CN114601019A - Application of placenta water-soluble extract as wall material for preparing probiotic microcapsules - Google Patents
Application of placenta water-soluble extract as wall material for preparing probiotic microcapsules Download PDFInfo
- Publication number
- CN114601019A CN114601019A CN202210324482.0A CN202210324482A CN114601019A CN 114601019 A CN114601019 A CN 114601019A CN 202210324482 A CN202210324482 A CN 202210324482A CN 114601019 A CN114601019 A CN 114601019A
- Authority
- CN
- China
- Prior art keywords
- placenta
- wall material
- soluble extract
- water
- microcapsule
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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Images
Classifications
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Abstract
The invention discloses application of a placenta water-soluble extract as a wall material to preparation of a probiotic microcapsule. The application comprises the following steps: dropwise adding a procyanidine solution into the placenta water-soluble extract solution, uniformly stirring, and performing thermal denaturation treatment to obtain a microcapsule wall material; mixing probiotic bacteria liquid with microcapsule wall materials, adding transglutaminase to uniformly disperse the transglutaminase, adding the mixed liquid into preheated soybean oil, and reacting the system under stirring; after the liquid drops are converted into gel particles under the action of enzyme, the microcapsule particles are centrifugally collected to obtain the probiotic microcapsule. The placenta water-soluble extract anthocyanin provided by the invention is used as a wall material, aiming at the problem of low survival rate of probiotics, the procyanidine is added, the survival rate of the probiotics in the preservation time and the bioavailability of the procyanidine are obviously improved, meanwhile, the activity of the placenta water-soluble extract can be exerted, the activity of a single wall material is increased, and the function of utilizing the biological activity of the wall material is improved.
Description
Technical Field
The invention relates to the technical field of microcapsules, in particular to application of a placenta water-soluble extract as a wall material to preparation of probiotic microcapsules.
Background
The microcapsule technology is a technology in which a solid, liquid or gaseous substance sensitive to the surrounding environment is embedded in a capsule and the release can be controlled in a specific environment. The size of the microcapsule particle generally varies with the processing technology, and the particle size range is generally 0.1 to 1000 μm. The microcapsule is divided into two parts of a wall material and a core material, wherein the substance embedded in the microcapsule is called the core material, the substance embedded outside the microcapsule is the wall material, and the wall material is generally a natural polymer material, a semi-synthetic polymer material, a fully synthetic polymer material and an inorganic material. The microcapsule technology has wide application in various fields and becomes a research hotspot all the time.
In recent years, with the increasing awareness of the environment, ecology, sustainable development and other problems, the use of environmentally friendly microcapsule wall materials is becoming an important research direction today. In the food industry, microcapsule technology has unique functions and performances and takes a remarkable advantage. The selection of the wall material plays a crucial role in the performance and the application of the microcapsule, and three types of wall materials which are widely applied nowadays are generally selected from degradable natural polymers, saccharides, proteins and emerging biopolymers. The wall material should have good film-forming properties, non-toxicity and good biocompatibility, while not affecting the properties of the core material.
The human placenta, also known as placenta hominis, is a traditional Chinese medicine in traditional Chinese medicine, has the functions of warming kidney, replenishing essence, benefiting qi and nourishing blood, and is often boiled to be eaten in folk traditional medicine so as to enhance the body resistance and treat various diseases such as body deficiency, asthma and the like.
The key to the role of probiotics in maintaining health and preventing diseases is the number of viable probiotics and the ability to reproduce after reaching a particular site, which determine the strength or effect of the benefit provided to the host. Since probiotics are sensitive to the external environment, many factors influence the survival rate of probiotics in the process from processing to final eating. More and more technologies are used for improving the tolerance of probiotics, the final product can ensure the stable metabolism and the activity of the probiotics, and after the probiotics pass through a digestive system on a host, a large amount of viable thalli can be obtained and released and added in the intestinal tract of the host, so that the benefits of the probiotics are brought into play, the functions of the probiotics are realized, and the problem can be well solved by a microcapsule technology.
