CN108741082A - A kind of preparation method of Sweet Potato Leaf anti-oxidation active substance - Google Patents
A kind of preparation method of Sweet Potato Leaf anti-oxidation active substance Download PDFInfo
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- CN108741082A CN108741082A CN201810354284.2A CN201810354284A CN108741082A CN 108741082 A CN108741082 A CN 108741082A CN 201810354284 A CN201810354284 A CN 201810354284A CN 108741082 A CN108741082 A CN 108741082A
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- sweet potato
- potato leaf
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- oxidation active
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- 235000002678 Ipomoea batatas Nutrition 0.000 title claims abstract description 93
- 230000003064 anti-oxidating effect Effects 0.000 title claims abstract description 39
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- 238000002360 preparation method Methods 0.000 title claims abstract description 21
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- 230000010148 water-pollination Effects 0.000 description 3
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- NPDBDJFLKKQMCM-SCSAIBSYSA-N (2s)-2-amino-3,3-dimethylbutanoic acid Chemical class CC(C)(C)[C@H](N)C(O)=O NPDBDJFLKKQMCM-SCSAIBSYSA-N 0.000 description 1
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 description 1
- 241000207782 Convolvulaceae Species 0.000 description 1
- 241000234273 Dioscorea Species 0.000 description 1
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- 235000006536 Dioscorea esculenta Nutrition 0.000 description 1
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- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000000908 anti-sweet Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
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Classifications
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- 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
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Nutrition Science (AREA)
- Molecular Biology (AREA)
- Food Science & Technology (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Sustainable Development (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of preparation methods of Sweet Potato Leaf anti-oxidation active substance, including：Degreasing, extraction, de- albumen, decoloration, first by ultrasonic wave added hot water extraction after Sweet Potato Leaf powder degreasing, then it is digested with papain, neutral proteinase, crude product is obtained after alcohol precipitation, medical active carbon powder is added in crude product solution to decolourize to get Sweet Potato Leaf anti-oxidation active substance with t- leucine tert-butyl esters.It has the beneficial effect that：This preparation method is simple for process, cheap environmental protection, higher with the Deproteinated efficiency of mode of action, removal of protein is thorough, it is excellent with decolorizing effect with t- leucine tert-butyl esters connection with medical active carbon powder, while the amount that polysaccharide is tightly held by activated carbon can be substantially reduced again, improve the content and purity of final product.
Description
Technical field
The present invention relates to the field of deep of sweet potato, more particularly, to a kind of preparation side of Sweet Potato Leaf anti-oxidation active substance
Method.
Technical background
Sweet potato（Scientific name：Dioscorea esculenta (Lour.) Burkill）：Also known as sweet potato, Convolvulaceae Dioscorea
Wind herbaceous species.Underground stem tuber top branch ends enlarge into the stem tuber of ovoid, and crust is faint yellow, smooth.Stem is left-handed, base portion
Spinosity, by T-shaped pubescence.Single leaf alternate, it is wealthy heart-shaped；Male inflorescence is spike, Dan Sheng, and male flower is extremely short without obstructing or having
Stalk；Bract is oval, and top is tapering；Perianth tazza shape, by pubescence；Capsule prismatic, top dimple, base portion section shape, per rib wing
Shape；Seed is round, has wing.Early summer at florescence.The short day crop of sweet potato genus light, property happiness temperature, can not resist cold, more drought-enduring.Main point
Cloth is on the south 40 ° of north latitude.Cultivated area is most with Asia, and Africa is taken second place, and America occupies the 3rd.Sweet potato list leaf alternate, it is leaf to divide
For heart, triangle and palmate etc., maximum blade is up to 15 centimetres, 17 centimetres wide, general length and it is wide be no more than 10 centimetres, leaf
Fate full edge, band tooth, it is shallow singly incise, it is shallow incise again and it is deep incise again, the anxious point in top, base portion is heart-shaped, and base goes out arteries and veins 9~13, by fourth
The long pubescence of font, it is especially more with the back side；Petiole grows 5~8 centimetres, and base portion color is divided into purple, brown, green etc., spinosity.Leaf color have it is green,
Light green and purple etc..Vacuum side of blade vein color is divided light red, red, purple, pale purple and green etc..
