CN103408617A - Preparation method of acanthopanax sessiliflorus berry anthocyanin with antioxidant activity - Google Patents
Preparation method of acanthopanax sessiliflorus berry anthocyanin with antioxidant activity Download PDFInfo
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Abstract
The invention relates to a method for extracting anthocyanin from plant berries and particularly relates to a preparation method of acanthopanax sessiliflorus berry anthocyanin with antioxidant activity. The preparation method comprises the following steps of (1) selecting acanthopanax sessiliflorus berries as raw materials; (2) extracting the acanthopanax sessiliflorus berry anthocyanin by using an enzyme freezing and thawing method: mixing the frozen raw materials and an extracting solution, unfreezing and breaking the frozen raw materials, and carrying out enzymolysis; (3) carrying out ultrasonic-microwave synergistic extraction on the acanthopanax sessiliflorus berry anthocyanin: carrying out ultrasonic-microwave synergistic extraction on enzymatic hydrolysate, and centrifuging to obtain a crude extracting solution; (4) purifying the crude extruding solution of the anthocyanin: absorbing the crude extruding solution by using macroporous resin, collecting an eluant, and carrying out vacuum concentration; (5) freezing and drying in vacuum; and (6) smashing the anthocyanin, and then, packaging the anthocyanin. The preparation method has the advantages of high extracting efficiency, short time, low energy consumption, small reagent dosage, stable properties of effective components and the like; and meanwhile, a vacuum freeze drying technology is adopted, so that a product has better biological activity, color, luster and solubility and the preparation method is suitable for industrial production.
Description
Technical field
The present invention relates to a kind of from the plant berry, extracting the method for anthocyanogen, specifically a kind of preparation method with Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity.
Background technology
In the prior art, research finds that long-term edible synthesized coloring matter is detrimental to health, and some countries have started the legislation restriction or banned use of synthetic colour.Meanwhile, people recognize from natural phant, extracting pigment not only nontoxic, harmless, and also have certain health protection effect.Therefore, developing natural pigment replaces synthetic colour to become the development trend of food service industry.
Anthocyanogen extensively is present in the enchylema of flower, fruit, stem, leaf and root organ of plant, is a kind of natural water colo(u)r, belongs to the flavones polyphenolic compound.The effects such as that anthocyanogen has is anti-oxidant, antitumor, anticancer, anti-inflammatory, prevent diabetes, fat-reducing, vision protection.As a kind of natural pigment, anthocyanogen is safe, nontoxic, and human body is had to many nourishing functions, all has broad application prospects in industries such as food, healthcare products, makeup and medicine.
Sessileflower Acanthopanax Bark is studies on plants of Acanthopanax Miq. in Araliaceae, and another name Radix Acanthopanacis Senticosi, thorn bent stick, mainly be distributed in the ground such as the Northeast and Hebei, Shanxi, Henan.Its effect characteristics and ginseng are basic identical, have the various health care functions such as immunomodulatory, antitumor, anti-ageing, radioprotective, anti-damage and antifatigue, deeply be subjected to consumers in general favor.The Sessileflower Acanthopanax Bark fruit is Berry-like drupe, and pulp is full, spherical in shape or avette, and 9-10 month maturation, become atropurpureus by green after maturation.The Sessileflower Acanthopanax Bark berry contains multiple essential trace element and the whole indispensable amino acids of human body such as Fe, Se, Cu, Mn, is the desirable resource of dietotherapy among the people.
At present, the extracting method of anthocyanogen mainly contains solvent extration, supercritical fluid extraction, ultrasonic-assisted extraction method, microwave-assisted extraction method and enzyme process etc.Wherein, solvent extration is the most frequently used extracting method, but have high, the consuming time length of temperature, the shortcoming such as energy consumption is large, organic solvent residual is large, easily cause the change of extract structure and character.Supercritical fluid extraction is particularly suitable for fat-soluble component, not good to water miscible anthocyanogen effect of extracting, and one-time investment is excessive, and the technique of extracting natural anthocyanogen is still immature and be not widely used.Single by means of ultrasonic wave, microwave, enzyme process assisted extraction anthocyanogen, there are the shortcomings such as energy consumption is large, extraction yield is low, temperature is high.As the preparation to black soybean seed capsule anthocyanin such as Zhang Mingwei (patent No. 200510036419.3), be studied, adopt the broken chrotoplast of planting of microwave frequency 2450MHz, microwave power 12-20kw, adopt again extraction using alcohol, extraction time 1-2 hour, extracting times 2-3 time.This method and the present invention compare and have the problems such as power is high, energy consumption is large, the time is long, and the too high material temperature that easily makes of power significantly raises, and cause the change of extract structure and character.
