CN104673497B - A kind of extraction process of plants essential oil, polysaccharide and flavones - Google Patents
A kind of extraction process of plants essential oil, polysaccharide and flavones Download PDFInfo
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Abstract
It is to make Extraction solvent with the ethanol water of cellulase to extract plants essential oil, flavones and polysaccharide 3 times under ultrasonic assistant the invention discloses the extraction process of a kind of plants essential oil, flavones and polysaccharide；Merge extract solution, essential oil crude product, aqueous phase and flavone aglycone crude product are centrifuged to obtain after being evaporated under reduced pressure recovery ethanol.Using n-butanol from aqueous phase extracting flavone glycosides, flavone aglycone is dissolved in the extraction phase containing flavonoid glycoside, is evaporated under reduced pressure recovery n-butanol, dry flavones crude product；Raffinate phase containing polysaccharide is through ethanol precipitation, filtering, dry polysaccharide crude.The extraction process makes plant material obtain relatively sufficiently utilization, has the advantages that simple, safety, efficiency of pcr product are high and activity is impaired small.
Description
Technical field
The present invention relates to the extraction process in natural active matter extraction field, more particularly to plants essential oil, polysaccharide and flavones.
Background technology
Plants essential oil is a kind of odorous volatility oily liquid substance, can be from the flower of plant, leaf, branch, root, skin, fruit
Obtained in real etc., there is antibacterial, anti-inflammatory, antitumor, antiviral, antipyretic-antalgic, expelling phlegm and arresting coughing and other effects.The extraction of plants essential oil
Method includes steam distillation extraction, solvent extraction, molecular distillation is extracted, squeezing is extracted, ultrasonic assistant extracts, enzyme auxiliary carries
Take, supercritical CO2Extraction, microwave radiation exaraction etc..Polysaccharide is one of natural macromolecular in body, by multiple identical or differ
Monose with glycosidic bond be connected and is formed, be widely present in high animals and plants, fungi etc. in vivo.Polysaccharide conduct always in cell
The role of energy stores molecule and structure composition, recently as going deep into for polysaccharide researches, the multiple biological activities of polysaccharide are sent out
It is existing, such as anti-oxidant, antitumor, moisturizing, protect liver, anti-inflammatory, and it is smaller to human normal cell's toxic side effect, is preferable medicine
Thing source.The extracting method of polysaccharide has water extraction, acid extraction, alkali carries to take, microwave radiation exaraction, enzyme assisted extraction, ultrasonic wave are auxiliary
Help extraction, super-pressure assisted extraction, supercritical CO2Extraction etc..Flavones is a kind of Polyphenols thing being widely present in plant
Matter, such compound have the multiple biological activities such as anti-oxidant, antitumor, antibacterial, analgesia, anticoagulation, anti-inflammatory, protection liver,
It can be applied to pharmacy and food processing etc..The extracting method of flavones has water extraction, organic solvent extraction, supercritical CO2Carry
Take, the extraction of microwave radiation exaraction, ultrasonic assistant, enzyme assisted extraction etc..
Ultrasonic assistant extraction natural active matter is a kind of physics shattering process, and principle is made using the cavitation of ultrasonic wave
With the cell for destroying plant tissue, solvent is easily penetrated into the cell, while more second order effects of ultrasonic wave, as mechanical shock,
Emulsify, spread, smashing, chemical effect etc., can also accelerate the diffusion of extract component, release and with solvent be sufficiently mixed and beneficial to carrying
Take, the technology energy consumption is low, investment is small, easy to operate, easily realizes industrial expanding production.Enzyme assisted extraction natural active matter is root
According to the composition of plant cell wall, the characteristics of having selectivity and high efficiency using enzyme reaction, selects corresponding enzyme, by cell membrane
Constituent (cellulose, hemicellulose and pectic substance etc.) hydrolyze or degraded, destroy cell wall structure, make it is intracellular into
Divide dissolving, suspension or peptization in solvent, so as to reach extraction purpose.There is the technology yield height, its lytic activity to be damaged small, ring
The advantages that guarantor.