Invention content
The purpose of the present invention is to provide a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance, this preparation method techniques
Simply, cheap environmental protection, higher with the Deproteinated efficiency of mode of action, removal of protein is thorough, with medical active carbon powder and t-
Leucine tert-butyl ester connection is excellent with decolorizing effect, while can substantially reduce the amount that polysaccharide is tightly held by activated carbon again, improves final product
Content and purity.
The present invention is directed to the problem of being mentioned in background technology, and the technical solution taken is：A kind of Sweet Potato Leaf antioxidant activity
The preparation method of substance, including：Degreasing, extraction, de- albumen, decoloration, specifically include following steps：
Degreasing：It chooses health, without rotten, not yellow sweet potato leaf, is dried after cleaning up, smash it through 100~140 mesh sieve, add
Enter methanol and carry out degreasing in 68~70 DEG C of 4~5h of reflux, takes filter residue after suction filtration, volatilize methanol naturally；According to solid-liquid ratio 1:60~
70 add ultra-pure water mixing, 1~1.5h of extraction time under conditions of microwave power is 420~450W, temperature is 60~63 DEG C to take
Filtrate is up to Sweet Potato Leaf solution；The content that anti-oxidation active substance in Sweet Potato Leaf solution is not only greatly improved with methanol degreasing, has
Conducive to its further extraction, it is also prevented from pollution and degradation of the lipid material to antioxidant；High solid-liquid ratio, microwave are auxiliary
Extraction is helped, is not only saved energy, and reduces the viscosity of extracting solution, anti-oxidation active substance is to solution diffusion velocity in leaf
Accelerate, it is advantageous to mass transfer, it is conducive to extraction；
Extraction：Take Sweet Potato Leaf solution, the extraction of setting supersonic frequency 30~35KHz, 350~400W of power, 48~52 DEG C of temperature
Condition, extracts 25~30min every time, and leaching merges, evaporates, concentrates filtrate afterwards three times；It is diluted and is stood with absolute ethyl alcohol after cooling
Overnight, precipitation is filtered out, Sweet Potato Leaf active material crude product is obtained after dry；It is molten that the cavitation effect of ultrasonic wave can strengthen Sweet Potato Leaf significantly
The dissociation organized in liquid can accelerate the diffusion of polysaccharide, polyphenol, flavones, protein etc., be stablized with ultrasonic wave assisted extraction process
Good, easy to operate, the at low cost, less energy consumption of property；
De- albumen：Sweet Potato Leaf active material crude product is pressed into solid-liquid ratio 1:20~22 are dissolved in distilled water, adjust pH be 6.5~
7.0,3200~3600U/g papains, 3800~4500U/g neutral proteinases is successively added, adds protease gross weight
1.2~1.3% pentyl p-hydroxybenzoate and 1.8~2 ‰ (-)-sparteine are measured, 2.5 are digested at a temperature of 48~50 DEG C
~3h, boiling water bath enzyme deactivation after enzymolysis, 12000~12500r/min, 10~15min of centrifugal treating take supernatant that ethyl alcohol is added
It to volume fraction up to 65~70%, stands overnight, centrifuges, precipitation is freeze-dried up to active material crude product；In papain
During the Combined Treatment of neutral proteinase, zymoprotein constantly acts on protein macromolecule, makes the protein breakdown of macromolecular
At the polypeptide even amino acid of small molecule, by processing, most free proteins and the albumen partly combined with polysaccharide
Matter is hydrolyzed, therefore protein content substantially reduces；However as the reduction of protein content in system, enzyme molecule and protein
The contact probability of molecule can taper into, and the catalytic action of enzyme also accordingly weakens, and reaches the time needed for certain hydrolysis effect
It is corresponding to extend, and pentyl p-hydroxybenzoate and (-)-sparteine are added in hydrolytic process can then keep higher hydrolysis
