CN103408610A - Method for extracting arbutin from pear leaves - Google Patents
Method for extracting arbutin from pear leaves Download PDFInfo
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Abstract
The invention discloses a method for extracting arbutin from pear leaves. Leaf juice containing arbutin extracted from the pear leaves is subjected to concentration, purification and recrystallization to obtain arbutin through. The method comprises the following steps: step 1, drying and grinding the pear leaves, impregnating with methanol, filtering, and concentrating the filtrate to obtain extract; step 2, putting resin for filling a column, dissolving the extract obtained in the step 1 in water, filtering to obtain filtrate with concentration of arbutin of 2 mg/mL, putting the filtrate for filling the column and performing purification to obtain crude extract of arbutin; step 3, putting ENV filler for filling a column, dissolving the crude extract of arbutin obtained in step 2, and putting the dissolved extract for filling the column and performing purification to a obtain crude product of arbutin; step 4, conducting crystallization on the crude product of arbutin to obtain arbutin; step 5, conducting recrystallization on the arbutin obtained in step 4. According to the invention, a raw material adopted in the method is a waste material generated after the harvest of fruit trees, the sources of the raw material are various, the raw material is reachable, and the cost is low; the extraction method provided by the invention is simple in technology and convenient to operate, and can obtain arbutin with high purity.
Description
Technical field
The invention belongs to arbutin extracting method technical field, relate to a kind of from leaf of pear tree, extracting the method for arbutin.
Background technology
Arbutin (arbutin) is a kind of natural active matter, can effectively suppress the biological tyrosinase activity in skin, block melanic formation, accelerate melanic decomposition and excretion, thereby minimizing Skin pigmentation, dispelling stain and freckle are the main active substances at present domestic and international skin-lightening cosmetic.In recent years, along with the development of Cosmetics in China industry, for complying with the trend of skin protection cosmetics " back to nature ", " the green whitening " of natural active matter is also day by day prevailing, and arbutin is as nontoxic, the obvious natural active matter of whitening effect, the market demand expanding day.
At present, the raw material that extracts arbutin is mainly fruit tree, as blue berry, and its operational path complexity, and because the fruit source is limited, the cost that therefore causes extracting arbutin is very high.
The concentration of the related reagent of this paper is volumetric concentration, and all the other related per-cents all refer to weight percent.
Summary of the invention
The object of the present invention is to provide a kind of from leaf of pear tree, extracting the method for arbutin, the complex process that the solution prior art exists, the problem that cost is high.
Technical scheme of the present invention, from leaf of pear tree, extracting the method for arbutin, from leaf of pear tree, extracting the leaf juice that contains arbutin, obtain arbutin through concentrated, purification and recrystallization.
Characteristics of the present invention also are, specifically comprise the following steps:
Step 1, after leaf of pear tree is dried, pulverized, with the methyl alcohol dipping, filter, by filtrate simmer down to medicinal extract;
Step 2, get resin dress post, by step 1 gained medicinal extract water dissolution, filters, and in filtrate, arbutin content is 2mg/mL, and column purification on filtrate obtains the arbutin crude extract;
Step 3, get ENV filler dress post, and after step 2 gained arbutin crude extract was dissolved, upper prop purified again, obtained the arbutin crude product;
Step 4, carry out crystallization to step 3 gained arbutin crude product, obtains arbutin.
Step 5, carry out recrystallization to step 4 gained arbutin.
In step 1, yield of extract is 7%～10%, and in medicinal extract, arbutin content approximately 6%～10%.
Resin in step 2 is the LSA-900B resin; Column purification on filtrate, with the drip washing of different concns methyl alcohol, 0.4～0.5 times of column volume of maintenance flow velocity/hour, obtaining scavenging solution, scavenging solution, after concentrated, obtains the arbutin crude extract of solid state, and wherein arbutin content 30%～34%.
