CN110302240A - The method for extracting polyphenol in walnut Diaphragma juglandis - Google Patents
The method for extracting polyphenol in walnut Diaphragma juglandis Download PDFInfo
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- CN110302240A CN110302240A CN201811242937.4A CN201811242937A CN110302240A CN 110302240 A CN110302240 A CN 110302240A CN 201811242937 A CN201811242937 A CN 201811242937A CN 110302240 A CN110302240 A CN 110302240A
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- diaphragma juglandis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/52—Juglandaceae (Walnut family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention discloses a kind of methods of polyphenol in extraction walnut Diaphragma juglandis, including pretreatment, extraction, purifying.The present invention introduces low-temp. extrusion expanding treatment technology and cryogenic high pressure homogeneous extractive technique in the extraction process of traditional solvent extraction method, by the low Extracting temperature of adjustable pressure drops, loss is reduced while improving polyphenol release, to improve polyphenol extraction efficiency.The present invention is suitable for extracting the polyphenol substance walnut Diaphragma juglandis.
Description
Technical field
The invention belongs to a kind of technology of effective component extracting from natural plants fruit, specifically a kind of extraction core
The method of polyphenol in peach Diaphragma juglandis.
Background technique
Diaphragma juglandis is the wooden diaphragm in walnut fruit stone, usually accounts for the 4 ~ 5% of walnut quality.Traditional medicine and modern study
Show Diaphragma juglandis have good medical value, can be used for treat invigorating the spleen reinforce the kidney, diuresis heat-clearing, treatment insomnia forgetfulness, leaching
The diseases such as sick hematuria, hot summer weather dysentery.Contain the functional phenolic material such as gallic acid, progallin A, Quercetin in Diaphragma juglandis
Matter has the plurality of health care functions such as anti-inflammatory, anti-oxidant, antibacterial, antiviral.But functional components in Diaphragma juglandis are extracted
Study it is less, it is more rare for the report of the extraction of polyphenols in Diaphragma juglandis.Also just because of the research to Diaphragma juglandis is opened
Send out insufficient, the functional raw material of this great Development volue caused not to be fully utilized, mostly as processing byproduct by with
Fuel or discarding are done, which results in the great wastings of resources.Therefore, using Diaphragma juglandis as research object, extract polyphenol therein at
Split pole has Development volue.
Traditional polyphenol extraction mostly uses solvent extraction method, and the cell wall of Diaphragma juglandis is thicker, and polyphenol substance is not easy to discharge, and mentions
Hot conditions are needed when taking, while the used time is longer.Due to the easier release of the higher polyphenol of temperature in technical process, but it is excessively high
Temperature can be such that the double bond in polyphenol ring is broken again, cause the degradation of polyphenol, polyphenol property be destroyed, to influence the extraction of polyphenol
Rate, therefore temperature can not be efficiently controlled in the prior art, not only guarantee the release of polyphenol, but also the drop of polyphenol can be effectively prevent
Solution.
The polyphenol in Diaphragma juglandis is extracted, needs to carry out broken wall pretreatment, traditional wall-breaking method has following several: (1) ball
Film machine broken wall, this wall-breaking method is more ancient, and sporoderm-broken rate is low, and effect is poor；(2) chemical-agent technique, this method can be to extracts
Matter is destroyed, and physical property is made to change；(3) biologic enzymolysis method, this method easily cause product inhibition, and lyase price
Height limits extensive utilization；(4) method is pulverized, this method is most common wall breaking technology, using shearing force to object
Matter is crushed, the disadvantage is that crushing is irregular, and needs to operate under the high temperature conditions, the double bond in polyphenol ring can be made to be broken, drawn
The degradation for playing polyphenol, to influence the recovery rate of polyphenol.Extruding and puffing technology is that the material that will be crushed inputs in extruding machine cavity,
By adjusting the active force between machine cavity, screw rod and material, so that the pressure of inside cavity gradually rises.Later, work as object
When material is released, pressure moment declines, and the moisture flash evaporation in material, structure changes.Extruding and puffing technology usually quilt
As food processing, using high temperature extrusion extruding to modified technique, in the process, the partially protein in material becomes
Property；Partial starch is degraded；Few fibers are broken.By low-temp. extrusion puffing technique to destroy cell wall application compared with
To be rare, which has destruction to plant-derived physical cell wall and cell membrane, can make the active material in cell
Preferably release is applied to the pretreatment stage of polyphenol extraction at a lower temperature, sporoderm-broken rate can be improved, greatly increase
The release rate of polyphenol.
