CN109503683A - The isolation and purification method of phloridzin in a kind of Malus spectabilis Aqueous extracts - Google Patents
The isolation and purification method of phloridzin in a kind of Malus spectabilis Aqueous extracts Download PDFInfo
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- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
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- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
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Abstract
The present invention provides a kind of isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts, and Malus spectabilis aqueous extract is specifically concentrated to give medicinal extract at 75-85 DEG C；It is cooled to room temperature to medicinal extract, is transferred to 0~5 DEG C of freezer and stands still for crystals 20-30h；Medicinal extract after crystallization is mixed with ice water, separation filters out mother liquor；After filtering out mother liquor, ice water cleaning is added, drying obtains the product containing phloridzin；At normal temperature by phloridzin crude product, ethyl alcohol dissolves, and filters, and is concentrated under reduced pressure, and receives cream, is added after hot water mixes and stands cooling, and 0~5 DEG C of freezer is transferred to after temperature drops to room temperature and is stood, crystallization 20-30h obtains phloridzin sterling.A kind of economic and practical, reliable and stable separation method for phloridzin in Hubei Chinese flowering crabapple aqueous extract is developed using common industrial equipment and solvent, and there is simple process, safe and reliable, easy to implement, the features such as rate of recovery is high, low in cost.
Description
Technical field
The present invention relates to a kind of isolation and purification method of phloridzin in Hubei Chinese flowering crabapple aqueous extract and its in industrialized production
In application
Background technique
Phloridzin is to extract and obtain from apple, apple bark and leaf etc. earliest, is combined by aglycon phloretin and glucose
Flavonoid glycoside compound, belong to dihydrochalcone glycosides.Current study show that prevention and treatment of the phloridzin to diabetes and its complication
There is effect unique, while having the effects that good anti-oxidant, anticancer, whitening.
Hubei Chinese flowering crabapple (Malus hupehensis (Pamp.) Rehd.), originate in Hubei, Hunan, Jiangxi, Jiangsu, Zhejiang,
The ground such as Anhui, Fujian, Guangdong, Gansu, Shaanxi, Henan, Shanxi, Shandong, Sichuan, Yunnan, Guizhou.Raw hillside or mountain valley jungle
In, 50-2900 meters of height above sea level.Contain phloridzin in Hubei Chinese flowering crabapple, each position content is differed 0.9~13%, wherein Hubei Chinese flowering crabapple
Content highest in spray and leaf, can be used as extract phloridzin source (Fang Rong, etc., phloridzin assay in Hubei Chinese flowering crabapple,
Food science and technology, 6 phases of volume 33 in 2008).Early-stage study is proved can be effectively by phloridzin from Hubei Chinese flowering crabapple spray by solvent of hot water
It is extracted in tender leaf, and the relevant technologies is applied for a patent into (Deng Gaigai, etc. the extraction side of phloridzin in Hubei Chinese flowering crabapple leaf
Method, the patent No.: ZL 201410346970.7).
It is extracted compared to by solvent of ethyl alcohol, taking water as a solvent the extraction for carrying out phloridzin has safety, low cost, work
The features such as skill is simple adapts to the requirement of industrialized production, but the polarity of water is big, and dissolved impurity is more when extraction, to phloridzin
Isolate and purify and cause certain difficulty.The present invention is directed to establish the simple and easy method of one kind to divide from Hubei Chinese flowering crabapple aqueous extract
Method from high-purity phlorizin.
Summary of the invention
More for impurity in Hubei Chinese flowering crabapple aqueous extract, viscosity is big, and the shortcoming being not readily separated, the purpose of the present invention exists
The isolation and purification method of phloridzin in the Hubei Chinese flowering crabapple aqueous extract for providing a kind of technical grade, it has simple process, safety
Reliably, easy to implement, the features such as rate of recovery is high, low in cost.