In step 3, on filtrate after column purification, collect effluent liquid, water wash with 2 times of column volumes, in the loading effluent liquid, arbutin content, lower than 0.005mg/mL, discards effluent liquid, then uses 10% methanol-eluted fractions of 10 times of column volumes, each elutriant of collecting 1 times of column volume, whole process flow velocity keeps 2～3mL/min; In first three time wash-out gained elutriant, arbutin content is the highest, is respectively 3.0mg/mL, 4.8mg/mL, 1.6mg/mL, collects the elutriant of first three time wash-out gained, concentrated, obtains solid state arbutin crude product, and its arbutin content is greater than 82%.
In step 4, the arbutin crude product is added to ethyl acetate, reflux 0.5～1h, filtered while hot, hold over night, separate out the white powder solid, filters, and drying, obtain the white powder solid, and in this pulverulent solids, arbutin content is greater than 95%.
In step 5, recrystallization can repeat repeatedly, and arbutin purity reaches more than 99%.
The present invention has following beneficial effect:
1, the present invention's raw material used is the waste material after economical fruit tree is gathered, and raw material sources are abundant, be convenient to obtain, and cost are low.
2, extracting method technique of the present invention is simple, easy to operate, and gained arbutin purity is high.
The accompanying drawing explanation
Fig. 1 is the infrared spectrogram of the arbutin that adopts the present invention and make from the method for extracting arbutin leaf of pear tree.
Embodiment
The present invention is further detailed explanation below in conjunction with embodiment and accompanying drawing.
From leaf of pear tree, extracting the method for arbutin, from leaf of pear tree, extracting the leaf juice that contains arbutin, obtain arbutin through concentrated, purification and recrystallization.Specifically comprise the following steps:
Step 1, after leaf of pear tree is dried, pulverized, with the methyl alcohol dipping, filter, by filtrate simmer down to medicinal extract; Yield of extract is 7%～10%, and in medicinal extract, arbutin content approximately 6%～10%.Yield of extract is the percentage composition that medicinal extract accounts for the quality of leaf of pear tree used.
Step 2, get resin dress post, by step 1 gained medicinal extract water dissolution, filters, and in filtrate, arbutin content is 2mg/mL, and column purification on filtrate obtains the arbutin crude extract; Resin is the LSA-900B resin; Column purification on filtrate, with the drip washing of different concns methyl alcohol, 0.4～0.5 times of column volume of maintenance flow velocity/hour, obtaining scavenging solution, scavenging solution, after concentrated, obtains the arbutin crude extract of solid state, and wherein arbutin content 30%～34%.
Step 3, get ENV filler dress post, and after step 2 gained arbutin crude extract was dissolved, upper prop purified again, obtained the arbutin crude product; On filtrate, after column purification, collect effluent liquid, with the water wash of 2 times of column volumes, in the loading effluent liquid, arbutin content, lower than 0.005mg/mL, discards effluent liquid, then uses 10% methanol-eluted fractions of 10 times of column volumes, each elutriant of collecting 1 times of column volume, whole process flow velocity keeps 2～3mL/min; In first three time wash-out gained elutriant, arbutin content is the highest, is respectively 3.0mg/mL, 4.8mg/mL, 1.6mg/mL, collects the elutriant of first three time wash-out, concentrated, obtains solid state arbutin crude product, and its arbutin content is greater than 82%.
Step 4, carry out crystallization to step 3 gained arbutin crude product, obtains arbutin.Be specially the arbutin crude product is added to ethyl acetate, reflux 0.5～1h, filtered while hot, hold over night, separate out the white powder solid, filters, and drying, obtain the white powder solid, and in this pulverulent solids, arbutin content is greater than 95%.
Step 5, carry out recrystallization to step 4 gained arbutin.Can repeat repeatedly recrystallization, arbutin purity reaches more than 99%.
Embodiment 1
Step 1, from leaf of pear tree, extracting the extract contain arbutin, its concrete steps are:
Step 1a: win the leaf of pear tree sample, dry in 40 ℃ of baking ovens, pulverize.
