CN105054074A - Surimi enzymatic hydrolysis peptide chelated iron (II) anti-oxidation microcapsule and preparation technology thereof - Google Patents
Surimi enzymatic hydrolysis peptide chelated iron (II) anti-oxidation microcapsule and preparation technology thereof Download PDFInfo
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- CN105054074A CN105054074A CN201510387030.7A CN201510387030A CN105054074A CN 105054074 A CN105054074 A CN 105054074A CN 201510387030 A CN201510387030 A CN 201510387030A CN 105054074 A CN105054074 A CN 105054074A
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- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 title claims abstract description 93
- 239000003094 microcapsule Substances 0.000 title claims abstract description 77
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 63
- 238000002360 preparation method Methods 0.000 title claims description 19
- 238000005516 engineering process Methods 0.000 title claims description 18
- 230000003064 anti-oxidating effect Effects 0.000 title abstract description 11
- 230000007071 enzymatic hydrolysis Effects 0.000 title abstract 7
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 title abstract 7
- 235000019465 surimi Nutrition 0.000 title abstract 7
- 239000013522 chelant Substances 0.000 claims abstract description 83
- 239000001814 pectin Substances 0.000 claims abstract description 33
- 235000010987 pectin Nutrition 0.000 claims abstract description 33
- 229920001277 pectin Polymers 0.000 claims abstract description 33
- 239000000463 material Substances 0.000 claims abstract description 24
- 239000000843 powder Substances 0.000 claims abstract description 18
- 150000001875 compounds Chemical class 0.000 claims description 74
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 50
- 241000251468 Actinopterygii Species 0.000 claims description 46
- 230000003078 antioxidant effect Effects 0.000 claims description 34
- 239000003963 antioxidant agent Substances 0.000 claims description 28
- 235000006708 antioxidants Nutrition 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000000839 emulsion Substances 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 235000012424 soybean oil Nutrition 0.000 claims description 9
- 239000003549 soybean oil Substances 0.000 claims description 9
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 108090000526 Papain Proteins 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 239000004365 Protease Substances 0.000 claims description 6
- 238000004945 emulsification Methods 0.000 claims description 6
- 230000001804 emulsifying effect Effects 0.000 claims description 6
- 229940055729 papain Drugs 0.000 claims description 6
- 235000019834 papain Nutrition 0.000 claims description 6
- 238000000265 homogenisation Methods 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 229960005070 ascorbic acid Drugs 0.000 claims description 4
- 235000010323 ascorbic acid Nutrition 0.000 claims description 4
- 239000011668 ascorbic acid Substances 0.000 claims description 4
- 239000004567 concrete Substances 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- 239000006185 dispersion Substances 0.000 claims description 2
- 238000001976 enzyme digestion Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 abstract description 16
- 230000000694 effects Effects 0.000 abstract description 9
- 229930003427 Vitamin E Natural products 0.000 abstract description 8
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 abstract description 8
- 235000019165 vitamin E Nutrition 0.000 abstract description 8
- 229940046009 vitamin E Drugs 0.000 abstract description 8
- 239000011709 vitamin E Substances 0.000 abstract description 8
- 230000007062 hydrolysis Effects 0.000 abstract description 7
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 7
- 230000009471 action Effects 0.000 abstract description 3
- 239000013543 active substance Substances 0.000 abstract 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract 1
- 229910052760 oxygen Inorganic materials 0.000 abstract 1
- 239000001301 oxygen Substances 0.000 abstract 1
- 239000011162 core material Substances 0.000 description 27
- 239000000047 product Substances 0.000 description 18
- 238000000034 method Methods 0.000 description 11
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 9
- 238000011160 research Methods 0.000 description 8
- 238000005213 imbibition Methods 0.000 description 7
- 229920001184 polypeptide Polymers 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 229910052742 iron Inorganic materials 0.000 description 5
- 239000003921 oil Substances 0.000 description 4
- 235000019198 oils Nutrition 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 239000011149 active material Substances 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 239000004519 grease Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 150000002978 peroxides Chemical class 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000005728 strengthening Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241000282994 Cervidae Species 0.000 description 1
- 206010011732 Cyst Diseases 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 240000007049 Juglans regia Species 0.000 description 1
- 235000009496 Juglans regia Nutrition 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 238000010923 batch production Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000007942 carboxylates Chemical class 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000005354 coacervation Methods 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000000748 compression moulding Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 208000031513 cyst Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229910001448 ferrous ion Inorganic materials 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000010200 folin Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002992 thymic effect Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 235000015099 wheat brans Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Medicinal Preparation (AREA)
