CN110179964A - A kind of preparation method of functional peanut small peptide carrier - Google Patents
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Abstract
The invention discloses a kind of preparation methods of functional peanut small peptide carrier, including, inner aqueous phase solution containing sucrose and functional peanut small peptide is mixed with the methylene chloride oil-phase solution containing soybean lecithin, cholesterol and olein, colostrum is made, then the outer aqueous phase solution containing glucose and L-lysine is added in colostrum, vortex concussion, which emulsifies, is made emulsion, and last rotary evaporation removal methylene chloride is up to functional peanut small peptide carrier.Preparation method simple possible provided by the present invention, process conditions are mild, and preparation flow is controllable and the time is short, and preparation cost is low, are that one kind had not only been able to achieve the functional activity of defencive function peanut oligopeptides but also had been able to achieve the preparation method of the carrier of sustained release；The final encapsulation rate of the preparation-obtained carrier of the present invention is 82.00 ± 0.25%, meets the requirement of National Pharmacopeia；Preparation method application easy to spread provided by the present invention, actual application prospect is wide, significant.
Description
Technical field
The invention belongs to Farm products processing technical fields, and in particular to a kind of system of functional peanut small peptide carrier
Preparation Method.
Background technique
Protein is the second largest nutrient in peanut, and according to ASSOCIATE STATISTICS, in the world, peanut is for extracting oil and eating
Ratio is respectively 54% and 36%, the rich content of protein resource in the grouts after oil expression, but it is by reasonable benefit
With to cause the waste of serious protein resource, or even causing certain destruction to environment.
With constantly exploding for world population, the diet structure of people is also constantly reformed and is adjusted, and causes the shortage of food
With the deficiency of protein supply, having become influences human health, stable and sustainable development serious problems.
Studies have shown that protein extracts etc. by enzymatic hydrolysis, fermentation or soda acid, to will form molecular weight after process different
Peptide fragment mixture, rich in nutrition content, amino acid classes are complete, it is necessary to which amino acid content is up to 38.29%, and has had
Complete independent mechanism of absorption and it is better than free amino acid, peptide is formed after different aminoacids permutation and combination has the function of different work
Property.Currently, more hypotensive activity and the antioxidation activity for functional peanut small peptide of research, some scholars report
The functional activities such as antibacterial/antibacterial of functional peanut small peptide, immunological regulation, but its research is relatively fewer.Development functionality peanut
Small peptide not only can solve the case where peanut protein wasting of resources, can also alleviate and even solve human foods and protein confession
The deficiency given.
Currently, numerous study the optimum preparation condition for concentrating on functional peanut small peptide, functional characteristic exploitation and structure effect
In the research and development of relationship, but after functional oligopeptides entrance in vivo, after the effect of direct oral cavity and gastrointestinal tract digestive ferment, knot
Structure can change, and functional activity is caused to change.
A kind of it can protect functional oligopeptides not by the delivery of the destructions such as oral cavity and gastrointestinal tract digestive ferment in conclusion establishing
Body, meanwhile, slow releasing function can be played, multiple medication is avoided to bring pain to sufferer, becomes the crucial skill for developing functional oligopeptides
Art problem.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of preparation sides of functional peanut small peptide carrier
Method.
In order to achieve the above objectives, the invention adopts the following technical scheme:
A kind of preparation method of functional peanut small peptide carrier, including, it will be containing sucrose and functional peanut small peptide
Inner aqueous phase solution is mixed with the methylene chloride oil-phase solution containing soybean lecithin, cholesterol and olein and is made just
Outer aqueous phase solution containing glucose and L-lysine is then added in colostrum by cream, and vortex concussion, which emulsifies, is made emulsion, most
Rotate evaporative removal methylene chloride afterwards up to functional peanut small peptide carrier.
In the above-mentioned technical solutions, in the methylene chloride oil-phase solution, the concentration of the soybean lecithin is 14-
17.5mg/mL, preferably 16mg/mL.
