CN104382020A - Preparation method of highland barley dietary fiber-polyphenol compounds - Google Patents
Preparation method of highland barley dietary fiber-polyphenol compounds Download PDFInfo
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- CN104382020A CN104382020A CN201410604115.1A CN201410604115A CN104382020A CN 104382020 A CN104382020 A CN 104382020A CN 201410604115 A CN201410604115 A CN 201410604115A CN 104382020 A CN104382020 A CN 104382020A
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- 235000007340 Hordeum vulgare Nutrition 0.000 title claims abstract description 80
- 235000005911 diet Nutrition 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 230000000378 dietary effect Effects 0.000 title abstract description 13
- 240000005979 Hordeum vulgare Species 0.000 title 1
- 241000209219 Hordeum Species 0.000 claims abstract description 79
- 239000007788 liquid Substances 0.000 claims abstract description 38
- 235000013312 flour Nutrition 0.000 claims abstract description 37
- 239000006228 supernatant Substances 0.000 claims abstract description 27
- 238000009777 vacuum freeze-drying Methods 0.000 claims abstract description 20
- 239000002244 precipitate Substances 0.000 claims abstract description 17
- 238000000034 method Methods 0.000 claims abstract description 8
- 238000001816 cooling Methods 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 68
- 238000006243 chemical reaction Methods 0.000 claims description 37
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 27
- 239000007983 Tris buffer Substances 0.000 claims description 24
- 238000013019 agitation Methods 0.000 claims description 22
- 102000004139 alpha-Amylases Human genes 0.000 claims description 22
- 108090000637 alpha-Amylases Proteins 0.000 claims description 22
- 229940024171 alpha-amylase Drugs 0.000 claims description 22
- 108010089934 carbohydrase Proteins 0.000 claims description 18
- 108091005804 Peptidases Proteins 0.000 claims description 15
- 239000004365 Protease Substances 0.000 claims description 15
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 15
- 239000003513 alkali Substances 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 230000037213 diet Effects 0.000 claims description 13
- 230000010355 oscillation Effects 0.000 claims description 11
- 238000001556 precipitation Methods 0.000 claims description 11
- 239000012535 impurity Substances 0.000 claims description 9
- 230000001105 regulatory effect Effects 0.000 claims description 8
- 108091005658 Basic proteases Proteins 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 239000000203 mixture Substances 0.000 abstract description 6
- 238000011161 development Methods 0.000 abstract description 5
- 238000012545 processing Methods 0.000 abstract description 4
- 230000002255 enzymatic effect Effects 0.000 abstract description 3
- 238000007710 freezing Methods 0.000 abstract 2
- 230000008014 freezing Effects 0.000 abstract 2
- 102000004190 Enzymes Human genes 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 1
- 239000007853 buffer solution Substances 0.000 abstract 1
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 235000013325 dietary fiber Nutrition 0.000 description 13
- 235000013339 cereals Nutrition 0.000 description 11
- 238000012216 screening Methods 0.000 description 7
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- 238000005516 engineering process Methods 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 230000003064 anti-oxidating effect Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
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- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
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- 208000010643 digestive system disease Diseases 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 230000002641 glycemic effect Effects 0.000 description 1
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- 208000019622 heart disease Diseases 0.000 description 1
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- 229930005346 hydroxycinnamic acid Natural products 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
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- 235000020824 obesity Nutrition 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
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- NGSWKAQJJWESNS-ZZXKWVIFSA-N trans-4-coumaric acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C=C1 NGSWKAQJJWESNS-ZZXKWVIFSA-N 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- 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
Abstract
The invention discloses a preparation method of highland barley dietary fiber-polyphenol compounds. The preparation method specifically comprises the following steps: dissolving highland barley flour in a buffer solution to obtain a mixture I, performing enzymolysis on the mixture I in a dark place, centrifuging while the enzymatic hydrolysate is hot to obtain precipitates which are highland barley insoluble dietary fiber-polyphenol compounds (IDF-PC), collecting and freezing the precipitates at low temperature, performing vacuum freeze drying on the frozen precipitates, and storing the vacuum freeze-dried precipitates at -20 DEG C; adding alcohol into centrifuged liquid supernatant to obtain a mixture II, standing and naturally cooling the mixture II to the room temperature, precipitating and staying the mixture II overnight at low temperature, taking a lower-layer muddy part and centrifuging to obtain precipitates which are highland barley soluble dietary fiber-polyphenol compounds (SDF-PC), collecting and freezing precipitates at low temperature, performing vacuum freeze drying on the frozen precipitates, and storing the vacuum freeze-dried precipitates at -20 DEG C. According to the preparation method of the highland barley dietary fiber-polyphenol compounds disclosed by the invention, the highland barley soluble dietary fiber-polyphenol compounds and the highland barley insoluble dietary fiber-polyphenol compounds are separated and prepared by utilizing an enzyme weight method, so that scientific basis is provided for developing and utilizing the highland barleys and a new thought is also provided for deeply processing the highland barleys, and therefore, the development of the highland barely industry is promoted.