The procyanidin has antioxidant and antithrombotic effects, and can regulate blood vessel cells. The performance of procyanidine is researched more at home and abroad, and the procyanidine is widely applied to the fields of foods, cosmetics, health-care products and the like. In addition, procyanidins have good water solubility, but poor stability, are easily oxidized, are sensitive to temperature and pH, and limit their use.
Patent CN106723233B discloses a method for preparing microcapsules by using rice bran protein-polysaccharide as probiotic bacteria of wall material. The wall material is as follows: the microcapsule product taking the rice bran protein aggregate-polysaccharide mixed gel as the wall material has good acid resistance and enteric solubility, and can delay the release of the core material in a simulated gastric digestion environment, namely, in the simulated gastric digestion environment, the polymer formed by adsorbing a layer of polysaccharide with negative electricity through the action of electrostatic attraction by protein with positive charge can effectively prevent pepsin from diffusing into the interior of the microcapsule, thereby playing a role in protecting certain active ingredients. However, the survival rate of bifidobacteria decreases slowly with time.
Disclosure of Invention
In order to overcome the defects and shortcomings of the prior art, the invention aims to provide a method for preparing probiotic microcapsules by using a placenta water-soluble extract as a wall material, so as to solve the problem of low survival rate after embedding probiotics in the prior art and provide a multi-activity embedded probiotic product.
Another object of the present invention is to provide probiotic microcapsules prepared by the above method.
The invention further aims to provide application of the probiotic microcapsule.
The purpose of the invention is realized by the following technical scheme:
a method for preparing probiotic microcapsules by using placenta water-soluble extract as a wall material comprises the following steps:
s1, preparing a placenta water-soluble extract: cleaning the obtained placenta, lyophilizing, grinding, dissolving in normal saline, centrifuging, collecting supernatant, filtering, and sterilizing to obtain placenta water soluble extract;
s2, preparing microcapsule wall materials: dropwise adding 2-4%, preferably 3% procyanidine solution into 8-12%, preferably 10% placenta water-soluble extract solution by mass, stirring and mixing uniformly, and performing thermal denaturation treatment to obtain microcapsule wall material;
s3, preparation of probiotic microcapsules: mixing probiotic bacteria liquid with microcapsule wall materials, adding transglutaminase (TGase) and uniformly dispersing, adding the mixed liquid into preheated soybean oil, and reacting the system under stirring; after the liquid drops are converted into gel particles under the action of enzyme, the microcapsule particles are centrifugally collected, and then the probiotic microcapsule is obtained.
Further, the specific operations of the cleaning in step S1 are: soaking the placenta in water, pinching the edge of the placenta with hands, extruding blood from the great vessel to the opening with umbilical cord, cleaning, and draining.
Further, the placenta described in step S1 is cut into small pieces of 0.5-1.5 cm before lyophilization.
Further, the dosage of the physiological saline in the step S1 is as follows: 1g of physiological saline: metering by 30-70 ml; preferably, the weight ratio of 1 g: and (4) counting by 50 ml.
Further, the specific operation of centrifuging and collecting the supernatant in step S1 is: centrifuging at 8000-9000 g for 25-35 min for the first time, and centrifuging at 27000-29000 g for 110-130 min for the second time to collect supernatant; preferably, the supernatant is collected by a first 7000g, 30min centrifugation and a second 28000g, 2h centrifugation.
Further, the filtration membrane used for the filtration described in step S1 had a pore size of 0.22. mu.m.
Further, the stirring described in step S2 is magnetic stirring.
Further, the heat denaturation treatment in the step S2 is carried out at a temperature of 75-85 ℃ for 20-40 min; preferably at a temperature of 80 ℃ for 30 min.
Further, the probiotics in step S3 are one or more of lactobacillus and bifidobacterium probiotics; wherein the lactobacillus probiotics comprise Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus jensenii, etc., and the bifidobacterium probiotics comprise Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium thermophilum, Bifidobacterium adolescentis, Bifidobacterium bifidum, etc.; preferably lactobacillus plantarum.
