CN109021125A - A kind of method that the co-precipitation of quinine quaternary ammonium salt prepares high-purity garlic polysaccharide - Google Patents
A kind of method that the co-precipitation of quinine quaternary ammonium salt prepares high-purity garlic polysaccharide Download PDFInfo
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Abstract
The invention belongs to technical field of phytochemistry, and in particular to a kind of method that the co-precipitation of quinine quaternary ammonium salt prepares high-purity garlic polysaccharide.The present invention forms sediment as precipitating reagent and garlic polysaccharide using quinine quaternary ammonium salt, sediment is dissociated in inorganic salt solution, and ultrafiltration obtains dissociation solution, is carried out alcohol to dissociation solution and is analysed to obtain high-purity garlic polysaccharide, molecular weight distribution is at normal distribution, average molecular weight 4380Da.Extracting method of the present invention is simple, easily controllable, and each process all has crucial qualifying point, is conducive to production amplification.
Description
Technical field
The invention belongs to technical field of phytochemistry, and in particular to a kind of quinine quaternary ammonium salt co-precipitation preparation high-purity
The method of garlic polysaccharide.
Background technique
Garlic polysaccharide is the highest healthcare function ingredient of content in garlic, accounts for the 15%~25% of fresh garlic, has proliferation intestines
Road probiotics assists digestion, strengthen immunity, prevents hepatic injury, anti-oxidant and antiviral etc. multiple efficacies.Garlic polysaccharide is being cured
The fields such as medicine, food have wide actual application prospect.
Currently, the extracting method of garlic polysaccharide mainly has hot water, dilute salt, diluted alkaline and diluted acid to extract, also at useful complex enzyme
Reason, ultrasonic wave added accelerate the extracting method of polysaccharide release.Hot water extraction is a kind of common method for extracting plant polyose,
Advantage is low in cost, simple process, without special instruments and equipment, can be used for being mass produced, the disadvantage is that when extraction process
Between it is long.Enzymatic Extraction is the method combined using enzyme with hot water extraction, there is combined-enzyme method and single enzyme process.Common enzyme has egg
White enzyme, cellulase, pectase etc..Enzymatic Extraction plant polyose is with reaction condition is mild, recovery rate is high, impurity easily removes, mentions
Take the time short, simple process, and it is able to maintain the natural activity of polysaccharide, but the price of some enzymes is higher, the condition used compares
It is harsh.Extraction be using diluted acid or diluted alkaline effect under, cell, the cell wall of plant or mushroom be sufficiently swollen so that
Rupture is increased the polysaccharide wherein to dissociate, is more readily dissolved in diluted acid or aqueous alkali, can be improved using this characteristic such more
The recovery rate of sugar.And traditional garlic polysaccharide extracting method generally requires multiple removing protein, except pectin, ethyl alcohol sedimentation, gel
The purification procedures of a series of complex such as column purification, dialysis can just prepare the garlic polysaccharide of high-purity, preparation section and its cumbersome.
Compared to traditional garlic polysaccharide extracting method, emerging in recent years some new technologies, such as supercritical CO2Extract skill
Art；1818045 A of CN discloses a kind of using supercritical CO2The method for extracting garlic oil and garlic polysaccharide simultaneously from fresh garlic,
Although the technological parameter is simple, technique is environmentally protective, and separation process needs to carry out under high pressure, equipment one-time investment mistake
Greatly, extraction can not be carried out continuously, and cause effective low yield of equipment.Water-insoluble is formed at salt using quaternary ammonium salt and acidic polysaccharose
Compound come carry out purifying be a kind of simplicity purification process, common quaternary ammonium salt be cetyl trimethylammonium bromide
(CTMAB) and its cetyltrimethylammonium hydroxide (CTA-OH) and cetylpyridinium chloride (CPC), but after it is at salt
It is not easy to dissociate, causes polysaccharide yield low.
Therefore, exploitation is concisely and efficiently, the garlic polysaccharide separation and Extraction method of industrialized production is suitble to be very important.
Summary of the invention
The purpose of the present invention is overcoming deficiency in the prior art, a kind of isolation and purification method of garlic polysaccharide is provided, originally
Invention uses quinine quaternary ammonium salt to form sediment as precipitating reagent and garlic polysaccharide for the first time, and sediment is in inorganic salt solution
In dissociated, ultrafiltration obtains dissociation solution, to dissociation solution carry out alcohol analyse to obtain high-purity garlic polysaccharide.Extracting method of the present invention is simple,
Easily controllable, each process all has crucial qualifying point, is conducive to production amplification.