In the above-mentioned technical solutions, in the methylene chloride oil-phase solution, the concentration of the cholesterol is 11.2-
13.5mg/mL, preferably 12mg/mL.
In the above-mentioned technical solutions, in the methylene chloride oil-phase solution, the concentration of the olein is
10.8-12.8mg/mL, preferably 12mg/mL.
Further, in the above-mentioned technical solutions, the inner aqueous phase solution the preparation method comprises the following steps: weighing sugarcane respectively according to the ratio
Sugar and functional peanut small peptide, are dissolved in deionized water for sucrose, stir evenly obtained sucrose solution, then that functional peanut is short
Peptide is added, and mixes to obtain the final product.
Preferably, in the above-mentioned technical solutions, the concentration of the sucrose solution is 3-5%w/v, preferably 5%w/v.
Still further, in the above-mentioned technical solutions, in the outer aqueous phase solution, the concentration of the glucose is 6.5-
8.8mg/mL, preferably 7.5%w/v.
Still further, in the above-mentioned technical solutions, in the outer aqueous phase solution, the concentration of the L-lysine is
32-45mmol/L, preferably 40mmol/L.
Further, in the above-mentioned technical solutions, in the preparation process of the colostrum, the methylene chloride oil mixes
The volume ratio of liquid and the inner aqueous phase solution is 1:(0.8-1.2).
Further, in the above-mentioned technical solutions, in the preparation process of the emulsion, the colostrum and the outer water
The volume ratio of phase solution is 2:(2.8-3.5).
Also further, in the above-mentioned technical solutions, the mixing method of the colostrum is high-speed stirred homogenate, and revolving speed is
18000-22000rpm, mixing time 9-12min.
Also further, in the above-mentioned technical solutions, the vortex concussion emulsification times of the emulsion are 8-15s, preferably
10s。
Still further preferably, in the above-mentioned technical solutions, the rotary evaporation is low speed medium temperature rotary evaporation, evaporation temperature
Degree and mixing speed are respectively 32-37.5 DEG C and 9-11.5rpm.
Another aspect of the present invention additionally provides the functional peanut small peptide carrier that above-mentioned preparation method is prepared.
Another aspect of the invention additionally provides above-mentioned functional peanut small peptide carrier and is preparing blood-pressure reducing health care product, antioxygen
Change the application in health care product bacteriostasis antibiosis health care product and immunological regulation health care product.
Advantages of the present invention:
(1) the preparation method simple possible of functional peanut small peptide carrier provided by the present invention, process conditions are mild,
Preparation flow is controllable and the time is short, and preparation cost is low, be a kind of functional activity for being not only able to achieve defencive function peanut oligopeptides but also
It is able to achieve the preparation method of the carrier of sustained release；
(2) the final encapsulation rate for the carrier that preparation method provided by the present invention is prepared is 82.00 ± 0.25%,
And meet requirement of the National Pharmacopeia for slow-releasing system；
(3) preparation method of functional peanut small peptide carrier provided by the present invention, method are simple, low in cost, easy
In popularization and application, actual application prospect is wide, and theoretical and practical significance is great.
Detailed description of the invention
Fig. 1 be the embodiment of the present invention provided by functional peanut small peptide carrier preparation method in technological parameter with
The relation curve of carrier encapsulation rate；
Fig. 2 be the embodiment of the present invention provided by functional peanut small peptide carrier preparation method in prescribed parameters with
The relation curve of carrier encapsulation rate；
Fig. 3 is the apparent and structural schematic diagram of functional peanut small peptide carrier provided by the embodiment of the present invention.
Specific embodiment
In the following with reference to the drawings and specific embodiments, specific embodiments of the present invention will be described in further detail.
The following examples are intended to illustrate the invention, but is not limited to protection scope of the present invention, protection model of the invention
It encloses and is subject to claims.