High pressure homogenization technique is that liquid material high speed is made to flow through narrow gap using high pressure, in powerful shearing force, because of impact
Equipment inner wall and generate powerful impact force, and because of static pressure bust and the comprehensive function of hole explosive force that is generated due to jumping
Under, the coarseer emulsion of former particle or suspension are processed into particle subtle stable emulsion or suspension, to make
Liquid material system is more stable.Extracting temperature is reduced using high pressure homogenization technique, it can be by the low boiling points such as ethyl alcohol, low cost
Organic solvent introduce this system, greatly reduce polyphenol loss so that recovery rate further increases.
Summary of the invention
The purpose of the present invention is to provide for a kind of method for extracting polyphenol in walnut Diaphragma juglandis, extracts in traditional solvent
Low-temp. extrusion expanding treatment technology and cryogenic high pressure homogeneous extractive technique are introduced on the basis of process, by adjusting pressure, reducing
Extracting temperature reduces loss while improving polyphenol release, to improve polyphenol extraction efficiency.
To achieve the goals above, used technical solution is as follows by the present invention,
A method of polyphenol in walnut Diaphragma juglandis being extracted, sequence carries out according to the following steps:
One, it pre-processes
Walnut Diaphragma juglandis is chosen, grinding and sieving, through low-temp. extrusion extruding, must divert one's attention wood powder A；
Two, it extracts
1) ratio for being 1kg:5 ~ 10L in mass volume ratio, will divert one's attention wood powder A and ethanol solution addition cryogenic high pressure homogenizer,
Cryogenic high pressure homogeneous is carried out, 2 ~ 4h is extracted, obtains the thick polyphenol suspension B of Diaphragma juglandis；Wherein, the concentration of the ethanol solution is 60
~80%；
2) the thick polyphenol suspension B of Diaphragma juglandis is centrifuged, centrifuge speed is 3500 ~ 4000r/min, gained supernatant
The as thick polyphenol extracting solution C of Diaphragma juglandis；
Three, it purifies
Polyphenol extracting solution C thick to Diaphragma juglandis carries out concentration, removes in the thick polyphenol extracting solution C of Diaphragma juglandis 10 ~ 30% ethyl alcohol,
The thick polyphenol concentrate D of Diaphragma juglandis, the thick polyphenol concentrate D of Diaphragma juglandis is injected into chromatographic column, then with ethanol elution, gained eluent
Ethyl alcohol, the Diaphragma juglandis polyphenol extract E purified are removed through vacuum distillation.
As a kind of restriction of the invention, Diaphragma juglandis polyphenol extract E is dried using freeze drier to get dividing
Heart wood polyphenol freeze-dried powder.
It is limited as another kind, 50 ~ 70 DEG C of the extruding jacket temperature of the low-temp. extrusion extruding, screw speed 180 ~
200r/min, 8 ~ 10nm of film hole diameter.
As the third restriction, the homogenization pressure of the cryogenic high pressure homogeneous is 100 ~ 200MPa, and homogenizing temperature is 20 ~ 40
℃。
The present invention is there are also a kind of restriction, the chromatographic column macroreticular resin model AB-8, and elution concentration of alcohol is 60% ~
80%。
Due to the adoption of the above technical solution, compared with prior art, acquired technological progress is the present invention:
1) invention introduces the extruding and puffing technologies for being usually only used to modified technique, because it is to plant-derived physical cell wall
The destruction having with cell membrane can be such that the active material in cell preferably discharges, be applied at a lower temperature
In the pretreatment stage that polyphenol extracts, the release rate of polyphenol in raw material is increased；
2) present invention further decreases Extracting temperature using high pressure homogenization technique, the organic solvents such as ethyl alcohol can be introduced this body
System greatly reduces polyphenol loss, so that recovery rate further increases；
3) the method for the present invention is simple and convenient to operate, and optimal conditions are relatively easily accessible, and is suitble to industrial production, preparation-obtained
Diaphragma juglandis polyphenol content is high, can enrich the type of domestic polyphenol product；
4) remaining byproduct can further extract wood polysaccharose of diverting one's attention etc. in the present invention, improve walnut utilization rate.