Description
Technical field
The invention belongs to food processing field, be specifically related to the preparation method of barley diet fiber-polyphenol complex.
Background technology
Whole grain food effectively can prevent metabolic syndrome, the incidence of disease of prevention and reduction angiocardiopathy, type II diabetes, obesity and some cancer.Cereal foods, as food source main in the world, provides the dietary fiber that 50% human body is taken in.Research finds, takes in dietary fiber and can regulate human body starvation, glycemic index, fat metabolism and inflammation, play prebiotic function, can effectively preventing chronic heart disease and disease of digestive system.Many researchers are studied for the polysaccharide component in cereal, but do not illustrate its wholesome biochemical mechanism so far.Because grain polysaccharide component contains a large amount of polyphenolic substances, the cereal polyphenol of 95% and cell wall polysaccharides are combined closely, researcher proposes " dietary-fiber antioxidant (dietary fibre-antioxidants) " and the concept of " dietary fiber-polyphenol complex (dietary fibre-phenoliccompounds; DF-PC) ", thinks that grain dietary fiber is the natural function composition transmitting polyphenolic substance in body.In cereal, dietary fiber comprises soluble dietary fiber (solubledietary fibre, SDF) and soluble dietary fiber (insoluble dietary fibre, IDF), the two example in cereal and grain species relation very large; Phenolic acid in cereal mainly hydroxycinnamic acid, wherein ferulaic acid content is the abundantest, is regarded as the topmost polyphenolic substance of cereal antioxidation activity.Dietary fiber and polyphenolic substance pass through, together with ester bond covalent bond, to form DF-PC compound.It is generally acknowledged, in cereal, SDF/IDF ratio is higher, and DF-PC compound antioxidant effect is better.
Highland barley, as traditional long-term cropping of Tibetan Autonomous Prefecture of Deqen of Yunnan Province, containing functional components such as abundant dietary fiber, polyphenolic substance and trace elements, has good Development volue.But highland barley deep processing degree is not dark at present, limits increasing peasant income, constrains the development of highland barley industry.This technology is by the preparation method of research highland barley DF-PC compound, and by studying wherein polyphenolic substance, Flavonoid Content and its antioxidation activity, for illustrating dietary fiber and the mechanism of action of polyphenol interaction to polyphenol bioavilability, improve biological effectiveness and the bioavilability of polyphenolic substance, continue to play antioxidation, prevent disease, maintain body health and scientific basis is provided, there is provided thinking for highland barley develops simultaneously, promote the development of Yunnan Province's highland barley industry.
Summary of the invention
The object of the present invention is to provide barley diet fiber-polyphenol complex preparation method, this law adopts enzymatic gravimetric method, barley diet fiber-polyphenol complex preparation method that preparation purity is high, performance good, cost is low.