At present, the method for extracting Sessileflower Acanthopanax Bark berry anthocyanogen about freezing to melt enzyme process and ultrasonic-microwave cooperating has no report.
Summary of the invention
The present invention seeks to provide for above-mentioned deficiency a kind of combined method of freezing to melt enzyme process and ultrasonic-microwave cooperating extraction Sessileflower Acanthopanax Bark berry anthocyanogen.The advantages such as the method has mild condition, target is single-minded, extraction yield is high, the time is short, energy consumption is low, reagent dosage is few, effective constituent stable in properties, can enhance productivity widely.Simultaneously, adopt Vacuum Freezing & Drying Technology, make product have biological activity, color and luster and solvability preferably, be applicable to suitability for industrialized production.
Technical solution of the present invention is: a kind of preparation method with Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity is characterized in that step is as follows:
(1) material choice and pre-treatment: the ripe Sessileflower Acanthopanax Bark fresh fruit of take is raw material, freezing after cleaning.
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry freezed, adding liquid ratio is the ethanolic soln of 8:1-15:1, pH4-5.5, volume fraction 40%-60%, thaw rear with the broken 3-10min of stamp mill, the cellulase that adds again massfraction 0.1%-1%, enzymolysis 10min-30min under the condition of 40 ℃-60 ℃.Cellulase can make the cellulose substances degraded in intercellular substance and cell walls, makes its short texture, is conducive to the stripping of anthocyanogen, improves extraction yield.By enzymolysis step, more be conducive to following supersound extraction.Be that in step (2), liquid ratio is preferably 10:1-15:1.
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH2-3 with acid, under ultrasonic power 50-100w, microwave power 100-150w, 40 ℃ of-60 ℃ of conditions, extract 3-8min.By extracting solution centrifugal 5-15min under 3000rpm-5000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract.Device name SL-SM50 type ultrasonic-microwave composite reaction system: the business of Nanjing following current Instrument Ltd. sells.
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 2h-5h, absorption flow velocity 1mL/min-2mL/min.Desorption time is 1h-3h, and with the anthocyanogen on pH2-3, volume fraction 40%-60% ethanol elution macroporous resin, flow velocity is 1mL/min-2mL/min.Collect elutriant, 30 ℃ of-60 ℃ of concentrating under reduced pressure 1-3h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution.
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 5-20 mm, condenser temperature-35 ℃～-50 ℃, vacuum freezing 2-5h, reach below-30 ℃ the material core temperature.25 ℃-55 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 25Pa-60Pa, dry 10-20h.
(6) Sessileflower Acanthopanax Bark berry anthocyanogen is pulverized, packs: the Sessileflower Acanthopanax Bark berry anthocyanogen after vacuum lyophilization is ground into rapidly 60～120 order powder, vacuum packaging.
The present invention also provides the application of described Sessileflower Acanthopanax Bark berry anthocyanogen in the antioxidation in vitro experiment.Experiment shows: Sessileflower Acanthopanax Bark berry anthocyanogen has certain Anti-oxidant ability, also has and removes preferably hydroxyl radical free radical, ultra-oxygen anion free radical, DPPH free radical and H
2O
2Ability.
Preferably: have the preparation method of the Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity, its step is as follows:
(1) material choice and pre-treatment: the Sessileflower Acanthopanax Bark fresh fruit ripe, after selecting of take is raw material, after cleaning, puts into-18 ℃ of refrigerator and cooled and freezes preservation, standby.
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry that 200g freezes, the ethanolic soln 2000ml that adds pH4.5, volume fraction 40%, thaw rear with tissue mashing crusher machine 5min, the cellulase (enzyme 50u/mg alive) that adds again 13.2g, enzymolysis 20min under the condition of 55 ℃.
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH2 with hydrochloric acid, under ultrasonic power 75w, microwave power 150w, 50 ℃ of conditions, extract 5min; By extracting solution centrifugal 10min under 4000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract.By the differential method, measure the content of anthocyanogen in the content of the total anthocyanogen of Sessileflower Acanthopanax Bark berry and crude extract, calculating the anthocyanogen extraction yield is 92.8%.