To achieve the goals above, the present invention is to realize by the following technical solutions: a kind of Hubei Chinese flowering crabapple water extracts
The process for separation and purification of skin glycosides in liquid root, it includes following content:
1. the isolation and purification method of phloridzin, includes the following steps: in a kind of Malus spectabilis Aqueous extracts
(1) it is concentrated under reduced pressure: Malus spectabilis aqueous extract being concentrated to give medicinal extract at 75-85 DEG C, collects medicinal extract, stands cooling；
(2) low temperature crystallization: being cooled to room temperature to medicinal extract, is transferred to 0~5 DEG C of freezer and stands still for crystals 20-30h, after being crystallized
Medicinal extract；
(3) it is centrifugated: the medicinal extract after crystallization being mixed with ice water, flat centrifuge is poured into and is separated by solid-liquid separation, filter out mother
Liquid；
(4) ice water washs: after mother liquor all filters out, adding ice water and cleans to the phloridzin that step (3) separates, after drying
Obtain the product containing phloridzin；
(5) crude product is dried: the product drying containing phloridzin being crushed, mixing obtains phloridzin content 65%~85%
Crude product, the rate of recovery of phloridzin is 85%~95% in Hubei Chinese flowering crabapple extracting solution；
(6) crude product purifies: at normal temperature by phloridzin crude product, being dissolved, is filtered using ethyl alcohol, is concentrated under reduced pressure, receive cream, hot water is added
Cooling is stood after mixing, and 0~5 DEG C of freezer is transferred to after temperature drops to room temperature and is stood, centrifugal filtration after 20-30h is crystallized, receives
Collect phloridzin wet product, dry, crushes, obtain high purity product of the phloridzin content 95%~99%.
In the step (1), Malus spectabilis aqueous extract is concentrated into after specific gravity reaches 1.05-1.30 at 80 DEG C and is collected clearly
Cream.
In the step (3), the volume ratio of clear cream and ice water is 1:0.9-1.2.
In the step (4), the volume ratio of the clear cream in ice water and step (3) is 1:0.9-1.2.
Drying is dried under the conditions of -0.09MPa, 55-65 DEG C in the step (5), in drying to phloridzin
Moisture content is lower than 6.0%.
Phloridzin crude product at normal temperature, uses mass concentration to carry out for 95% or more ethyl alcohol in the step (6)
The mass ratio of dissolution, ethyl alcohol and phloridzin crude product that wherein mass concentration is 95% or more is 9-12:1.
Being concentrated under reduced pressure in the step (6) is to be concentrated into after specific gravity reaches 1.4-1.6 to receive cream at 45-50 DEG C.
After receiving cream in the step (6), standing is down to room temperature after 90-98 DEG C of hot water mixing is added, and is then transferred to 4 DEG C
Freezer stand, low temperature crystallization 24 hours, obtain phloridzin wet product.
Drying described in the step (6) is dried under the conditions of -0.09MPa, 45-50 DEG C, dry to root
Moisture content is lower than 6.0% in skin glycosides.
Beneficial effects of the present invention: it is developed using common industrial equipment and solvent a kind of economic and practical, reliable and stable
The separation method for phloridzin in Hubei Chinese flowering crabapple aqueous extract, and there is simple process, safe and reliable, it is easy to implement, return
High income, it is low in cost the features such as.
Specific embodiment
Below by embodiment, the invention will be further described, but does not therefore limit the present invention to the implementation
In example.
Embodiment 1
Hubei Chinese flowering crabapple aqueous extract is concentrated under reduced pressure, temperature control is at 80 DEG C hereinafter, being concentrated into specific gravity reaches 1.15 (80
DEG C) after, cream is received, clear cream is fitted into the open plastic barrel of 35L or so, stands cooling.
After Hubei Chinese flowering crabapple clear cream temperature is down to room temperature, it is transferred to 0~5 DEG C of freezer and stands, low temperature crystallization 24 hours, remove
It is never acutely shaken when dynamic.
0 DEG C of ice water mixing by the medicinal extract after sufficient crystallising and in equal volume, pours into flat centrifuge and is separated by solid-liquid separation,
Filter out mother liquor.
After mother liquor all filters out, the 0 DEG C ice water isometric with former medicinal extract is added to phloridzin crude product in centrifuge
It is cleaned, collects phloridzin crude product in filter cloth after drying.