Efficiency, pentyl p-hydroxybenzoate will produce synergistic effect with (-)-sparteine, and the two acts synergistically on and activates pawpaw egg
The sulfenyl of white enzyme active center cysteine substantially increases to reduce the energy barrier between Papain enzyme-to-substrate
The hydrolysis efficiency of papain, reduces enzymolysis time, has saved energy loss and time cost；
Decoloration：Sweet Potato Leaf active material crude product is dissolved in 100~120 times of distilled water, mixed solution 1.05~1.2% is added
Medical active carbon powder and 0.5~0.8 ‰ t- leucine tert-butyl esters, wherein in t- leucine tert-butyl esters containing 11.2~
11.5% L-t- leucine tert-butyl esters；8~10min of oscillation mixing, decoloration under the conditions of 28~32 DEG C, 3000~4500r/min
After the completion, with 12500~13000r/min, 30~45min of high speed centrifugation, take supernatant water-soluble up to the active material after decolourizing
Liquid, up to Sweet Potato Leaf anti-oxidation active substance after low temperature drying；Sweet Potato Leaf polyoses content in Sweet Potato Leaf anti-oxidation active substance
It is 55~60%, Polyphenols content is 18.5~25.8%；Medical active carbon powder belongs to lipophile material, utilizes active carbon powder
In the adsorption principle of capillary adsorb organic pigment, preferable decoloration, turbidity removal effect can be reached, meanwhile, capillary
The adsorption of pipe acts on Sweet Potato Leaf polysaccharide, the Polyphenols isoreactivity substance of meeting absorbed portion, the L-t- in polysaccharide solution
Leucine tert-butyl ester can form total solution system with D-t- leucine tert-butyl esters with sweet potato leaf polyose, polyphenol, by a series of all
As hydrogen bond, reunion, absorption the effects that can substantially reduce polysaccharide, polyphenol surface polarity, improve polysaccharide, polyphenol hydrophily,
Make polysaccharide, polyphenol and capillary micropore of its aggregation far from activated carbon, to substantially reduce polysaccharide, polyphenol is tightly held by activated carbon
Amount, improve final product potato leaf anti-oxidation active substance in polysaccharide, Polyphenols content；Sweet potato leaf polyose is mostly sulfur-rich acidic group
Polysaccharide, has and removes free radical, anti-oxidant, anti-aging, antiphlogistic antibacterial and other effects, and polyphenols has very strong anti-oxidant
Effect, therefore Sweet Potato Leaf active material can be used as a kind of novel efficient green nutriment.
Compared with the prior art, the advantages of the present invention are as follows：
1）The synergistic effect that hydrolysis takes off the pentyl p-hydroxybenzoate and (-)-sparteine that are added in protein process can activate
The sulfenyl of papain activity center cysteine greatly improves pawpaw to reduce the energy barrier between enzyme-to-substrate
The hydrolysis efficiency of protease reduces enzymolysis time, energy saving loss and time cost；
2）L-t- leucine tert-butyl esters in polysaccharide solution can be formed altogether with D-t- leucine tert-butyl esters with sweet potato leaf polyose
Solution system can substantially reduce polysaccharide, the surface polarity of polyphenol, raising by a series of the effects that such as hydrogen bonds, reunion, absorption
Hydrophily makes polysaccharide, polyphenol and its sugared capillary micropore of the aggregation far from activated carbon, to substantially reduce polysaccharide, polyphenol quilt
The amount of activated carbon adsorption, improve final product Sweet Potato Leaf anti-oxidation active substance in polysaccharide, Polyphenols content；
3）Sweet potato leaf polyose is mostly sulfur-rich acidic group polysaccharide, has and removes the work(such as free radical, anti-oxidant, anti-aging, antiphlogistic antibacterial
Effect, polyphenols has very strong antioxidation, therefore Sweet Potato Leaf active material can be as a kind of novel efficient green
Color nutriment uses.
Description of the drawings
Fig. 1 is removing result schematic diagram of the active material in the embodiment of the present invention 3 to DPPH free radicals and hydroxy radical.