To sum up, the present invention is further processed as freeze-dried powder, simultaneously suitable for extracting to the polyphenol walnut Diaphragma juglandis
It can be applied in the polyphenol extraction of a variety of raw material, development prospect with higher.
Specific embodiment
Below with reference to embodiment, the invention will be further described, it is understood by those skilled in the art that of the invention
It is not limited to following embodiment, any improvements and changes made on the basis of the specific embodiment of the invention are all in right of the present invention
Within the scope of claimed.
Reagent used in following embodiment is unless otherwise required common agents in the art.
Method used in following embodiment is unless otherwise required conventional method in the art.
A kind of method for extracting polyphenol in walnut Diaphragma juglandis of embodiment 1
The present embodiment sequentially carries out according to the following steps:
One, it pre-processes
The Diaphragma juglandis in the walnut of Hebei province's Xingtai City Lincheng County production is chosen, crushed 200 meshes, inputs extruding machine cavity
It is interior, via low-temp. extrusion extruding, obtain the wood powder A1 that diverts one's attention.Condition for expansion are as follows: 70 DEG C of extruding jacket temperature, screw speed 200r/
Min, film hole diameter 8nm.It carries out by adjusting the active force between machine cavity, screw rod and material, so that the pressure of inside cavity
Power gradually rises.Later, when material is released, pressure moment declines, and the moisture flash evaporation in material, structure changes
Become.By cryogenic high pressure extruding, the active material in cell can be made preferably to discharge, be applied at a lower temperature more
The pretreatment stage of phenol extraction, the release rate for the polyphenol being significantly greatly increased.
The crushing of Diaphragma juglandis, to fineness, there is no limit, but granularity is smaller, and the recovery rate of polyphenol is higher, and opposite cost is got over
It is low.
Two, it extracts
1) by 5kg divert one's attention wood powder A1 and 50L concentration be 70% ethanol solution after mixing, be placed in cryogenic high pressure homogenizer,
Processing condition are as follows: homogenization pressure 180MPa, extracts 2h, obtains the thick polyphenol suspension B1 of Diaphragma juglandis by 40 DEG C of homogenizing temperature.High pressure is equal
Matter technology is that liquid material high speed is made to flow through narrow gap using high pressure, in powerful shearing force, is generated because of impact device inner wall
Powerful impact force, and because static pressure bust under the comprehensive function of hole explosive force that is generated due to jumping, by former particle ratio
Coarseer emulsion or suspension is processed into particle subtle stable emulsion or suspension, to make liquid material system more
Add stabilization.
Divert one's attention the ratio of wood powder and ethanol solution, the ethanol solution of addition of being subject to is enough the wood powder submergence that will divert one's attention, and second
The additional amount of alcoholic solution is higher, and the recovery rate of polyphenol can more be guaranteed.
2) the thick polyphenol suspension B1 of Diaphragma juglandis is centrifuged, centrifugal rotational speed 3500r/min, gained supernatant is
For the thick polyphenol extracting solution C1 of Diaphragma juglandis.
Three, it purifies
Polyphenol extracting solution C1 thick to gained Diaphragma juglandis carries out concentration, removes in the thick polyphenol extracting solution C1 of Diaphragma juglandis 10% second
Alcohol obtains the thick polyphenol concentrate D1 of Diaphragma juglandis, is concentrated using the thick polyphenol of chromatographic column absorption Diaphragma juglandis of macroreticular resin model AB-8
Polyphenol component in liquid D1, then with the ethanol elution of 70% concentration, eluent flow rate 200mL/h, the vacuum distillation of gained eluent
Ethyl alcohol is removed, Diaphragma juglandis polyphenol extract E1 is obtained.
