The method that stalk pigment is extracted using steam explosion technology
Technical field
The invention belongs to stalk comprehensive utilization technical field, and in particular to a kind of utilization steam explosion technology extracts stalk pigment
Method.
Background technology
According to statistics, China can harvest more than 900,000,000 tons of various crop materials every year.Stalk described here, including wheat, corn,
Remain after the herbage harvesting seed such as the crop such as paddy rice, cotton, Chinese sorghum, crudefiber crop, soybean, peanut, potato class, melon and clover, prairie milk vetch
Remaining stem, leaf, skin(Shell), climing rattan(Seedling)Deng and sugarcane(Slag), reed, bamboo etc..It is different with water content during stalk cutting,
Stalk is divided into into following two class:First kind stalk, moisture content more than 35%, such as:Melon seedling(Moisture content is more than 80%), part it is beautiful
Rice kind(Moisture content more than 50%, harvest season are still green), sugarcane(Moisture content more than 50%), herbage(Moisture content 70% with
On)Deng;Equations of The Second Kind stalk, water content below 35%, including most of stalk, receiving by such as wheat straw, straw, corn, cotton etc.
Obtain the stalk of season collection.
Containing abundant pigment in stalk.Crop material pigment includes chloroplast pigment and cell liquid pigment, and the former is present
It is in chloroplaset, relevant with photosynthesis, such as chlorophyll, lutein;The latter is present in vacuole, such as Flavonoid substances(Including
Anthocyanidin, isoflavones etc.).Stalk pigment is distributed mainly on the positions such as the bar of stalk, stem, rattan, seedling, Ye, Miho.
The function value of natural colour chlorins compound, action value are increasingly paid close attention to by people.Flavones is a big class day
Right pigment family, its most essential physiological function are antioxidation.Flavone compound is got over as the effect of functional component
Come more important, its Main Function to the mankind has:Antitumor, antiallergy, antiviral, strengthen immunity, improve cardiovascular and cerebrovascular, adjust
Section endocrine, anti-aging etc..Most well known in flavones is isoflavones.Crop flavones content highest at the maturity period,
Thus crop material is the good raw material for preparing flavones.
And carotenoid and chloroplast pigment are industrially a kind of colouring agents of important foods and cosmetics.It is existing
In technology, from stalk extracting flavones, carotenoid and chloroplast pigment simultaneously does not also have feasible solution, it is impossible to give full play to
The value of stalk.
The content of the invention
In order to make up the deficiencies in the prior art, the present invention provides one kind and can effectively extract Flavonoid substances and class in stalk
The method of carrotene and chloroplast pigment.
The technical scheme is that:
A kind of method that utilization steam explosion technology extracts stalk pigment, comprises the following steps:
The extraction of A flavones
A1)Stalk fragment is placed in steam-explosion jar, and inert gas is passed through in the steam-explosion jar will seal after air emptying in tank;
Then 1-5 MPa are forced into, pressurize was spurted after 20-120 minutes, obtained stalk slurry;
A2)The stalk slurry is placed in steam-explosion jar, ether is added, inert gas is passed through in the steam-explosion jar by tank
Seal after air emptying;Then intermittently inert gas is passed through into tank, to maintain pressure inside the tank as 1-5 Mpa, temperature is
16-20 DEG C, pressurize was spurted after 20-180 minutes;
A3)Separation of solid and liquid, obtains filter residue one and filtrate one;Filter residue one is washed using ether, the ether solution after washing is closed with filtrate one
And, obtain amalgamation liquid one;
A4)Solid phase particles in removing amalgamation liquid one, remove the amalgamation liquid after solid phase particles and sequentially pass through ultrafiltration, reverse osmosis membrane point
From, obtain molecular weight between 300-1000 dalton flavones clarified solution and molecular weight more than 1000 dalton chloroplast pigment
Clarified solution;
A5)Evaporation removes the solvent ether in the flavones clarified solution, residue ethanol repeated washing, drying obtain flavones into
Product;
The preparation of B carotenoid