Unless otherwise specified, experiment reagent used in the embodiment of the present invention and material etc. are commercially available, if not having
Body indicates, the conventional means that technological means used in the embodiment of the present invention is well known to the skilled person.
Soybean lecithin employed in the embodiment of the present invention, cholesterol, olein and L-lysine are purchased from
Beijing Suo Laibao Science and Technology Ltd.
The measuring method of encapsulation rate is as follows in the embodiment of the present invention:
Functional peanut small peptide obtained delivery liquid solution is transferred in 50mL centrifuge tube, 4 DEG C in refrigerated centrifuge
It is centrifuged, is centrifuged 10min under 2500rpm；Liquid is discarded supernatant, then is centrifuged again after washing precipitating with above-mentioned outer aqueous phase solution, is collected
It is centrifuged the substance of bottom of the tube, and the suitable 10% broken capsule of Triton X-100 ethanol solution is added, and on vortex suspension instrument
Oscillation a moment keeps brokenly capsule abundant；After the completion, using UPLC method measurement function peanut oligopeptides content.
Computational envelope rate according to the following formula:
Apparent and structure observation using optical microscopy (Fig. 3 a), laser confocal microscope (Fig. 3 b) and electron cryo-microscopy into
Row (Fig. 3 c).
The measuring method of permeability is as follows in the embodiment of the present invention:
Prepare sample according to optimization formula, be put in 4 DEG C and at room temperature respectively, in after a certain period of time (12h, for 24 hours, 48h,
72h, 96h, 120h and 144h) measurement liposome encapsulation variation, calculate the percolation ratio of drug, draw percolation ratio and become at any time
The curve of change.
The percolation ratio of drug is calculated as follows:
The measuring method of sustained release performance is as follows in the embodiment of the present invention:
Three batches of liposomes are prepared according to prescription, liposome is resuspended with appropriate physiological saline for centrifuging and taking precipitating；In low speed magnetic
It under power stirring, is taken in 5mL Liposomal suspensions to the centrifuge tube for the 15mL for having numbered (1-15) respectively, separately adds 5mL physiology salt
Water is as dissolution medium；Centrifuge tube is put in 37 DEG C, in the shaking table of 30rpm, respectively at setting time point (0.5h, 1h, 2h,
4h, 8h, 12h, for 24 hours, 36h, 48h, 60h, 72h, 84h, 96h, 108h and 120h) take out the 15mL centrifuge tube of same number,
It is resuspended after 2500rpm centrifugation 10min with physiological saline, again centrifuge washing；Liquid is discarded supernatant, is added 5% (w/v's)
TritonX-100 ethanol solution breaks capsule, surveys its light absorption value at 663nm, calculates release percentage and draws release profiles.
The preparation (%) of drug is calculated as follows:
Wherein:
Q0, the dose of release beginning proliposome encapsulating；
QT, remaining dose in liposome when being discharged into t.
The multivesicular liposome of different cholesterol dosages (4mg/mL, 12mg/mL, 20mg/mL) is prepared respectively, investigates cholesterol
Influence of the dosage to liposome release performance；The multivesicular liposome of different olein dosages is prepared respectively, investigates three oil
Influence of the acid glyceride dosage (4mg/mL, 12mg/mL, 20mg/mL) to liposome release performance.
Embodiment 1 compares influence of the different technical parameters to carrier encapsulation rate
Specific step is as follows:
S1, recipe quantity sucrose is dissolved in deionized water, is stirred well to and is completely dissolved, prepared sucrose solution, then add
Enter recipe quantity functional peanut small peptide, above-mentioned mixed liquor is known as inner aqueous phase；
S2, recipe quantity glucose, L-lysine (assistant for emulsifying agent) are dissolved in deionized water, are stirred well to completely molten
Solution, prepares glucose and L-lysine, above-mentioned mixed liquor are known as outer aqueous phase；
S3, the soybean lecithin, cholesterol, olein of recipe quantity are dissolved in dichloromethane solution, configuration is big
Beans lecithin, cholesterol, olein, above-mentioned solution are known as oily phase；
S4, the mixing (grease phase 1:1) by the oil-phase solution 3mL in the inner aqueous phase solution 3mL and step S3 in step S1,
It is stirred on high-speed homogenization machine, preparation acquired solution is known as colostrum；
S5, (colostrum and outer aqueous phase volume ratio in gained colostrum is added in step S4 in outer aqueous phase solution obtained by 6mL step S2
2:3), vortex concussion emulsification, preparation acquired solution are known as emulsion；
S6, gained emulsion in step S5 is placed in Rotary Evaporators, removes methylene chloride, the final acquired solution for preparing is function
It can property peanut oligopeptides carrier.
