METHOD FOR EMBEDDING A LOAD BASED ON GEL HIGH HYDROSTATIC PRESSURE LIQUEFACTION

Abstract

The invention relates to a method for embedding a load based on gel high hydrostatic pressure liquefaction. Using the phenomenon that the physical gel is liquefied under high pressure, the vacuum-packaged high-methoxyl pectin gel is treated under a pressure of 400-600 MPa for 5-30 min, mixed with the load, and then subjected to a pressure of 400-600 MPa for homogenization treatment for 5 to 30 min After pressure relief, the liquefied gel is poured into a mold for reshaping, followed by removal of free water and coating treatment. This method combines the advantages of high hydrostatic pressure technology in modification and sterilization. It has mild embedding conditions and wide sources of raw materials to prepare the carrier, which has excellent biocompatibility and biodegradability. It can be widely used for embedding microorganisms, enzymes, proteins and small molecular substances. The loaded gel prepared by the method has high microbial safety, can effectively maintain the activity of the load. The load distribution is uniform, and the load amount is much larger than the traditional adsorption load.

Claims

1. A method for embedding a load based on gel high hydrostatic pressure liquefaction, wherein comprising the following steps: (1) preparation of pectin gel: a certain amount of high-methoxyl pectin is dissolved in distilled water, stirred at room temperature for 8-12 h and then placed in a water bath at 80-90 C. After the temperature is stable, a certain amount of soluble solids is added, and the mixed solution is obtained after stirring the mixed solution is then poured into the soft package, vacuum-packaged and fully cooled; the mass concentrations of high-methoxyl pectin and soluble solids in the mixture are 0.5-2% and 50-70% respectively; (2) integrated treatment of high hydrostatic pressure gel liquefaction and sterilization: the vacuum-packaged gel in step (1) is placed in the pressure vessel filled with liquid pressure transmitting medium and treated at the pressure of 400-600 MPa for 5-30 min; (3) embedding of the load: after high pressure treatment, the pectin gel is converted into a sol state, and the load and the sol are mixed with fully stirred; the operation is completed within 10 to 20 min after pressure relief; (4) the mixture thoroughly stirred in step (3) is poured into a soft package and vacuum-packaged; the vacuum-packaged mixture is placed in the pressure vessel filled with liquid pressure transmitting medium and treated at the pressure of 400-600 MPa for 5-30 min, after pressure relief, the mixture is quickly poured into the mold and formed at suitable temperature for 1-3 days; or, the mixture of the loaded material and the sol after full stirred in step (3) is directly poured into the mold for forming; (5) removal of the free water: the formed gel prepared by step (4) is dried to constant weight at suitable temperature; (6) coating of the gel: solution A: a certain amount of low-methoxyl pectin is dissolved in distilled water, stirred at room temperature for 8 to 12 h, after which the pH is adjusted, the mass concentration of the low-methoxyl pectin is 2 to 5%, and the pH of the solution is 8 to 12; solution B: a calcium salt solution having a Ca.sup.2+ concentration of 1 to 5 mmol/L is prepared; the dry gel prepared by step (5) is infiltrated in Solution B and transferred rapidly to Solution A, and then rapidly transferred from Solution A to Solution B.

2. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: the degree of methoxylation of the high-methoxyl pectin described in step (1) is 50-80%.

3. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: the soluble solids described in step (1) include at least one of sucrose, fructose or glucose.

4. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: the loads mentioned in step (3) include at least one of microorganisms, enzymes, proteins and small molecular substances.

5. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: in step 4, for the embedding of small molecule substances, the high hydrostatic pressure homogenization treatment is adopted; while for the embedding of microorganisms and macromolecular substances, the mixture of load and sol which is sufficiently stirred in the step (3) is directly poured into a mold for forming.

6. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: the suitable temperature mentioned in the steps (4) and (5) refers to the optimum temperature for maintaining the activity of the loaded substance.

7. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: the degree of methoxylation of the low-methoxyl pectin mentioned in step (6) is 10-30%.

8. The method for embedding a load based on gel high hydrostatic pressure liquefaction according to claim 1, wherein: in step (6), the dry gel is infiltrated in the Solution B and rapidly transferred to the Solution A, and then rapidly transferred from the Solution A to the Solution B, the operation is repeated 1-2 times.

Description

DRAWINGS

[0025] FIG. 1 is the section morphology of the high-methoxyl apple pectin gel without high hydrostatic pressure treatment.

[0026] FIG. 2 is the section morphology of the high-methoxyl apple pectin gel after treatment at 400 MPa.

[0027] FIG. 3 is the sol-gel transition kinetics curve of high-methoxyl apple pectin after treatment at different pressures.

[0028] FIG. 4 is a release kinetic curve of procyanidins from a loaded gel prepared according to the steps described in embodiment 2.

DETAILED DESCRIPTION

[0029] The following is further explained with specific embodiments. It should be understood that these embodiments are used only to illustrate the present invention and not to limit the scope of application of the present invention. In addition, after reading the contents of the present invention, skilled personnel in the field may make various modifications to the present invention, which fall within the scope of claims attached to this application.