Abstract
The invention discloses a surimi enzymatic hydrolysis peptide chelated iron (II) anti-oxidation microcapsule. The surimi enzymatic hydrolysis peptide chelated iron (II) anti-oxidation microcapsule is composed of surimi enzymatic hydrolysis peptide chelated iron (II) powder, SPI and pectin, the surimi enzymatic hydrolysis peptide chelated iron (II) powder is adopted as a core, and the SPI and pectin are adopted as wall materials to realize double layer embedding of the inside and the outside of the core, wherein the SPI is the internal layer wall material, and pectin is the external layer wall material. The surimi enzymatic hydrolysis peptide chelated iron (II) anti-oxidation microcapsule has a high embedding rate (reaching 82%), low hydrolysis degree (being 10.97% at 90min and being 13.71% after 24h), and has strong anti-oxidation activity as oil, and the anti-oxidation effect is basically same to that of vitamin E. The surimi enzymatic hydrolysis peptide chelated iron (II) anti-oxidation microcapsule also has slow release ability, can effectively perform active substances of the core and prolong the action time, and can resist influences of oxygen molecules in environment on the core chelate in order to improve the product stability and utilization efficiency.
Description
Technical field
The present invention relates to the rotten anti-oxidant microcapsules of Peptides ferrous chelate compound of a kind of fish and preparation technology thereof.
Background technology
Oceanic biological active peptides (Marinebiologicalactivepeptide), as most important constituent in current marine natural products active material, becomes the focus of biologically active peptide research gradually because it has unique physiological function.Adopt the marine protein range of hydrolysed peptides that complex enzyme zymohydrolysis legal system is standby, its output is large, cost is low, and reaction substrate, reactant and reaction environment are all safe from harm, the little peptide of part generated also has higher nutritive value and higher biologically active, is applied [1] at value segments such as medicine, cosmetics, health products at present.
Marine organisms range of hydrolysed peptides can a large amount of ferro element of complexing, the chelating carrier being ferro element with marine protein range of hydrolysed peptides makes medical food, can be easy to effectively be absorbed by enteron aisle through intestinal mucosa, it not only has the high usage of iron and high foodsafety, also has biologically active and proteinaceous nutrient functional characteristics such as improving hypoferric anemia, growth promoting effects, delay senility.Therefore, with marine protein enzymolysis polypeptide for primary raw material, prepare range of hydrolysed peptides-beneficial metallic elements chelate by the composite modified technology of iron, and be applied in medical food, the present situation generally lacked for nutrients such as solving iron has important promotion meaning.But the easily oxidizable of ferrous ion is the batch production of this product and preserves transport and all bring tremendous influence, and this just constrains greatly, and rotten Peptides ferrous chelate compound (calling ferrous chelate compound in the following text) of fish is more effective is applied to field of medicaments.
Microcapsules technology refers to the core substances such as macromolecule filming material embedding solid, liquid, the gas utilizing natural or synthesis, forming diameter is the particle with semi permeability or sealing cyst membrane from tens microns to several thousand microns, and this particle can in the technology of slowly-releasing under given conditions.Current Chinese scholars has successfully achieved the microencapsulation to soybean function polypeptide, wheat bran peptide, deer placenta polypeptide, walnut protein polypeptide, thymic peptide and reduced glutathione etc.Temporarily yet there are no the report of ferrous chelate compound microencapsulation.Microencapsulation provides a kind of useful technology to protect product from environmental condition, extends the shelf life.Ferrous chelate compound microencapsulation effectively can be solved ferrous oxidizable problem, a new direction is opened up in the development for this product.
Summary of the invention
The invention provides that a kind of embedding rate is high, solubility is low, water imbibition is low, anti-oxidant friendship fruit is good, the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish that slow-release capability is strong;
Another object of the present invention is to provide the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of above-mentioned fish.
Object of the present invention carrys out specific implementation by the following technical programs:
The anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of a kind of fish, be made up of fish rotten Peptides ferrous chelate compound powder, SPI and pectin, fish rotten Peptides ferrous chelate compound powder is for making core, SPI and pectin as wall material by double-layer embedment inside and outside core, wherein, SPI is inner layer wall material, and pectin is outer wall material.
