CN103333519A - Hickory nut shell brown pigment and preparation method thereof - Google Patents
Hickory nut shell brown pigment and preparation method thereof Download PDFInfo
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Abstract
The invention provides a method of extracting hickory nut shell brown pigment with high performance and high stability and relates to a natural pigment processing technology. The method comprises the following steps: the petroleum ether is adopted to wash hickory nut shells 10 to 15 minutes during pre-processing, so that subsequent enzymolysis is promoted and the extraction efficiency of ultrasonic wave is improved greatly; alcohol-citric acid system prepared at a specific ratio is regarded as efficient extraction solvent; the solvent (alcohol-citric acid system), specific compound enzyme and ultrasonic wave are effectively combined in a creative way and reacted mutually, so that secondary digestion and extraction to residues are not required, the residues is only required to be digested once, and influences to pigment activity caused by secondary extraction are effectively avoided; the whole processing is carried out at the temperature of 35 DEG C to 38 DEG C, so that the pigment activity and stability of the activity are guaranteed, and unexpected technical effects to the inventor are obtained. The method of extracting hickory nut shell brown pigment with high performance and high stability has the advantages that the technology is simple, the operation is easy, the period is short, the complicated equipment is not required, and the product performance is improved obviously; the hickory nut shell brown pigment with high performance and high stability is suitable for industrialized production.
Description
Technical field
It is good to the invention provides a kind of purity, and performance height, the brown pigmentation that stability is high relate to the natural pigment processing technology, belong to the food chemistry processing technique field.
Background technology
Edible natural pigment refers to the food dye that obtains from natural resource, has plenty of from the edibility extracting section of long-term edible organism; Have plenty of and extract root from medicine food dual purpose plant, stem, leaf, the fruit; Have plenty of from the shell of organism or pericarp and extract.From the source of natural pigment as can be seen, most of natural pigments are safe, and human body is almost had no adverse effects, and can be used as the healthy eating pigment.
Heilongjiang Province's forest land resource is very luxuriant, and forest land area is 2427.6 ten thousand hectares, ranks first in the country, and accounts for 51.3% of the whole province's soil total area.Mountain special product product kind is various, standing stock are big, and Semen Caryae Cathayensis is one of wherein important nut just.Along with the expansion year by year of walnut product amount of finish, the main byproduct nut-shell of walnut processing has also caused people's attention.The past nut-shell is dropped or is burned, and has caused the significant wastage of resource.Make fuel after having occurred in recent years nut-shell is processed as gac, improved the added value of nut-shell.Further investigation to nut-shell is found, contain natural brown pigmentation in the nut-shell, if this natural pigment can be extracted, and for food-processing, both avoided the waste of nut-shell resource, satisfy the demand of people to healthy, safe natural pigment again, embodied the value of nut-shell comprehensively fully.
Northeast Semen Caryae Cathayensis is because unique region, other place of production of hardness ratio of pecan shell higher, draw world elite, contained pigment quality height, and all directly adopt solvent lixiviate or hyperacoustic independent method to extract in the prior art usually, exist extraction time long, obtain pigment product poor quality, active shortcoming such as low.At present, the research of pecan shell brown pigmentation extraction aspect, domestic northeast, pay attention to the pigment crude product of high yield, biological activity, purity and the extraction efficiency of having ignored product, and all be the secondary lixiviate usually, time is long, increases production cost and process complexity, and quality is low can't to satisfy on the market demand to high-end pigment product.There is not the prozyme of zymin, especially specific proportioning to be used in the pigment extraction of northeast pecan shell yet.Also there are not at present a kind of effective ways that northeast Semen Caryae Cathayensis pigment extracts that are exclusively used in.Therefore, for the special raw material resources of high-quality, provide a kind of effective extracting method, just become one of present industry problem demanding prompt solution.
Also Shang Weiyou unites solvent-ultrasonic wave-prozyme the report that uses in the pecan shell pigment extraction northeastward at present.
Summary of the invention
Purpose of the present invention: provide a kind of and have that technology is simple, easy handling, cycle are short, need not complex apparatus, obviously enhance product performance, and are fit to the extracting method of the pecan shell brown pigmentation of advantages such as suitability for industrialized production, effectively utilize the nut-shell resource.