Embodiment 1

[0030] A method for embedding Lactobacillus acidophilus based on gel high hydrostatic pressure liquefaction, which comprises the following steps:

[0031] (1) preparation of pectin gel: 3 g of high-methoxyl pectin (from apple, the degree of methoxylation of 66%) was weighed in a beaker, and 117 g of distilled water was added, then stirred at room temperature for 12 h, dissolved thoroughly, placed in a water bath at 90 C., 180 g of glucose was added after the temperature is stable, after stirring and dissolving, the water lost by evaporation was replenished, the mixture was poured into a soft food package while hot, and stored it in a refrigerator at 4 C. for 30 h after vacuum packaging to obtain a pectin gel;

[0032] (2) integrated treatment of high hydrostatic pressure gel liquefaction and sterilization: the vacuum-packaged gel in step (1) was placed in the pressure vessel filled with water and treated at 600 MPa for 5 min, and then the pectin sol was obtained after pressure relief;

[0033] (3) embedding of Lactobacillus acidophilus: 1 ml of the bacterial suspension (100 cfu/ml, containing the nitrogen source, inorganic elements and growth factors necessary for the growth of Lactobacillus acidophilus) and the sol obtained in the step (2) were mixed within 10 min after pressure relief, and thoroughly stirred, then poured into a mold and stored at 37 C. for 72 h;

[0034] (4) coating of the gel: Solution A: 5 g of low-methoxyl pectin (from apple, the degree of methoxylation of 14%) was dissolved in distilled water, stirred at room temperature for 8 h, pH was adjusted with 0.05 g/ml NaOH solution, the final mass concentration of low-methoxyl pectin was 5%, and the final pH of the solution was 9; Solution B: 5 g of anhydrous calcium chloride was weighed and diluted to 100 ml with distilled water to obtain a calcium chloride solution of 0.05 g/ml; the shaped gel prepared in the step (3) was infiltrated in the solution B and rapidly transferred to the solution A, and then rapidly transferred from the solution A to the solution B, and repeated twice.

[0035] The loaded gel prepared in this embodiment can culture Lactobacillus acidophilus while fixing it, and the glucose in the gel provides a natural carbon source for the growth of Lactobacillus acidophilus. The embedding conditions are mild to effectively maintain the activity of Lactobacillus acidophilus and a high cell density (269 cfu/g) can be obtained; in addition, the gel can carry the Lactobacillus acidophilus to the gastrointestinal tract smoothly, thereby exerting its biological effects.

Embodiment 2

[0036] A method for embedding proanthocyanidins based on gel high hydrostatic pressure liquefaction includes the following steps:

[0037] (1) preparation of pectin gel: 6 g of high-methoxyl pectin (from citrus, the degree of methoxylation of 69%) was weighted in a beaker, 144 g of distilled water was added, stirred at room temperature for 8 h, dissolved thoroughly, placed in a water bath at 80 C., 150 g of sucrose was added after temperature stabilization, after stirring and dissolving, the water lost by evaporation was replenished, the mixture was poured into a soft food package while hot, and stored it in a refrigerator at 4 C. for 24 h after vacuum packaging to obtain a pectin gel;

[0038] (2) integrated treatment of high hydrostatic pressure gel liquefaction and sterilization: the vacuum-packaged gel in step (1) was placed in the pressure vessel filled with water and treated at 400 MPa for 20 min, and then the pectin sol is obtained after pressure relief;

[0039] (3) embedding of proanthocyanidins: the proanthocyanidin powder was mixed with the sol obtained in step (2) within 10 min after pressure relief, then fully stirred and poured into the soft food package for vacuum packaging;

[0040] (4) high hydrostatic pressure homogenization treatment: the vacuum-packaged mixture in step (3) was placed in a pressure vessel and treated at 600 MPa for 5 min After pressure relief, the mixture was quickly poured into the mold and formed in a refrigerator at 4 C. for 24 h;

[0041] (5) removal of the free water: the gel prepared in step (4) was placed in a desiccator and dried at room temperature for 8 h;

[0042] (6) coating of the gel: Solution A: 4 g of low-methoxyl pectin (from citrus, the degree of methoxylation of 29%) was dissolved in distilled water, stirred at room temperature for 12 h, pH was adjusted with 0.05 g/ml NaOH solution, the final mass concentration of low-methoxyl pectin was 4%, and the final pH of the solution was 12; Solution B: 4 g of anhydrous calcium chloride was weighed and diluted to 100 ml with distilled water to obtain a calcium chloride solution of 0.04 g/ml; the dry gel prepared by step (5) was infiltrated in Solution B and transferred rapidly to Solution A, and then transferred rapidly from Solution A to Solution B, and repeated twice.