The present invention is achieved through the following technical solutions:
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after highland barley removal of impurities, cross 50 mesh sieves, the lower highland barley flour of sieve is for subsequent use;
2) take the lower highland barley flour of sieve, dissolve highland barley flour with the MES-TRIS cushioning liquid of pH6.25, concentration 0.05mol/L, magnetic agitation is until sample is dispersed in cushioning liquid completely;
3) lucifuge adds Thermostable α-Amylase (adding Thermostable α-Amylase by 165U/g highland barley flour) solution, is placed in 95 DEG C of constant temperature oscillation water-bath Keep agitation, when the temperature of solution is increased to 95 DEG C of beginning timing, reaction 30min;
4) after having reacted, treat that solution is cooled to 45 DEG C, regulate pH value of solution: regulate pH to 10.5 by NaOH solution, add alkali protease (adding alkali protease by 750U/g highland barley flour) solution in the solution, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started, Keep agitation, reaction 30min when solution temperature reaches 45 DEG C;
5) after reaction terminates, regulate pH value of solution: regulate pH to 4.25 with HCl solution, in solution, add carbohydrase (adding carbohydrase by 600U/g highland barley flour) while stirring, Keep agitation, starts timing when temperature reaches 61 DEG C, reaction 30min;
6) by enzymolysis liquid while hot in the centrifugal 15min of 4000r/min, be precipitated as IDF-PC, freeze, vacuum freeze drying 4d after collection at-18 DEG C, load in valve bag ,-18 DEG C of lucifuges store;
7) add 60 DEG C of 95% ethanolic solution of 4 times of volumes in supernatant, leave standstill, naturally cool to room temperature, in 0-4 DEG C of refrigerator, precipitate more than 12 hours;
8) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store.
During preparation method's step (3) Thermostable α-Amylase enzymolysis of the present invention, the Thermostable α-Amylase added with now taking Thermostable α-Amylase, dissolves obtained with the MES-TRIS cushioning liquid of pH=6.25, concentration 0.05mol/L by existing.
During preparation method's step (4) cooling of the present invention, directly beaker is put into water at low temperature and cool.
During preparation method's step (5) alkali protease enzymolysis of the present invention, the alkaline protease solution added dissolves obtained by the MES-TRIS cushioning liquid now with the alkali protease pH=10.5 now taken, concentration 0.05mol/L.
Preparation method's step (8) of the present invention ensures that ethanolic solution temperature is 60 DEG C, and when temperature does not reach, precipitation is incomplete, and supernatant is more muddy, treats that temperature is cooled to room temperature and puts into refrigerator again.
Beneficial effect of the present invention is: the technology of the present invention enzymatic gravimetric method carries out separation preparation to soluble dietary fiber-polyphenol complex, insoluble dietary fiber-polyphenol complex in highland barley, for the exploitation of highland barley provide scientific basis, also for the intensive processing of highland barley provides new approaches, the development of highland barley industry is promoted.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described further, the following stated, only to preferred embodiment of the present invention, not do other forms of restriction to the present invention, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed to the Equivalent embodiments of equal change.Everyly do not depart from the present invention program's content, according to technical spirit of the present invention to any simple modification made for any of the above embodiments or equivalent variations, all drop in protection scope of the present invention.
The dietary fibre materials used in following examples is highland barley, and other are containing the raw material of dietary fiber, as barley, wheat etc. also can be prepared by method of the present invention.
Embodiment 1
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) claim screenings highland barley flour 10g, be placed in 500mL, add 400mL MES-TRIS (pH=6.25,0.05mol/L) cushioning liquid, heating water bath to 95 DEG C, add 330u/mL Thermostable α-Amylase 5mL;
3) after adding Thermostable α-Amylase, temperature is increased to 95 DEG C of beginning timing, reaction 30min, and constantly stirs;
4) beaker is taken out, be cooled to 45 DEG C;
5) pH value of solution is regulated: regulate the pH value of solution to 10.5 in beaker with 6mol/L NaOH, add MES-TRIS (pH=10.5,0.05mol/L) the 7500U/mL alkaline protease solution 1mL of cushioning liquid configuration, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started, Keep agitation reaction 30min when temperature in beaker reaches 45 DEG C;
6) after reaction terminates, by beaker as in 61 DEG C of water-baths, regulate pH value of solution: regulate pH to 4.25 with 6mol/LHCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 30 μ L are added while stirring in beaker, Keep agitation, timing is started, reaction 30min as the solution Nei Wenduda in beaker 61 DEG C;
7) enzymolysis liquid is placed in Buchner funnel, filter paper is fast quantification filter paper (9cm), 0.08MP suction filtration, the rate of filtration is slower, cannot extract in a large number, by enzymolysis liquid centrifugal 15min under 4000r/min condition, result can abundant separatin non-soluble dietary fiber-polyphenol complex (IDF-PC);
8) collecting precipitation IDF-PC, after freeze at-18 DEG C, vacuum freeze drying 4d, loads in valve bag, and-18 DEG C of lucifuges store;
9) supernatant is transferred in another beaker, adds 95% ethanolic solution equaling supernatant 4 times of volumes, leaves standstill, in 0-4 DEG C of refrigerators, precipitate more than 12 hours;
10) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store.