In recent years, the report of relevant natural active matter extraction is gradually increased, but it is many exist product is single, cost is high,
Raw material availability is low, has used toxic larger or the shortcomings of inflammable and explosive reagent.
The content of the invention
The technical problem to be solved in the present invention is overcome the deficiencies in the prior art, there is provided a kind of utilization rate for improving raw material
And efficiency of pcr product, the extraction process of the plants essential oil of extraction process safety and environmental protection, flavones and polysaccharide.
In order to solve the above technical problems, technical scheme proposed by the present invention is：
The extraction process of a kind of plants essential oil, flavones and polysaccharide, concrete operation step are as follows：
（1）Dry and crush：By raw material cleaning, drying, 10 ~ 100 mesh sieves are crossed after crushing, it is standby；
（2）Immersion：By step（1）Standby raw material soaks 1 ~ 15 h with ethanol water；
（3）Once extract：By step（2）In raw material and ethanol water be put into extraction vessel, toward Extraction solvent second
In alcohol solution add cellulase after under the conditions of 25 DEG C ~ 65 DEG C the min of ultrasonication 10 ~ 120, filtered after extraction,
Collect filtrate, filter residue；
（4）Second extraction：To step（3）Filter residue add ethanol water, soak repeat step after 10 ~ 120 min
（3）Extraction and filtration step；
（5）Extract three times：Repeat step（4）Process；
（6）Once it is evaporated under reduced pressure：By step（3）、（4）、（5）In filtrate merge, under the conditions of 45 ~ 60 DEG C depressurize steam
Recovery ethanol is evaporated, removes part water；
（7）Centrifuge：Using centrifuge by step（6）Remaining filtrate centrifuges 10 ~ 60 min afterwards, obtain essential oil crude product,
Aqueous phase and flavone aglycone crude product；
（8）Extract and separate：Using n-butanol from step（7）Extracting flavone glycosides is to n-butanol phase in the intermediate water phase of gained
Without obvious color, the extraction phase containing flavonoid glycoside and the raffinate phase containing polysaccharide are obtained；
（9）Alcohol precipitation：Toward step（8）In the raffinate containing polysaccharide be added to the absolute ethyl alcohols of 4 ~ 10 times of volumes, stand 12 ~ 30
H, filtering, collect filtrate, filter residue；
（10）Secondary pressure is distilled and dried：By step（7）Obtained flavone aglycone crude product adds step（8）In contain flavones
In the extraction phase of glycosides, under the conditions of 70 DEG C ~ 80 DEG C be evaporated under reduced pressure recovery n-butanol, remaining extraction phase be dried under reduced pressure flavones is thick
Product；By step（9）Obtained filtrate is evaporated under reduced pressure recovery ethanol under the conditions of 45 ~ 60 DEG C, by step（9）Obtained filter residue decompression
Dry polysaccharide crude；
（11）Molecular distillation refines：By step（7）Obtained essential oil crude product refines through molecular distillation technique；
（12）Detect essential oil crude product and the quality after molecular distillation technique is refined；Detect polysaccharide crude and contained more saccharics
Amount；Detect flavones crude product and contained flavones quality.
Further improvement as above-mentioned technical proposal：
Preferably, the step（2）, step（3）, step（4）, step（5）Middle ethanol water is that ethanol configures with water,
Volume fraction is 30% ~ 70%.
Preferably, the step（2）, step（4）, step（5）The amount of the added etoh solvent aqueous solution, its volume and raw material
Mass ratio is 5 ~ 25: 1 mL/g.
Preferably, the step（3）, step（4）, step（5）In, the concentration for adding cellulase is 0.3 ~ 4.0 U/mL
Extraction solvent.
Preferably, the power of the ultrasonic wave is 200 W, and frequency is 25 kHz.
Preferably, the rotating speed of the centrifuge is 3 000 ~ 5 000 rpm.