Specific implementation mode
The present invention program is described further below by embodiment：
Embodiment 1：
A kind of preparation method of Sweet Potato Leaf anti-oxidation active substance, includes the following steps：
1）Choose health, without rotten, not yellow sweet potato leaf, dried after cleaning up, smash it through 100 mesh sieve, be added methanol in
68 DEG C of reflux 4h carry out degreasing, take filter residue after suction filtration, volatilize methanol naturally；According to solid-liquid ratio 1:60 add ultra-pure water mixing, micro-
Extraction time 1h under conditions of wave power is 420W, temperature is 60 DEG C, takes filtrate up to Sweet Potato Leaf solution；2）Take Sweet Potato Leaf molten
Liquid sets the extraction conditions of supersonic frequency 30KHz, power 350W, 48 DEG C of temperature, extracts 25min every time, and leaching is closed afterwards three times
And it evaporates, concentration filtrate；It is stood overnight with absolute ethyl alcohol dilution after cooling, filters out precipitation, Sweet Potato Leaf active material is obtained after dry
Crude product；3）Sweet Potato Leaf active material crude product is pressed into solid-liquid ratio 1:20 are dissolved in distilled water, and it is 6.5 to adjust pH, is successively added
3200U/g papains, 3800U/g neutral proteinases, add the pentyl p-hydroxybenzoate of protease total weight 1.2%
(-)-sparteine with 1.8 ‰, digests 2.5h at a temperature of 48 DEG C, boiling water bath enzyme deactivation after enzymolysis, 12000r/min from
The heart handles 10min, takes supernatant that ethyl alcohol is added to volume fraction up to 65%, stands overnight, centrifuge, precipitation is freeze-dried to obtain the final product
Active material crude product；4）Sweet Potato Leaf active material crude product is dissolved in 100 times of distilled water, the medicine of mixed solution 1.05% is added
With active carbon powder and 0.5 ‰ t- leucine tert-butyl esters, 11.2% L-t- leucines are wherein contained in t- leucine tert-butyl esters
The tert-butyl ester；Oscillation mixing 8min under the conditions of 28 DEG C, 3000r/min, after the completion of decoloration, with 12500r/min high speed centrifugations
30min takes supernatant up to the active material aqueous solution after decoloration, up to Sweet Potato Leaf antioxidant activity object after low temperature drying
Matter；Sweet Potato Leaf polyoses content is 55~60% in Sweet Potato Leaf anti-oxidation active substance, and Polyphenols content is 18.5~25.8%.
Embodiment 2：
A kind of preparation method of Sweet Potato Leaf anti-oxidation active substance, includes the following steps：
1）Degreasing：It chooses health, without rotten, not yellow sweet potato leaf, is dried after cleaning up, smash it through 140 mesh sieve, first is added
Alcohol carries out degreasing in 70 DEG C of reflux 5h, takes filter residue after suction filtration, volatilizes methanol naturally；According to solid-liquid ratio 1:70 add ultra-pure water mixing,
Extraction time 1.5h under conditions of microwave power is 450W, temperature is 63 DEG C, takes filtrate up to Sweet Potato Leaf solution；It is de- with methanol
Fat not only greatly improves the content of anti-oxidation active substance in Sweet Potato Leaf solution, is conducive to its further extraction, can also prevent
Only pollution and degradation of the lipid material to antioxidant；
2）Extraction：Sweet Potato Leaf solution is taken, the extraction conditions of supersonic frequency 35KHz, power 400W, 52 DEG C of temperature is set, carries every time
30min, leaching is taken to merge afterwards three times, evaporate, concentrate filtrate；It is stood overnight with absolute ethyl alcohol dilution after cooling, filters out precipitation, done
Sweet Potato Leaf active material crude product is obtained after dry；The cavitation effect of ultrasonic wave can strengthen the dissociation organized in Sweet Potato Leaf solution significantly,
The diffusion that can accelerate polysaccharide, polyphenol, flavones, protein etc., good, easy to operate with ultrasonic wave assisted extraction process stability,
At low cost, less energy consumption；
3）De- albumen：Sweet Potato Leaf active material crude product is pressed into solid-liquid ratio 1:22 are dissolved in distilled water, and it is 7.0 to adjust pH, successively
3600U/g papains, 3800~4500U/g neutral proteinases is added, 3h is digested at a temperature of 50 DEG C, is boiled after enzymolysis
Water-bath enzyme deactivation, 12500r/min centrifugal treating 15min take supernatant that ethyl alcohol is added to volume fraction up to 70%, stand overnight, from
The heart, precipitation are freeze-dried up to active material crude product；During the Combined Treatment of papain and neutral proteinase, enzyme
Albumen constantly acts on protein macromolecule, makes the protein breakdown of macromolecular at the polypeptide even amino acid of small molecule, passes through
The protein that processing, most free proteins and part are combined with polysaccharide is hydrolyzed, therefore protein content substantially reduces；
4）Decoloration：Sweet Potato Leaf active material crude product is dissolved in 120 times of distilled water, the medical active of mixed solution 1.2% is added
Carbon powder, oscillation mixing 10min under the conditions of 32 DEG C, 4500r/min, after the completion of decoloration, with 13000r/min high speed centrifugations
45min takes supernatant up to the active material aqueous solution after decoloration, up to Sweet Potato Leaf antioxidant activity object after low temperature drying
Matter；Sweet Potato Leaf polyoses content is 55~60% in Sweet Potato Leaf anti-oxidation active substance, and Polyphenols content is 18.5~25.8%；It is medicinal
Active carbon powder belongs to lipophile material, and organic color is adsorbed using the adsorption principle of the capillary in active carbon powder
Element can reach preferable decoloration, turbidity removal effect, and sweet potato leaf polyose is mostly sulfur-rich acidic group polysaccharide, have and remove free radical, resist
Oxidation, anti-aging, antiphlogistic antibacterial and other effects, polyphenols have very strong antioxidation, therefore Sweet Potato Leaf active material
It can be used as a kind of novel efficient green nutriment.