Embodiment 2
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) claim screenings highland barley flour 5g, be placed in 500mL, add 200mL MES-TRIS (pH=6.25,0.05mol/L) cushioning liquid;
3) heating water bath to 95 DEG C, adds the Thermostable α-Amylase 2.5mL of 330u/mL, starts timing when temperature in beaker reaches 95 DEG C, reaction 30min, and constantly stirs;
4) beaker is taken out, be cooled to 45 DEG C;
5) pH value of solution is regulated: regulate the pH value of solution to 10.5 in beaker with 6mol/L NaOH, add MES-TRIS (pH=10.5,0.05mol/L) the 7500U/mL alkaline protease solution 0.5mL of cushioning liquid configuration, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started when temperature in beaker reaches 45 DEG C, Keep agitation, reaction 30min;
6) after reaction terminates, by beaker as in 61 DEG C of water-baths, regulate pH value of solution: regulate pH to 4.25 with 6mol/LHCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 15 μ L are added while stirring in beaker, Keep agitation, timing is started, reaction 30min as the solution Nei Wenduda in beaker 61 DEG C;
7) enzymolysis liquid is placed in the centrifugal 15min of 4000r/min, sediment fraction is IDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store;
8) supernatant is transferred in another beaker, adds preheating 60 DEG C of 95% ethanolic solution equaling supernatant 4 times of volumes, leaves standstill, naturally cools to room temperature, precipitates overnight in 0-4 DEG C of refrigerator;
9) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained sediment fraction is SDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store.
Embodiment 3
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) claim screenings highland barley flour 100g, be placed in 5L Plastic Drum, add 2L MES-TRIS (pH=6.25,0.05mol/L) cushioning liquid;
3) heating water bath to 95 DEG C, adds the Thermostable α-Amylase 50mL of 330u/mL, starts timing, and constantly stir when temperature in beaker reaches 95 DEG C, reaction 30min;
4) taken out by Plastic Drum, cold bath is cooled to 45 DEG C;
5) pH value of solution is regulated: regulate the pH value of solution to 10.5 in beaker with 6mol/L NaOH, add MES-TRIS (pH=10.5,0.05mol/L) the 7500U/mL alkaline protease solution 10mL of cushioning liquid configuration, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started when temperature in beaker reaches 45 DEG C, Keep agitation, reaction 30min;
6) after reaction terminates, by beaker as in 61 DEG C of water-baths, regulate pH value of solution: regulate pH to 4.25 with 6mol/LHCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 600 μ L are added while stirring in beaker, timing is started as the solution Nei Wenduda in beaker 61 DEG C, Keep agitation, reaction 30min;
7) enzymolysis liquid is placed in while hot the centrifugal 15min of 4000r/min, is precipitated as IDF-PC, freeze, vacuum freeze drying 4d after collection at-18 DEG C, load in valve bag ,-18 DEG C of lucifuges store;
8) supernatant is transferred in another beaker, add equal supernatant 4 times of volumes be preheating to 60 DEG C of 95% ethanolic solution, leave standstill, naturally cool to room temperature, precipitates overnight in 0-4 DEG C of refrigerator;
9) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store;
Embodiment 4
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) claim screenings highland barley flour 300g, be placed in 5L Plastic Drum, add 4.5LMES-TRIS (pH=6.25,0.05mol/L) cushioning liquid;
3) heating water bath to 95 DEG C, add with 150mL MES-TRIS (pH=6.25,0.05mol/L) Thermostable α-Amylase (adding Thermostable α-Amylase by the 165U/g highland barley flour) 12.375g of cushioning liquid dissolving, timing is started when temperature in beaker reaches 95 DEG C, and constantly stir, reaction 30min;
4) taken out by Plastic Drum, cold bath is cooled to 45 DEG C;
5) pH value of solution is regulated: regulate the pH value of solution to 10.5 in beaker with 6mol/L NaOH, add with 30mL MES-TRIS (pH=10.5,0.05mol/L) alkali protease (adding alkali protease by the 750U/g highland barley flour) 1.875g of cushioning liquid dissolving, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started when temperature in beaker reaches 45 DEG C, Keep agitation, reaction 30min;
6) after reaction terminates, by beaker as in 61 DEG C of water-baths, regulate pH value of solution: regulate pH to 4.25 with 6mol/LHCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 1.8mL is added while stirring in beaker, timing is started as the solution Nei Wenduda in beaker 61 DEG C, Keep agitation, reaction 30min;
7) enzymolysis liquid is placed in while hot the centrifugal 15min of 4000r/min, is precipitated as IDF-PC, freeze, vacuum freeze drying 4d after collection at-18 DEG C, load in valve bag ,-18 DEG C of lucifuges store;
8) supernatant is transferred in another beaker, add equal supernatant 4 times of volumes be preheating to 60 DEG C of 95% ethanolic solution, leave standstill, naturally cool to room temperature, precipitates overnight in 0-4 DEG C of refrigerator;
9) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store.