The principle of the present invention：
Cellulase is one group of general name for being capable of the glucogenic enzyme of degraded cellulose, its wide material sources, such as software
Animal, insect, protozoan, bacterium, actinomyces, fungi etc. can all produce cellulase.Water is renewable, pollution-free, and ethanol is early
Be used as the Extraction solvent of active material, can have been mixed in any proportion with water, have pollute it is small, toxicity is low, it is easy recovery etc. it is excellent
Point, under the auxiliary of ultrasonic wave and cellulase, essential oil, flavones and polysaccharide can be extracted from plant using ethanol water.
N-butanol is similar to ethanol, and the product that the crops such as grain, corncob are obtained by fermenting, the micro presence in white wine,
Also allow to use as food additives.
Essential oil is insoluble in water, is slightly soluble in ethanol, and the boiling point of essential oil is higher than water and ethanol.Polysaccharide is large biological molecule, readily soluble
Yu Shui, it is insoluble in the organic solvents such as ethanol.Flavone aglycone is soluble in ethanol, is insoluble in water, and flavonoid glycoside is soluble in water and n-butanol,
N-butanol is slightly soluble in water；Using these characteristics by after ethanol rotary evaporation in extract solution, essential oil crude product, aqueous phase are obtained by centrifugation
With flavone aglycone crude product, using n-butanol, extracting flavone glycosides separates polysaccharide and flavonoid glycoside from aqueous phase, and extraction finishes, and utilizes alcohol
It is heavy that polysaccharide in raffinate phase is purified, flavone aglycone crude product is added in extraction phase, rotated evaporation recovery n-butanol, subtracted
Press dry dry flavones crude product.
Compared with prior art, the advantage of the invention is that：
1. the multiple products such as ultrasonic synergistic cellulase assisted extraction plants essential oil, polysaccharide and flavones
Ultrasonic synergistic cellulase assisted extraction process, from once feeding intake while plants essential oil, polysaccharide can be extracted
With the multiple product such as flavones, efficiency of pcr product and raw material availability can be improved, reduces cost, and its lytic activity is damaged low, thermal sensitivity
Composition wrecks small.
2. product concentrates and solvent recovery technology is simple and environmentally-friendly, efficiency high
Be soluble in ethanol using flavone aglycone, be insoluble in water and essential oil, and the boiling point of essential oil and water is higher than ethanol, essential oil with
These immiscible characteristics of water, recovery ethanol is evaporated under reduced pressure for the first time, is removed after the water of part by centrifuging essential oil, flavonoid glycoside
Member and aqueous phase；It is soluble in water using polysaccharide, n-butanol is insoluble in, flavonoid glycoside is soluble in water and n-butanol, n-butanol are slightly soluble in water
These characteristics, using n-butanol from aqueous phase extracting flavone glycosides separating flavone glycosides and polysaccharide, by flavone aglycone crude product add containing Huang
The extraction phase of ketoside, it is evaporated under reduced pressure after recovery n-butanol by being dried under reduced pressure to obtain flavones crude product；Raffinate phase containing polysaccharide through alcohol precipitation,
Filter and be dried under reduced pressure and to obtain polysaccharide crude.Solvent has the advantages that nontoxic, pollution is small, easily reclaimed.
Embodiment
For the ease of understanding the present invention, present invention work more comprehensively, is meticulously described below in conjunction with preferred embodiment,
But protection scope of the present invention is not limited to embodiment in detail below.