In step S4, influence of the different homogenate speed for encapsulation rate as a result, seeing Fig. 1 a.
By, it can be seen that revolving speed is within the scope of 12000-25000rpm, encapsulation rate increases with the increase of revolving speed in Fig. 1 a
Greatly, but after 20000rpm not there is significant difference in ascendant trend, so selecting revolving speed in step S4 is 20000rpm.
In step S4, influence of the different high-speed homogenization times for encapsulation rate as a result, seeing Fig. 1 b.
By it can be seen that Homogenization time is out of 5-25min, encapsulating takes the lead in reducing after increasing in Fig. 1 b, when 10min, reaches
To maximum, so selecting step S4 high speed Homogenization time is 10min.
In step S5, influence of the different vortexs concussion times for encapsulation rate as a result, seeing Fig. 1 c.
By, it can be seen that vortex shakes the time out of 5-30s, encapsulating takes the lead in reducing after increasing, and when 10s reaches in Fig. 1 c
To maximum, so selecting the vortex concussion time in step S5 is 10s.
Embodiment 2 compares influence of the different prescribed parameters to carrier encapsulation rate
Specific step is as follows:
S1, recipe quantity sucrose is dissolved in deionized water, is stirred well to and is completely dissolved, prepared sucrose solution, then add
Enter recipe quantity functional peanut small peptide, above-mentioned mixed liquor is known as inner aqueous phase；
S2, recipe quantity glucose, L-lysine (assistant for emulsifying agent) are dissolved in deionized water, are stirred well to completely molten
Solution, prepares glucose and L-lysine, above-mentioned mixed liquor are known as outer aqueous phase；
S3, the soybean lecithin, cholesterol, olein of recipe quantity are dissolved in dichloromethane solution, configuration is big
Beans lecithin, cholesterol, olein, above-mentioned solution are known as oily phase；
S4, by the mixing (grease phase 1:1) of solution 3mL in solution 3mL in step S1 and step S3, stirred on high-speed homogenization machine
It mixes, preparation acquired solution is known as colostrum；
S5, rotation in colostrum obtained by step S4 (colostrum and outer aqueous phase volume ratio 2:3) is added in 6mL step S2 acquired solution
Whirlpool concussion emulsification, preparation acquired solution are known as emulsion；
S6, gained emulsion in step S5 is placed in Rotary Evaporators, removes methylene chloride, the final acquired solution for preparing is function
It can property peanut oligopeptides carrier.
In step S2, influence of the dosage of different L-lysines for encapsulation rate as a result, seeing Fig. 2 a.
By, it can be seen that the content of L-lysine is within the scope of 20-100mmol/L, encapsulating takes the lead in dropping after increasing in Fig. 2 a
It is low, but 40mmol/L reaches maximum value, so selecting L-lysine content in step S2 is 40mmol/L.
In step S3, influence of the dosage of different soybean lecithins, cholesterol and olein for encapsulation rate
As a result, seeing Fig. 2 b-2d.
By in Fig. 2 b-2d it can be seen that selection encapsulation rate highest point, so, soybean lecithin in step S3, cholesterol and
The dosage of olein is respectively as follows: 16mg/mL, 12mg/mL and 12mg/mL.