Embodiment 5
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) claim screenings highland barley flour 500g, be placed in Plastic Drum, add 7.5L MES-TRIS (pH=6.25,0.05mol/L) cushioning liquid;
3) heating water bath to 95 DEG C, add with 250mL MES-TRIS (pH=6.25,0.05mol/L) Thermostable α-Amylase (adding Thermostable α-Amylase by the 165U/g highland barley flour) 20.625g of cushioning liquid dissolving, timing is started when temperature in beaker reaches 95 DEG C, and constantly stir, reaction 30min;
4) taken out by Plastic Drum, cold bath is cooled to 45 DEG C;
5) pH value of solution is regulated: regulate the pH value of solution to 10.5 in beaker with 6mol/L NaOH, add with 50mL MES-TRIS (pH=10.5,0.05mol/L) alkali protease (adding alkali protease by the 750U/g highland barley flour) 3.125g of cushioning liquid dissolving, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started when temperature in beaker reaches 45 DEG C, Keep agitation, reaction 30min;
6) after reaction terminates, by beaker as in 61 DEG C of water-baths, regulate pH value of solution: regulate pH to 4.25 with 6mol/LHCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 3mL is added while stirring in beaker, timing is started as the solution Nei Wenduda in beaker 61 DEG C, Keep agitation, reaction 30min;
7) enzymolysis liquid is placed in while hot the centrifugal 15min of 4000r/min, is precipitated as IDF-PC, freeze, vacuum freeze drying 4d, obtain IDF-PC133.75g after collection at-18 DEG C, yield is 26.75%, loads in valve bag, and-18 DEG C of lucifuges store;
8) supernatant is transferred in another beaker, add equal supernatant 4 times of volumes be preheating to 60 DEG C of 95% ethanolic solution, leave standstill, naturally cool to room temperature, precipitates overnight in 0-4 DEG C of refrigerator;
9) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freeze at-18 DEG C after collection, vacuum freeze drying 4d, obtain SDF-PC35.97g, yield is 7.20%, loads in valve bag, and-18 DEG C of lucifuges store.
Embodiment 6
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) claim screenings highland barley flour 250g, be placed in Plastic Drum, add 3.75L MES-TRIS (pH=6.25,0.05mol/L) cushioning liquid;
3) heating water bath to 95 DEG C, add with 125mL MES-TRIS (pH=6.25,0.05mol/L) Thermostable α-Amylase (adding Thermostable α-Amylase by the 165U/g highland barley flour) 10.3125g of cushioning liquid dissolving, timing is started when temperature in beaker reaches 95 DEG C, and constantly stir, reaction 30min;
4) taken out by Plastic Drum, cold bath is cooled to 45 DEG C;
5) pH value of solution is regulated: regulate the pH value of solution to 10.5 in beaker with 6mol/L NaOH, add with 25mL MES-TRIS (pH=10.5,0.05mol/L) alkali protease (adding alkali protease by the 750U/g highland barley flour) 1.5625g of cushioning liquid dissolving, be placed in the constant temperature oscillation water-bath of 45 DEG C, timing is started when temperature in beaker reaches 45 DEG C, Keep agitation, reaction 30min;
6) after reaction terminates, by beaker as in 61 DEG C of water-baths, regulate pH value of solution: regulate pH to 4.25 with 6mol/LHCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 1.5mL is added while stirring in beaker, timing is started as the solution Nei Wenduda in beaker 61 DEG C, Keep agitation, reaction 30min;
7) enzymolysis liquid is placed in while hot the centrifugal 15min of 4000r/min, is precipitated as IDF-PC, freeze, vacuum freeze drying 4d, obtain IDF-PC51.41g after collection at-18 DEG C, yield is 20.56%, loads in valve bag, and-18 DEG C of lucifuges store;
8) supernatant is transferred in another beaker, add equal supernatant 4 times of volumes be preheating to 60 DEG C of 95% ethanolic solution, leave standstill, naturally cool to room temperature, precipitates overnight in 0-4 DEG C of refrigerator;
9) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freeze at-18 DEG C after collection, vacuum freeze drying 4d, obtain SDF-PC24.95g, yield is 9.98%, loads in valve bag, and-18 DEG C of lucifuges store.