Embodiment 1：
It dry, pulverize after radix valerianae is cleaned, cross 100 mesh sieves, choose 100 g powder and make raw material.With 1.5 L 50% second
Alcohol solution（That is Extraction solvent）After soaking the h of raw material 12, it is put into extraction vessel, and adds cellulase, it is dense in cellulase
Spend under the conditions of 2.4 U/mL Extraction solvents, 55 DEG C of Extracting temperature, ultrasonic wave（Power：200 W, frequency：25 kHz）Processing 35
Min, filtered after extraction, collect filtrate, filter residue, completion is once extracted；The 50% of 1.5 L is added to the filter residue once extracted
Ethanol water soaks 60 min, the operation such as extraction and filtering for being repeated once in extraction, collects filtrate, filter residue, completes secondary
Extraction；1.5 L 50% ethanol water is added to the filter residue of second extraction, repeats immersion, extraction and filtering in second extraction
Deng operation, completion is extracted three times.The filtrate being filtrated to get three times is merged again, carries out being evaporated under reduced pressure recovery second under the conditions of 45 DEG C
Alcohol, part water is removed, afterwards centrifuge（Rotating speed：4000 r/min）15 min, obtain upper strata essential oil crude product, intermediate water phase
With bottom flavone aglycone crude product.N-butanol is extracted to from intermediate water phase mutually to finish without obvious color, extraction, receive using n-butanol
Phase more than extraction phase of the collection containing flavonoid glycoside and the extraction containing polysaccharide.Flavone aglycone crude product is added after the extraction phase containing flavonoid glycoside 75
Rotary evaporation reclaims n-butanol under the conditions of DEG C, and remaining extraction phase is dried under reduced pressure to obtain flavones crude product；It is added more than extraction containing polysaccharide
Enter the absolute ethyl alcohol of 6 times of volumes, filtered after standing 12h, collect filter residue, filtrate, filter residue is filtered through being dried under reduced pressure to obtain polysaccharide crude
Liquid is evaporated under reduced pressure recovery ethanol under the conditions of 55 DEG C；Essential oil crude product is refined using molecularly distilled.It is thick finally to respectively obtain essential oil
The g of product 0.94,0.81 g after refining；The g of polysaccharide crude 7.46, containing the g of polysaccharide 2.87；The g of flavones crude product 8.09, containing flavones 2.31
g。
Embodiment 2：
It dry, pulverize after camphortree leaf is cleaned, cross 10 mesh sieves, choose 100 g powder and be tested raw material.With 1.8 L's
50% ethanol water（That is Extraction solvent）After soaking the h of raw material 15, it is put into extraction vessel, and adds cellulase, in fiber
Plain enzyme concentration is ultrasonic wave under the conditions of 2.1 U/mL Extraction solvents, 50 DEG C of Extracting temperature（Power：200 W, frequency：25 kHz）
25 min are handled, are filtered after extraction, collect filtrate, filter residue, completion is once extracted；1.8 are added to the filter residue once extracted
L 50% ethanol water soaks 100 min, the operation such as extraction and filtering for being repeated once in extraction, collects filtrate, filter residue, complete
Into second extraction；1.8 L 50% ethanol water is added to the filter residue of second extraction, repeats immersion, extraction in second extraction
Extracted three times with operation, completions such as filterings.The filtrate being filtrated to get three times is merged again.It is evaporated under reduced pressure under the conditions of 50 DEG C
Reclaim ethanol, remove part water, afterwards centrifuge（Rotating speed：4000 r/min）20 min, obtain upper strata essential oil crude product, in
Between aqueous phase and bottom flavone aglycone crude product.N-butanol is extracted to mutually without obvious color, extraction from intermediate water phase using n-butanol
Finish, collect phase more than the extraction phase containing flavonoid glycoside and the extraction containing polysaccharide.Flavone aglycone crude product is added into the extraction containing flavonoid glycoside
Rotary evaporation reclaims n-butanol under the conditions of 75 DEG C after phase, and remaining extraction phase is dried under reduced pressure to obtain flavones crude product；Extraction containing polysaccharide
Remainder is added to the absolute ethyl alcohol of 4 times of volumes, is filtered after standing 24 h, collects filter residue, filtrate, filter residue is through being dried under reduced pressure much
Sugared crude product, filtrate are evaporated under reduced pressure recovery ethanol under the conditions of 55 DEG C；Essential oil crude product is refined using molecularly distilled.Respectively finally
To the g of essential oil crude product 2.69,2.29 g after refining；The g of polysaccharide crude 38.90, containing the g of polysaccharide 11.89；The g of flavones crude product 14.84,
Containing the g of flavones 3.25.