Preferably, the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of above-mentioned fish, wall core weight ratio is more than 2:1; Preferably, wall core weight ratio is 2:1; Further preferred, SPI: the mass ratio of pectin is 45:55.
The preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of above-mentioned fish, with fish rotten Peptides ferrous chelate compound powder for core, embed SPI and pectin respectively in powder particle skin, the average grain diameter of these microcapsules is 65 μm.
Above-mentioned technique, specifically comprises the steps:
1) preparation of the rotten Peptides ferrous chelate compound of fish
Obtain enzymolysis liquid with after the gruel of papain enzymolysis fish, then add ClFe
2carry out chelatropic reaction, obtain the rotten Peptides ferrous chelate compound of fish;
2) double-layer embedment microcapsules preparation
A.SIP emulsifying soln
By step 1) in the fish that obtains rotten Peptides ferrous chelate compound powder dispersion in soybean oil, then with SIP emulsifying soln, obtain O/W emulsion;
B. embed
Poured into by pectin in the emulsion obtained in step a, stir 20-30min mixing, after adjust ph to 4.4, store overnight at 7 DEG C, refreezes into freeze-dried powder again, can obtain the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish.
Preferably, described step 1) in enzyme digestion reaction concrete operations as follows:
First fish gruel is pressed 50g:500ml with water to mix, regulate pH to be 6.0, at 45 DEG C, be incubated 10min, then add papain in the ratio of 2000U/g, enzymolysis 8h at 45 DEG C.
Preferably, described step 1) in chelatropic reaction specific as follows:
By centrifugal for enzymolysis liquid go precipitation, adjust pH be 6.5, add ascorbic acid, then liquid weight 1%-3% add FeCl
2, chelating 30min in constant temperature oscillator, then Vacuum Concentration, freeze drying adult fish rotten Peptides ferrous chelate compound powder.
Preferably, described step 2) step a in, concrete operations are:
With ferrous chelate compound: the weight ratio of soybean oil be the ratio of 1:3 by ferrous ferrous chelate compound emulsification, obtain the emulsion of W/O; Then by the sample that processed and SIP emulsifying soln, O/W emulsion is obtained;
Preferably, the concentration of SIP solution is 20-25%, and the best is 25%;
Preferred further, the PH of SIP solution is 8.0.
Preferably, two step emulsifications all realize in high speed homogenization device, and preferably, high speed homogenization device rotating speed is 2000rmp, maintain 2min; .
Preferably, described step 2) step b in, slowly add the aqueous pectin solution that concentration is 20-25%, pH5.0-5.5, and control emulsion temperature and be always 40 DEG C.
Preferably, described step 2) step b in, with the phosphoric acid of 1mol/L, ph value of emulsion is transferred to 4.4; Preferably, freezing employing-18 DEG C is carried out.
The anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish provided by the invention, embedding rate high (can 82% be reached), degree of hydrolysis is low, and (90min degree of hydrolysis is 10.97%, be 13.71% after 24h), the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish are that the non-oxidizability making grease has stronger antioxidation activity, the substantially same vitamin E of its effect.And, the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish have slow-release capability simultaneously, effectively can play the active material of its core, and extend action time, oxygen molecule to be resisted in environment on the impact of core chelate, thus improve stability and the utilization ratio of product.
Accompanying drawing explanation
Accompanying drawing is used to provide a further understanding of the present invention, and forms a part for description, together with embodiments of the present invention for explaining the present invention, is not construed as limiting the invention.In the accompanying drawings:
Fig. 1 is the non-chelate of enzymolysis polypeptide and ferrous chelate compound photo and corresponding infrared spectrogram;
Fig. 2 is that ferrous chelate compound under light microscope is at microencapsulated forms (coomassie brilliant blue staining) photo (eyepiece 100 ×) being wall material with SPI and pectin;
Fig. 3 shows ferrous chelate compound aggregate under light microscope;
Fig. 4 is the impact of wall core comparison embedding rate;
Fig. 5 is that SPI and pectin ratio are on the impact of embedding rate;
Fig. 6 is that the antioxidant effect of sample to soybean oil compares.