Embodiment 7
The preparation method of barley diet fiber-polyphenol complex, comprises the following steps:
1) pulverize after the removal of impurities of cloud black highland barley, cross 50 mesh sieves, extracting screen underflow is for subsequent use;
2) screenings highland barley flour 100g is claimed, be placed in 28cm electromagnetic oven and be suitable for flat steamer, in flat steamer, add 1500mL MES-TRIS (pH=6.25,0.05mol/L) cushioning liquid, magnetic agitation is until sample is dispersed in cushioning liquid completely.
3) after reaction terminates, add with 50mL MES-TRIS (pH=6.25,0.05mol/L) Thermostable α-Amylase (adding Thermostable α-Amylase by the 165U/g highland barley flour) 4.1498g of cushioning liquid dissolving, the 40cm electromagnetic oven being placed in boiling is suitable for flat slaughterhouse Keep agitation, timing is started, reaction 30min when the 28cm electromagnetic oven temperature be suitable in flat steamer is increased to 95 DEG C;
4) 28cm electromagnetic oven is suitable for flat steamer to take out, in cold bath, is cooled to 45 DEG C;
5) pH value of solution is regulated: be suitable for the pH value of solution to 10.5 in flat steamer with 2mol/L NaOH adjustment 28cm electromagnetic oven, add with 10mL MES-TRIS (pH=10.5,0.05mol/L) alkali protease (adding alkali protease by the 750U/g highland barley flour) 0.3882g of cushioning liquid dissolving, be placed in the constant temperature oscillation water-bath of 45 DEG C, start timing when 28cm electromagnetic oven is suitable for when temperature reaches 45 DEG C in flat steamer, continue jolting reaction 30min;
6) after reaction terminates, regulate pH value of solution: regulate pH to 4.25 with 6mol/L HCl, 100000U/mL carbohydrase (adding carbohydrase by 600U/g highland barley flour) 600 μ L are added while stirring in the solution that 28cm electromagnetic oven is suitable in flat steamer, continue jolting, timing is started, reaction 30min when being suitable for the solution Nei Wenduda 61 DEG C in flat steamer when 28cm electromagnetic oven;
7) by enzymolysis liquid while hot in the centrifugal 15min of 4000r/min, be precipitated as IDF-PC, IDF-PC weight is 152.9467g, yield is 1.53g/g (can obtain 1.53gIDF-PC in every gram of highland barley flour), freeze at-18 DEG C after collection, vacuum freeze drying 4d, loads in valve bag, and-18 DEG C of lucifuges store;
8) supernatant is transferred in another beaker, adds 60 DEG C of 95% ethanolic solution equaling supernatant 4 times of volumes, leaves standstill, naturally cools to room temperature, in 0-4 DEG C of refrigerator, precipitate more than 12 hours;
9) supernatant is poured out gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, SDF weight is 34.5342g, yield is 0.34g/g (can obtain 0.34gSDF-PC in every gram of highland barley flour), freezes, vacuum freeze drying 4d after collection at-18 DEG C, load in valve bag ,-18 DEG C of lucifuges store.