Embodiment 3：
50 mesh sieves are crossed after dry, pulverize after Pomelo peel is cleaned, 100 g powder is chosen and is tested raw material.With 2.0 L
45% ethanol water（That is Extraction solvent）After soaking the h of raw material 10, it is put into extraction vessel, and adds cellulase, in fibre
Under the conditions of plain enzyme concentration is tieed up as 2.2 U/mL Extraction solvents, 50 DEG C of Extracting temperature, ultrasonic wave（Power：200 W, frequency：25
kHz）30 min are handled, are filtered after extraction, collect filtrate, filter residue, completion is once extracted；Added to the filter residue once extracted
2.0 L 45% ethanol water soaks 30 min, the operation such as extraction and filtering for being repeated once in extraction, collects filtrate, filter
Slag, complete second extraction；2.0 L 45% ethanol water is added to the filter residue of second extraction, repeats the leaching in second extraction
The operations such as bubble, extraction and filtering, completion are extracted three times.The filtrate being filtrated to get three times is merged again.Carried out under the conditions of 60 DEG C
It is evaporated under reduced pressure recovery ethanol, removes part water, afterwards centrifuge（Rotating speed：4000 r/min）30 min, obtain upper strata essence
Oily crude product, intermediate water phase and bottom flavone aglycone crude product.N-butanol is extracted to mutually without obvious from intermediate water phase using n-butanol
Color, extraction finish, and collect phase more than the extraction phase containing flavonoid glycoside and the extraction containing polysaccharide.Flavone aglycone crude product is added and contains flavones
Rotary evaporation reclaims n-butanol under the conditions of 75 DEG C after the extraction phase of glycosides, and remaining extraction phase is dried under reduced pressure to obtain flavones crude product；Extraction
Remainder is added to the absolute ethyl alcohol of 5 times of volumes, is filtered after standing 24 h, collects filter residue, filtrate, filter residue is through being dried under reduced pressure much
Sugared crude product, filtrate are evaporated under reduced pressure recovery ethanol under the conditions of 55 DEG C；Essential oil crude product is refined using molecularly distilled.Respectively finally
To the g of essential oil crude product 2.42,2.11 g, the g of polysaccharide crude 27.21 after refining, containing the g of polysaccharide 7.49；The g of flavones crude product 6.84, contains
The g of flavones 1.61.
Embodiment 4：
It dry, pulverize after onion is cleaned, cross 50 mesh sieves, choose 100 g powder and be tested raw material.With the 50% of 1.8 L
Ethanol water（That is Extraction solvent）After soaking the h of raw material 1, it is put into extraction vessel, and adds cellulase, in cellulase
Concentration is ultrasonic wave under the conditions of 1.6 U/mL Extraction solvents, 45 DEG C of Extracting temperature（Power：200 W, frequency：25 kHz）Processing
25 min, filtered after extraction, collect filtrate, filter residue, completion is once extracted；Add 1.8 L's to the filter residue once extracted
50% ethanol water soaks 20 min, the operation such as extraction and filtering for being repeated once in extraction, collects filtrate, filter residue, completes two
Secondary extraction；1.8 L 50% ethanol water is added to the filter residue of second extraction, repeats immersion, extraction and mistake in second extraction
The operation such as filter, completion are extracted three times.The filtrate being filtrated to get three times is merged again.Vacuum distillation recovery is carried out under the conditions of 50 DEG C
Ethanol, part water is removed, afterwards centrifuge（Rotating speed：4000 r/min）10 min, obtain upper strata essential oil crude product, middle water
Phase and bottom flavone aglycone crude product.N-butanol is extracted to using n-butanol from intermediate water phase mutually to finish without obvious color, extraction,
Phase more than extraction phase of the collection containing flavonoid glycoside and the extraction containing polysaccharide.By flavone aglycone crude product add the extraction phase containing flavonoid glycoside after
Rotary evaporation reclaims n-butanol under the conditions of 75 DEG C, and remaining extraction phase is dried under reduced pressure to obtain flavones crude product；Phase more than extraction containing polysaccharide