Phloridzin is layered in stainless steel pallet, is placed in vacuum oven, is dried under the conditions of -0.09MPa, 60 DEG C
It is dry, it is dry to be lower than 6.0% to moisture content in phloridzin, crush, mixing, can obtain phloridzin content 65%~85% it is thick
Product, the rate of recovery of phloridzin is between 85%~95% in Hubei Chinese flowering crabapple extracting solution.
Phloridzin crude product is sufficiently stirred at normal temperature with 10 times of 95% edible ethanols of amount, is dissolved, the filtration of 250 mesh takes filter
Liquid is concentrated under reduced pressure at 50 DEG C or less, recycles edible ethanol, is concentrated into specific gravity and reaches 1.5 (50 DEG C), receive cream, 5 times of weights of medicinal extract are added
95 DEG C of hot water of amount mix well, barrelling, stand cooling, and 0~5 DEG C of freezer is transferred to after temperature drops to room temperature and is stood, low
Temperature crystallization 24 hours, centrifugal filtration, collect phloridzin wet product, be placed in vacuum oven, under the conditions of -0.09MPa, 50 DEG C into
Row drying, it is dry to be lower than 6.0% to moisture content in phloridzin, it crushes, mixing can obtain phloridzin content 95%~99%
High purity product.
Solubility study in the aqueous solution of phloridzin at different temperatures
Phloridzin sample, Natural products research are made by oneself with using key lab, Hubei Province (SanXia University), through detecting phloridzin
Content is 98.13%.
Instrument
3000 high performance liquid chromatograph of Ultimate, Dai An (Shanghai) Analytical Instrument Co., Ltd；SB-100 Rotary Evaporators,
Japanese EYELA company；Analyze electronic balance, Shanghai Yue Ping scientific instrument Co., Ltd；C18Chromatographic column (250 × 4.6mm), Beijing
Di Ma Science and Technology Ltd.；Digital display thermostat water bath, changzhou Zhi Bo instrument manufacturing Co., Ltd；Conversion hysteria freezing and refrigeration
Case, Jiangsu Bai Xue limited liability company.
Standard items and reagent
Phloridzin standard items (ANPEL company, Lot No:94990025)；Acetonitrile (chromatographically pure, Tedia company, the U.S.).
Sample preparation methods
It accurately weighs 6 parts of 2.00g phloridzin samples to be respectively placed in 10ml tool plug teat glass, 5ml distillation is added in every test tube
Water is placed in 90 DEG C of heating water baths in thermostat water bath, is ultrasonically treated 5 minutes per hour, 5h is handled repeatedly, to root in test tube
Skin glycosides is not redissolved, by 6 parts of samples be respectively placed in 4 DEG C, 15 DEG C, 30 DEG C, 50 DEG C, 70 DEG C, in 90 DEG C of water bath with thermostatic control environment
It stands for 24 hours, supernatant liquor is taken to carry out phloridzin assay as sample.
Detection method
Chromatographic condition: chromatographic column C18Chromatographic column (250mm × 4.6mm), the acetonitrile-water (volume ratio) that mobile phase is 27%, into
Sample amount is 10 μ l, and flow velocity 1.0ml/min, column temperature is 25 DEG C, Detection wavelength 285nm.
The preparation of reference substance solution: precision weighs reference substance phloridzin 10mg, sets in 10ml volumetric flask, simultaneously with methanol dissolution
Be diluted to scale, shake up to obtain 1mg/ml reference substance solution, then be diluted to various concentration to get.
Accurate 10 μ l of pipette samples injection liquid chromatogram measures corresponding chromatographic peak area, then establishing criteria product calculated by peak area
Phloridzin content in sample.
Experimental result
Phloridzin content data in the aqueous solution of different temperatures is shown in Table 1.
1 phloridzin of table content in the aqueous solution of different temperatures
| Aqueous temperature | Phloridzin content in aqueous solution |
| 4℃ | 0.05% |
| 15℃ | 0.08% |
| 30℃ | 0.17% |
| 50℃ | 0.54% |
| 70℃ | 2.51% |
| 90℃ | 24.48% |
When 1 data of table are shown in 4 DEG C, the phloridzin content in phloridzin aqueous solution is 0.05%, in 70 DEG C of aqueous solutions below
The content of phloridzin is not very high, and the dissolubility of phloridzin significantly increases in 70 DEG C of aqueous solutions up, at 90 DEG C
Phloridzin content has reached 24.48%, 490 when being 4 DEG C times.