B1)Evaporation removes the solvent ether in the chloroplast pigment clarified solution, residue solid-liquid after ethanol repeated washing point
From, filter residue two and filtrate two are obtained, filter residue two is carotenoid;
B2)Using after petroleum ether dissolution, evaporation removes solvent petroleum ether to filter residue two, residue with after petroleum ether repeated washing class
The petroleum ether solution of carrotene;
B3)The petroleum ether solution of the carotenoid is separated into alpha-carotene, beta carotene, γ-Hu using fibre column chromatography
Radish three kinds of pigments of element, are eluted with ethanol-petroleum ether mixed solvent, and 3 kinds of pigments are collected separately washing in different time sections
In de- liquid;
B4)Evaporation respectively removes the vehicle of ethanol-petroleum ether mixed solvent of three kinds of eluents, residue ethanol-oil
Ether mixed solvent repeated washing, drying obtain alpha-carotene, beta carotene, gamma carotene finished product;
The preparation of C chloroplast pigments
C1)Vaporization removal step B1)Solvent in gained filtrate two, residue with after ethanol repeated washing chloroplast pigment
Ethanol solution;
C2)The ethanol solution of the chloroplast pigment is separated into chlorophyll a, chlorophyll b, lutein using fibre column chromatography, uses
Ethanol-petroleum ether mixed solvent is eluted, and 3 kinds of pigments are collected separately in the eluent of different time sections;
C3)Evaporation respectively removes the vehicle of ethanol-petroleum ether mixed solvent of three kinds of eluents, and the repetition of residue ethanol is washed
Wash, drying obtains chlorophyll a, chlorophyll b, lutein finished product.
Preferably, step A1)Described in the obtaining step of stalk fragment be:Fresh straw is harvested, sandstone, miscellaneous is removed
The fragment of 1-3cm is washed, is cut into after thing with clear water.
Preferably, step A1)5-50% of the amount in middle stalk fragment addition steam-explosion jar for steam explosion tank volume.
Further, step A1)Described in stalk segment process isolation air, flavones is easily oxidized, and completely cuts off air-treatment,
Can ensure that the integrality of flavones.
Further, step A2)Middle addition ether volume is 1-20 times of stalk slurry.
Preferably, step A2)In middle steam-explosion jar, temperature is by inert gas control, the holding vessel of the inert gas
It is externally provided with interlayer, interlayer and is full of Cryogenic air, it is 16-18 DEG C that Cryogenic air controls inert gas temperature in holding vessel.Using
Lower temperature and inert gas environment purpose are to avoid decomposition, oxidation of pigment component molecule etc. from reacting.
Further, step A2)In be passed through the volume of inert gas in steam-explosion jar for 1-10 times of steam explosion tank volume.
Preferably, step B3)In in ethanol-petroleum ether mixed solvent used ethanol with the volume ratio of petroleum ether be
(1-3):(1-9).
Preferably, step C2)In in ethanol-petroleum ether mixed solvent used ethanol with the volume ratio of petroleum ether be
(1-7):(1-9).
Preferably, step A5)、C1)、C3)Described in ethanol volume fraction be 95%.
Beneficial effects of the present invention are:
The present invention obtains high-purity flavones, carotenoid and chloroplast pigment simultaneously by stalk, by cryogenic high pressure, steam explosion skill
The measures such as art, inert environments, organic solvent have refined out high-purity, high-quality above-mentioned stalk pigment component.
The present invention, can be with newly-built when implementing, it is also possible to makes full use of existing numerous papermaking for or frequently facing bankruptcy
Slurrying enterprise, on the basis of its workshop, equipment, facility etc., slightly transformation can be produced, and put into little, cycle is short, easily promote.
Specific embodiment
Embodiment 1
A kind of method that utilization steam explosion technology extracts stalk pigment, comprises the following steps:
The extraction of A flavones
A1)Maize straw fragment is placed in steam-explosion jar, and stalk fragment addition reaches the 15% of tank volume, to the steam-explosion jar
In be passed through inert gas will after air emptying in tank seal;Then 1.5 MPa are forced into, pressurize was spurted after 100 minutes, obtained stalk
Slurry;Load hermetic bag, be filled with inert gas, put shady place storage, it is standby.