According to an aspect of the present invention, this hair provides a kind of quinine quaternary ammonium salt co-precipitation and prepares high-purity garlic
The method of polysaccharide, comprising the following steps:
1) degreasing pre-treatment: to drying and crushing after garlic peeling, using the mixed liquor of acetone and tetrahydrofuran to crushing after
Garlic carry out ungrease treatment obtain primary garlic slag；
2) low temperature ultrasonic removes small molecular sugar: by the primary garlic slag Jing Guo ungrease treatment in the ethanol water of 90%V
Ultrasound removal small molecular sugar obtains desugar second level garlic slag；
3) grading extraction of garlic polysaccharide: desugar second level garlic slag is successively carried out in neutrality, faintly acid, weakly alkaline water
The extracting solution of garlic polysaccharide must be contained by extracting garlic polysaccharide；
4) garlic polysaccharide crude product the separation of garlic polysaccharide crude product: is analysed to obtain using alcohol to the extracting solution containing garlic polysaccharide；
5) purifying of garlic polysaccharide crude product: garlic polysaccharide crude product is successively decolourized, is dissolved in purified water and being added after de- albumen
Quinine quaternary ammonium salt forms insoluble matter precipitating, is centrifuged to obtain sediment, sediment is dissociated in inorganic salt solution, ultrafiltration
Dissociation solution is obtained, alcohol is carried out to dissociation solution and analyses to obtain high-purity garlic polysaccharide.
Extracting method according to the present invention, specific steps are as follows:
1) degreasing pre-treatment: through being dried under vacuum to constant weight at 35-40 DEG C after garlic peeling, then shredding to partial size is 200-
300 mesh obtain garlic granule, and garlic granule is placed in high-pressure closed vessel, and the mixed liquor of acetone and tetrahydrofuran is added, is dispersed with stirring 10-
Then 20min is added the dry ice that partial size is 0.5-1cm from feed opening, closes feed opening, be stirred at room temperature, in kettle
When temperature warms naturally to 0-5 DEG C, stop stirring；Filters pressing removes filtrate, carries out being dried under vacuum to constant weight at 35-40 DEG C to filter cake
Obtain degreasing primary garlic slag；It in the mixed liquor of the acetone and tetrahydrofuran, is calculated according to volume ratio, acetone/tetrahydrofuran=95:
5；The weight ratio of the mixed liquor and dry ice of the acetone and tetrahydrofuran is 1:1, the garlic granule and the acetone and tetrahydrofuran
Mixed liquor weight ratio be 1:5；
(35-40 DEG C) removes garlic internal moisture to the present invention in the case where nearly room temperature first, then in the super of dry ice
The fat in garlic is removed under low temperature environment, is usually associated with part sugar in common process during removing fat
The removing of substance, has lost par-tial polysaccharide；The present invention is in order to avoid carbohydrate is in the loss of skimming processes polysaccharide, using ultralow temperature
(dry ice and acetone mixing, warm naturally to 0-5 DEG C from -78 DEG C) removing fat, in addition as dry ice is constantly by Solid State Transformation
Gaseous state, the CO of generation2Molecule has not only acted as the other stirring action of molecular level, and reacting kettle inner pressure constantly increases, in height
Pressure carries out degreasing, accelerates degreasing process and degreasing degree；
2) low temperature ultrasonic removes small molecular sugar: the ethyl alcohol for the 90%V that degreasing primary garlic slag is added to 5-6 times of weight is water-soluble
In liquid, ultrasound 6-8h at 10-20 DEG C, centrifugal filtration removes filtrate, must take off to constant weight is dried under vacuum at 35-40 DEG C of filter residue progress
Sugared second level garlic slag；The present invention is before extracting polysaccharide, using the ethanol water of 90%V as solvent, using the method for ultrasound
Remove the small molecules carbohydrate impurity such as the monosaccharide and disaccharide in garlic slag；
3) grading extraction of garlic polysaccharide: desugar second level garlic slag is placed in purified water, 60-70 DEG C of ultrasonic extraction is warming up to