Experimental result shows that dissolubility of the phloridzin in cold water is poor, but has preferable dissolution in high temperature aqueous solution
Property, this is also that the hot water of phloridzin in Hubei Chinese flowering crabapple leaf extracts, the technology path of crystallisation by cooling separation is laid a good foundation.Phloridzin
490 times when meltage in 90 DEG C of hot water is 4 DEG C, Hubei Chinese flowering crabapple extracting solution stands cooling through a degree of concentration
Phloridzin in aqueous solution can be precipitated out, to achieve the purpose that separate phloridzin, and phloridzin in 4 DEG C of aqueous solutions
Content be only 0.05%, can theoretically recycle 99.95% of phloridzin in extracting solution, illustrate in Hubei Chinese flowering crabapple Aqueous extracts
Phloridzin can be effectively separated by the method for concentration, low-temperature precipitation, be recycled.
The research of phloridzin Crystallization Separation condition
Experimental material
Raw material
Hubei Chinese flowering crabapple leaf raw material is provided by five Feng Lvrun Biotechnology Co., Ltd, referring to a kind of patent (Hubei Chinese flowering crabapple leaf raw material
Preparation method and its extracting the application on phloridzin or phloretin, application number: 201711366683.2) technology provided into
Row preparation.
Hubei Chinese flowering crabapple aqueous extract, according to patent (extracting method of phloridzin, the patent No.: ZL in Hubei Chinese flowering crabapple leaf
201410346970.7) prepared using pilot plant.
Instrument and equipment
Densimeter, the Yuyao City port Huang Jia glass instrucment and meter plant；Thermometer, the Yuyao City port Huang Jia glass instrucment and meter plant；0.5m3It is multi-functional to mention
Take tank, Wuhan Pharmaceutical Machinery Factory；150 type outer circulation inspissators, Wuhan Pharmaceutical Machinery Factory；Flat centrifuge, China, Zhangjagang City
Glad Machinery Manufacturing Co., Ltd.；Conversion hysteria fridge-freezer, Jiangsu Bai Xue limited liability company.
Test method
Influence of the concentrate specific gravity to crystallization
Extracting solution is concentrated into the medicinal extract that 80 DEG C of specific gravity are 1.05,1.10,1.15,1.20,1.25,1.30 respectively and carries out crystallization ratio
Compared with comparing phloridzin crystallization effect under the conditions of different specific weight, obtain best specific gravity condition.
The research of phloridzin crystallization time
Select the medicinal extract 2L of best specific gravity be respectively placed in 6h, 12h in 4 DEG C of refrigerator-freezers, 18h, for 24 hours, 30h separated, filters out knot
Brilliant product, drying weighing, compares crude yield, obtains best crystallization time.
Crude product centrifuge separation plus the research of water multiple
The medicinal extract that finishes of crystallization is separately added into 0 times of volume, 0.5 times of volume, 1.0 times of volumes, 1.5 times of volumes, 2.0 times of volumes
Ice water is poured into centrifuge after being sufficiently mixed and is centrifuged, shutdown observation separating effect after centrifugation 15 minutes.
The research of wash number in crude product centrifuge
The ice water that the crude product that centrifugation finishes is separately added into former medicinal extract volume is subjected to cleaning 1 time, 2 times, 3 times, after observation cleaning
Phloridzin crude product color and viscosity, with tack-free, facilitating paving disk dry is optimal cleaning effect.
Influence result of study of the concentrate specific gravity to crystallization
The concentrate crystallization effect comparison result of 1 different specific weight of table
1.10 or less specific gravity of medicinal extract can be because phloridzin content degree of super saturation be inadequate in solution as the result is shown described in table 1, it is difficult to
Nucleus is formed, crystallization is slowly and particle is tiny, is unfavorable for improving yield and later separation；Specific gravity crystal form at 1.15 is preferable, benefit
In centrifuge separation；And after specific gravity continues increase, preferable crystal form can not be formed because phloridzin settles out too fast, led
It causes filter effect to be deteriorated, is unfavorable for subsequent centrifuge separation.