Four, dry
Using freeze drier, Diaphragma juglandis polyphenol extract E1 is dried to get Diaphragma juglandis polyphenol freeze-dried powder.
The method that embodiment 2-8 extracts polyphenol in walnut Diaphragma juglandis
Embodiment 2-8 be respectively it is a kind of extraction walnut Diaphragma juglandis in polyphenol method, their extraction step with 1 phase of embodiment
Together, only the operating parameter in extraction process is different, and concrete operations parameter is as shown in table 1:
Table 1
Embodiment 9 extracts the extraction effect of the method for polyphenol in walnut
The present embodiment is the extraction effect measurement for being used to extract the method for polyphenol in walnut to embodiment 1-8.
The polyphenol content in extract is measured using Folin-Ciocalteu method, specific detecting step is as follows:
Gallic acid titer is prepared, draws the titer 1.0mL of each concentration respectively, each sodium carbonate liquor that 3mL7.5% is added
It develops the color with 1mL forint phenol reagent, adds distilled water to be settled to 10mL, shake up, after avoid light place 2h, measured at 765nm wavelength every
The absorbance value of a standard solution.The absorbance y surveyed is drawn into standard curve, recurrence side with corresponding gallic acid concentration x
Journey are as follows: y=13.24x+0.0833, R2=0.997。
After the Diaphragma juglandis freeze-dried powder in embodiment 1-8 is diluted to extracting solution respectively, respectively take 1.0mL extracting solution, respectively plus
Sodium carbonate liquor and the colour developing of 1mL forint phenol reagent for entering 3mL7.5%, add distilled water to be settled to 10mL, shake up, avoid light place 2h
Afterwards, the absorbance value of each solution is measured at 765nm wavelength.Substituting into standard curve, (y=13.24x+0.0833, y are absorbance, x
For gallic acid concentration), find out polyphenol concentration.
Polyphenol recovery rate is calculated according to the following formula:
Polyphenol recovery rate (%)=X × V/ M × 100%
In formula: X- polyphenol concentration (g/mL)；Extracting liquid volume (mL) after V- dilution；M- sample Diaphragma juglandis quality (g).
Testing result is as shown in table 2:
Table 2
Comparative example
In order to detect the extraction effect of this method, diverting one's attention in the walnut of same batch Hebei province Xingtai City Lincheng County production is used
Wood is compared using Conventional solvents extraction method and ultrasound assisted extraction method with embodiment 1.The detection side of polyphenol content used
Method is the Folin-Ciocalteu detection method in embodiment 9.
Conventional solvents extraction method is as follows using technique:
One, it pre-processes
Walnut Diaphragma juglandis is chosen, crushed 200 meshes.
Two, it extracts
1) 5kg is divert one's attention into ethanol solution that wood powder A1 and 50L concentration is 70% after mixing, is placed under 70 DEG C of water-baths to flow back and mentions
When taking 2h(using ultrasound assisted extraction method, ultrasonic wave added power is 300W), obtain the thick polyphenol suspension of Diaphragma juglandis；
2) the thick polyphenol suspension of Diaphragma juglandis obtained in step 1) is centrifuged, gained supernatant is that Diaphragma juglandis is slightly more
Phenol extraction liquid.
Three, it purifies
Concentration is carried out to the thick polyphenol extracting solution of gained Diaphragma juglandis, in the thick polyphenol extracting solution of Diaphragma juglandis 10% ethyl alcohol is removed, obtains
The thick polyphenol concentrate of Diaphragma juglandis, using more in the chromatographic column absorption thick polyphenol concentrate of Diaphragma juglandis of macroreticular resin model AB-8
Phenol ingredient, then with the ethanol elution of 70% concentration, eluent flow rate 200mL/h, gained eluent carries out vacuum distillation again and removes
Ethyl alcohol is removed, Diaphragma juglandis polyphenol extract is obtained.