Detailed description of the invention
First it may be noted that embodiments of the invention only disclose preferred embodiment several, not should be appreciated that paired restriction of the invention process, protection scope of the present invention is still as the criterion with the content disclosed in claims.
1 material and instrument
Material:
Fish gruel (industrial Co., Ltd provides by Zhejiang)
Papain (Nanning Pang Bo bioengineering Co., Ltd)
Ascorbic acid, soybean protein isolate, pectin, vitamin E (traditional Chinese medicines group is analysis pure)
Soybean oil (food grade, supermarket is bought)
Instrument:
SHAB constant temperature oscillator (Changzhou Guo Hua Co., Ltd)
Rotary Evaporators (the triumphant instrument and equipment Co., Ltd in Shanghai one)
DECTA320A/CpH counts (Mei Teletuo benefit company)
90-2 type constant temperature blender with magnetic force (Shanghai lover Instrument Ltd.)
U-2800 ultraviolet specrophotometer (FDAC Co., Ltd)
Freeze drier (Shanghai De Lefu)
2 methods
The preparation of the rotten Peptides ferrous chelate compound of 2.1 fishes
50g fish is rotten to be mixed with 500ml water, adjusts PH6.0, to being incubated 10min at 45 DEG C, then add papain in the ratio of 2000U/g, enzymolysis eight hours at same temperature, then enzymolysis liquid is centrifugal goes precipitation, adjust PH to 6.5, add ascorbic acid, then add FeCl in proportion
2, chelating 30min in constant temperature oscillator, last Vacuum Concentration, freeze drying adult fish rotten Peptides ferrous chelate compound powder is for subsequent use.
2.2 double-layer embedment microcapsules preparations
First, with ferrous chelate compound: the ratio of soybean oil (1:3), by ferrous ferrous chelate compound emulsification, obtains the emulsion of W/O; Then by the sample processed and SIP solution (PH is 8.0) emulsification, obtain O/W emulsion, above two steps are all in the inner realization of high speed homogenization device (2000rmp, 2min).
Aqueous pectin solution is slowly poured in this emulsion, mix under magnetic stirrer.Then with the phosphoric acid of 1mol/L, emulsion PH is transferred to 4.4 (temperature remains on 40 DEG C).
By the material store overnight at 7 DEG C handled well, then inner freezing at general refrigerator (-18 DEG C), last freeze drying powdered.
The mensuration of 2.3 microcapsule embedded rates
Embedding rate measures the most important index of microcapsule product.Account form is formula (1)
Use aas determination iron concentration.Measure the embedding rate of microcapsule product, first prepared ferrous chelate compound microcapsules with distilled water cyclic washing, washed away the chelate of surface of microcapsule, suction filtration, dry.Precision takes dry microcapsules 100mg in 100ml tool plug conical flask again, adds the sodium citrate solution 50ml of 0.5mol/l, in 40 DEG C, the centrifugal 10min of 50r/min jolting 6h, 5000r/min, gets filtrate 1ml and be diluted to 10ml.More than step is played empty and is treated to contrast, and use aas determination iron concentration, right back-pushed-type (1) calculates microcapsule embedded rate.
The mensuration of 2.4 microcapsules solubility
Solid sample (10 mg/ml) is dissolved in (pH value 7.0) in distilled water.At room temperature constantly stir, 90 minutes and 24 hours.Then centrifugal 5 minutes of 6000r at 15 DEG C.Measure from supernatant lyolysis degree and use Lowry-Folin method, calculate from the microcapsules concentration of solution, spectrophotometer measures absorbance under 750nm, with reference to formula (2).
Linear and the ascent of formula (2) is from ferrous chelate compound concentration (mg/ml) and absorbance.Formula (3) shows the formula of ferrous chelate compound concentration (mg/ml).
Solubility ferrous chelate compound solubility is expressed as every 100 grams of g total protein.Microcapsules solubility carry out measuring be dissolved in 90 minutes with their behavior of sample observation after 24 hours short with expansion contact with water.
2.2.5 water content and absorptive mensuration
The moisture determination of microcapsules, the encapsulation dried bean noodles of 0.2g is at 100 DEG C of dry and cooling culture dishes in advance in 24 hours.Then moisture is calculated according to formula (4).
The mensuration of water absorption rate, sample thief (2g) is placed in culture dish, puts into containing Na
2so
4airtight plastic containers in allow it permeate, after one week, the quality that sample increases is the water imbibition of sample.