Embodiment 3：
A kind of preparation method of Sweet Potato Leaf anti-oxidation active substance, including：Degreasing, extraction, de- albumen, decoloration, specifically include with
Lower step：
Degreasing：It chooses health, without rotten, not yellow sweet potato leaf, is dried after cleaning up, smash it through 120 mesh sieve, methanol is added
The 4h that flows back in 68 DEG C carries out degreasing, takes filter residue after suction filtration, volatilizes methanol naturally；According to solid-liquid ratio 1:65 add ultra-pure water mixing,
Extraction time 1h under conditions of microwave power is 440W, temperature is 62 DEG C, takes filtrate up to Sweet Potato Leaf solution；Not with methanol degreasing
The content for only greatly improving anti-oxidation active substance in Sweet Potato Leaf solution is conducive to its further extraction, is also prevented from fat
Pollution and degradation of the substance to antioxidant；High solid-liquid ratio, microwave radiation exaraction, not only save energy, and reduce
The viscosity of extracting solution, anti-oxidation active substance is accelerated to solution diffusion velocity in leaf, advantageous to mass transfer, is conducive to extraction；
Extraction：Sweet Potato Leaf solution is taken, the extraction conditions of supersonic frequency 33KHz, power 360W, temperature 50 C is set, extracts every time
25min, leaching merge, evaporate, concentrate filtrate afterwards three times；It is stood overnight with absolute ethyl alcohol dilution after cooling, filters out precipitation, it is dry
Sweet Potato Leaf active material crude product is obtained afterwards；The cavitation effect of ultrasonic wave can strengthen the dissociation organized in Sweet Potato Leaf solution significantly, can
To accelerate the diffusion of polysaccharide, polyphenol, flavones, protein etc., it is good, easy to operate with ultrasonic wave assisted extraction process stability, at
This low, less energy consumption；
De- albumen：Sweet Potato Leaf active material crude product is pressed into solid-liquid ratio 1:21 are dissolved in distilled water, and it is 6.8 to adjust pH, is successively added
Enter 3500U/g papains, 4000U/g neutral proteinases, adds the P-hydroxybenzoic acid penta of protease total weight 1.2%
Ester and 2 ‰ (-)-sparteine, digest 2.5h at a temperature of 50 DEG C, boiling water bath enzyme deactivation after enzymolysis, 12000r/min from
The heart handles 14min, takes supernatant that ethyl alcohol is added to volume fraction up to 68%, stands overnight, centrifuge, precipitation is freeze-dried to obtain the final product
Active material crude product；During the Combined Treatment of papain and neutral proteinase, zymoprotein constantly acts on protein
Macromolecular makes the protein breakdown of macromolecular at the polypeptide even amino acid of small molecule, by processing, most floating preteins
The protein that matter and part are combined with polysaccharide is hydrolyzed, therefore protein content substantially reduces；However as protein in system
The contact probability of the reduction of content, enzyme molecule and protein molecule can taper into, and the catalytic action of enzyme also accordingly weakens, and reaches
Time needed for certain hydrolysis effect also accordingly extends, and pentyl p-hydroxybenzoate and (-)-goldspink are added in hydrolytic process
Flower alkali can then keep higher hydrolysis efficiency, and pentyl p-hydroxybenzoate and (-)-sparteine will produce synergistic effect, and two
Person act synergistically on and veraldon active site cysteine sulfenyl, to reduce Papain enzyme-to-substrate it
Between energy barrier, substantially increase the hydrolysis efficiency of papain, reduce enzymolysis time, saved energy loss and when
Between cost；