Step A1)The obtaining step of middle maize straw fragment is:Fresh straw is harvested in the fall, is used after removing sandstone, debris
Clear water washing, the fragment of 2-3cm is cut into, then moves into freezer storage, stand-by;This kind corn is feed form corn variety,
It is still green when autumn harvests.
The characteristics of having oxidizable due to phytochrome according to the present invention, thus will be made in inert gas environment
It is standby, to ensure the integrality of pigment composition.
Step A1)Stalk cell wall is broken through using the impulsive force from inside to outside that gases at high pressure are produced during spurting,
Make stalk cellular content all " broken wall and go out ", the Component seperation and refining beneficial to next step;Because the present invention needs to prepare
Flavonoids product, and these compositions almost exist only in it is intracellular, thus must broken wall.
A2)The stalk slurry is placed in steam-explosion jar, 8 times of volume ether is added, is passed through in the steam-explosion jar lazy
Property gas will seal after air emptying in tank, it is 4 times of tank volume to be passed through inert gas volume;Then intermittently into tank
Inert gas is passed through, and pressure inside the tank is maintained as 1.5 Mpa, temperature is 16-20 DEG C, and pressurize was spurted after 120 minutes.
Step A2)Lower temperature and nitrogen environment are adopted in steam explosion separation process, it is therefore intended that avoid pigment component
The reaction such as the decomposition of molecule, oxidation;Using high pressure(By being passed through nitrogen realization)Ensure that being fully contacted for pigment and solvent
With dissolution.
Temperature in tank is controlled by the temperature of nitrogen.The holding vessel of nitrogen is externally provided with interlayer, inside has Cryogenic air, thus
Nitrogen temperature in holding vessel is between 16-18 DEG C.
A3)Press master shove is separated, and obtains filter residue one and filtrate one, and pigment, lipid of dissolution etc., filter are in filtrate one
Slag one is the slurry containing starch, protein, fibrous fraction etc.;One twice of filter residues, the second after washing are washed using 3 times of ether
Ether liquid is merged with filtrate one, obtains amalgamation liquid one;Filter residue one can further be separated and prepare other components.
A4)Solid phase particles in removing amalgamation liquid one, remove the amalgamation liquid after solid phase particles and sequentially pass through ultrafiltration, counter-infiltration
UF membrane, obtain molecular weight between 300-1000 dalton flavones clarified solution and molecular weight more than 1000 dalton chloroplaset
Pigment clarified solution.
A5)It is evaporated in vacuo the solvent ether removed in the flavones clarified solution, residue 95% ethanol repeated washing 3 times,
It is freeze-dried to obtain flavones finished product.
After testing, the quality of gained flavones for maize straw butt quality 1.7%.
The preparation of B carotenoid
B1)Evaporation removes the solvent ether in the chloroplast pigment clarified solution, residue absolute ethanol washing 3 times, repeats
This step, spiral shell centrifugation of crouching obtain filter residue two and filtrate two, and filter residue two is carotenoid.
B2)Filter residue two is washed using removing solvent petroleum ether, the repetition of residue petroleum ether after petroleum ether dissolution, is evaporated in vacuo
Wash 3 times, after repeating this step, obtain the petroleum ether solution of carotenoid.
B3)The petroleum ether solution of the carotenoid using fibre column chromatography be separated into alpha-carotene, beta carotene,
Three kinds of pigments of gamma carotene, are eluted with ethanol-petroleum ether mixed solvent, in ethanol-petroleum ether mixed solvent ethanol with
The volume ratio of petroleum ether is 1:9,3 kinds of pigments are collected separately in the eluent of different time sections.
B4)Vehicle of ethanol-petroleum ether the mixed solvent for removing three kinds of eluents, residue second are evaporated in vacuo respectively
Alcohol-petroleum ether mixed solvent repeated washing 3 times, repeats this step, washes away impurity, freeze-dried to obtain alpha-carotene, β-Hu
Radish element, gamma carotene finished product.
After testing, the quality of gained carotenoid for maize straw butt quality 1.3%.