Technical scheme of the present invention: a kind of extracting method of pecan shell brown pigmentation is characterized in that preparing according to following steps:
(1) with the shell decon of Semen Caryae Cathayensis, with petroleum ether 1 time, washing time 10-15min, clear water or alcohol flushing are at room temperature placed 1h, treat that unrecovered oil ether volatilizees fully after, under 35-38 ℃ of temperature, continue oven dry, it is standby to be ground into 40-80 purpose powder;
(2) according to 1: 30-1: 35 solid-liquid ratio (w/v) adds ethanol-citric acid system with powder, fully soak wetting, in extracting solution, add the prozyme of being formed by cellulase, hemicellulase and polygalacturonase again, the addition of enzyme is the 0.01-0.03% of walnut powder quality, and wherein cellulase, hemicellulase and polygalacturonase ratio are 4: 2: 1; Wherein, ethanol-citric acid system is: as solute, the aqueous ethanolic solution with 50% is solvent with citric acid, and being mixed with concentration by the quality volume percent is citric acid-ethanolic soln of 0.1%.
(3) ultrasonication: the mixture in the step 2 is carried out ultrasonication, and ultrasonic power is 300w-350w, and ultrasonic temperature is 35 ℃-38 ℃, extracts 50min-80min, gets crude extract;
(4) crude extract filters, and filtrate centrifugation under 5000r/min, 5min condition gets centrifugate, and the centrifugal cryogenic vacuum that carries out is concentrated, and pressure is 0.085-0.090MPa, and temperature is 35-38 ℃, is concentrated into the 1/2-1/3 of original volume, gets concentrated solution;
(5) with the polar macroporous resin column of NKA-9 on the concentrated solution, last sample flow velocity is 1mL/min-3mL/min, and is saturated to the macroporous resin adsorption brown pigments;
(6) wash-out: use the washing that is no less than 2 times of column volumes to remove debris and the more weak water-soluble impurity of adsorptive power that is not adsorbed, use aqueous ethanolic solution (V/V) the flush away middle polarity impurity of the 15-30% that is no less than 3 times of column volumes again, use citric acid-ethanol elution brown pigmentation of 0.1% at last, elution flow rate is 0.5mL/min-2.0mL/min;
(7) elutriant is under 0.085-0.090MPa, 35-38 ℃ condition, and vacuum concentration becomes the brown pigments concentrated solution;
(8) with concentrated solution under-50-38 ℃ condition, lyophilize 25-32h gets the brown pigments powder.
Semen Caryae Cathayensis is the Wild hickory on Changbai Mountain, northeast.Preferred other technical scheme, namely, in step (2), add the prozyme of being formed by cellulase, hemicellulase, polygalacturonase and Phospholipid hydrolase, the addition of enzyme is the 0.01-0.03% of walnut powder quality, and wherein cellulase, hemicellulase, polygalacturonase and Phospholipid hydrolase ratio are 5: 4: 4: 1.
PH adjusts or remains on 3-6 in the whole system of preferred steps (2), and pigment can be used in the preparation cola.
Wherein, the step of (5), (6) and (7) three purifying can be omitted, and namely the technical scheme of step (1), (2), (3), (4) and (8) formation is also within protection domain; Maybe can use other conventional purification process to substitute.
Beneficial effect of the present invention:
Only need adopt petroleum ether 10-15min when (1) the present invention's innovation is pre-treatment, the time is short, and the fatty substance of washing surface and impurity just can promote follow-up enzymolysis greatly, improve hyperacoustic extraction efficiency.
(2) be not raw material with the pecan shell also in the prior art, use solvent (ethanol-citric acid system)-prozyme-ultrasonic wave combining method to carry out the trial that pigment extracts, far super other place of production walnut of northeast Semen Caryae Cathayensis shell hard and compact degree, this method is applied to needs to overcome many technical barriers on the pecan shell of northeast, for example extract choosing of solvent, the kind of enzyme and proportioning, how the addition of enzyme effectively works in coordination with numerous factors etc.