6-8h is then centrifuged for, and obtains neutral filtrate and A grades of filter residues；A grades of filter residues are placed in the phosphate-buffered aqueous solution of pH=5.7, are risen
Temperature is then centrifuged for 60-70 DEG C of ultrasonic extraction 3-4h, obtains faintly acid filtrate and B grades of filter residues；B grades of filter residues are placed in pH=7.6's
In phosphate-buffered aqueous solution, it is warming up to 60-70 DEG C of ultrasonic extraction 3-4h, is then centrifuged for, obtains alkalescent filtrate and C grades of filter residues；
C grades of filter residues of reject, merging neutral filtrate, faintly acid filtrate and alkalescent filtrate must contain the extracting solution of garlic polysaccharide；
The present invention is reformed in the extraction process of garlic polysaccharide, in order to improve the extract yield of garlic polysaccharide, this
Invention carries out neutral, faintly acid and alkalescent multistage is extracted, extracted under more multiple neutrallty condition garlic polysaccharide extracted amount have compared with
It is big to improve；In addition, in order to increase extraction efficiency, the present invention uses ultrasound assisted extraction technique, increases extraction efficiency；
4) separation of garlic polysaccharide crude product: the extracting solution containing garlic polysaccharide is concentrated under reduced pressure at 45-50 DEG C, when dense
Being reduced to density is 1.18-1.13g/cm3When, stop concentration, then handling material is down to 40-45 DEG C, use is compacted into stirred tank
The ethanol water of 90%V is added dropwise in dynamic pump into stirred tank with the charging rate of 2-3L/h, gradually there is solubility during being added dropwise
Soluble solids is precipitated；Every the 1h filtering with microporous membrane that feeding liquid is 0.2 micron by aperture from stirred tank, filtrates tested density
For 1.02-1.03g/cm3When stop be added dropwise 90%V ethanol water；2-5 DEG C is cooled to the rate of temperature fall of 2-5 DEG C/min
Garlic polysaccharide crude product is filtered to obtain after standing 6-8h；Density of the present invention each means the density at 25 DEG C；
The present invention striking point that garlic polysaccharide is monitored using density domination method creative when using ethyl alcohol sedimentation
With the terminal that 90%V ethanol water is added dropwise, traditional direct dropwise addition fixed amount ethyl alcohol has been abandoned into system, has taken density control
System is more scientific, is in addition also easier to realize in production amplification process, avoid in production amplification process due to enlarge-effect
Generate mass deviation.
5) purifying of garlic polysaccharide crude product: garlic polysaccharide crude product is successively decolourized, is dissolved in purified water and being added after de- albumen
Quinine quaternary ammonium salt forms insoluble matter precipitating, and mixture is centrifuged to obtain sediment and filtrate after heat preservation is stood；Filtrate uses ethyl alcohol
Settle to obtain neutral garlic polysaccharide；Sediment is dissociated in inorganic salt solution, and ultrafiltration obtains dissociation solution, carries out alcohol to dissociation solution
Analyse to obtain high-purity garlic polysaccharide；
The present invention forms this property of sediment using quinine quaternary ammonium salt and acid garlic polysaccharide, and acid garlic is more
Sugar is separated with neutral garlic polysaccharide, and the precipitating of formation is then carried out dissociation using inorganic salts and releases acid garlic polysaccharide, from
And play the purpose for purifying acid garlic polysaccharide；
Preferably, the quinine quaternary ammonium salt is N- (9- anthracene methyl) bromination quinine, N- Benzylphosphonium Bromide quinine
Alkali or O- allyl-N- (9- anthracene methyl) bromination quinine, further preferably N- Benzylphosphonium Bromide quinine；In order to improve
The purity of acid garlic polysaccharide, the present invention has screened a large amount of quaternary ammonium salts and has been used to form sediment with acid garlic polysaccharide, final to select
Select quinine quaternary ammonium salt；Quinine quaternary ammonium salt and cetyl trimethylammonium bromide (CTMAB) and its cetyl front three
Base ammonium hydroxide (CTA-OH) is compared with cetylpyridinium chloride (CPC), and the garlic polysaccharide prepared either purity is still
Yield is above conventional quaternary ammonium salts.