Step 1b: the leaf of pear tree sample 400g after claim pulverizing, add the pure methyl alcohol of 4L, under normal temperature, flood 3, filter, filtrate is rotated to evaporation, concentrated removal methyl alcohol, obtain containing the medicinal extract of arbutin, yield of extract 7%, in medicinal extract, arbutin content approximately 6%.
Step 2, the standby arbutin crude extract of use LSA-900B resin:
Step 2a: the screening of resin:
Adopt the Static Adsorption test to be studied the adsorption effect of 4 kinds of different model resins, 4 kinds of resins selecting are LSA-900B, LSA-900C, LX-200 and LX-400, accurately take each model resin 1g, after pre-treatment, add on request in tool plug Erlenmeyer flask, get the 0.1g leaf of pear tree and extract medicinal extract, add the 40mL water dissolution, in every kind of resin, add the 10mL medicinal extract aqueous solution, Static Adsorption 1h under room temperature, get the supernatant liquid of every kind of resin, (measuring method is shown in document: Zhao Jie by the HPLC method, to measure arbutin content, He Qiang, Yao Binghua, Deng, in leaf of pear tree, the conclusive evidence of effective constituent arbutin and HPLC detect, Chinese Journal of Modern Applied Pharmacy, 2011), with arbutin content in initial soln, compare, calculate the Static Adsorption rate of various resins to arbutin, result is as shown in table 1, by as seen from Table 1, the Static Adsorption rate of LSA-900B type resin is the highest, so select LSA-900B type resin to carry out rough purification.
Table 1 different model resin is to arbutin Static Adsorption rate
| The resin model | LSA-900B | LSA-900C | LX-200 | LX-400 |
| Adsorption rate (%) | 62.8 | 9.4 | 20.8 | 8.5 |
Step 2b: obtain the arbutin crude extract by LSA-900B type resin isolation:
Get LSA-900B type resin 300g dress post (the about 450mL of column volume), activation on request, get the 8g leaf of pear tree and extract medicinal extract, use the 240mL water dissolution, filter, in filtrate, arbutin content is 2mg/mL, column purification on filtrate, 0.4 times of column volume of maintenance flow velocity/hour, collect loading effluent liquid (numbering 0), use successively again the distilled water (numbering 1) of 1 times of column volume, 10% methyl alcohol of 1 times of column volume (numbering 2), 20% methyl alcohol of 1 times of column volume (numbering 3), 30% methyl alcohol of 1 times of column volume (numbering 4), 1 times of pure methyl alcohol of column volume (numbering 5), 1 times of pure methyl alcohol of column volume (numbering 6), collect the leacheate in each stage, detect arbutin content in leacheate, result is as shown in table 2.
Arbutin content in the different elutriants of table 2
As can be seen from Table 2, No. 3 elutriants (20% methyl alcohol) start to have more arbutin to be eluted, No. 5 elutriants (pure methyl alcohol) are basic complete by the arbutin wash-out adsorbed on resin, so, according to the experimental result in table 2, determine after loading the water wash of first using 1 times of column volume, use again 10% methyl alcohol drip washing of 1 times of column volume, discard leacheate, elutriant is through follow-up optimization, and the pure methanol-based of 1.4 times of column volumes originally can be complete by the arbutin wash-out adsorbed on resin.Meoh eluate is concentrated into dry, obtains the about 1.2g of arbutin crude extract, wherein arbutin content approximately 30%.
Step 3, use ENV polymerization filling purify the arbutin crude extract.
Through a large amount of preliminary experiments, select the ENV adsorption stuffing further to purify the arbutin crude extract.The ENV filler is a kind of polystyrene-divinylbenzene copolymer material, arbutin in the aqueous solution is had to adsorption preferably, and the large polar materials such as carbohydrate are not had to absorption substantially, and the arbutin be adsorbed on ENV is easily eluted by 10% methanol solution, other impurity that are adsorbed on the ENV filler need the methyl alcohol of higher concentration just can elute, thereby reach good decontamination effect improving.