Phloridzin crystallization time result of study
2 phloridzin crystallization time result of study of table
| Crystallization time<h> | Crude product weight (g) |
| 6 | 24.6 |
| 12 | 160.7 |
| 18 | 204.4 |
| 24 | 211.8 |
| 30 | 212.4 |
Described in table 2 as the result is shown when 6h, crystallization just starts, this still needs after may being put into cold sentence with solution
A period of time cools down related, is standing to after for 24 hours, and solution crystallizes completion substantially, then extends crystallization time and can not also increase production
Amount, so optimal crystallization time of repose is in left and right for 24 hours.
The result of study of crude product centrifuge separation plus water multiple
The result of study of the centrifuge separation of 3 crude product of table plus water multiple
Described in table 3 as the result is shown in the case where being added without ice water, a large amount of carbohydrate and protein are contained in the mother liquor after crystallization
Substance, viscosity is very big, is easy blocking filter cloth, it is not easy to filter, be added after suitable ice water, solution can be effectively reduced
Viscosity, and too many phloridzin is not lost, it can carry out effectively being centrifuged after the ice water that former 1 times of volume of medicinal extract is added and divide
From can dry mother liquor as far as possible, reduce the impurity content in crude product phloridzin, be conducive to drying.Although the ice water being added is got over
The water that the effect more being centrifugated is better but addition is excessive is centrifuged, and portioned product can be taken away, and yield is reduced, so with just
The water that can satisfy centrifuging process, the ice water that 1 times of former clear cream volume is mixed into before centrifuge separation mix.
The result of study of wash number in crude product centrifuge
The result of study of wash number in 4 crude product centrifuge of table
| Number of water | Crude product state after cleaning |
| 1 | Colour cast is yellow, tack-free, loose |
| 2 | Colour cast is greyish white, tack-free, loose |
| 3 | Colour cast is greyish white, tack-free, loose |
The ice water cleaning for carrying out 1 time described in table 4 after centrifugation as the result is shown, it is residual can to effectively remove phloridzin crude product surface
The sticky contaminant stayed is centrifuged after cleaning 1 time, and crude product seems that canescence is presented after drying cleaning solution, more loose,
It is easy drying.Increasing wash number will increase the clean level of product, washes out more foreign pigments, product is made to seem purer
Only, but excessive wash number will increase the loss of phloridzin, and the influence to product purity is also not very big, thus after cleaning with
Tack-free, facilitating paving disk dry is optimal cleaning effect, is cleaned 1 time.
The aqueous extract of Hubei Chinese flowering crabapple leaf not only contains target component phloridzin, also contains a large amount of carbohydrate and protide object
Matter, carbohydrate and protein substance are affected to the separating effect of phloridzin, and the program utilizes phloridzin in the hot water molten
Solution degree is big, and insoluble,practically characteristic carries out Crystallization Separation to product in ice water, while carbohydrate and protein substance are cold
Changes in solubility in hot water is little, and the separation of solid and liquid of product is realized under conditions of effectively control solution concentration, removes impurity,
Realize the separating-purifying of product.
The concentration of solution namely the specific gravity control of medicinal extract when the key point of the technology controlling and process is to crystallize, test discovery is only
Have and the specific gravity control of medicinal extract be can be only achieved into optimal crystallization effect when 1.15 (80 DEG C) left and right, concentration is too high or too low
Ideal crystallization effect cannot all be reached.Crude product after crystallization is since, containing a large amount of sugar and protein, viscosity is very in mother liquor
Greatly, it needs to increase part ice water to reduce solution viscosity, improves filter effect, add base after the ice water of 1 times of medicinal extract volume
Originally it can effectively filter, but the Hubei Chinese flowering crabapple leaf sugar content of Various Seasonal is not quite similar, the raw material harvested after August needs
To increase the volume of part ice water, suitably to accelerate filter effect.Water cleaning effect on the rocks is wanted again after mother liquor centrifugation finishes
It is better than the dosage that ice water is increased directly in mother liquor, water on the rocks cleans 1 time and can have after centrifuge is by mother liquor all drying
If effect elutes the coloring matter on crude product, but addition mother liquor is centrifuged together, the foreign pigment on crude product surface is difficult once to remove,
Therefore the technique that selection increases by 1 cleaning.The number of cleaning also need not be more, and increased number effect is not obvious, and also will increase
The loss of product.