The preparation of C chloroplast pigments
C1)Vaporization removal step B1)Solvent in gained filtrate two, residue repeat this with after 95% ethanol repeated washing 3 times
Step, washes away impurity, obtains the ethanol solution of chloroplast pigment.
C2)It is yellow that the ethanol solution of the chloroplast pigment is separated into chlorophyll a, chlorophyll b, leaf using fibre column chromatography
Element, is eluted with ethanol-petroleum ether mixed solvent, and in ethanol-petroleum ether mixed solvent, ethanol with the volume ratio of petroleum ether is
5:5,3 kinds of pigments are collected separately in the eluent of different time sections.
C3)Evaporation respectively removes the vehicle of ethanol-petroleum ether mixed solvent of three kinds of eluents, and residue is used respectively
95% ethanol repeated washing 3 times, repeats this step, washes away impurity, it is freeze-dried obtain chlorophyll a, chlorophyll b, lutein into
Product.
After testing, the chlorophyllous quality of gained for maize straw butt quality 1.1%, the quality of lutein is corn stalk
The 0.7% of stalk butt quality.
Embodiment 2
A kind of method that utilization steam explosion technology extracts stalk pigment, comprises the following steps:
The extraction of A flavones
A1)Watermelon seedling fragment is placed in steam-explosion jar, and watermelon seedling fragment addition reaches the 20% of tank volume, to the steam-explosion jar
In be passed through inert gas will after air emptying in tank seal;Then 2 MPa are forced into, pressurize was spurted after 90 minutes, obtain melon seedling slurry
Shape thing;Load hermetic bag, be filled with inert gas, put shady place storage, it is standby.
Step A1)The obtaining step of Chinese and Western melon seedling fragment is:Fresh watermelon seedling is harvested in the fall, is used after removing sandstone, debris
Clear water washing, the fragment of 2-3cm is cut into, then moves into freezer storage, stand-by.
The characteristics of having oxidizable due to phytochrome according to the present invention, thus will be made in inert gas environment
It is standby, to ensure the integrality of pigment composition.
Step A1)Melon seedling cell membrane is broken through using the impulsive force from inside to outside that gases at high pressure are produced during spurting,
Make watermelon seedling cellular content all " broken wall and go out ", the Component seperation and refining beneficial to next step;Because the present invention needs system
Standby flavonoids product, and these compositions almost exist only in it is intracellular, thus must broken wall.
A2)The melon seedling slurry is placed in steam-explosion jar, 9 times of volume ether is added, is passed through in the steam-explosion jar lazy
Property gas will seal after air emptying in tank, it is 5 times of tank volume to be passed through inert gas volume;Then intermittently into tank
Inert gas is passed through, and pressure inside the tank is maintained as 2 Mpa, temperature is 16-20 DEG C, and pressurize was spurted after 110 minutes.
Step A2)Lower temperature and nitrogen environment are adopted in steam explosion separation process, it is therefore intended that avoid pigment component
The reaction such as the decomposition of molecule, oxidation;Using high pressure(By being passed through nitrogen realization)Ensure that being fully contacted for pigment and solvent
With dissolution.
Temperature in tank is controlled by the temperature of nitrogen.The holding vessel of nitrogen is externally provided with interlayer, inside has Cryogenic air, thus
Nitrogen temperature in holding vessel is between 16-18 DEG C.
A3)Press master shove is separated, and obtains filter residue one and filtrate one, and pigment, lipid of dissolution etc., filter are in filtrate one
Slag one is the slurry containing starch, protein, fibrous fraction etc.;One twice of filter residues, the second after washing are washed using 3 times of ether
Ether liquid is merged with filtrate one, obtains amalgamation liquid one;Filter residue one can further be separated and prepare other components.
A4)Solid phase particles in removing amalgamation liquid one, remove the amalgamation liquid after solid phase particles and sequentially pass through ultrafiltration, counter-infiltration
UF membrane, obtain molecular weight between 300-1000 dalton flavones clarified solution and molecular weight more than 1000 dalton chloroplaset
Pigment clarified solution.
A5)It is evaporated in vacuo the solvent ether removed in the flavones clarified solution, residue 95% ethanol repeated washing 3 times,
It is freeze-dried to obtain flavones finished product.