Concrete experimental procedure: get 20g ENV filler dress post (the about 56mL of column volume), use successively the methyl alcohol of 1 times of column volume, the water activation pillar of 1 times of column volume, get the 1.2g arbutin crude extract that rough purification obtains through LSA-900B, use the 50mL water dissolution, filter, filtrate is crossed column purification, collect effluent liquid, water wash with 2 times of column volumes, in the loading effluent liquid, arbutin content is all lower than 0.005mg/mL, discard effluent liquid, use again 10% methanol-eluted fractions of 10 times of column volumes, each elutriant (being numbered 1～10) of collecting 1 times of column volume, whole process flow velocity keeps 2mL/min, in first three time wash-out gained elutriant, arbutin content is the highest, be respectively 3.0mg/mL, 4.8mg/mL, 1.6mg/mL, in the 4th wash-out gained elutriant, arbutin content is lower, be only 0.06mg/mL, from the 5th elutriant, in elutriant, be mainly other composition.So according to detection case, collect first three time gained elutriant, concentrated, obtain the arbutin crude product that about 0.3g purity is higher (for solid state), after testing, arbutin content is about 82%.
Step 4, recrystallization obtain the arbutin monomer
Get the arbutin crude product that about 0.3g is purer, add the 10mL ethyl acetate, reflux 0.5h, filtered while hot, hold over night, separate out the white powder solid, filter, drying, obtain the about 0.2g of white powder solid, and in this pulverulent solids, arbutin content is about 95%, if need the higher arbutin of preparation purity, can repeat recrystallization, recrystallization 3 times, the arbutin purity is more than 99%.
The conclusive evidence of step 5, arbutin
The arbutin that separation is obtained, adopt high performance liquid chromatography, mass spectrum, infrared spectra and nucleus magnetic resonance H spectrum and C spectrum to prove conclusively, wherein separate the infrared spectra of the arbutin obtained as shown in Figure 1, consistent with the standard infrared spectrogram of arbutin in the standard spectrum storehouse.
Shaanxi Province is the large province of Production of fruit, and wherein the cultivated area of pear tree is nearly 1,000,000 mu, and leaf of pear tree, as the by product of pear tree plantation, is originated very abundant, and cost is low.
Embodiment 2
Step 1, from leaf of pear tree, extracting the extract contain arbutin, its concrete steps are:
Step 1a: win the leaf of pear tree sample, dry in 50 ℃ of baking ovens, pulverize.
Step 1b: the leaf of pear tree sample 450g after claim pulverizing, add the pure methyl alcohol of 4.5L, under normal temperature, flood 4, filter, filtrate is rotated to evaporation, concentrated removal methyl alcohol, obtain containing the medicinal extract of arbutin, yield of extract 8%, in medicinal extract, arbutin content approximately 7%.
Step 2, the standby arbutin crude extract of use LSA-900B resin:
Step 2a: the screening of resin:
Adopt the Static Adsorption test to be studied the adsorption effect of 4 kinds of different model resins, 4 kinds of resins selecting are LSA-900B, LSA-900C, LX-200 and LX-400, accurately take each model resin 3g, after pre-treatment, add on request in tool plug Erlenmeyer flask, get the 0.3g leaf of pear tree and extract medicinal extract, add the 120mL water dissolution, in every kind of resin, add the 30mL medicinal extract aqueous solution, Static Adsorption 1h under room temperature, get the supernatant liquid of every kind of resin, (measuring method is shown in document: Zhao Jie by the HPLC method, to measure arbutin content, He Qiang, Yao Binghua, Deng, in leaf of pear tree, the conclusive evidence of effective constituent arbutin and HPLC detect, Chinese Journal of Modern Applied Pharmacy, 2011), with arbutin content in initial soln, compare, calculate the Static Adsorption rate of various resins to arbutin, result is as shown in table 3, by as seen from Table 3, the Static Adsorption rate of LSA-900B type resin is the highest, so select LSA-900B type resin to carry out rough purification.