Further, the concentration of the probiotic bacteria liquid in step S3 is 108～1011CFU/mL; preferably 109CFU/mL。
Further, the ratio of the probiotic bacteria liquid, the microcapsule wall material, the transglutaminase and the soybean oil in step S3 is 2 mL: 18-22 mL: 0.1-0.3 g: 140-160 g; preferably 2 mL: 20mL of: 0.2 g: 150 g.
Further, the transglutaminase described in step S3 is used in an amount of
Further, the preheating temperature of the soybean oil in the step S3 is 38-42 ℃; preferably 40 deg.c.
Further, the rotating speed of stirring in the step S3 is 800-1000 r/min; preferably 900 r/min.
Further, the conditions of the reaction described in step S3 are: the temperature is 38-42 ℃, and the time is 170-190 min; preferably, the temperature is 40 ℃ and the time is 180 min.
Further, the conditions of centrifugation described in step S3 are: the rotating speed is 400-600 r/min, and the time is 55-65 s; preferably, the rotating speed is 500r/min and the time is 60 s.
Further, after the probiotic microcapsules are obtained in step S3, the ringer' S reagent is added, and the mixture is centrifuged at 700r/min for 5min, and the colloidal particles are collected again for washing.
A probiotic microcapsule is prepared by the above preparation method.
The probiotic microcapsule is applied to preparation of food, feed, medicines, cosmetics or daily necessities.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the placenta water-soluble extract anthocyanin provided by the invention is used as a wall material, aiming at the problem of low survival rate of probiotics, procyanidine is added, and the survival rate of the probiotics in the preservation time and the bioavailability of procyanidine are obviously improved.
The invention adopts the placenta water-soluble extract polymer, can exert the activity of the placenta water-soluble extract, increase the activity of a single wall material and improve the function of utilizing the biological activity of the wall material.
Drawings
Figure 1 effect of different microcapsules on the storage stability of bifidobacteria.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples, but the present invention is only illustrated by the reference examples and is not limited to the scope of the present invention. Other embodiments of the invention based on the present invention, which can be made by a person skilled in the art without inventive step, belong to the scope of protection of the present invention.
Unless otherwise specified, the reagents and materials used in the present invention are commercially available products or products obtained by a known method.
The preparation of the water-soluble extract of placenta used in the following examples was as follows: the preparation method comprises the steps of obtaining sterile placenta tissues, immersing the placenta in water, pinching the edge of the placenta with hands, extruding blood of a large blood vessel to an opening with an umbilical cord cut off along the direction of the large blood vessel, cleaning, draining the cleaned placenta, cutting into small blocks of about 1cm, freeze-drying, grinding, dissolving 5g of the freeze-dried placenta powder in 250ml of normal saline, centrifuging for the first time at 8500g for 30min, and collecting supernatant. 28000g, centrifuging for 2h, collecting supernatant, and filtering and sterilizing with 0.22um filter membrane to obtain placenta water soluble extract.
Procyanidins used in the following examples were obtained from Shanghai Allantin Biotech Co., Ltd, under the CAS designation 4852-22-6.
MRS liquid medium used in the following examples was purchased from Kyork Microbiology, Inc., product number SR 0370.
Transglutaminase (TGase, enzyme activity 1000U/g) used in the following examples was purchased from California Biotech Inc., Suzhou, under the trade name: 80146-85-6
Example 1
1. Strain culture and sample preparation
Inoculating Bifidobacterium (Bifidobacterium bifidum) into sterile MRS liquid culture medium at a ratio of 1: 10(V/V), and anaerobically culturing at 37 deg.C for 24 hr. After 3 times of activation, the mixture is inoculated into a sterile MRS liquid culture medium according to the inoculation amount of 1: 10(V/V) and is subjected to anaerobic culture at the constant temperature of 37 ℃ for 24 hours. Centrifuging at 4000r/min for 5min, collecting thallus, washing with sterile physiological saline (0.85% NaCl) for 2 times, resuspending, and adjusting the concentration of concentrated bacterial liquid to 109CFU/mL。
2. Preparation of microcapsule wall material
Dropwise adding 3% procyanidine solution into 10% placenta water-soluble extract solution, and stirring and mixing uniformly in a magnetic stirrer;
3. preparation of microcapsules
3.1 preparation of placental extract microcapsules
Mixing 2mL of concentrated bacterial liquid with 20mL of heat-denatured (80 ℃ and 0.5h) microcapsule wall material (10% of placenta water-soluble extract and 3% of procyanidine), quickly adding 0.2g of TGase (10U/g) and carrying out vortex oscillation to uniformly disperse the mixture, adding the mixed solution into a 250mL conical flask, containing 150g of preheated (40 ℃) soybean oil, reacting the system at 900r/min and 40 ℃ for 180min under the action of a magnetic stirrer, centrifuging (500r/min and 1min) after liquid drops are converted into gel particles under the action of enzyme, collecting the microcapsule particles, washing the microcapsule particles for 2 times by using a ringer's reagent, centrifuging for 5min under the condition of 700r/min, collecting the colloid particles again, and storing the microcapsule particles at 4 ℃ for later use.