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 2h, absorption flow velocity 2mL/min; Desorption time is 1h, and with the anthocyanogen on pH2, volume fraction 40% ethanol elution macroporous resin, flow velocity is 2mL/min; Collect elutriant, 50 ℃ of concentrating under reduced pressure 2h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution.
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 10mm, condenser temperature-40 ℃, vacuum freezing 3h, reach below-30 ℃ the material core temperature; 50 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 50Pa, dry 10h.
(6) dried Sessileflower Acanthopanax Bark berry anthocyanogen is ground into rapidly 80 order powder, vacuumizes packing, obtains Sessileflower Acanthopanax Bark berry anthocyanogen.
Sessileflower Acanthopanax Bark berry anthocyanogen after vacuum lyophilization is loose, the nothing caking.For preventing its moisture absorption, be ground into rapidly 80 order powder, vacuumize packing, obtain having the Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity, its look valency is 196, through HPLC, detects Anthocyanin content and reaches 52.5%.
Advantage of the present invention is: 1, fresh Sessileflower Acanthopanax Bark berry is freezed, solved the problems such as raw-material storing is difficult, production is subject to seasonal restrictions, the supply whole year that has guaranteed to produce Raw.
2, after the Sessileflower Acanthopanax Bark berry freezed melts, be conducive to the fragmentation of cell walls, accelerate the stripping of anthocyanogen.
3, adopt enzymolysis step, improved the extraction yield of anthocyanogen.In the situation that ultrasonic-microwave cooperating extraction conditions is constant, carry out the enzymolysis test, as can be seen from Table 1, add enzyme can improve the extraction yield of anthocyanogen, but polygalacturonase is effective not as cellulase.This is because cellulase acts on Sessileflower Acanthopanax Bark berry cell, make the mass degradation such as Mierocrystalline cellulose, hemicellulose in intercellular substance and cell walls, the short texture of cell walls and intercellular substance, expansion, reduce anthocyanogen from born of the same parents to the resistance of solvent diffusion, improve extraction yield.Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen mild condition, specificity are high.
The impact of the different enzymes of table 1 on the anthocyanogen extraction yield
| Method | Test conditions | Extraction yield |
| Do not add enzyme | Liquid ratio 10:1, pH4.5, volume fraction 40% ethanol, 55 ℃, extraction 60min | 60.2% |
| Cellulase | Liquid ratio 10:1, pH4.5, volume fraction 40% ethanol, 55 ℃, enzyme dosage 0.6%, enzymolysis 20min | 92.8% |
| Polygalacturonase | Liquid ratio 10:1, pH4.5, volume fraction 40% ethanol, 55 ℃, enzyme dosage 0.6%, enzymolysis 20min | 77.3% |
4, above-mentioned cellulase degradation liquid carries out respectively alcohol extracting, ultrasonic assisted extraction, microwave-assisted extracts and ultrasonic-microwave cooperating extracts test.As can be seen from Table 2, ultrasonic-microwave cooperating extracts has the advantages such as temperature is low, energy consumption is low, the time is short, and extraction yield is the highest.This is the instantaneous penetration heat effect due to hyperacoustic judder, cavitation effect, stirring action and microwave, pass cell walls, touch intracellular matter, after the absorption energy in cell, high-pressure phenomena appears, cell rises brokenly, reduce the bonding force of anthocyanogen and matrix, accelerate anthocyanogen and enter solvent from matrix.
The impact of table 2 Different Extraction Method on the anthocyanogen extraction yield
| Method | Test conditions | Extraction yield |
| Alcohol extracting | Liquid ratio 10:1, pH2, volume fraction 40% ethanol, 70 ℃, extraction 2h | 58.7% |
| Ultrasonic assisted extraction | Liquid ratio 10:1, pH2, volume fraction 40% ethanol, 70 ℃, ultrasonic power 170w, extraction 30min | 70.6% |
| Microwave-assisted extracts | Liquid ratio 10:1, pH2, volume fraction 40% ethanol, 70 ℃, microwave power 200w, extraction 10min | 78.5% |
| Ultrasonic-microwave cooperating extracts | Liquid ratio 10:1, pH2, volume fraction 40% ethanol, 50 ℃, ultrasonic power 75w, microwave power 150w, extract 5min | 92.8% |
5, as can be seen from Table 3, different resins is different with resolution factor to the absorption of anthocyanogen, and AB-8 macroporous resin adsorption and desorption performance are best.With AB-8 macroporous resin purification Sessileflower Acanthopanax Bark berry anthocyanogen crude extract, have that adsorptive capacity is large, resolution factor is high, good selective.Simultaneously, AB-8 macroporous resin repeat performance is good, can effectively reduce production costs.