Preferably, the step 5) inorganic salts are ammonium hydrogen sulfate, ammonium sulfate, ammonium dihydrogen phosphate.The selection of inorganic salts and season
The selection of ammonium salt be it is mutually matched, the two plays synergistic effect；Even if using certain quaternary ammonium salt can to greatest extent with acid
Property garlic polysaccharide formed sediment, however sediment hardly result in subsequent processing dissociation can not also go to actually use；This hair
It is bright to have attempted NaCl, KCl, CaCl2Equal neutral salt, test result discovery neutral salt are unfavorable for the sediment of quinine formation
Dissociation, causes the yield of acid garlic polysaccharide relatively low；Test discovery can greatly increase the solution of sediment using faintly acid ammonium salt
From obtaining the garlic polysaccharide of high yield.
Preferably, decoloration described in step 5) of the present invention refers to garlic polysaccharide crude product is dissolved in water after, adjusted using ammonium hydroxide
System pH to 7.8-7.9, is then added the hydrogen peroxide of 10%wt to system solution into off-white color, heat preservation decoloration 30- at 35-40 DEG C
The garlic polysaccharide solution that 60min must decolourize；The DEAE-cellulose (DEAE cellulose) of weak base type can also be used
It decolourizes to garlic polysaccharide；Due to charcoal-stripped relatively strong, and dosage is larger, is lost using active carbon decoloring garlic polysaccharide
It measures larger, therefore should not be decolourized to garlic polysaccharide crude product of the present invention using active carbon.
Preferably, step 5) the de- albumen is directed to that three chloroethenes of 6-8%wt are added dropwise in the garlic polysaccharide solution of decoloration
Aqueous acid is stopped that trichloroacetic acid is added dropwise when no longer being changed using in-line turbidimeter monitoring system turbidity, 1- deoxidation-is then added
1- (methylamino)-D-glucitol stands the Deproteinated garlic polysaccharide solution being centrifuged after 6-8h at 20-30 DEG C；The present invention is in egg
Trichloroacetic acid method is used in the removal of white matter, it is few compared with traditional Sevage method consumption of organic solvent, but trichloroacetic acid can make part
Garlic polysaccharide degradation, declines the recovery rate of garlic polysaccharide；Present invention adds a small amount of 1- deoxidation -1- (methylamino)-D- sorbs
Alcohol, discovery are able to suppress the degradation of garlic polysaccharide, improve the recovery rate of garlic polysaccharide.
Compared with prior art, the present invention has the advantage that
1) it is big to purify to form sediment as precipitating reagent and garlic polysaccharide using quinine quaternary ammonium salt for the first time by the present invention
Garlic polysaccharide, quinine quaternary ammonium salt and cetyl trimethylammonium bromide (CTMAB) and its cetyltrimethylammonium hydroxide
(CTA-OH) it is compared with cetylpyridinium chloride (CPC), the garlic polysaccharide prepared either purity or yield is above
Conventional quaternary ammonium salts；
2) present invention is when garlic polysaccharide crude product removes removing protein, while using trichloroacetic acid as remover, addition portion
Divide 1- deoxidation -1- (methylamino)-stabilizer of the D-glucitol as garlic polysaccharide, avoids garlic polysaccharide and trichloroacetic acid acts on
When cause garlic polysaccharide to decompose, influence product purity and yield.
Detailed description of the invention
Fig. 1 is High Performance Gel Permeation chromatography (the High Performance that the present invention prepares high-purity garlic polysaccharide
GelPermeation Chromatography, HPGPC) figure；
Fig. 2 is the infrared spectrogram that the present invention prepares high-purity garlic polysaccharide.
Specific embodiment
In order to make the objectives, technical solutions and advantages of the present invention clearer, With reference to embodiment, to this
Invention is further described.It should be understood that these descriptions are merely illustrative, and it is not intended to limit the scope of the invention.