3.2 preparation of sodium alginate microcapsules
Firstly, 2mL of concentrated bacterial liquid is mixed with 20mL of 2.0g/100mL sodium alginate solution, and then CaCO3The microcrystals are uniformly dispersed in the mixed solution, wherein CaCO3The addition amount of the sodium alginate is 7.3 percent of the mass of the sodium alginate, then the mixed solution is emulsified into 100mL of Span80 soybean oil (300r/min) with the volume fraction of 1.0 percent, after 15min, 20mL of soybean oil containing glacial acetic acid, glacial acetic acid and CaCO are added3The mass ratio of the substances is 3.5, stirring is continuously carried out to promote the contact reaction of CaCO3 microcrystals and glacial acetic acid, after 30min, 120mL of acetate solution (pH 5.5) is added into the system and stirred slowly, after all the microcapsules formed by gel are settled to the bottom of the acetate solution, the oil phase is sucked off, AM is collected by filtration, and the residual oil phase is removed by washing for 2 times. Finally, the microcapsules collected by filtration were stored in a 0.2g/100mL NaCl solution at 4 ℃.
4. Enumeration of embedded probiotics
Accurately weighing 1g each of 3 kinds of wet microcapsule samples, and adding 9mL of solution for dissolving the capsule (NaHCO solution)3Dissolving in 0.1mol/L Phosphate Buffer Solution (PBS) at pH 7.0 (NaHCO)30.2mol/L), filtering and sterilizing), and shaking for 10min until complete capsulization. Taking a certain amount of liquid, diluting, coating on MRS agar culture medium, culturing the plate at 37 ℃ under anaerobic condition for 48h, counting, and calculating the embedding efficiency. Embedding efficiency calculation formula: N/N0X 100%. In the formula: n is the total number of living cells/CFU embedded in the microcapsule; n0 represents the total number of bacteria/CFU in the concentrated bacterial solution.
5. Particle size analysis of microcapsules
The particle size distribution of the microcapsules was measured by a laser particle size analyzer, and the average particle size is expressed as a volume average particle size D, which is a weighted average of the microcapsule particle size to volume.
6. Survival of probiotic bacteria in simulated gastric fluid and high bile salt solution
Preparing simulated gastric juice: adjusting pH value of 0.2g/100mL NaCl solution to 2.0 with concentrated hydrochloric acid solution, adding a certain amount of pepsin to make its final mass concentration be 0.3g/L, and filtering and sterilizing with 0.22 μm membrane; preparing a high-bile salt solution: bile salts were homogeneously dispersed in a phosphate buffer solution at a final mass concentration of 4.5g/L, and the mixture was adjusted to pH 7.4 with 0.1mol/L NaOH solution and sterilized.
0.5g of microcapsules was added to 4.5mL of simulated gastric fluid or high bile salt solution preheated at 37 ℃ and sufficiently shaken with a vortex shaker for 10s, followed by incubation at 37 ℃ and 100r/min for 2 h. At 5, 30, 60, 120min, the sample solution was disrupted with a high speed homogenizer and counted as described above.