2.2.6 the mensuration of microcapsules antioxidant effect
Anti-oxidant method for measuring: take certain quantity of fresh soybean oil, put into beaker, add a certain amount of antioxidant, stir, be placed in 60 ± 0.5 DEG C of insulating box strengthenings to preserve, stir once every 24h, and exchange its position in insulating box, the peroxide value (POV value) in results of regular determination oil sample.
3. result and discussion
The description of 3.1 ferrous chelate compounds
As shown in Figure 1, enzymolysis polypeptide is pale yellow powder shape without the product of ferrous chelating.The ferrous chelate products of enzymolysis protein, in white, is also Powdered.
Adopt fourier transform infrared spectroscopy to carry out the initial analysis of chelate structure: take a morsel sample, adds KBr and make diluent, in agate mortar, be ground to particle tiny and even, compression molding.With Fourier transformation infrared spectrometer test, resolution ratio is set to 4cm
-1, scanning times 40 times.The results are shown in Figure 2 and Fig. 3.
Infrared spectrogram in Fig. 1 is the infrared spectrogram before and after Peptides and ferrous chelating respectively, 2901.23cm in Fig. 2
-1be caused by N-H stretching vibration, and in figure 3, this wave band become 3011.67cm
-1, move 110 wave numbers to high band, become ammonium salt NH
4+characteristic frequency, therefore can judge that the amino in fish rotten albumen complex enzyme hydrolysis peptide take part in ferrous chelating; 1202.12cm simultaneously in Fig. 2
-11254.36cm in figure 3
-1, move 52 wave numbers to Gao Bowei, become the characteristic frequency of the carboxylate that COO-stretching vibration causes.Therefore can judge, carboxyl also take part in the chelatropic reaction of fish rotten albumen complex enzyme hydrolysis peptide-ferrous iron.
3.2 microencapsulated forms
Fig. 2 represents that ferrous chelate compound is at the microencapsulated forms being wall material with SPI and pectin, and coomassie brilliant blue staining demonstrates them and defines different layers.Have research display, the form of microcapsules can present solid form, namely if there is similar form, means and so uses double-layer embedment (complex coacervation) to prepare microcapsules success.Fig. 3 clearly show the existence of oil droplet, also has microcapsules and the irregular gathering of surrounding ferrous chelate compound.
3.3 Elements research relevant with embedding rate
In experiment process, several factors can affect the embedding rate of microcapsules, and we are contrasted by document here, chooses and does experiment of single factor to ratio (SPI/ pectin) and the embedding time of the larger wall core ratio of embedding rate impact, wall material material.
3.2.1 the impact of wall core comparison embedding rate
Getting SPI with the ratio of pectin is 1:1, embedding time 20min, and other factors are according to reference in experiment flow, and design wall core ratio is respectively 1:3,1:2,1:1,2:1,3:1,5:1, measures embedding rate.Result shows below:
Fig. 4 result shows, and be that embedding rate can rise when wall material material is greater than core material gradually, wall core reaches more satisfactory level than when 2:1, even if the quality of wall material is not remarkable especially on the impact of embedding rate higher than core quality far away yet afterwards.Wall core ratio is fixed on 2:1 by two experiment of single factor next.
3.2.2SPI with the impact of the comparison embedding rate of pectin quality
Get wall core than being 2:1, the embedding time is decided to be 20min, and the mass ratio of design SPI and pectin is respectively 30%:70%, 40%:60%, 45%:55%, 50%:50%, 60%:40%, measures embedding rate result as follows:
Fig. 5 shows, when the mass fraction of pectin is a bit larger tham SPI, embedding rate can reach more satisfactory level, when SPI and pectin mass ratio are 45%:55%, embedding rate reaches optimum level, and this result is also similar to the result of study of the people such as DeboraV, namely in double-layer embedment method, the mass fraction of outer wall material, slightly larger than the mass fraction of inner layer wall material, is conducive to product and improves embedding rate.