Decoloration：Sweet Potato Leaf active material crude product is dissolved in 110 times of distilled water, the medicinal carbon of mixed solution 1.1% is added
Powder and 0.6 ‰ t- leucine tert-butyl esters, 11.4% L-t- leucine tert-butyl esters are wherein contained in t- leucine tert-butyl esters；
Oscillation mixing 8min, after the completion of decoloration, with 12500r/min high speed centrifugation 40min, takes under the conditions of 30 DEG C, 4000r/min
Clear liquid is up to the active material aqueous solution after decoloration, up to Sweet Potato Leaf anti-oxidation active substance after low temperature drying；Sweet Potato Leaf is anti-
Sweet Potato Leaf polyoses content is 55~60% in active oxide material, and Polyphenols content is 18.5~25.8%；Medical active carbon powder
Belong to lipophile material, organic pigment is adsorbed using the adsorption principle of the capillary in active carbon powder, can reach
Preferable decoloration, turbidity removal effect, meanwhile, Sweet Potato Leaf polysaccharide, the Polyphenols of the adsorption effect meeting absorbed portion of capillary
Isoreactivity substance, L-t- leucine tert-butyl esters in polysaccharide solution and D-t- leucine tert-butyl esters can with sweet potato leaf polyose,
Polyphenol forms total solution system, and polysaccharide, polyphenol surface can be substantially reduced by a series of the effects that such as hydrogen bonds, reunion, absorption
Polarity, improve the hydrophily of polysaccharide, polyphenol, make polysaccharide, polyphenol and capillary micropore of its aggregation far from activated carbon, to
It substantially reduces polysaccharide, the amount that polyphenol is tightly held by activated carbon, improves polysaccharide, Polyphenols in final product potato leaf anti-oxidation active substance
Content；Sweet potato leaf polyose is mostly sulfur-rich acidic group polysaccharide, has and removes the work(such as free radical, anti-oxidant, anti-aging, antiphlogistic antibacterial
Effect, polyphenols has very strong antioxidation, therefore Sweet Potato Leaf active material can be as a kind of novel efficient green
Color nutriment uses.
Embodiment 4：
The Sweet Potato Leaf anti-oxidation active substance chosen in embodiment 3 carries out free radical scavenging activity measurement：
1）DPPH free radical scavenging activities measure：Compound concentration is the work of 5mg/mL, 10mg/mL, 25mg/mL, 50mg/mL respectively
Property each 2.0mL of substance solution, branch be added 2.0mL0.04mg/mLDPPH solution（Absolute ethyl alcohol is as solvent）, after mixing well
It is protected from light 30min at room temperature, its absorbance A is measured at wavelength 517nm1, while measuring absolute ethyl alcohol（2.0mL）With DPPH
The absorbance A of (2.0mL) mixed liquorC, absolute ethyl alcohol（2.0mL）With the absorbance A of sample liquid (2.0mL) mixed liquor2, at other
Under condition is constant, active material is replaced as positive control using VC.Then DPPH free radical scavenging activities calculation formula is：
；
2）Scavenging action to hydroxyl free radical measures：Take the FeSO of 1mL4Solution（10mmol/L）In test tube, the salicylic acid-of 1mL is added
Ethanol solution（10mmol/L）, the work that 1mL concentration is respectively 5mg/mL, 10mg/mL, 25mg/mL, 50mg/mL is added after shaking up
Property substance solution, is eventually adding 8.8mmol/LH2O2Solution 1mL starts reaction, and 37 DEG C, water-bath 1h is cooled to room temperature, in wavelength
510nm surveys absorbance.In the case where other conditions are constant, active material is replaced as positive control using VC, then Scavenging action to hydroxyl free radical
Calculation formula is：
,
In formula：B1Active material is replaced to do blank by distilled water；B2For sample liquid；BCH is replaced by distilled water2O2Active material
Background solution.