After testing, the quality of gained flavones for watermelon seedling butt quality 1.2%.
The preparation of B carotenoid
B1)Evaporation removes the solvent ether in the chloroplast pigment clarified solution, residue absolute ethanol washing 3 times, repeats
This step, spiral shell centrifugation of crouching obtain filter residue two and filtrate two, and filter residue two is carotenoid.
B2)Filter residue two is washed using removing solvent petroleum ether, the repetition of residue petroleum ether after petroleum ether dissolution, is evaporated in vacuo
Wash 3 times, after repeating this step, obtain the petroleum ether solution of carotenoid.
B3)The petroleum ether solution of the carotenoid using fibre column chromatography be separated into alpha-carotene, beta carotene,
Three kinds of pigments of gamma carotene, are eluted with ethanol-petroleum ether mixed solvent, in ethanol-petroleum ether mixed solvent ethanol with
The volume ratio of petroleum ether is 2:8,3 kinds of pigments are collected separately in the eluent of different time sections.
B4)Vehicle of ethanol-petroleum ether the mixed solvent for removing three kinds of eluents, residue second are evaporated in vacuo respectively
Alcohol-petroleum ether mixed solvent repeated washing 3 times, repeats this step, washes away impurity, freeze-dried to obtain alpha-carotene, β-Hu
Radish element, gamma carotene finished product.
After testing, the quality of gained carotenoid for watermelon seedling butt quality 1.8%.
The preparation of C chloroplast pigments
C1)Vaporization removal step B1)Solvent in gained filtrate two, residue repeat this with after 95% ethanol repeated washing 3 times
Step, washes away impurity, obtains the ethanol solution of chloroplast pigment.
C2)It is yellow that the ethanol solution of the chloroplast pigment is separated into chlorophyll a, chlorophyll b, leaf using fibre column chromatography
Element, is eluted with ethanol-petroleum ether mixed solvent, and in ethanol-petroleum ether mixed solvent, ethanol with the volume ratio of petroleum ether is
4:6,3 kinds of pigments are collected separately in the eluent of different time sections.
C3)Evaporation respectively removes the vehicle of ethanol-petroleum ether mixed solvent of three kinds of eluents, and residue is used respectively
95% ethanol repeated washing 3 times, repeats this step, washes away impurity, it is freeze-dried obtain chlorophyll a, chlorophyll b, lutein into
Product.
After testing, the chlorophyllous quality of gained for watermelon seedling butt quality 1.3%, the quality of lutein are dry for watermelon seedling
The 0.6% of matrix amount.
Embodiment 3
A kind of method that utilization steam explosion technology extracts stalk pigment, comprises the following steps:
The extraction of A flavones
A1)Sugarcane fragment is placed in steam-explosion jar, and sugarcane fragment addition reaches the 25% of tank volume, is led to in the steam-explosion jar
Enter inert gas to seal after air emptying in tank;Then 2.5 MPa are forced into, pressurize was spurted after 85 minutes, obtained sugarcane pulpous state
Thing;Load hermetic bag, be filled with inert gas, put shady place storage, it is standby.
Step A1)The obtaining step of middle sugarcane fragment is:Fresh cane is harvested, washed with clear water after removing sandstone, debris, cut
Into the fragment of 2-3cm, then move into freezer storage, it is stand-by.
The characteristics of having oxidizable due to phytochrome according to the present invention, thus will be made in inert gas environment
It is standby, to ensure the integrality of pigment composition.
Step A1)Sugarcane cell wall is broken through using the impulsive force from inside to outside that gases at high pressure are produced during spurting,
Make sugarcane cell content all " broken wall and go out ", the Component seperation and refining beneficial to next step;Because the present invention needs to prepare
Flavonoids product, and these compositions almost exist only in it is intracellular, thus must broken wall.
A2)The sugarcane slurry is placed in steam-explosion jar, 10 times of volume ether is added, is passed through in the steam-explosion jar lazy
Property gas will seal after air emptying in tank, it is 6 times of tank volume to be passed through inert gas volume;Then intermittently into tank
Inert gas is passed through, and pressure inside the tank is maintained as 2 .5Mpa, temperature is 16-20 DEG C, and pressurize was spurted after 100 minutes.