It is illustrated in figure 3 preparation-obtained functional peanut small peptide carrier in optimal conditions of the embodiment of the present invention
Apparent and structural schematic diagram；Wherein, as can be seen that preparing resulting carrier in egg shape or circular shape, internal more capsules from Fig. 3 a
A large amount of functional peanut small peptides are encapsulated in bubble structure, internal compartment is clear in structure, and olein is presented bright at tie point
Spot；In Fig. 3 b, carrier particle is clear, and obvious phosphatide fragment does not occur in surrounding, and the double layerings of external phosphatide cause dense and uniform；Fortune
For carrier integral particle structure by Fig. 3 c as it can be seen that wherein internal voids are that sublimation ribbon walks moisture gap, diamond shape, phosphorus is presented in internal compartment
The double layering gaps of rouge are filled by olein, ensure that the stabilization of integral skeleton structure.
Finally, being not intended to limit the scope of the present invention the above is only preferred embodiment of the invention.It is all this
Within the spirit and principle of invention, any modification, equivalent replacement, improvement and so on should be included in protection model of the invention
Within enclosing.
Claims (10)
1. a kind of preparation method of functional peanut small peptide carrier, which is characterized in that including sucrose and functionality flower will be contained
The inner aqueous phase solution of raw small peptide is mixed with the methylene chloride oil-phase solution containing soybean lecithin, cholesterol and olein
Colostrum is made, then the outer aqueous phase solution containing glucose and L-lysine is added in colostrum, vortex concussion emulsification is made
Emulsion, last rotary evaporation removal methylene chloride is up to functional peanut small peptide carrier.
2. preparation method according to claim 1, which is characterized in that in the methylene chloride oil-phase solution:
The concentration of the soybean lecithin is 14-17.5mg/mL, preferably 16mg/mL；
And/or the concentration of the cholesterol is 11.2-13.5mg/mL, preferably 12mg/mL；
And/or the concentration of the olein is 10.8-12.8mg/mL, preferably 12mg/mL.
3. preparation method according to claim 1 or 2, which is characterized in that the inner aqueous phase solution the preparation method comprises the following steps: pressing
Proportion weighs sucrose and functional peanut small peptide respectively, and sucrose is dissolved in deionized water, stirs evenly obtained sucrose solution, then
Functional peanut small peptide is added, is mixed to obtain the final product.
4. preparation method according to claim 3, which is characterized in that the concentration of the sucrose solution is 3-5%w/v, excellent
It is selected as 5%w/v.
5. preparation method according to claim 1-4, which is characterized in that in the outer aqueous phase solution:
The concentration of the glucose is 6.5-8.8mg/mL, preferably 7.5%w/v；
And/or the concentration of the L-lysine is 32-45mmol/L, preferably 40mmol/L.
6. preparation method according to claim 1-5, which is characterized in that
In the preparation process of the colostrum, the volume ratio of the methylene chloride oil-phase solution and the inner aqueous phase solution is 1:
(0.8-1.2)；
And/or in the preparation process of the emulsion, the volume ratio of the colostrum and the outer aqueous phase solution is 2:(2.8-
3.5)。
7. preparation method according to claim 1-6, which is characterized in that
The mixing method of the colostrum is high-speed stirred homogenate, revolving speed 18000-22000rpm, mixing time 9-12min；
And/or the vortex concussion emulsification times of the emulsion are 8-15s, preferably 10s.
8. preparation method according to claim 1-7, which is characterized in that the rotary evaporation is the rotation of low speed medium temperature
Turn evaporation, evaporating temperature and mixing speed are respectively 32-37.5 DEG C and 9-11.5rpm.
9. the functional peanut small peptide carrier that the described in any item preparation methods of claim 1-8 are prepared.
10. functional peanut small peptide carrier as claimed in claim 9 is to prepare blood-pressure reducing health care product, antioxidant health-care product antibacterial
Application in antibiotic health care product and immunological regulation health care product.
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2019
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|---|---|---|---|---|
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