Embodiment 1
1) degreasing pre-treatment: through being dried under vacuum to constant weight at 35-40 DEG C after garlic peeling, then shredding to partial size is 200-
300 mesh obtain garlic granule 5.0kg, and garlic granule is placed in 100L stainless steel high-pressure closed vessel, and 25kg acetone and tetrahydrofuran is added
Mixed liquor (calculates, acetone/tetrahydrofuran=95:5) according to volume ratio, is dispersed with stirring 10-20min, is then added from feed opening
25kg partial size is the dry ice of 0.5-1cm, closes feed opening, is stirred at room temperature, warms naturally to 0-5 DEG C to temperature in the kettle
When, stop stirring；Filters pressing removes filtrate, obtains 4.91kg degreasing primary garlic to constant weight is dried under vacuum at 35-40 DEG C of filter cake progress
Slag；
2) low temperature ultrasonic removes small molecular sugar: 4.91kg degreasing primary garlic slag is added to the ethanol water of the 90%V of 27kg
In solution, ultrasound 6-8h at 10-20 DEG C, centrifugal filtration removes filtrate, obtains to constant weight is dried under vacuum at 35-40 DEG C of filter residue progress
4.75kg desugar second level garlic slag；
3) grading extraction of garlic polysaccharide: 4.75 desugar second level garlic slags are placed in 29kg purified water, are warming up to 60-70 DEG C
Ultrasonic extraction 6-8h, is then centrifuged for, and obtains neutral filtrate and A grades of filter residues；The phosphate that A grades of filter residues are placed in 28kgpH=5.7 is delayed
It washes by water in solution, is warming up to 60-70 DEG C of ultrasonic extraction 3-4h, is then centrifuged for, obtain faintly acid filtrate and B grades of filter residues；By B grades of filter residues
It is placed in the phosphate-buffered aqueous solution of 26kgpH=7.6, is warming up to 60-70 DEG C of ultrasonic extraction 3-4h, is then centrifuged for, obtains weak base
Property filtrate and C grades of filter residues；C grades of filter residues of reject merge neutral filtrate, faintly acid filtrate and alkalescent filtrate and obtain 82.6kg containing garlic
The extracting solution of polysaccharide；
4) separation of garlic polysaccharide crude product: extracting solution of the 82.6kg containing garlic polysaccharide is carried out depressurizing at 45-50 DEG C dense
Contracting is 1.18-1.13g/cm when being concentrated into density3When (concentrate is taken to measure density at 25 DEG C), stop concentration, handling material
Into stirred tank, it is then down to 40-45 DEG C, peristaltic pump is used to be added dropwise 90%V's into stirred tank with the charging rate of 2-3L/h
Ethanol water gradually has soluble soluble solids to be precipitated during being added dropwise；Every 1h, feeding liquid by aperture is from stirred tank
0.2 micron of filtering with microporous membrane, filtrates tested density are 1.02-1.03g/cm3Stop drop when (measuring density at 25 DEG C)
Add the ethanol water of 90%V；Be cooled to after 2-5 DEG C of standing 6-8h with the rate of temperature fall of 2-5 DEG C/min filter 0.26kg is big
Garlic polysaccharide crude (raw sugar recovery rate is 5.2%)；
Garlic polysaccharide primary passivation: to the decoloration of garlic polysaccharide crude product, removing protein:
Decoloration: (using polysaccharide quality in sulfuric acid-phynol method detection solution after taking garlic polysaccharide crude product 100g to be dissolved in 0.9L water
100%) concentration is denoted as, using ammonia water conditioning system pH to 7.8-7.9, the hydrogen peroxide of 10%wt is then added to system solution
At off-white color, the garlic polysaccharide solution that heat preservation decoloration 30-60min must decolourize at 35-40 DEG C (is detected molten using sulfuric acid-phynol method
Polysaccharide mass concentration in liquid, for 96.6%)；The polysaccharide concentration of decoloration front and back garlic polysaccharide has dropped about 3.4%, is in reasonable
In loss range；
Removing protein: the trichloroacetic acid solution of 7%wt is added dropwise in the garlic polysaccharide solution 1L of decoloration, is supervised using in-line turbidimeter
Stop that trichloroacetic acid solution is added dropwise when survey system turbidity no longer changes, 3.5g 1- deoxidation -1- (methylamino)-D- is then added
After sorbierite stirring and dissolving, 6-8h is stood at 20-30 DEG C, the Deproteinated garlic polysaccharide solution of centrifugation (uses sulfuric acid-phynol method
Polysaccharide mass concentration in solution is detected, for 89.2%)；Then ethyl alcohol is added and carries out alcohol analysis, filtering, dry off-white color are to pale yellow
The garlic polysaccharide particle 71.2g (being defined as Pre- garlic polysaccharide) of color.