Step A2)Lower temperature and nitrogen environment are adopted in steam explosion separation process, it is therefore intended that avoid pigment component
The reaction such as the decomposition of molecule, oxidation;Using high pressure(By being passed through nitrogen realization)Ensure that being fully contacted for pigment and solvent
With dissolution.
Temperature in tank is controlled by the temperature of nitrogen.The holding vessel of nitrogen is externally provided with interlayer, inside has Cryogenic air, thus
Nitrogen temperature in holding vessel is between 16-18 DEG C.
A3)Press master shove is separated, and obtains filter residue one and filtrate one, and pigment, lipid of dissolution etc., filter are in filtrate one
Slag one is the slurry containing starch, protein, fibrous fraction etc.;One twice of filter residues, the second after washing are washed using 3 times of ether
Ether liquid is merged with filtrate one, obtains amalgamation liquid one;Filter residue one can further be separated and prepare other components.
A4)Solid phase particles in removing amalgamation liquid one, remove the amalgamation liquid after solid phase particles and sequentially pass through ultrafiltration, counter-infiltration
UF membrane, obtain molecular weight between 300-1000 dalton flavones clarified solution and molecular weight more than 1000 dalton chloroplaset
Pigment clarified solution.
A5)It is evaporated in vacuo the solvent ether removed in the flavones clarified solution, residue 95% ethanol repeated washing 3 times,
It is freeze-dried to obtain flavones finished product.
After testing, the quality of gained flavones for sugarcane butt quality 1.4%.
The preparation of B carotenoid
B1)Evaporation removes the solvent ether in the chloroplast pigment clarified solution, residue absolute ethanol washing 3 times, repeats
This step, spiral shell centrifugation of crouching obtain filter residue two and filtrate two, and filter residue two is carotenoid.
B2)Filter residue two is washed using removing solvent petroleum ether, the repetition of residue petroleum ether after petroleum ether dissolution, is evaporated in vacuo
Wash 3 times, after repeating this step, obtain the petroleum ether solution of carotenoid.
B3)The petroleum ether solution of the carotenoid using fibre column chromatography be separated into alpha-carotene, beta carotene,
Three kinds of pigments of gamma carotene, are eluted with ethanol-petroleum ether mixed solvent, in ethanol-petroleum ether mixed solvent ethanol with
The volume ratio of petroleum ether is 2:8,3 kinds of pigments are collected separately in the eluent of different time sections.
B4)Vehicle of ethanol-petroleum ether the mixed solvent for removing three kinds of eluents, residue second are evaporated in vacuo respectively
Alcohol-petroleum ether mixed solvent repeated washing 3 times, repeats this step, washes away impurity, chilled freeze-drying obtain alpha-carotene,
Beta carotene, gamma carotene finished product.
After testing, the quality of gained carotenoid for sugarcane butt quality 1.9%.
The preparation of C chloroplast pigments
C1)Vaporization removal step B1)Solvent in gained filtrate two, residue repeat this with after 95% ethanol repeated washing 3 times
Step, washes away impurity, obtains the ethanol solution of chloroplast pigment.
C2)It is yellow that the ethanol solution of the chloroplast pigment is separated into chlorophyll a, chlorophyll b, leaf using fibre column chromatography
Element, is eluted with ethanol-petroleum ether mixed solvent, and in ethanol-petroleum ether mixed solvent, ethanol with the volume ratio of petroleum ether is
4:6,3 kinds of pigments are collected separately in the eluent of different time sections.
C3)Evaporation respectively removes the vehicle of ethanol-petroleum ether mixed solvent of three kinds of eluents, and residue is used respectively
95% ethanol repeated washing 3 times, repeats this step, washes away impurity, it is freeze-dried obtain chlorophyll a, chlorophyll b, lutein into
Product.
After testing, the chlorophyllous quality of gained for sugarcane butt quality 1.2%, the quality of lutein is sugarcane butt matter
The 0.7% of amount.