(3) on the basis of early-stage Study, the contriver creatively carries out effective combination with solvent (ethanol-citric acid system)-prozyme-ultrasonic wave, synergy makes residue not need secondary lixiviate or extraction mutually, effectively avoids second extraction to the active influence that produces of pigment; Ultrasonic wave does not need repetition yet; The anti-oxidant activity height, the anti-oxidant activity high stability is also high.Wherein, the effect of prozyme: cellulase and hemicellulase are with the Mierocrystalline cellulose in the mode hydrolysis shell that goes forward one by one, nut-shell also belongs to vegetable cell, the contriver innovates the trial polygalacturonase, finds that unexpectedly in cellulase, hemicellulase combining pectin enzyme better changes the permeability of cell walls, makes the histocyte inclusion fully contact with solvent, make the quick solution transfer of effective constituent, thereby change the permeability of cell walls, accelerate the release of pigment, thereby improve extraction yield.Mass ratio is cellulase, hemicellulase and the polygalacturonase of 0.01%-0.03%, can be conducive to the stripping of pigment more; Cellulase, hemicellulase and polygalacturonase ratio have novelty at 4: 2: 1, do not use three kinds of enzymes in the prior art and in described ratio the pecan shell pigment is extracted, enzyme with strong points, more preferably add Phospholipid hydrolase again, cellulase, hemicellulase, polygalacturonase, the usage ratio of Phospholipid hydrolase is 5: 4: 4: 1, the phosphatide that dissociates out from nut-shell in the enzymolysis process is effectively removed in the adding of Phospholipid hydrolase, phosphatide is oxidized and corrupt easily, if remain in the quality that will have a strong impact on the nut-shell pigment in the nut-shell pigment, also avoid the nut-shell pigment to use chemical reagent sherwood oil repetitive scrubbing, guaranteed performance and the quality of nut-shell pigment.Ultrasonic wave is a kind of elastic mechanical ripple, swirling effect, cavitation effect and the heat effect of utilizing the ultrasonic wave radiation pressure to produce changes material weave construction, state, function or accelerates the process of these changes, can make and produce circulation in cell peripheral and the cell, thereby improved the permeability of cell walls and cytolemma, played the effect of strengthening extraction process, be conducive to the extraction of effective constituent in the cell, shorten extraction time, extraction rate is fast, economizes solvent, save energy.Solvent (ethanol-citric acid system)-prozyme-ultrasonic wave is carried out effective combination, and synergy has produced greater than solvent+prozyme+hyperacoustic technique effect mutually.
(4) ethanol-citric acid system with specific proportioning is solvent, effectively soaks into and softens, and also is beneficial to extraction, also for cellulase, hemicellulase and polygalacturonase provide suitable pH, adjusts pH between 3-6; Simultaneously, the trier finds that unexpectedly in the technology before lyophilize, consistent 35 ℃ of-38 ℃ of low temperature of whole process using, gentle temperature are carried out, and have guaranteed the activity of pigment, and active stability, have obtained the technique effect that the contriver is difficult to expect.
Each performance of product all is improved significantly, and the target product that the present invention obtains is the synergistic result of each processing parameter, has produced unforeseeable technique effect.
The product property evaluation method is as follows, (also can adopt other ordinary method of this area to estimate): controlled trial adopts uses solvent or ultrasonic method to extract, also can be contrast with solvent and ultrasonic wave coupling separately.It also can be other method in contrast or the prior art with the traditional method.
1, the extraction yield of nut-shell brown pigments: (yield) %=brown pigments powder (g)/nut-shell powder (g) * 100%.Yield of the present invention is at 7-8%, and extraction yield is up to 4-5% in the different controlled trial.
2, the spectral response curve of nut-shell brown pigments
The brown pigments solution of 50 μ g/ml is measured its characteristic absorption spectrum with ultraviolet-visible pectrophotometer in the 250-400nm wavelength region, be contrast with distilled water.The aqueous solution of nut-shell pigment has maximum absorption band at the 289nm place, so test and Selection 289nm is as the foundation of quantitative measurment.
3, potential of hydrogen is to the influence of brown pigments
The pigment solution of 6 parts of 25ml50 μ g/ml is joined in the solution of the different pH value that 10ml NaOH or hydrochloric acid is made into, vibration, after leaving standstill 2h, with the pH test paper record that pH value of solution is respectively 2～3,5-6,7～8,9-10,11-12,13-14, measure its light absorption value.