Table 3 different model resin is to arbutin Static Adsorption rate
| The resin model | LSA-900B | LSA-900C | LX-200 | LX-400 |
| Adsorption rate (%) | 60.6 | 8.3 | 18.1 | 7.4 |
Step 2b: obtain the arbutin crude extract by LSA-900B type resin isolation:
Get LSA-900B type resin 350g dress post (the about 530mL of column volume), activation on request, get the 9g leaf of pear tree and extract medicinal extract, use the 320mL water dissolution, filter, in filtrate, arbutin content is 2mg/mL, column purification on filtrate, 0.4 times of column volume of maintenance flow velocity/hour, collect loading effluent liquid (numbering 0), use successively again the distilled water (numbering 1) of 1 times of column volume, 10% methyl alcohol of 1 times of column volume (numbering 2), 20% methyl alcohol of 1 times of column volume (numbering 3), 30% methyl alcohol of 1 times of column volume (numbering 4), 1 times of pure methyl alcohol of column volume (numbering 5), 1 times of pure methyl alcohol of column volume (numbering 6), collect the leacheate in each stage, detect arbutin content in leacheate, result is as shown in table 4.
Arbutin content in the different elutriants of table 4
As can be seen from Table 4, No. 3 elutriants (20% methyl alcohol) start to have more arbutin to be eluted, No. 5 elutriants (pure methyl alcohol) are basic complete by the arbutin wash-out adsorbed on resin, so, according to the experimental result in table 2, determine after loading the water wash of first using 1 times of column volume, use again 10%～15% methyl alcohol drip washing of 1 times of column volume, discard leacheate, elutriant is through follow-up optimization, and the pure methanol-based of 1.4～1.5 times of column volumes originally can be complete by the arbutin wash-out adsorbed on resin.Meoh eluate is concentrated into dry, obtains the about 2.1g of arbutin crude extract, wherein arbutin content approximately 30%.
Step 3, use ENV polymerization filling purify the arbutin crude extract.
Through a large amount of preliminary experiments, select the ENV adsorption stuffing further to purify the arbutin crude extract.The ENV filler is a kind of polystyrene-divinylbenzene copolymer material, arbutin in the aqueous solution is had to adsorption preferably, and the large polar materials such as carbohydrate are not had to absorption substantially, and the arbutin be adsorbed on ENV is easily eluted by 10% methanol solution, other impurity that are adsorbed on the ENV filler need the methyl alcohol of higher concentration just can elute, thereby reach good decontamination effect improving.
Concrete experimental procedure: get 22g ENV filler dress post (the about 60mL of column volume), use successively the methyl alcohol of 1～1.5 times of column volume, the water activation pillar of 1～2 times of column volume, get the 2.1g arbutin crude extract that rough purification obtains through LSA-900B, use the 55mL water dissolution, filter, filtrate is crossed column purification, collect effluent liquid, water wash with 2 times of column volumes, in the loading effluent liquid, arbutin content is all lower than 0.005mg/mL, discard effluent liquid, use again 10% methanol-eluted fractions of 10 times of column volumes, each elutriant (being numbered 1～10) of collecting 1 times of column volume, whole process flow velocity keeps 3mL/min, in first three time gained elutriant, arbutin content is the highest, be respectively 2.8mg/mL, 4.5mg/mL, 1.7mg/mL, in the 4th gained elutriant, arbutin content is lower, be only 0.05mg/mL, from the 5th gained elutriant, in elutriant, be mainly other composition.So according to detection case, collect the elutriant of first three time wash-out gained, concentrated, obtain the arbutin crude product that about 0.44g purity is higher (for solid state), after testing, arbutin content is about 85%.