The process route effectively can separate phloridzin from Hubei Chinese flowering crabapple Aqueous extracts, and obtained crude product phloridzin is in low temperature
Between 65%~85%, the fluctuation of content is mainly influenced by material quality general content after dry, and Hubei Chinese flowering crabapple leaf is former
Phloridzin content is high in material, then the content of the crude product phloridzin obtained is also high, conversely, then due in raw material impurity it is too many, obtain
Product content also can be lower, further purification & isolation is needed to obtain the phloridzin of high-purity.
Phloridzin purification Technology's Study
Experimental material
Raw material
Phloridzin sample, Natural products research are made by oneself with using key lab, Hubei Province (SanXia University), through detecting phloridzin
Content is 98.13%.Phloridzin crude product, Natural products research are made by oneself with using key lab, Hubei Province (SanXia University), warp
Detecting phloridzin content is 72.58%.
Instrument
3000 high performance liquid chromatograph of Ultimate, Dai An (Shanghai) Analytical Instrument Co., Ltd；SB-100 Rotary Evaporators,
Japanese EYELA company；Analyze electronic balance, Shanghai Yue Ping scientific instrument Co., Ltd；C18Chromatographic column (250 × 4.6mm), Beijing
Di Ma Science and Technology Ltd.；Digital display thermostat water bath, changzhou Zhi Bo instrument manufacturing Co., Ltd；Conversion hysteria freezing and refrigeration
Case, Jiangsu Bai Xue limited liability company.
Standard items and reagent
Phloridzin standard items (ANPEL company, Lot No:94990025)；Acetonitrile (chromatographically pure, Tedia company, the U.S.).
Test method
Solubility study of the phloridzin in high-concentration and low-concentration ethanol solution
Sample preparation
5 parts of 2.00g phloridzin samples are accurately weighed to be respectively placed in 10ml tool plug teat glass, test tube number is 1,2,3,4,5,
Wherein No. 1 and No. 2 test tubes are separately added into 5% ethanol solution of 5ml, and No. 3 and No. 4 test tubes are separately added into 10% ethanol solution of 5ml,
95% ethanol solution of 5m is added in No. 5 test tubes, and all test tubes are placed in 90 DEG C of heating water baths in thermostat water bath, per hour at ultrasound
Reason 5 minutes, handles 5h repeatedly, is not redissolved to phloridzin in test tube, No. 1 and No. 2 test tubes are placed in 4 DEG C of refrigerators, 3,
4, No. 5 test tubes are placed in 25 DEG C of water bath with thermostatic control environment, are stood respectively for 24 hours, are taken supernatant liquor to carry out phloridzin as sample and are contained
It is fixed to measure.
Detection method
Chromatographic condition: chromatographic column C18Chromatographic column (250mm × 4.6mm), the acetonitrile-water (volume ratio) that mobile phase is 27%, into
Sample amount is 10 μ l, and flow velocity 1.0ml/min, column temperature is 25 DEG C, Detection wavelength 285nm.
The preparation of reference substance solution: precision weighs reference substance phloridzin 10mg, sets in 10ml volumetric flask, simultaneously with methanol dissolution
Be diluted to scale, shake up to obtain 1mg/ml reference substance solution, then be diluted to various concentration to get.
Accurate 10 μ l of pipette samples injection liquid chromatogram measures corresponding chromatographic peak area, then establishing criteria product calculated by peak area
Phloridzin content in sample.