[0043] The loaded gel prepared in this embodiment can effectively prevent oxidation of proanthocyanidins, and the proanthocyanidins in the gel can be slowly diffused after administration to facilitate absorption by the human body. The release kinetics curve of proanthocyanidins from the loaded gel prepared by this step is shown in FIG. 4. The release conditions are as follows:

[0044] (1) the gel mass was 4 g, and the gel shape was 25 mm in diameter and 5 mm in height cylinder;

[0045] (2) the release environment was 100 ml deionized water, the release temperature was 37 C., and the shaker speed is 100 rpm.

Embodiment 3

[0046] A method for embedding bovine serum albumin (BSA) based on gel high hydrostatic pressure liquefaction includes the following steps:

[0047] (1) preparation of pectin gel: 4.5 g high-methoxyl pectin (from apple, the degree of methoxylation of 79%) was weighed in beaker, 130.5 g distilled water was added, stirred for 10 h at room temperature, fully dissolved and put in a water bath at 85 C., after the temperature is stable, 165 g soluble solids were added, after stirring and dissolving, the water lost by evaporation was added, the mixture was poured into a soft food package while hot, and stored in a refrigerator at 4 C. for 48 h, and then the pectin gel was obtained;

[0048] (2) integrated treatment of high hydrostatic pressure gel liquefaction and sterilization: the vacuum-packaged gel in step (1) was placed in the pressure vessel filled with water and treated at 500 MPa for 30 min, and then the pectin sol was obtained after pressure relief;

[0049] (3) embedding of BSA: the BSA powder was mixed with the sol obtained in step (2) within 15 min after pressure relief, fully stirred, and poured into the mold, and stored at 4 C. for 48 h for forming;

[0050] (4) removal of the free water: the gel prepared in step (3) was placed in a desiccator and dried at room temperature for 12 h;

[0051] (5) coating of the gel: Solution A: 3 g of low-methoxyl pectin (from grapefruit, the degree of methoxylation of 25%) was dissolved in distilled water, stirred at room temperature for 12 h, and pH was adjusted with 0.05 g/ml NaOH solution, the final mass concentration of low-methoxyl pectin was 3%, and the final pH of the solution was 11; Solution B: 3 g of anhydrous calcium chloride was weighed and diluted to 100 ml with distilled water to obtain a calcium chloride solution of 0.03 g/ml; the dry gel prepared by step (4) was infiltrated in Solution B and transferred rapidly to Solution A, and then rapidly transferred from Solution A to Solution B.

[0052] Because of the hydrogen bond interaction between high-methoxyl pectin and BSA, the binding effect of carrier on drug molecules is enhanced and the release rate of BSA is reduced.

Embodiment 4

[0053] A method for embedding vitamin C by gel high hydrostatic pressure liquefaction includes the following steps:

[0054] (1) preparation of pectin gel: 6 g of high-methoxyl pectin (from citrus, the degree of methoxylation of 71%) was weighed in a beaker, 114 g of distilled water was added, stirred at room temperature for 9 h, dissolved well, placed in a water bath at 80 C., 180 g of soluble solids was added after the temperature is stable, after stirring and dissolving, the water lost by evaporation was replenished, the mixture was poured into a soft food package while hot, and stored it in a refrigerator at 4 C. for 32 h after vacuum packaging to obtain the pectin gel;

[0055] (2) integrated treatment of high hydrostatic pressure gel liquefaction and sterilization: the vacuum-packaged gel in step (1) was placed in the pressure vessel filled with water and treated at 550 MPa for 15 min, and then the pectin sol was obtained after pressure relief;

[0056] (3) embedding of vitamin C: the vitamin C powder was mixed with the sol obtained in step (2) within 10 min after pressure relief, fully stirred and poured into the soft food package for vacuum packaging;

[0057] (4) high hydrostatic pressure homogenization treatment: the vacuum-packaged mixture in step (3) was placed in a pressure vessel and treated at 500 MPa for 15 min After pressure relief, the mixture was quickly poured into the mold and formed in a refrigerator at 4 C. for 32 h;

[0058] (5) removal of the free water: the gel prepared in step (4) was placed in a desiccator and dried at room temperature for 24 h;

[0059] (6) coating of the gel: Solution A: 4.5 g of low-methoxyl pectin (from lemon, the degree of methoxylation of 21%) was dissolved in distilled water, stirred at room temperature for 12 h, pH was adjusted with 0.05 g/ml NaOH solution, the final mass concentration of low-methoxyl pectin was 4.5%, and the final pH of the solution was 10; Solution B: 3.5 g of anhydrous calcium chloride was weighed and diluted to 100 ml with distilled water to obtain a calcium chloride solution of 0.35 g/ml; the dry gel prepared by step (5) was infiltrated in Solution B and transferred rapidly to Solution A, and then transferred rapidly from Solution A to Solution B, and repeated twice.

[0060] The invention combines the advantages of high hydrostatic pressure sterilization and modification, and the prepared gel has good microbiological safety. Its raw materials are widely used, and the embedding operation can be carried out at room temperature, which retains the biological activity of the carrier. The method can be widely used for embedding microorganisms, enzymes, proteins and small molecular substances. The load distribution is uniform and the load amount is much larger than the traditional adsorption load.