3.2.3 the time that embeds is on the impact of embedding rate
By above two experiment of single factor, we determine, and SPI and pectin mass ratio are 45%:55%, wall core than being 2:1, arranging the embedding time is respectively 10min, 20min, 30min, 40min, 50min, 60min.Measure embedding rate respectively, result shows below:
The impact of the embedding time that shows on embedding rate of above visual result, the embedding time 20 to 30min time reach maximum, but, when the time more than 30min after, embedding rate is almost unchanged, mean that 30min is the time of embedding microcapsules the best, time expand, effectively can not improve embedding rate, also can cause the wasting of resources.
Embedding rate affects by several factors, in our study, determined that the optimum process condition of the best embedding microcapsules is: wall core is 45%:55% than being 2:1, SPI and pectin mass ratio, the embedding time is 30 minutes, farthest can use our core ferrous chelate compound.
3.4 solubility studies
The assessment carrying out solubility is to observe the behavior of microcapsules in water, and this is to verify that ferrous chelate compound hydrolysate will be released in this medium.As the cohesion that the expection of microcapsules produces, if solubility is very low in water (even if soaking after 24 hours), then confirm the maintenance of microcapsules integrality.Therefore can know that using method and encapsulating material can obtain highly stable microcapsules, there is good controlled release characteristics.
Solubility increases directly relevant with the increase of the hydrolysis degree of microcapsules.Within 90 minutes and 24 hours at sample solubility, constantly increase.This result can relate to such fact: the embedding rate of hydrolysis higher concentration is lower.Therefore, solable matter is do not form microcapsules by ferrous chelate compound and SPI certainly, and table 1 also show similar result.Therefore, the result of solubility be due to ferrous chelate compound and the huge affinity of water.Certainly, certain error is had, because can not suppose that microcapsules do not dissolve completely here with solubility assessment embedding rate.But solubility also must be an effectively evaluation aspect.
3.5 microcapsules water content and absorptive research
The research display moisture of the people such as DeboraV and water imbibition are also significantly to evaluate the index of microcapsules effect.Study with other and compare, sample moisture content is in critical field, and moisture content is also the reference index of microbial stability.By ferrous chelate compound and the absorptive comparison of its microencapsulated products, the water imbibition of ferrous chelate compound has been (table 1) about the absorptive twice of microcapsule product, and namely the water imbibition of microcapsule product is significantly lower than water imbibition under its core free state.
Table 1: ferrous chelate compound and its microcapsule product characteristic research result
mean value ± the standard deviation of numeral replication
sample comes from after Optimum Experiment, determines that SPI and pectin mass ratio are that 45%:55% embeds time 30min, gets its wall core ratio and be respectively 2:1,1:1,1:2, numbered samples 1,2,3.
3.6 microcapsules Study of Antioxidation
Get four 100ml beakers and add 50g fresh soyabean oil respectively, be numbered 1,2,3,4.Antioxidant is not added in beaker 1, as blank test, the ferrous chelate compound that 0.5g does not embed is added in 2,0.5g ferrous chelate compound microcapsules are added in 3, add 0.5g in 4 and enter vitamin E, stir, be placed in 60 ± 0.5 DEG C of insulating box strengthenings and preserve, stir once every 24h, and exchange its position in insulating box.Then according to the peroxide value (POV value) of GB/T5009.37-1996 method results of regular determination oil sample, carry out contrast and experiment and see Fig. 6.
Shown by Fig. 6 result, ferrous microcapsules are have stronger antioxidation activity as the non-oxidizability of grease, and its effect is same vitamin E substantially, illustrate that our ferrous microcapsules are feasible as the antioxidant of grease.Fig. 6 also demonstrates, initial several days of experiment, add the soybean oil POV value of ferrous microcapsules a little more than the ferrous chelate compound not having to embed, but 4 days are starkly lower than the ferrous chelate compound do not embedded later, illustrate that ferrous microcapsules have certain slow-release capability, oxygen molecule can also to be resisted in environment on the impact of core chelate.So need the regular hour due to the release of microcapsule product core, so before not reaching release balance, the ferrous chelate compound thing of its antioxidant effect not as not embedding; And after reaching release balance, its antioxidant effect is brought into play gradually.
And the ferrous chelate compound never embedded compares with the antioxidant effect of vitamin E, can find, early stage, the ferrous chelate compound antioxidant effect that do not embed was a little more than vitamin E, but caused final result to be not so good as vitamin E ideal due to the unstability of self.So the microcapsule product of ferrous chelate compound, because microcapsules characteristic, make its slow releasing antioxidant content in the sample to which, thus improve stability and the utilization ratio of product.