The comparison of the absorption of table 3 different resins and resolution factor
| The resin model | Adsorption rate % | Desorption efficiency % |
| AB-8 | 88.1 | 70.2 |
| S-8 | 52.3 | 65.3 |
| HDP-100 | 75.4 | 58.6 |
| HDP-300 | 80.5 | 52.7 |
6, as can be seen from Table 4, the different dry drying method is larger on the impact of anthocyanogen product quality, and the vacuum lyophilization effect is best.Adopting Vacuum Freezing & Drying Technology, is water vapour by the moisture in Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution by the ice distillation, preserves to greatest extent the biological activity of anthocyanogen, makes product have color and luster and solvability preferably.Product is atropurpureus, and the smell of fruits is very sweet, without bad smell.
Table 4 different dry drying method affects the anthocyanogen product quality
| Index | Warm air drying | Vacuum lyophilization | Spraying drying |
| Smell | Be charred flavor | The smell of fruits is very sweet, without bad smell | Fruital is lightly seasoned, without bad smell |
| Color and luster | Black | Atropurpureus | Atropurpureus |
| Solvability | Poorly soluble, speed is slow | Solvability is good, and speed is fast | Solvability is better, speed |
The advantages such as 7, compared with prior art, the present invention has mild condition, target is single-minded, extraction yield is high, the time is short, energy consumption is low, reagent dosage is few, effective constituent stable in properties.The smell of fruits is very sweet for prepared Sessileflower Acanthopanax Bark berry anthocyanogen, bright in colour, solvability is good, stability is strong, anti-oxidant activity is high, with other synthetic colours, antioxidant, compares safely, is applicable to suitability for industrialized production.
8, can improve the added value of Sessileflower Acanthopanax Bark berry, enlarge the raw material sources of natural pigment, natural antioxidants.
Below in conjunction with embodiment, embodiments of the present invention are described in further detail.
Embodiment
Embodiment 1
(1) material choice and pre-treatment: the Sessileflower Acanthopanax Bark fresh fruit ripe, that deceive without scab, free from insect pests, color and luster purple of take is raw material, after cleaning, puts into-18 ℃ of refrigerator and cooled and freezes preservation, standby.
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry that 200g freezes, the ethanolic soln 2000ml that adds pH4.5, volume fraction 40%, thaw rear with tissue mashing crusher machine 5min, the cellulase (enzyme 50u/mg alive) that adds again 13.2g, enzymolysis 20min under the condition of 55 ℃.
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH2 with hydrochloric acid, under ultrasonic power 75w, microwave power 150w, 50 ℃ of conditions, extract 5min; By extracting solution centrifugal 10min under 4000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract; By the differential method, measure the content of anthocyanogen in the content of the total anthocyanogen of Sessileflower Acanthopanax Bark berry and crude extract, calculating the anthocyanogen extraction yield is 92.8%.
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 2h, absorption flow velocity 2mL/min; Desorption time is 1h, and with the anthocyanogen on pH2, volume fraction 40% ethanol elution macroporous resin, flow velocity is 2mL/min; Collect elutriant, 50 ℃ of concentrating under reduced pressure 2h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution.
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 10mm, condenser temperature-40 ℃, vacuum freezing 3h, reach below-30 ℃ the material core temperature; 50 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 50Pa, dry 10h.
(6) Sessileflower Acanthopanax Bark berry anthocyanogen is pulverized, packs: the Sessileflower Acanthopanax Bark berry anthocyanogen after vacuum lyophilization is loose, the nothing caking.For preventing its moisture absorption, be ground into rapidly 80 order powder, vacuumize packing, obtain having the Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity, its look valency is 196, through HPLC, detects Anthocyanin content and reaches 52.5%.
Embodiment 2
(1) material choice and pre-treatment: the Sessileflower Acanthopanax Bark fresh fruit ripe, that deceive without scab, free from insect pests, color and luster purple of take is raw material, after cleaning, puts into-18 ℃ of refrigerator and cooled and freezes preservation, standby.
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry that 200g freezes, the ethanolic soln 3000ml that adds pH5, volume fraction 50%, thaw rear with tissue mashing crusher machine 3min, the cellulase (enzyme 50u/mg alive) that adds again 6.4g, enzymolysis 30min under the condition of 50 ℃.
