| The water control group | 11 | 23.47±8.67 |
| Embodiment 1 | 11 | 47.00±11.62 |
| Embodiment 2 | 11 | 43.40±10.75 |
| Embodiment 3 | 11 | 45.40±10.16 |
| Embodiment 4 | 11 | 40.36±9.57 |
| Embodiment 5 | 11 | 38.22±10.34 |
| Embodiment 6 | 11 | 46.13±11.27 |
| Comparative Examples 1 | 11 | 37.57±9.46 |
| Comparative Examples 2 | 11 | 40.12±10.39 |
| Comparative Examples 3 | 11 | 36.25±11.08 |
| Comparative Examples 4 | 11 | 27.31±10.85 |
By as seen from Table 6, per os gave each embodiment of mouse or the prepared Bifidus factor oral liquid of Comparative Examples 30 days, between its NK cytoactive and control group significant difference is arranged more all, namely this oral liquid can strengthen the NK cytoactive of mouse, and embodiment 1-3 is better than other embodiment and Comparative Examples, and embodiment 1 best results.
Can be found out by above experimental example, the Bifidus factor oral liquid that the present invention makes can strengthen the mouse lymphocyte conversion capability, strengthen mouse antibodies cellulation number, improve the mice serum hemolysin level, strengthen the NK cytoactive of mouse, have conveniently and take, product purity is high, the characteristic of being convenient to absorb, and have the enhancing immunity of organism, the endotrophic effect of balance.
Claims (2)
1. Bifidus factor oral liquid, it is characterized in that, made by the supplementary material of following weight proportioning: rainbow conk 10-20g, composite aminophenol powder 60-80g, xylo-oligosaccharide 45-55g, taurine 7.5-8.5g, zinc gluconate 5-8g, Aspartame 4-6g, potassium sorbate 1-3g;
The preparation method is:
One, the extraction of coriolan:
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with the 80-95% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 3-5g, in 55 ℃ of enzymolysis 15-30min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 10-30min in the microwave of 300-500w;
E, alcohol precipitation: filter to get filtrate, filtrate is with 50-70% ethanol precipitation, and the sediment vacuum drying gets coriolan a;
Two, the edulcoration purification of xylo-oligosaccharide and coriolan:
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution;
Three, the supplementary material homogeneous is rare joins:
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 50-80Mpa 15-30min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid pH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
Wherein, composite aminophenol powder comprises: L-Leu, ILE, L lysine HCL, L-Phe, L-threonine, Valine, L-Trp, L-Methionine, and L-Liang An Suan ︰ L-Yi Liang An Suan ︰ 1B Yan Suan Yan ︰ L-Ben Bing An Suan ︰ L-Su An Suan ︰ L-Xie An Suan ︰ L-Se An Suan ︰ L-Methionine=18:6:25:5:4:7:5:18.
2. Bifidus factor oral liquid as claimed in claim 1, it is characterized in that, made by the supplementary material of following weight proportioning: rainbow conk 20g, composite aminophenol powder 60g, xylo-oligosaccharide 50g, taurine 8g, zinc gluconate 6g, Aspartame 5g, potassium sorbate 2g;
The preparation method is:
One, the extraction of coriolan:
A, pulverizing: cross 80 mesh sieves after manyzoned polypore sporophore is pulverized;
B, degreasing: reflux 3 times with 90% alcohol degreasing, filter merging filtrate;
C, except cell membrane: filtrate adds cellulase 5g, in 55 ℃ of enzymolysis 20min, has reacted the rear boiling water enzyme 5min that goes out;
D, microwave treatment: add again the distilled water of 20 times of rainbow conk weight, in 90 ℃, process 15min in the microwave of 400w;
E, alcohol precipitation: filter to get filtrate, filtrate is precipitated with 60% ethanol, and the sediment vacuum drying gets coriolan a;
Two, the edulcoration purification of xylo-oligosaccharide and coriolan:
A, dissolving: by the accurate weighing xylo-oligosaccharide of prescription, the coriolan a that extracts among xylo-oligosaccharide and () is dissolved in the 20ml sodium phosphate buffer;
B, DEAE-cellulose chromatography purifying: with DEAE-cellulose column on the above-mentioned buffer solution, with flow velocity 3BV/h wash-out, collect eluent b with 0.3mol/LNaCl solution;
Three, the supplementary material homogeneous is rare joins:
A, by prescription accurate weighing other supplementary materials, with purified water dense join to fill be heated to 80 ℃, add supplementary material and eluent b stirring and dissolving;
B, high-pressure homogeneous: in the high-pressure homogeneous processing of 60Mpa 20min, make the supplementary material refinement, evenly mix, thus the absorptivity of raising human body;
C, liquid is filled into rare filling of joining by filter; The NaOH adjusting liquid pH value that adds amount of preparation 0.3% in dilute preparing tank is 7.5, and adds purified water to amount of preparation, the packing sterilization;
Wherein, composite aminophenol powder comprises: L-Leu, ILE, L lysine HCL, L-Phe, L-threonine, Valine, L-Trp, L-Methionine, and L-Liang An Suan ︰ L-Yi Liang An Suan ︰ 1B Yan Suan Yan ︰ L-Ben Bing An Suan ︰ L-Su An Suan ︰ L-Xie An Suan ︰ L-Se An Suan ︰ L-Methionine=18:6:25:5:4:7:5:18.
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| CN101249259A (en) * | 2008-03-04 | 2008-08-27 | 上海师范大学 | High content and high activity oral polysaccharide-peptide and preparing method and application of the same |
| CN101336661A (en) * | 2008-07-14 | 2009-01-07 | 西北农林科技大学 | Non-specific complex polysaccharide immunity milk and preparation method thereof |
| CN101861959A (en) * | 2010-01-21 | 2010-10-20 | 海南京润珍珠生物技术股份有限公司 | Pearl nutritional vinegar oral liquid |
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| CN101249259A (en) * | 2008-03-04 | 2008-08-27 | 上海师范大学 | High content and high activity oral polysaccharide-peptide and preparing method and application of the same |
| CN101336661A (en) * | 2008-07-14 | 2009-01-07 | 西北农林科技大学 | Non-specific complex polysaccharide immunity milk and preparation method thereof |
| CN101861959A (en) * | 2010-01-21 | 2010-10-20 | 海南京润珍珠生物技术股份有限公司 | Pearl nutritional vinegar oral liquid |
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| 益生元―双歧杆菌生长促进因子;胡学智;《工业微生物》;20050630(第02期);第50-60页 * |
| 胡学智.益生元―双歧杆菌生长促进因子.《工业微生物》.2005,(第02期), |
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Inventor after: Lou Xiuyu Inventor after: Zhou Longcai Inventor after: Wu Yunjiang Inventor after: Lou Genfu Inventor before: Lou Xiuyu Inventor before: Zhou Longcai Inventor before: Wu Yunjiang |
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