DPPH free radicals and Scavenging action to hydroxyl free radical measurement result are as shown in Figure 1, as shown in Figure 1, the embodiment of the present invention 3
In Sweet Potato Leaf anti-oxidation active substance more apparent scavenging effect, while its are all had to DPPH free radicals and hydroxy radical
Dose dependent is presented in free radical scavenging activity.
Routine operation in operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance, including：Degreasing, extraction, de- albumen, decoloration, feature exist
In：The de- albumen step is：Sweet Potato Leaf active material is dissolved in distilled water, papain, neutral egg is successively added
White enzyme adds pentyl p-hydroxybenzoate and is digested with (-)-sparteine, boiling water bath enzyme deactivation after enzymolysis, centrifuging and taking supernatant
Ethyl alcohol is added in liquid, stands overnight, and centrifuges, and precipitation is freeze-dried up to active material crude product.
2. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 1, it is characterised in that：It is described de-
In albumen step, the additive amount of papain is 3200~3600U/g, and the additive amount of neutral proteinase is 3800~4500U/
G, the additive amount of pentyl p-hydroxybenzoate are the 1.2~1.3% of protease total weight, and the additive amount of (-)-sparteine is egg
1.8~the 2 ‰ of white enzyme total weight.
3. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 1, it is characterised in that：It is described de-
In albumen step, enzymolysis solid-liquid ratio is 1:20~22, pH be 6.5~7.0, temperature be 48~50 DEG C, enzymolysis time be 2.5~
3h。
4. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 1, it is characterised in that：It is described de-
Ethyl alcohol is added in albumen step, in supernatant to volume fraction of ethanol up to 65~70%.
5. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 1, it is characterised in that：It is described de-
Fat step is：Choose health, without rotten, not yellow sweet potato leaf, dried after cleaning up, grinding and sieving, be added methanol in return
Degreasing takes filter residue after suction filtration, volatilizes methanol naturally；According to solid-liquid ratio 1:60~70 add ultra-pure water mixing, Microwave Extraction to take filtrate
Up to Sweet Potato Leaf solution.
6. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 1, it is characterised in that：It is described to carry
The step is taken to be：Sweet Potato Leaf solution, ultrasonic extraction, extraction is taken to merge afterwards three times, evaporate, concentrate filtrate；Absolute ethyl alcohol is used after cooling
Dilution is stood overnight, and filters out precipitation, Sweet Potato Leaf active material crude product is obtained after dry.
7. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 6, it is characterised in that：It is described to carry
Take in step, supersonic frequency be 30~35KHz, power be 350~400W, temperature be 48~52 DEG C, every time extract 25~
30min。
8. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 1, it is characterised in that：It is described de-
Color step is：Sweet Potato Leaf active material crude product is dissolved in 100~120 times of distilled water, medical active carbon powder and t- is added
Leucine tert-butyl ester, oscillation mixing, after the completion of decoloration, high speed centrifugation take supernatant up to decolourize after active material aqueous solution,
Up to Sweet Potato Leaf anti-oxidation active substance after low temperature drying.
9. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 8, it is characterised in that：It is described de-
In color step, the additive amount of medical active carbon powder is mixed solution 1.05~1.2%, and the additive amount of t- leucine tert-butyl esters is
Contain 11.2~11.5% L-t- leucine tert-butyl esters in 0.5~0.8 ‰, t- leucine tert-butyl esters.
10. a kind of preparation method of Sweet Potato Leaf anti-oxidation active substance according to claim 8, it is characterised in that：It is described
In decolorization process, in Sweet Potato Leaf anti-oxidation active substance Sweet Potato Leaf polyoses content be 55~60%, Polyphenols content be 18.5~
25.8%。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109874839A (en) * | 2019-04-11 | 2019-06-14 | 广西壮族自治区农业科学院 | A kind of processing method of Sweet Potato Leaf biscuit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109874839A (en) * | 2019-04-11 | 2019-06-14 | 广西壮族自治区农业科学院 | A kind of processing method of Sweet Potato Leaf biscuit |
| CN109874839B (en) * | 2019-04-11 | 2023-02-21 | 广西壮族自治区农业科学院 | Processing method of sweet potato leaf biscuits |
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