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH3 with hydrochloric acid, under ultrasonic power 100w, microwave power 100w, 40 ℃ of conditions, extract 6min; By extracting solution centrifugal 5min under 5000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract; By the differential method, measure the content of anthocyanogen in the content of the total anthocyanogen of Sessileflower Acanthopanax Bark berry and crude extract, calculating the anthocyanogen extraction yield is 90.8%.
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 3h, absorption flow velocity 1mL/min; Desorption time is 2h, and with the anthocyanogen on pH3, volume fraction 50% ethanol elution macroporous resin, flow velocity is 1mL/min; Collect elutriant, 60 ℃ of concentrating under reduced pressure 1h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution.
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 15mm, condenser temperature-45 ℃, vacuum freezing 4h, reach below-30 ℃ the material core temperature; 40 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 60Pa, dry 15h.
(6) Sessileflower Acanthopanax Bark berry anthocyanogen is pulverized, packs: the Sessileflower Acanthopanax Bark berry anthocyanogen after vacuum lyophilization is loose, the nothing caking.For preventing its moisture absorption, be ground into rapidly 100 order powder, vacuumize packing, obtain having the Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity, its look valency is 190, through HPLC, detects Anthocyanin content and reaches 51.6%.
Embodiment 3
(1) material choice and pre-treatment: the Sessileflower Acanthopanax Bark fresh fruit ripe, that deceive without scab, free from insect pests, color and luster purple of take is raw material, after cleaning, puts into-18 ℃ of refrigerator and cooled and freezes preservation, standby.
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry that 200g freezes, the ethanolic soln 1600ml that adds pH5.5, volume fraction 60%, thaw rear with tissue mashing crusher machine 10min, the cellulase (enzyme 50u/mg alive) that adds again 18g, enzymolysis 10min under the condition of 60 ℃.
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH2.5 with hydrochloric acid, under ultrasonic power 50w, microwave power 125w, 60 ℃ of conditions, extract 3min; By extracting solution centrifugal 15min under 3000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract; By the differential method, measure the content of anthocyanogen in the content of the total anthocyanogen of Sessileflower Acanthopanax Bark berry and crude extract, calculating the anthocyanogen extraction yield is 87.9%.
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 5h, absorption flow velocity 1.5mL/min; Desorption time is 3h, and with the anthocyanogen on pH2.5, volume fraction 60% ethanol elution macroporous resin, flow velocity is 1.5mL/min; Collect elutriant, 30 ℃ of concentrating under reduced pressure 3h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution.
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 5mm, condenser temperature-35 ℃, vacuum freezing 2h, reach below-30 ℃ the material core temperature; 30 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 30Pa, dry 20h.
(6) Sessileflower Acanthopanax Bark berry anthocyanogen is pulverized, packs: the Sessileflower Acanthopanax Bark berry anthocyanogen after vacuum lyophilization is loose, the nothing caking.For preventing its moisture absorption, be ground into rapidly 60 order powder, vacuumize packing, obtain having the Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity, its look valency is 181, through HPLC, detects Anthocyanin content and reaches 50.1%.
Experimental example 1
Take the lard of 7 part of 50 g, 1 part adds 0.02%BHT, and 5 parts add respectively 0.05 %, 0.1 %, 0.2 %, 0.3 %, 0.4 % Sessileflower Acanthopanax Bark berry anthocyanogen powder, and 1 part does not add additive and makes blank.By baking oven high temperature induction lard generation peroxidation, measure every three days peroxide value one time.Experimental result shows, Sessileflower Acanthopanax Bark berry anthocyanogen has certain antioxygenation to grease.Along with the increase of Sessileflower Acanthopanax Bark berry anthocyanin content, its antioxygenation to lard strengthens.And the time is longer, effect is more obvious.
Experimental example 2
In reaction system, contain 8.8mmo1/L H
2O
21mL, 10mmol/LFeSO
41 mL, 10mmol/L Whitfield's ointment one ethanolic soln 1mL, add respectively different concns Sessileflower Acanthopanax Bark berry anthocyanogen solution 1mL, finally adds H
2O
2Start reaction, 37 ℃ of reaction 30min, make reference with distilled water, under 525nm, measures the absorbancy of each concentration.Experimental result shows, along with the increase of Sessileflower Acanthopanax Bark berry anthocyanogen concentration, removes the ability enhancing of hydroxyl radical free radical.And, when Sessileflower Acanthopanax Bark berry anthocyanogen concentration is higher, remove the ability of hydroxyl radical free radical higher than xitix.