Best crystalline ethanol solution concentration is investigated
Phloridzin crude product is dissolved at room temperature with 95% ethyl alcohol, filters out all insoluble matters, collects ethanol solution, second is added
The distilled water of alcoholic solution volume 10% is concentrated under reduced pressure into no alcohol taste, and phloridzin powder is collected in freeze-drying.Weigh above-mentioned phloridzin powder
Last every part of 200.0g, configuring above-mentioned phloridzin powder quality score with 5%, 10%, 30%, 50%, 95% ethyl alcohol respectively is
30% solution, heating dissolve it all, are placed in 4 DEG C of refrigerators and stand for 24 hours, observe phloridzin crystalline state, filter, dry
It is dry, phloridzin crystal weight is weighed, and detect the content of phloridzin crystal, compares crystallization effect.
Solubility study result of the phloridzin in high-concentration and low-concentration ethanol solution
Phloridzin content data in the aqueous solution of different temperatures is shown in Table 1.
1 phloridzin of table content in the aqueous solution of different temperatures
| Ethanol solution concentration and temperature | Phloridzin content in ethanol solution |
| 4 DEG C, 5% ethyl alcohol | 0.05% |
| 4 DEG C, 10% ethyl alcohol | 0.06% |
| 25 DEG C, 5% ethyl alcohol | 0.31% |
| 25 DEG C, 10% ethyl alcohol | 0.42% |
| 25 DEG C, 95% ethyl alcohol | 12.91% |
When 1 data of table are shown in 4 DEG C, the phloridzin content in 5% ethanol solution of phloridzin is containing in 0.05%, with aqueous solution
Quite, content in 10% ethanol solution also only has 0.06% to amount, more slightly higher than the phloridzin content in 5% ethanol solution；25℃
When 95% ethanol solution in phloridzin content can achieve 12.91%, illustrate at room temperature can using ethyl alcohol to thick
Product are dissolved, and the method after concentration again using low temperature crystallization purifies phloridzin crude product, according to phloridzin in room temperature
Under the conditions of solubility parameter in 95% ethyl alcohol it can be calculated that in 95% ethyl alcohol that 6~10 times of phloridzin crude product weight is added
Phloridzin can all be dissolved out after solution.
Best crystalline ethanol solution concentration investigates result
The result that phloridzin crystallizes in the ethanol solution of various concentration such as table 2.
The result that 2 phloridzin of table crystallizes in the ethanol solution of various concentration
Phloridzin can form good crystallization in 5% and 10% ethanol solution as the result is shown described in table 2, and yield is higher,
When concentration of alcohol is more than the yield decline 50% or so of phloridzin after standing for 24 hours, when ethanol solution to 50% after 30%
After, for phloridzin then than being less easily precipitated from solution, yield is lower, and crystal form is also bad.Crystallization in 10% ethanol solution
Though quantity is less than 5% ethanol solution, content wants higher, and the color of crystal is also better than the knot in 5% ethanol solution solution
Crystalline substance, the yield difference for being converted into phloridzin is little, so the condition of crystallization selects 10% or so ethanol solution.
Phloridzin is soluble in the ethanol solution of high concentration, and the impurity inside crude product phloridzin is mostly carbohydrate and protide
Compound, the solubility in high concentration ethanol solution is extremely low, this is just that the purification of phloridzin creates condition.
It is safer in order to produce, using high concentration ethyl alcohol when phloridzin crude product is dissolved at normal temperature, filter
Insoluble matter is removed, sugar and protein-based compound in most of phloridzin crude product can be removed, then with 10% or so ethyl alcohol
Solution is recrystallized, the impurity that removal part not can be removed with high concentration ethanol again, and then obtains content 95% or more
High-purity phlorizin.If obtaining the phloridzin of higher purity, it can be repeated a number of times recrystallization, to obtain 99%
Above root skin glycoside product.
By confirming repeatedly in experimentation, 95% ethanol solution of phloridzin is concentrated to the leaching of density 1.5 (50 DEG C)
After cream, the water of 5 times of medicinal extract volumes or so is added, the ethanol solution concentration of adjusting is just 10% or so, because phloridzin is cold
Solubility in water is low, and cold water, which is added, so that phloridzin is precipitated rapidly, is mingled with a large amount of impurity, the effect of separation is not achieved, so
What is be added is 95 DEG C of hot water, prevents phloridzin from precipitating suddenly, extends crystallization time, facilitates the promotion of product purity, in order to
The convenience of large-scale production, process conditions are produced as middle control condition, simple and practical, convenient for large-scale production.