Add 10 times of volumes absolute ethyl alcohol, stand 48 h after filter, collect filter residue, filtrate, filter residue through be dried under reduced pressure polysaccharide is thick
Product, filtrate are evaporated under reduced pressure recovery ethanol under the conditions of 55 DEG C；Essential oil crude product is refined using molecularly distilled.Finally respectively obtain essence
The oily g of crude product 0.61,0.51 g, the g of polysaccharide crude 13.96 after refining, containing the g of polysaccharide 4.62；The g of flavones crude product 18.53, flavones
4.83 g。
Claims (6)
1. the extraction process of a kind of plants essential oil, flavones and polysaccharide, concrete operation step are as follows：
（1）Dry and crush：By raw material cleaning, drying, 10 ~ 100 mesh sieves are crossed after crushing, it is standby；
（2）Immersion：By step（1）Standby raw material soaks 1 ~ 15 h with ethanol water；
（3）Once extract：By step（2）In raw material and ethanol water be put into extraction vessel, toward Extraction solvent ethanol water
In solution add cellulase after under the conditions of 25 DEG C ~ 65 DEG C the min of ultrasonication 10 ~ 120, filtered after extraction, collect
Filtrate, filter residue；
（4）Second extraction：To step（3）Filter residue add ethanol water, soak repeat step after 10 ~ 120 min（3）'s
Extraction and filtration step；
（5）Extract three times：Repeat step（4）Process；
（6）Once it is evaporated under reduced pressure：By step（3）、（4）、（5）In filtrate merge, be evaporated under reduced pressure back under the conditions of 45 ~ 60 DEG C
Ethanol is received, removes part water；
（7）Centrifuge：Using centrifuge by step（6）Remaining filtrate centrifuges 10 ~ 60 min afterwards, obtains essential oil crude product, aqueous phase
With flavone aglycone crude product；
（8）Extract and separate：Using n-butanol from step（7）In the intermediate water phase of gained extracting flavone glycosides to n-butanol mutually without bright
Aobvious color, obtains the extraction phase containing flavonoid glycoside and the raffinate phase containing polysaccharide；
（9）Alcohol precipitation：Toward step（8）In the raffinate containing polysaccharide be added to the absolute ethyl alcohols of 4 ~ 10 times of volumes, stand 12 ~ 48 h,
Filtering, collect filtrate, filter residue；
（10）Secondary pressure is distilled and dried：By step（7）Obtained flavone aglycone crude product adds step（8）In containing flavonoid glycoside
In extraction phase, recovery n-butanol is evaporated under reduced pressure under the conditions of 70 DEG C ~ 80 DEG C, remaining extraction phase is dried under reduced pressure to obtain flavones crude product；
By step（9）Obtained filtrate is evaporated under reduced pressure recovery ethanol under the conditions of 45 ~ 60 DEG C, by step（9）Obtained filter residue decompression is dry
It is dry to obtain polysaccharide crude；
（11）Molecular distillation refines：By step（7）Obtained essential oil crude product refines through molecular distillation technique；
（12）Detect essential oil crude product and the quality after molecular distillation technique is refined；Detect polysaccharide crude and contained polysaccharide quality；
Detect flavones crude product and contained flavones quality.
2. the extraction process of plants essential oil according to claim 1, flavones and polysaccharide, it is characterised in that：The step
（2）, step（3）, step（4）, step（5）Middle ethanol water is that ethanol configures with water, and volume fraction is 30% ~ 70%.
3. the extraction process of plants essential oil according to claim 1, flavones and polysaccharide, it is characterised in that：The step
（2）, step（4）, step（5）The amount of the added etoh solvent aqueous solution, its volume and material quality ratio are 5 ~ 25: 1 mL/g.
4. the extraction process of plants essential oil according to claim 1, flavones and polysaccharide, it is characterised in that：The step
（3）, step（4）, step（5）In, the concentration for adding cellulase is 0.3 ~ 4.0 U/mL Extraction solvents.
5. the extraction process of plants essential oil according to claim 1, flavones and polysaccharide, it is characterised in that：The ultrasonic wave
Power be 200 W, frequency is 25 kHz.
6. the extraction process of plants essential oil according to claim 1, flavones and polysaccharide, it is characterised in that：The centrifuge
Rotating speed be 3 000 ~ 5 000 rpm.
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