Claims (3)
1. preparation method with Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity is characterized in that step is as follows:
(1) material choice and pre-treatment: the ripe Sessileflower Acanthopanax Bark fresh fruit of take is raw material, freezing after cleaning;
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry freezed, adding liquid ratio is the ethanolic soln of 8:1-15:1, pH4-5.5, volume fraction 40%-60%, thaw rear with the broken 3-10min of stamp mill, the cellulase that adds again mass concentration 0.1%-1%, enzymolysis 10min-30min under the condition of 40 ℃-60 ℃;
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH2-3 with acid, under ultrasonic power 50-100w, microwave power 100-150w, 40 ℃ of-60 ℃ of conditions, extract 3-8min, by extracting solution centrifugal 5-15min under 3000rpm-5000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract;
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 2h-5h, absorption flow velocity 1mL/min-2mL/min; Desorption time is 1h-3h, and with the anthocyanogen on pH2-3, volume fraction 40%-60% ethanol elution macroporous resin, flow velocity is 1mL/min-2mL/min; Collect elutriant, 30 ℃ of-60 ℃ of concentrating under reduced pressure 1-3h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution;
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 5-20 mm, condenser temperature-35 ℃～-50 ℃, vacuum freezing 2-5h, reach below-30 ℃ the material core temperature; 25 ℃-55 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 25Pa-60Pa, dry 10-20h;
(6) Sessileflower Acanthopanax Bark berry anthocyanogen is pulverized, packs: the Sessileflower Acanthopanax Bark berry anthocyanogen after vacuum lyophilization is ground into rapidly 60～120 order powder, vacuum packaging.
2. according to the preparation method with Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity claimed in claim 1, it is characterized in that in step (2), liquid ratio is 10:1-15:1.
3. according to the described preparation method with Sessileflower Acanthopanax Bark berry anthocyanogen of anti-oxidant activity of claim 1 or 2, it is characterized in that step is as follows:
(1) material choice and pre-treatment: the Sessileflower Acanthopanax Bark fresh fruit ripe, after selecting of take is raw material, after cleaning, puts into-18 ℃ of refrigerator and cooled and freezes preservation, standby;
(2) freeze to melt Enzymatic Extraction Sessileflower Acanthopanax Bark berry anthocyanogen: get the Sessileflower Acanthopanax Bark berry that 200g freezes, the ethanolic soln 2000ml that adds pH4.5, volume fraction 40%, thaw rear with tissue mashing crusher machine 5min, then add the cellulase of 13.2g, enzymolysis 20min under the condition of 55 ℃;
(3) ultrasonic-microwave cooperating extracts Sessileflower Acanthopanax Bark berry anthocyanogen: above-mentioned enzymolysis solution is adjusted to pH2 with hydrochloric acid, under ultrasonic power 75w, microwave power 150w, 50 ℃ of conditions, extract 5min; By extracting solution centrifugal 10min under 4000rpm, collect supernatant liquor
,Obtain Sessileflower Acanthopanax Bark berry anthocyanogen crude extract;
(4) Sessileflower Acanthopanax Bark berry anthocyanogen crude extract purifying: above-mentioned crude extract is adsorbed with the AB-8 macroporous resin, and the static adsorption equilibria time is 2h, absorption flow velocity 2mL/min; Desorption time is 1h, and with the anthocyanogen on pH2, volume fraction 40% ethanol elution macroporous resin, flow velocity is 2mL/min; Collect elutriant, 50 ℃ of concentrating under reduced pressure 2h, what obtain atropurpureus, thickness has an anti-oxidant activity anthocyanogen concentrated solution;
(5) vacuum lyophilization Sessileflower Acanthopanax Bark berry anthocyanogen: Sessileflower Acanthopanax Bark berry anthocyanogen concentrated solution is poured into to the dry machine charging tray of vacuum freezing, thickness 10mm, condenser temperature-40 ℃, vacuum freezing 3h, reach below-30 ℃ the material core temperature; 50 ℃ of sublimation drying stepwise heating plate temperatures, vacuum tightness 50Pa, dry 10h;
(6) dried Sessileflower Acanthopanax Bark berry anthocyanogen is ground into rapidly 80 order powder, vacuumizes packing, obtains Sessileflower Acanthopanax Bark berry anthocyanogen.
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