If not adding 1- deoxidation -1- (methylamino)-D-glucitol during removing protein as polysaccharide protective agent, egg is taken off
To be only 71.3% (fall to 71.3 by 96.6 after decolourizing, loss is tight to the mass concentration of polysaccharide in white garlic polysaccharide solution
Weight), so Deproteinated while being added part 1- deoxidation -1- (methylamino)-D-glucitol plays prevents garlic polysaccharide from existing
The lower effect degraded of trichloroacetic acid effect.
Embodiment 2
Pre- garlic polysaccharide 1.0g is taken respectively, 50ml water is added to dissolve, and then adds the different quaternary ammonium salts that concentration is 2%wt
Aqueous solution 50ml stands overnight (8-12h) at 40 DEG C, is then cooled to room temperature, centrifugation, and the matter of sediment is detected after dry
Amount, the results are shown in Table 1:
Co-precipitation effect of the different quaternary ammonium salts of table 1 to garlic polysaccharide
The above result shows that being used to be co-precipitated with garlic polysaccharide using quinine quaternary ammonium salt, high yield can be obtained
Sediment, more traditional cetyl trimethylammonium bromide (CTMAB) and its cetyltrimethylammonium hydroxide (CTA-OH)
Yield is compared with cetylpyridinium chloride (CPC) significantly to be promoted.
Embodiment 3
The sediment that the present invention is formed respectively with N- (9- anthracene methyl) bromination quinine and N- Benzylphosphonium Bromide quinine
Each 1g carries out dissociation 4-6h at 35-40 DEG C using the different inorganic salt solutions of 160ml 2mol/L, is centrifuged to obtain filtrate, then
Dehydrated alcohol to no solid is added dropwise into filtrate to be precipitated, solid is collected by filtration, is dried in vacuo weighing at 40-45 DEG C, obtains different nothings
The high-purity garlic polysaccharide of machine salt dissociation, different inorganic salts gained garlic polysaccharide weight are as shown in table 2:
The weight of the different inorganic salts dissociation gained garlic polysaccharides of table 2
The above result shows that the dissociation effect of different inorganic salts is different, wherein the dissociation effect with ammonium hydrogen sulfate is best；No
Also not identical in the degree of dissociation of same inorganic salts with quaternary ammonium salt, the sediment that N- Benzylphosphonium Bromide quinine is formed is compared with N- (9-
Anthracene methyl) bromination quinine is easy dissociation, so final choice N- Benzylphosphonium Bromide quinine of the present invention is precipitation and complexation agent,
Selection is dissociated in the aqueous solution of ammonium hydrogen sulfate.
The sediment that N- Benzylphosphonium Bromide quinine is formed is taken and carries out dissociation acquisition in the aqueous solution of ammonium hydrogen sulfate
High-purity garlic polysaccharide carries out molecular weight test by HPGPC, is characterized by infrared spectroscopy to its structure, as a result such as Fig. 1
With shown in Fig. 2.
Molecular weight characterization: High Performance Gel Permeation chromatography (High Performance Gel Permeation is used
Chromatography, HPGPC) measurement garlic polysaccharide molecular weight: principle is by various glucans, the grape of standard molecular weight
Garlic polysaccharide sugared and prepared by the present invention is made into 0.2%wt solution respectively, various using the hplc determination with gel column
The retention time of sugar calculates garlic polysaccharide molecular weight according to the parameters such as various gel columns and known molecular amount polysaccharide.Calculate to obtain this
The molecular weight for inventing the garlic polysaccharide of preparation is M=4380Da, and high-purity garlic polysaccharide molecular weight prepared by the present invention is at just
State distribution, is shown in Fig. 1.
Infrared structure characterization: garlic polysaccharide is in 3358cm-1(stretching vibration absworption peak of OH), 2943cm-1、2887cm-1
(the C-H vibration absorption peak of methyl and methylene) 1650cm-1(carbonyl absorption peak), 1027cm-1And 1131cm-1(C-O-C is flexible
Vibration), see Fig. 2.
Although embodiments of the present invention are described in detail, it should be understood that, without departing from of the invention
In the case where spirit and scope, embodiments of the present invention can be made with various changes, replacement and change.