The present invention in pH7～8 absorbancy maximums, still has greatlyyer in absorbancy when pH2～3,11-12, product resistance to acids and bases of the present invention is stronger.The light absorption value general trend is to increase with the pH value, and light absorption value descends.
4, light stability
Get pigment solution 10ml Continuous irradiation 25d under daylight of 1 part of 50 μ g/ml, measure its light absorption value every day, analyze illumination to the influence of this brown pigments solution.
The present invention is along with the growth of sunshine duration, and the absorbancy of brown pigments solution when 289nm reduces gradually, and color is thin out but do not have precipitation and generate behind the 25d.; And the color of control group is obviously thin out, and has precipitation to generate.
5, thermostability
Get the pigment solution of 5 part of 50 μ g/ml, heating 1h in the time of 20 ℃, 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃, 90 ℃, 100 ℃ measures its light absorption value respectively.Analysis temperature is to the influence of this brown pigments solution.
The present invention is in the better heat stability below 90 ℃, even can tolerate 100 ℃ of short period of time, and all becomes unstable during 80 ℃ of control groups.
6, Oxidizing and Reducing Agents is to the influence of brown pigments
Get three parts of the brown pigments solution of 50 μ g/ml, add 0%, 0.1%, 0.4%, 0.6%, 0.8% H respectively
2O
2Solution behind the placement 2h, is measured its light absorption value.Other gets three parts of the brown pigments solution of 50 μ g/ml, adds 0%, 0.4%, 0.6%, 0.8% Na respectively
2SO
3Solution is placed the light absorption value of measuring pigment solution behind the 2h respectively.
The present invention is along with the increase of Oxidizing and Reducing Agents concentration, and the absorbancy of nut-shell brown pigments slightly reduces, but changes not quite, illustrates that certain density Oxidizing and Reducing Agents is less to the influence of brown pigments, and anti-oxidant reducing power is stronger; And control group is bigger to the influence of the absorbance of nut-shell brown pigments along with the increase of Oxidizing and Reducing Agents concentration, illustrate that its anti-oxidant reducing power a little less than.
7, metal ion is to the influence of brown pigments
Prepare the brown pigments solution of 6 parts of 10ml50 μ g/ml, add Na respectively
+, Fe
3+, Ca
2+, Mg
2+, Cu
2+, Al
3+10mg and 10ml distilled water are measured its light absorption value respectively.
Metal ion Na of the present invention
+, Fe
3+, Mg
2+, Ca
2+, Al
3+, Cu
2+Dilute solution be more stable, these ions within the specific limits can stable existence; And control group also is more stable, but Cu
2+, Fe
3+Influential to brown pigments.
8. anti-oxidant activity
Adopt Tripotassium iron hexacyanide reduction method to measure the brown pigments that the present invention obtains, its anti-oxidant activity can reach the 25%-30% of Vc, compares with control group or traditional method, and reducing power can improve 5%-10%.
The true qualities disposition can be high, stable can being used for the stronger cola of the fastidious property of pigment; Usually, in the pigment application process, should avoid adding phenylformic acid, and nut-shell pigment of the present invention also there is certain resistance to the phenylformic acid that should not add.