The method only uses market to be easy the edible ethanol bought and carries out the purification production of phloridzin as solvent, and exists mostly
It is operated under room temperature, reduces the volatilization of ethyl alcohol, promote safety, and ethyl alcohol may be reused, low energy consumption, and production equipment is simple,
Invest small, economic benefit is obvious.
Basic implementation method, main feature and advantages of the present invention of the invention has been shown and described above.
It should be understood by those skilled in the art that the present invention is not limited to the above embodiments, above-described embodiment and explanation
It is merely illustrated the principles of the invention described in book, without departing from the spirit and scope of the present invention, the present invention also has
Various changes and modifications, these changes and improvements all fall within the protetion scope of the claimed invention.The claimed scope of the invention
It is defined by the appending claims and its equivalent thereof.
Claims (9)
1. the isolation and purification method of phloridzin in a kind of Malus spectabilis Aqueous extracts, which comprises the steps of:
(1) it is concentrated under reduced pressure: Malus spectabilis aqueous extract being concentrated to give medicinal extract at 75-85 DEG C, collects medicinal extract, stands cooling；
(2) low temperature crystallization: being cooled to room temperature to medicinal extract, is transferred to 0~5 DEG C of freezer and stands still for crystals 20-30h, after being crystallized
Medicinal extract；
(3) it is centrifugated: the medicinal extract after crystallization being mixed with ice water, flat centrifuge is poured into and is separated by solid-liquid separation, filter out mother
Liquid；
(4) ice water washs: after mother liquor all filters out, adding ice water and cleans to the phloridzin that step (3) separates, after drying
Obtain the product containing phloridzin；
(5) crude product is dried: the product drying containing phloridzin being crushed, mixing obtains phloridzin crude product；
(6) crude product purifies: at normal temperature by phloridzin crude product, being dissolved, is filtered using ethyl alcohol, is concentrated under reduced pressure, receive cream, hot water is added
Cooling is stood after mixing, and 0~5 DEG C of freezer is transferred to after temperature drops to room temperature and is stood, centrifugal filtration after 20-30h is crystallized, receives
Collect phloridzin wet product, dry, crushes, obtain phloridzin sterling.
2. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (1)
In, Malus spectabilis aqueous extract is concentrated into after specific gravity reaches 1.05-1.30 at 80 DEG C and collects clear cream.
3. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (3)
In, the volume ratio of clear cream and ice water is 1:0.9-1.2.
4. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (4)
In, the volume ratio of the clear cream in ice water and step (3) is 1:0.9-1.2.
5. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (5)
Described in drying be to be dried under the conditions of -0.09MPa, 55-65 DEG C, it is dry to be lower than to moisture content in phloridzin
6.0%。
6. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (6)
Middle phloridzin crude product at normal temperature, uses mass concentration to be dissolved for 95% or more ethyl alcohol, and wherein mass concentration is 95%
Or more ethyl alcohol and phloridzin crude product mass ratio be 9-12:1.
7. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (6)
Middle reduced pressure is to be concentrated into after specific gravity reaches 1.4-1.6 to receive cream at 45-50 DEG C.
8. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (6)
After middle receipts cream, standing is down to room temperature after 90-98 DEG C of hot water mixing is added, and is then transferred to 4 DEG C of freezer and stands, low temperature crystallization 24
Hour, obtain phloridzin wet product.
9. the isolation and purification method of phloridzin in Malus spectabilis Aqueous extracts according to claim 1, which is characterized in that step (6)
Described in drying be to be dried under the conditions of -0.09MPa, 45-50 DEG C, it is dry to be lower than to moisture content in phloridzin
6.0%。
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Cited By (1)
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| CN111393491A (en) * | 2020-05-07 | 2020-07-10 | 江西省科学院应用化学研究所 | Method for green preparation of high-purity phlorizin from wild Jinchai |
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