Claims (9)
1. a kind of method that the co-precipitation of quinine quaternary ammonium salt prepares high-purity garlic polysaccharide, comprising the following steps:
1) degreasing pre-treatment: to drying and crushing after garlic peeling, using the mixed liquor of acetone and tetrahydrofuran to smashed big
Garlic carries out ungrease treatment and obtains primary garlic slag；
2) low temperature ultrasonic removes small molecular sugar: by the primary garlic slag Jing Guo ungrease treatment, ultrasound is gone in the ethanol water of 90%V
Except small molecular sugar obtains desugar second level garlic slag；
3) grading extraction of garlic polysaccharide: desugar second level garlic slag is successively extracted in neutrality, faintly acid, weakly alkaline water
Garlic polysaccharide must contain the extracting solution of garlic polysaccharide；
4) garlic polysaccharide crude product the separation of garlic polysaccharide crude product: is analysed to obtain using alcohol to the extracting solution containing garlic polysaccharide；
It is characterized by also including following steps:
5) purifying of garlic polysaccharide crude product: garlic polysaccharide crude product is successively decolourized, is dissolved in purified water after de- albumen golden pheasant is added
Alkali quaternary ammonium salt of receiving forms insoluble matter precipitating, is centrifuged to obtain sediment, sediment is dissociated in inorganic salt solution, and ultrafiltration must solve
Chaotropic carries out alcohol to dissociation solution and analyses to obtain high-purity garlic polysaccharide.
2. according to the method described in claim 1, it is characterized by: specific steps are as follows:
1) degreasing pre-treatment: through being dried under vacuum to constant weight at 35-40 DEG C after garlic peeling, then shredding to partial size is 200-300
Mesh obtains garlic granule, and garlic granule is placed in high-pressure closed vessel, and the mixed liquor of acetone and tetrahydrofuran is added, is dispersed with stirring 10-
Then 20min is added the dry ice that partial size is 0.5-1cm from feed opening, closes feed opening, be stirred at room temperature, in kettle
When temperature warms naturally to 0-5 DEG C, stop stirring；Filters pressing removes filtrate, carries out being dried under vacuum to constant weight at 35-40 DEG C to filter cake
Obtain degreasing primary garlic slag；It in the mixed liquor of the acetone and tetrahydrofuran, is calculated according to volume ratio, acetone/tetrahydrofuran=95:
5；The weight ratio of the mixed liquor and dry ice of the acetone and tetrahydrofuran is 1:1, the garlic granule and the acetone and tetrahydrofuran
Mixed liquor weight ratio be 1:5；
2) low temperature ultrasonic removes small molecular sugar: in the ethanol water for the 90%V that degreasing primary garlic slag is added to 5-6 times of weight,
Ultrasound 6-8h at 10-20 DEG C, centrifugal filtration remove filtrate, obtain desugar two to constant weight is dried under vacuum at 35-40 DEG C of filter residue progress
Grade garlic slag；
3) grading extraction of garlic polysaccharide: desugar second level garlic slag is placed in purified water, 60-70 DEG C of ultrasonic extraction 6- is warming up to
8h is then centrifuged for, and obtains neutral filtrate and A grades of filter residues；A grades of filter residues are placed in the phosphate-buffered aqueous solution of pH=5.7, are heated up
It to 60-70 DEG C of ultrasonic extraction 3-4h, is then centrifuged for, obtains faintly acid filtrate and B grades of filter residues；B grades of filter residues are placed in the phosphorus of pH=7.6
In hydrochlorate buffered aqueous solution, it is warming up to 60-70 DEG C of ultrasonic extraction 3-4h, is then centrifuged for, obtains alkalescent filtrate and C grades of filter residues；It abandons
Except C grades of filter residues, merging neutral filtrate, faintly acid filtrate and alkalescent filtrate must contain the extracting solution of garlic polysaccharide；
4) separation of garlic polysaccharide crude product: being concentrated under reduced pressure the extracting solution containing garlic polysaccharide at 45-50 DEG C, when being concentrated into
Density is 1.18-1.13g/cm3When, stop concentration, then handling material is down to 40-45 DEG C into stirred tank, using peristaltic pump
The ethanol water of 90%V is added dropwise with the charging rate of 2-3L/h into stirred tank, gradually there is soluble soluble solids during being added dropwise
It is precipitated；Every the 1h filtering with microporous membrane that feeding liquid is 0.2 micron by aperture from stirred tank, filtrates tested density is
1.02-1.03 g/cm3When stop be added dropwise 90%V ethanol water；With the rate of temperature fall of 2-5 DEG C/min be cooled to 2-5 DEG C it is quiet
Garlic polysaccharide crude product is filtered to obtain after setting 6-8h；
5) purifying of garlic polysaccharide crude product: garlic polysaccharide crude product is successively decolourized, is dissolved in purified water after de- albumen golden pheasant is added
Alkali quaternary ammonium salt of receiving forms insoluble matter precipitating, and mixture is centrifuged to obtain sediment and filtrate after heat preservation is stood；Filtrate is settled using ethyl alcohol
Obtain neutral garlic polysaccharide；Sediment is dissociated in inorganic salt solution, and ultrafiltration obtains dissociation solution, is carried out alcohol to dissociation solution and is analysed
High-purity garlic polysaccharide.