7. Effect of different microcapsules on the storage stability of bifidobacteria
Placing the freeze-dried microcapsules into a sterile penicillin bottle, then carrying out heat sealing under the condition of vacuumizing, placing the sealed penicillin bottle at the normal temperature for storage for 1 month, sampling from the penicillin bottle every 15 days, and counting the bifidobacteria.
The experimental results are as follows:
1. particle size and embedding efficiency of bifidobacterium microcapsules with different wall materials
The particle size and the embedding efficiency are important indexes for evaluating the microcapsules, and different wall materials have obvious influence on the particle size and the embedding efficiency of the microcapsules. As can be seen from Table 1, the particle size of the microcapsule prepared from sodium alginate is 125.2 μm, and the embedding efficiency of the microcapsule on Bifidobacterium is 58.6%, while the particle size of the microcapsule prepared from placenta extract is increased by 76.3 μm, and the embedding efficiency on Bifidobacterium is as high as 86.4%.
TABLE 1 particle size and embedding efficiency of Bifidobacterium microcapsules with different wall materials
2. Effect of different embedding wall materials on bifidobacteria protection effect
2.1 Effect of different microcapsules on the protective Effect of bifidobacteria in simulated gastric fluid
TABLE 2 survival of Bifidobacterium in simulated gastric fluid (pH 2.0, 37 ℃, 2h)
D value (time required to kill 90% of bacteria under certain treatment conditions)
As can be seen from table 2, the survival rate of bifidobacterium in simulated gastric juice was increased to different degrees compared with that of non-embedded bifidobacterium, and the survival rate of bifidobacterium embedded in placenta extract was increased by 7(lg (CFU/g). the D value shows that the protection effect of placenta extract (D value of 114.4min) on bifidobacterium is better than the effect of sodium alginate (D value of 20.3min) on the protection effect of bifidobacterium in high-bile-salt environment by 2.2 different microcapsules
TABLE 3 survival of Bifidobacterium in a model hyperbilirubine salt solution (pH 7.4, 37 ℃, 2h)
As is clear from Table 3, after 2 hours of treatment in a high-bile-salt atmosphere, about 3(lg (CFU/mL)) of Bifidobacterium died without embedding treatment, while about 1(lg (CFU/mL)) of Bifidobacterium died with embedding treatment. Sodium alginate and placenta extract have no significant difference in the protective effect on bifidobacteria in a high-bile-salt environment, and the D values of the sodium alginate and the placenta extract are at the same significant level.
3. Effect of different microcapsules on the storage stability of bifidobacteria
As can be seen from fig. 1, after storage for 30 days under ambient temperature conditions, the survival amount of bifidobacteria in all three microcapsules was reduced to different extents, and the tendency of sodium alginate microcapsules to be reduced was more pronounced when the placenta extract microcapsules were reduced from 7.59(lg (CFU/g)) to 6.71(lg (CFU/g)), and the lethal amount of bifidobacteria was close to 2(lg (CFU/g)) during the storage period of 1 month. The storage stability of the bifidobacterium embedded in the placenta extract microcapsule is better than that of the bifidobacterium embedded in the sodium alginate microcapsule, and the storage stability of the bifidobacterium embedded in the placenta extract microcapsule is the best when the placenta extract microcapsule is combined with the anthocyanin.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A method for preparing probiotic microcapsules by using placenta water-soluble extract as a wall material is characterized by comprising the following steps: the method comprises the following steps:
s1, preparing a placenta water-soluble extract: cleaning the obtained placenta, lyophilizing, grinding, dissolving in normal saline, centrifuging, collecting supernatant, filtering, and sterilizing to obtain placenta water soluble extract;
s2, preparation of microcapsule wall materials: dropwise adding 3% procyanidine solution into the placenta water-soluble extract solution with the mass percentage of 8% -12%, stirring and uniformly mixing, and performing thermal denaturation treatment to obtain a microcapsule wall material;
s3, preparation of probiotic microcapsules: mixing probiotic bacteria liquid with microcapsule wall materials, adding transglutaminase to uniformly disperse the transglutaminase, adding the mixed liquid into preheated soybean oil, and reacting the system under stirring; and after the liquid drops are converted into gel particles under the action of enzyme, centrifuging and collecting the microcapsule particles to obtain the probiotic microcapsule.
2. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1, characterized in that:
the probiotics in the step S3 are one or more of lactobacillus and bifidobacterium probiotics; wherein the Lactobacillus probiotics comprise Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus jensenii, and the Bifidobacterium probiotics comprise Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium thermophilum, Bifidobacterium adolescentis and Bifidobacterium bifidum.
3. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1 or 2, characterized in that:
the dosage of the physiological saline in the step S1 is as follows: 1g of physiological saline: metering by 30-70 ml;
the specific operation of centrifugally collecting the supernatant described in step S1 is: centrifuging at 8000-9000 g for 25-35 min for the first time, and centrifuging at 27000-29000 g for 110-130 min for the second time to collect the supernatant.
4. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1 or 2, characterized in that:
the specific operation of the cleaning in step S1 is: soaking the placenta in water, pinching the edge of the placenta with hands, extruding blood from the great vessel to the opening with umbilical cord, cleaning, and draining;
the placenta obtained in the step S1 is cut into small blocks of 0.5-1.5 cm before freeze-drying;
the dosage of the physiological saline in the step S1 is determined according to the ratio of 1 g: metering by 50 ml;
the specific operation of centrifugally collecting the supernatant described in step S1 is: centrifuging at 7000g for 30min for the first time, and centrifuging at 28000g for 2h for the second time to collect the supernatant;
the filtration membrane used for the filtration described in step S1 had a pore size of 0.22. mu.m.
5. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1 or 2, characterized in that:
the conditions of the heat denaturation process described in step S2 are: the temperature is 75-85 ℃ and the time is 20-40 min.
6. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1 or 2, characterized in that:
the concentration of the placenta water-soluble extract solution in the step S2 is 10%, and the concentration of the procyanidine solution is 3%;
the stirring in the step S2 is magnetic stirring;
the conditions of the thermal denaturation treatment described in step S2 were 80 ℃ for 30 min.
7. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1 or 2, characterized in that:
the concentration of the probiotic bacteria liquid in the step S3 is 108～1011CFU/mL；
The ratio of the probiotic bacteria liquid, the microcapsule wall material, the transglutaminase and the soybean oil in the step S3 is 2 mL: 18-22 mL: 0.1-0.3 g: 140-160 g;
the preheating temperature of the soybean oil in the step S3 is 38-42 ℃;
the rotating speed of stirring in the step S3 is 800-1000 r/min;
the reaction conditions described in step S3 are: the temperature is 38-42 ℃, and the time is 170-190 min;
the conditions of centrifugation described in step S3 are: the rotating speed is 400-600 r/min, and the time is 55-65 s.
8. The method for preparing probiotic microcapsules using water-soluble extract of placenta as wall material according to claim 1 or 2, characterized in that:
the concentration of the probiotic bacteria liquid in the step S3 is 109CFU/mL；
The ratio of the probiotic bacteria liquid, the microcapsule wall material, the transglutaminase and the soybean oil in the step S3 is 2 mL: 20mL of: 0.2 g: 150g of the total weight of the mixture;
the preheating temperature of the soybean oil in the step S3 is 40 ℃;
the stirring rotating speed in the step S3 is 900 r/min;
the reaction conditions described in step S3 are: the temperature is 40 ℃ and the time is 180 min;
the conditions of centrifugation described in step S3 are: the rotating speed is 500r/min, and the time is 60 s;
and (S3) adding a ringer reagent after the probiotic microcapsules are obtained, centrifuging for 5min at the speed of 700r/min, and collecting colloid particles again for cleaning.
9. A probiotic microcapsule obtained by the preparation method as set forth in any one of claims 1 to 8.
10. Use of probiotic microcapsules according to claim 9, characterized in that: can be used for preparing food, feed, medicine, cosmetic or daily necessities.
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| 李作美，等: "双歧杆菌微胶囊的制备及其理化性能", 《食品与发酵工业》 * |
| 林旭明，等: "原花青素交联的支架材料的生物相容性研究", 《眼科新进展》 * |
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