Embodiment
Embodiment 1
A kind of extracting method of pecan shell brown pigmentation is characterized in that preparing according to following steps:
(1) with the shell decon of Semen Caryae Cathayensis, with petroleum ether 1 time, washing time 12min, clear water or alcohol flushing are at room temperature placed 1h, treat that unrecovered oil ether volatilizees fully after, under 36 ℃ of temperature, continue oven dry, it is standby to be ground into 60 purpose powder;
(2) according to 1: 33 solid-liquid ratio (w/v) powder is added ethanol-citric acid system, fully soak wetting, in extracting solution, add the prozyme of being formed by cellulase, hemicellulase and polygalacturonase again, the addition of enzyme is 0.02% of walnut powder quality, and wherein cellulase, hemicellulase and polygalacturonase ratio are 4: 2: 1; Wherein, ethanol-citric acid system is: as solute, the aqueous ethanolic solution with 50% is solvent with citric acid, and being mixed with concentration by the quality volume percent is citric acid-ethanolic soln of 0.1%.Preferred system pH3-6;
(3) ultrasonication: the mixture in the step 2 is carried out ultrasonication, and ultrasonic power is 330w, and ultrasonic temperature is 36 ℃, extracts 70min, gets crude extract;
(4) crude extract filters, and filtrate centrifugation under 5000r/min, 5min condition gets centrifugate, centrifugate is carried out cryogenic vacuum concentrate, and pressure is 0.085-0.090MPa, and temperature is 36 ℃, is concentrated into the 1/2-1/3 of original volume, gets concentrated solution;
(5) absorption: with the polar macroporous resin column of NKA-9 on the concentrated solution, last sample flow velocity is 1mL/min-3mL/min, and is saturated to the macroporous resin adsorption brown pigments;
(6) wash-out: use the washing that is no less than 2 times of column volumes to remove debris and the more weak water-soluble impurity of adsorptive power that is not adsorbed, use aqueous ethanolic solution (V/V) the flush away middle polarity impurity of the 15-30% that is no less than 3 times of column volumes again, use citric acid-ethanol elution brown pigmentation of 0.1% at last, elution flow rate is 0.5mL/min-2.0mL/min;
(7) elutriant is under 0.085-0.090MPa, 36 ℃ of conditions, and vacuum concentration becomes the brown pigments concentrated solution;
(8) with concentrated solution under-50 ℃ of-38 ℃ of conditions, lyophilize 25-32h gets the brown pigments powder.
Embodiment 2
A kind of extracting method of pecan shell brown pigmentation is characterized in that preparing according to following steps:
(1) with the pecan shell decon, with petroleum ether 1 time, washing time 12min, clear water or alcohol flushing are at room temperature placed 1h, treat that unrecovered oil ether volatilizees fully after, under 36 ℃ of temperature, continue oven dry, it is standby to be ground into 60 purpose powder;
(2) according to 1: 33 solid-liquid ratio (w/v) powder is added ethanol-citric acid system, fully soak wetting, in extracting solution, add the prozyme of being formed by cellulase, hemicellulase, polygalacturonase and Phospholipid hydrolase again, the addition of enzyme is 0.02% of walnut powder quality, and wherein cellulase, hemicellulase, polygalacturonase and Phospholipid hydrolase ratio are 5: 4: 4: 1; Wherein, ethanol-citric acid system is: as solute, the aqueous ethanolic solution with 50% is solvent with citric acid, and being mixed with concentration by the quality volume percent is citric acid-ethanolic soln of 0.1%.Preferred system pH3-6.
(3) ultrasonication: the mixture in the step 2 is carried out ultrasonication, and ultrasonic power is 330w, and ultrasonic temperature is 36 ℃, extracts 70min, gets crude extract;
(4) crude extract filters, and filtrate centrifugation under 5000r/min, 5min condition gets centrifugate, centrifugate is carried out cryogenic vacuum concentrate, and pressure is 0.085-0.090MPa, and temperature is 36 ℃, is concentrated into the 1/2-1/3 of original volume, gets concentrated solution;
(5) absorption: with the polar macroporous resin column of NKA-9 on the concentrated solution, last sample flow velocity is 1mL/min-3mL/min, and is saturated to the macroporous resin adsorption brown pigments;
(6) wash-out: use the washing that is no less than 2 times of column volumes to remove debris and the more weak water-soluble impurity of adsorptive power that is not adsorbed, use aqueous ethanolic solution (V/V) the flush away middle polarity impurity of the 15-30% that is no less than 3 times of column volumes again, use citric acid-ethanol elution brown pigmentation of 0.1% at last, elution flow rate is 0.5mL/min-2.0mL/min;
(7) elutriant is under 0.085-0.090MPa, 36 ℃ of conditions, and vacuum concentration becomes the brown pigments concentrated solution;
(8) with concentrated solution under-50 ℃ of-38 ℃ of conditions, lyophilize 25-32h gets the brown pigments powder.