Four, dry
Diaphragma juglandis polyphenol extract after purification is dried to get Diaphragma juglandis polyphenol freeze-dried powder using freeze drier.
Ultrasound assisted extraction method is similar to Conventional solvents extraction method using processing step, is a difference in that in step 2
Ultrasonic wave added, ultrasonic power 300W are added when step 1) refluxing extraction.
Polyphenol recovery rate is 40.1% in embodiment 1, and the polyphenol purity of products obtained therefrom is 90.1%.
Traditional handicraft polyphenol recovery rate is 30.1%, and the polyphenol purity of products obtained therefrom is 89.8%.
It is 35% via polyphenol recovery rate after ultrasound assisted extraction, the polyphenol purity of products obtained therefrom is 90%.
Polyphenol recovery rate of the invention comparison traditional handicraft rises 5.1% ~ 10% known to comparison, improves the utilization of raw material
Rate has preferable development prospect.
Finally, it should be noted that the foregoing is only a preferred embodiment of the present invention, it is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
To modify the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in of the invention
Within protection scope.
Claims (7)
1. a kind of method for extracting polyphenol in walnut Diaphragma juglandis, which is characterized in that sequence carries out according to the following steps:
One, it pre-processes
Walnut Diaphragma juglandis is chosen, grinding and sieving, through low-temp. extrusion extruding, must divert one's attention wood powder A；
Two, it extracts
1) ratio for being 1kg:5 ~ 10L in mass volume ratio, will divert one's attention wood powder A and ethanol solution addition cryogenic high pressure homogenizer,
Cryogenic high pressure homogeneous is carried out, 2 ~ 4h is extracted, obtains the thick polyphenol suspension B of Diaphragma juglandis；Wherein, the concentration of the ethanol solution is 60
~80%；
2) the thick polyphenol suspension B of Diaphragma juglandis is centrifuged, centrifuge speed is 3500 ~ 4000r/min, gained supernatant
The as thick polyphenol extracting solution C of Diaphragma juglandis；
Three, it purifies
Polyphenol extracting solution C thick to Diaphragma juglandis carries out concentration, removes in the thick polyphenol extracting solution C of Diaphragma juglandis 10 ~ 30% ethyl alcohol,
The thick polyphenol concentrate D of Diaphragma juglandis, the thick polyphenol concentrate D of Diaphragma juglandis is injected into chromatographic column, then with ethanol elution, gained eluent
Ethyl alcohol, the Diaphragma juglandis polyphenol extract E purified are removed through vacuum distillation.
2. the method according to claim 1 for extracting polyphenol in walnut Diaphragma juglandis, it is characterised in that: use freeze drier
Diaphragma juglandis polyphenol extract E is dried to get Diaphragma juglandis polyphenol freeze-dried powder.
3. the method according to claim 1 or 2 for extracting polyphenol in walnut Diaphragma juglandis, it is characterised in that: the low temperature squeezes
The extruding jacket temperature for pressing extruding is 50 ~ 70 DEG C, 180 ~ 200r/min of screw speed, 8 ~ 10nm of film hole diameter.
4. the method according to claim 1 or 2 for extracting polyphenol in walnut Diaphragma juglandis, it is characterised in that: the low temperature is high
The homogenization pressure for pressing homogeneous is 100 ~ 200MPa, and homogenizing temperature is 20 ~ 40 DEG C.
5. the method according to claim 3 for extracting polyphenol in walnut Diaphragma juglandis, it is characterised in that: the cryogenic high pressure is equal
The homogenization pressure of matter is 100 ~ 200MPa, and homogenizing temperature is 20 ~ 40 DEG C.
6. the method according to claim 1 or 2 for extracting polyphenol in walnut Diaphragma juglandis, it is characterised in that: the chromatographic column
Macroreticular resin model AB-8, elution concentration of alcohol are 60% ~ 80%.
7. the method according to claim 3 for extracting polyphenol in walnut Diaphragma juglandis, it is characterised in that: the chromatographic column macropore
Resin model is AB-8, and elution concentration of alcohol is 60% ~ 80%.
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