3. according to the method described in claim 2, it is characterized by: quinine quaternary ammonium salt described in step 5) is N- (9- anthracene first
Base) bromination quinine, N- Benzylphosphonium Bromide quinine or O- allyl-N- (9- anthracene methyl) bromination quinine.
4. according to the method described in claim 3, it is characterized by: quinine quaternary ammonium salt described in step 5) is N- Benzylphosphonium Bromide
Quinine.
5. according to the method described in claim 2, it is characterized by: inorganic salts described in step 5) are acid ammonium salt.
6. according to the method described in claim 5, it is characterized by: acid ammonium salt described in step 5) is ammonium hydrogen sulfate, ammonium sulfate
Or ammonium dihydrogen phosphate.
7. according to the method described in claim 6, it is characterized by: acid ammonium salt described in step 5) is ammonium hydrogen sulfate.
8. according to the method described in claim 2, garlic polysaccharide crude product is dissolved in it is characterized by: decolourizing to refer to described in step 5)
After water, using ammonia water conditioning system pH to 7.8-7.9, then the hydrogen peroxide of addition 10%wt is to system solution at off-white color, 35-
The garlic polysaccharide solution that heat preservation decoloration 30-60min must decolourize at 40 DEG C.
9. according to the method described in claim 2, it is characterized by: taking off the garlic polysaccharide that albumen is directed to decoloration described in step 5)
The trichloroacetic acid solution of 6-8%wt is added dropwise in solution, stops being added dropwise when no longer changing using in-line turbidimeter monitoring system turbidity
Then 1- deoxidation -1-(methylamino is added in trichloroacetic acid solution)-D-glucitol, stand at 20-30 DEG C be centrifuged after 6-8h it is de-
The garlic polysaccharide solution of albumen.
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Citations (4)
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|---|---|---|---|---|
| CN1239719A (en) * | 1998-06-24 | 1999-12-29 | 中山市健康科技产业基地保健品研究所 | Extraction process of garlic polysaccharide |
| EP2778177A1 (en) * | 2011-11-07 | 2014-09-17 | Shenyang Kesi High-technology Co. Ltd. | Method for extracting polysaccharides from higher plants and fungi through microwave chemical treatment |
| CN104177507A (en) * | 2014-07-22 | 2014-12-03 | 江苏伟楼生物科技有限公司 | Preparation method of garlic polysaccharide |
| CN104356250A (en) * | 2014-11-27 | 2015-02-18 | 秦富英 | Method for extracting garlic crude polysaccharide |
-
2018
- 2018-06-27 CN CN201810680770.3A patent/CN109021125A/en not_active Withdrawn
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1239719A (en) * | 1998-06-24 | 1999-12-29 | 中山市健康科技产业基地保健品研究所 | Extraction process of garlic polysaccharide |
| EP2778177A1 (en) * | 2011-11-07 | 2014-09-17 | Shenyang Kesi High-technology Co. Ltd. | Method for extracting polysaccharides from higher plants and fungi through microwave chemical treatment |
| CN104177507A (en) * | 2014-07-22 | 2014-12-03 | 江苏伟楼生物科技有限公司 | Preparation method of garlic polysaccharide |
| CN104356250A (en) * | 2014-11-27 | 2015-02-18 | 秦富英 | Method for extracting garlic crude polysaccharide |
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| 梁丽军: "大蒜多糖的提取纯化及生物功能活性研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
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