4. conclusion
In this research, be the microcapsules that ferrous chelate compound prepared by wall material with SPI and pectin, pass through experiment of single factor, determining the optimum process condition preparing microcapsules is that wall core is than being 2:1, SPI and pectin mass ratio are 45%:55%, the embedding time is 30 minutes, the researchs such as solubility water imbibition are carried out to the finished product of microcapsules, and the mensuration of antioxidation activity, determine that microcapsules effectively play the effect of protection core, and due to the slow releasing function of microcapsules, effectively can play the active material of its core, and extend action time, for effective utilization of little peptide metallo-chelate provides effective reference frame.
Finally being also pointed out that any entity or individual use or implement technical scheme of the present invention is all to infringement of the present invention, and any entity or individual, without the permission of the applicant, can not implement this patent separately.And any entity or individual are subject to inspiration of the present invention or implement through simple adjustment, also should think the protection domain of this patent.
Claims (10)
1. the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish, it is characterized in that: be made up of fish rotten Peptides ferrous chelate compound powder, SPI and pectin, fish rotten Peptides ferrous chelate compound powder is for making core, SPI and pectin as wall material by double-layer embedment inside and outside core, wherein, SPI is inner layer wall material, and pectin is outer wall material.
2. the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 1, is characterized in that: wall core weight ratio is more than 2:1; Preferably, wall core weight ratio is 2:1; Further preferred, SPI: the mass ratio of pectin is 45:55.
3. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 1, it is characterized in that: with fish rotten Peptides ferrous chelate compound powder for core, embed SPI and pectin respectively in powder particle skin, the average grain diameter of these microcapsules is 65 μm.
4. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 3, is characterized in that: comprise the steps:
1) preparation of the rotten Peptides ferrous chelate compound of fish
Obtain enzymolysis liquid with after the gruel of papain enzymolysis fish, then add Fe
2cl carries out chelatropic reaction, obtains the rotten Peptides ferrous chelate compound of fish;
2) double-layer embedment microcapsules preparation
A.SIP emulsifying soln
By step 1) in the fish that obtains rotten Peptides ferrous chelate compound powder dispersion in soybean oil, then with SIP emulsifying soln, obtain O/W emulsion;
B. embed
Poured into by pectin in the emulsion obtained in step a, stir 20-30min mixing, after adjust ph to 4.4, store overnight at 7 DEG C, refreezes into freeze-dried powder again, can obtain the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish.
5. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 4, is characterized in that: described step 1) in enzyme digestion reaction concrete operations as follows:
First fish gruel is pressed 50g:500ml with water to mix, regulate pH to be 6.0, at 45 DEG C, be incubated 10min, then add papain in the ratio of 2000U/g, enzymolysis 8h at 45 DEG C.
6. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 4, is characterized in that: described step 1) in chelatropic reaction specific as follows:
By centrifugal for enzymolysis liquid go precipitation, adjust pH be 6.5, add ascorbic acid, more on weight of solution 1%-3% add FeCl
2, chelating 30min in constant temperature oscillator, then Vacuum Concentration, freeze drying adult fish rotten Peptides ferrous chelate compound powder.
7. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 4, is characterized in that: described step 2) step a in, concrete operations are:
With ferrous chelate compound: the weight ratio of soybean oil be the ratio of 1:3 by ferrous chelate compound emulsification, obtain the emulsion of W/O; Then by the sample that processed and SIP emulsifying soln, O/W emulsion is obtained;
Preferably, the concentration of SIP solution is 20-25%, and the best is 25%;
Preferred further, the PH of SIP solution is 8.0.
8. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 7, it is characterized in that: two step emulsifications all realize in high speed homogenization device, preferably, high speed homogenization device rotating speed is 2000rmp, maintains 2min.
9. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 6, it is characterized in that: described step 2) step b in, slowly add the aqueous pectin solution that concentration is 20-25%, pH5.0-5.5, and control emulsion temperature and be always 40 DEG C.
10. the preparation technology of the anti-oxidant microcapsules of the rotten Peptides ferrous chelate compound of fish according to claim 6, is characterized in that: described step 2) step b in, with the phosphoric acid of 1mol/L, ph value of emulsion is transferred to 4.4; Preferably, freezing employing-18 DEG C is carried out.
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