Claims (6)
1. the extracting method of a pecan shell brown pigmentation is characterized in that preparing according to following steps:
(1) with the shell decon of Semen Caryae Cathayensis, with petroleum ether 1 time, washing time 10-15min, the washing back is with clear water or alcohol flushing, at room temperature place 1h, after treating that unrecovered oil ether volatilizees fully, dry under 35-38 ℃ of temperature, it is standby to be ground into 40-80 purpose powder then;
(2) according to 1: 30-1: 35 solid-liquid ratio (w/v) adds ethanol-citric acid system with powder, fully soak wetting, add the prozyme of being formed by cellulase, hemicellulase and polygalacturonase again, the addition of enzyme is the 0.01-0.03% of walnut shell flour quality, and wherein cellulase, hemicellulase and polygalacturonase ratio are 4: 2: 1; Wherein, ethanol-citric acid system is: as solute, the aqueous ethanolic solution with 50% is solvent with citric acid, and being mixed with concentration by the quality volume percent is citric acid-ethanolic soln of 0.1%;
(3) ultrasonication: the mixture of step (2) is carried out ultrasonication, and ultrasonic power is 300w-350w, and ultrasonic temperature is 35 ℃-38 ℃, extracts 50min-80min, gets crude extract;
(4) crude extract filters, and filtrate centrifugation under 5000r/min, 5min condition gets centrifugate, and the centrifugal cryogenic vacuum that carries out is concentrated, and pressure is 0.085～0.090MPa, and temperature is 35-38 ℃, is concentrated into the 1/2-1/3 of original volume, gets concentrated solution;
(5) with the polar macroporous resin column of NKA-9 on the concentrated solution, last sample flow velocity is 1-3mL/min, and is saturated to the macroporous resin adsorption brown pigments;
(6) wash-out: use the washing that is no less than 2 times of column volumes to remove debris and the more weak water-soluble impurity of adsorptive power that is not adsorbed, use aqueous ethanolic solution (V/V) the flush away middle polarity impurity of the 15-30% that is no less than 3 times of column volumes again, use citric acid-ethanol elution brown pigmentation of 0.1% at last, elution flow rate is 0.5-2.0mL/min;
(7) elutriant is under 0.085-0.090MPa, 35-38 ℃ condition, and vacuum concentration becomes the brown pigments concentrated solution;
(8) lyophilize: preferred: under-50-38 ℃ condition, lyophilize 25-32h gets the brown pigments powder with concentrated solution.
2. the described method of claim 1 is characterized in that, Semen Caryae Cathayensis is the Wild hickory on Changbai Mountain, northeast.
3. claim 1 or 2 described methods, it is characterized in that, in step (2), add the prozyme of being formed by cellulase, hemicellulase, polygalacturonase and Phospholipid hydrolase, the addition of enzyme is the 0.01-0.03% of walnut powder quality, and wherein cellulase, hemicellulase, polygalacturonase and Phospholipid hydrolase ratio are 5: 4: 4: 1; Namely replace the different prozymes of forming.
4. the described method of claim 1 is characterized in that, pH adjusts or remain on 3-6 in the whole system of step (2).
5. the application of pigment in the preparation cola for preparing by the described method of claim 1-4.
6. a pecan shell brown pigmentation is characterized in that preparing according to the described method of claim 1-3.
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| CN104497623A (en) * | 2014-12-04 | 2015-04-08 | 常州大学 | Method for extracting natural pigment from fruit shell |
| CN107163616A (en) * | 2017-05-22 | 2017-09-15 | 郭良 | A kind of method and its purification process for extracting walnut green husk pigment |
| CN113801493A (en) * | 2021-10-14 | 2021-12-17 | 锦柯草木染(上海)纺织科技有限公司 | Method for extracting natural dye from outer skin of pecan |
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| CN103073915A (en) * | 2013-02-07 | 2013-05-01 | 湖南威嘉生物科技有限公司 | Process for extracting and separating capsanthin and capsaicin by using biological enzyme |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104497623A (en) * | 2014-12-04 | 2015-04-08 | 常州大学 | Method for extracting natural pigment from fruit shell |
| CN107163616A (en) * | 2017-05-22 | 2017-09-15 | 郭良 | A kind of method and its purification process for extracting walnut green husk pigment |
| CN113801493A (en) * | 2021-10-14 | 2021-12-17 | 锦柯草木染(上海)纺织科技有限公司 | Method for extracting natural dye from outer skin of pecan |
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