Claims (5)
1. the preparation method of barley diet fiber-polyphenol complex, is characterized in that: comprise the following steps:
1) pulverize after highland barley removal of impurities, cross 50 mesh sieves, the lower highland barley flour of sieve is for subsequent use;
2) take the lower highland barley flour of sieve, dissolve highland barley flour with MES-TRIS cushioning liquid, magnetic agitation is until sample is dispersed in cushioning liquid completely;
3) lucifuge adds Thermostable α-Amylase solution, is placed in 95 DEG C of constant temperature oscillation water-bath Keep agitation, when the temperature of solution is increased to 95 DEG C of beginning timing, reaction 30min;
4) after having reacted, treat that solution is cooled to 45 DEG C, regulate pH value of solution: regulate pH to 10.5 by NaOH solution, add alkaline protease solution in the solution, be placed in the constant temperature oscillation water-bath of 45 DEG C, start timing when solution temperature reaches 45 DEG C, Keep agitation, reaction 30min;
5) after reaction terminates, pH value of solution is regulated: regulate pH to 4.25 with HCl solution, in solution, add carbohydrase while stirring, Keep agitation, start timing when temperature reaches 61 DEG C, reaction 30min;
6) by enzymolysis liquid while hot in the centrifugal 15min of 4000r/min, be precipitated as IDF-PC, freeze, vacuum freeze drying 4d after collection at-18 DEG C, load in valve bag ,-18 DEG C of lucifuges store;
7) add 60 DEG C of 95% ethanolic solution of 4 times of volumes in supernatant, leave standstill, naturally cool to room temperature, in 0-4 DEG C of refrigerator, precipitate more than 12 hours;
8) pour out supernatant gently after precipitation, take off the muddy part of layer in the centrifugal 15min of 4000r/min, gained is precipitated as SDF-PC, freezes, vacuum freeze drying 4d after collection at-18 DEG C, loads in valve bag, and-18 DEG C of lucifuges store.
2. method according to claim 1, it is characterized in that: in step (3) during Thermostable α-Amylase enzymolysis, the Thermostable α-Amylase added is by now with now taking, Thermostable α-Amylase pH is 6.25, the MES-TRIS cushioning liquid of concentration 0.05mol/L dissolves obtained, and every gram of highland barley flour adds 165U Thermostable α-Amylase.
3. method according to claim 1, is characterized in that: in step (4) during cooling, directly solution is put into water at low temperature and cool.
4. method according to claim 1, it is characterized in that: during step (4) alkali protease enzymolysis, the alkaline protease solution added dissolves obtained by the MES-TRIS cushioning liquid now with the alkali protease pH=10.5 now taken, concentration 0.05mol/L, and every gram of highland barley flour adds 750U alkali protease.
5. method according to claim 1, is characterized in that: step (5) is added carbohydrase and added 600U by every gram of highland barley flour.
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| CN105795482A (en) * | 2016-03-29 | 2016-07-27 | 上海交通大学 | Water extraction method of highland barley dietary fibers by optimized by response surface method |
| CN107212424A (en) * | 2016-05-03 | 2017-09-29 | 上海交通大学 | A kind of extracting method of water-soluble barley diet fiber |
| CN109222096A (en) * | 2018-09-17 | 2019-01-18 | 南昌大学 | The extracting method of polyphenol is combined in carrot dietary fiber |
| CN110250444A (en) * | 2019-07-26 | 2019-09-20 | 福建农林大学 | It is a kind of to contain bamboo shoot fiber-polyphenol complex sausage and preparation method thereof |
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| CN105795482A (en) * | 2016-03-29 | 2016-07-27 | 上海交通大学 | Water extraction method of highland barley dietary fibers by optimized by response surface method |
| CN107212424A (en) * | 2016-05-03 | 2017-09-29 | 上海交通大学 | A kind of extracting method of water-soluble barley diet fiber |
| CN109222096A (en) * | 2018-09-17 | 2019-01-18 | 南昌大学 | The extracting method of polyphenol is combined in carrot dietary fiber |
| CN110250444A (en) * | 2019-07-26 | 2019-09-20 | 福建农林大学 | It is a kind of to contain bamboo shoot fiber-polyphenol complex sausage and preparation method thereof |
| CN111088317A (en) * | 2019-12-30 | 2020-05-01 | 云南农业大学 | Method for determining bacteriostatic activity of coarse polyphenol of small coarse cereals and application |
| CN111088317B (en) * | 2019-12-30 | 2024-02-20 | 云南农业大学 | Method for measuring antibacterial activity of coarse polyphenol of minor cereals and application |
| CN113244657A (en) * | 2021-06-18 | 2021-08-13 | 四川省农业科学院农产品加工研究所 | Echelon extraction method of functional components of purple highland barley bran |
| CN115251383A (en) * | 2022-07-27 | 2022-11-01 | 劲牌持正堂药业有限公司 | Method for continuously preparing highland barley flavone extracts with two different purposes from highland barley and application |
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