Step 4, recrystallization obtain the arbutin monomer
Get the arbutin crude product that about 0.44g is purer, add the 30mL ethyl acetate, reflux 1h, filtered while hot, hold over night, separate out the white powder solid, filter, drying, obtain the about 0.3g of white powder solid, and in this pulverulent solids, arbutin content is greater than 95%, if need the higher arbutin of preparation purity, can repeat recrystallization, recrystallization 3 times, the arbutin purity is more than 99%.
The conclusive evidence of step 5, arbutin
To the arbutin that separation obtains, adopt high performance liquid chromatography, mass spectrum, infrared spectra and nucleus magnetic resonance H spectrum and C spectrum to prove conclusively, all can be corresponding with the standard spectrum storehouse.
Shaanxi Province is the large province of Production of fruit, and wherein the cultivated area of pear tree is nearly 1,000,000 mu, and leaf of pear tree, as the by product of pear tree plantation, is originated very abundant, and cost is low.
Embodiment 3
Step 1, from leaf of pear tree, extracting the extract contain arbutin, its concrete steps are:
Step 1a: win the leaf of pear tree sample, dry in 60 ℃ of baking ovens, pulverize.
Step 1b: the leaf of pear tree sample 500g after claim pulverizing, add the pure methyl alcohol of 5L, under normal temperature, flood 5, filter, filtrate is rotated to evaporation, concentrated removal methyl alcohol, obtain containing the medicinal extract of arbutin, yield of extract 10%, in medicinal extract, arbutin content approximately 10%.
Step 2, the standby arbutin crude extract of use LSA-900B resin:
Step 2a: the screening of resin:
Adopt the Static Adsorption test to be studied the adsorption effect of 4 kinds of different model resins, 4 kinds of resins selecting are LSA-900B, LSA-900C, LX-200 and LX-400, accurately take each model resin 5g, after pre-treatment, add on request in tool plug Erlenmeyer flask, get the 0.5g leaf of pear tree and extract medicinal extract, add the 200mL water dissolution, in every kind of resin, add the 50mL medicinal extract aqueous solution, Static Adsorption 1h under room temperature, get the supernatant liquid of every kind of resin, (measuring method is shown in document: Zhao Jie by the HPLC method, to measure arbutin content, He Qiang, Yao Binghua, Deng, in leaf of pear tree, the conclusive evidence of effective constituent arbutin and HPLC detect, Chinese Journal of Modern Applied Pharmacy, 2011), with arbutin content in initial soln, compare, calculate the Static Adsorption rate of various resins to arbutin, result is as shown in table 5, by as seen from Table 5, the Static Adsorption rate of LSA-900B type resin is the highest, so select LSA-900B type resin to carry out rough purification.
Table 5 different model resin is to arbutin Static Adsorption rate
| The resin model | LSA-900B | LSA-900C | LX-200 | LX-400 |
| Adsorption rate (%) | 69.7 | 10.2 | 21.1 | 9.3 |
Step 2b: obtain the arbutin crude extract by LSA-900B type resin isolation:
Get LSA-900B type resin 400g dress post (the about 600mL of column volume), activation on request, get the 10g leaf of pear tree and extract medicinal extract, use the 500mL water dissolution, filter, in filtrate, arbutin content is 2mg/mL, column purification on filtrate, 0.4 times of column volume of maintenance flow velocity/hour, collect loading effluent liquid (numbering 0), use successively again the distilled water (numbering 1) of 1 times of column volume, 10% methyl alcohol of 1 times of column volume (numbering 2), 20% methyl alcohol of 1 times of column volume (numbering 3), 30% methyl alcohol of 1 times of column volume (numbering 4), 1 times of pure methyl alcohol of column volume (numbering 5), 1 times of pure methyl alcohol of column volume (numbering 6), collect the leacheate in each stage, detect arbutin content in leacheate, result is as shown in table 6.
Arbutin content in the different elutriants of table 6
As can be seen from Table 2, No. 3 elutriants (20% methyl alcohol) start to have more arbutin to be eluted, No. 5 elutriants (pure methyl alcohol) are basic complete by the arbutin wash-out adsorbed on resin, so, according to the experimental result in table 2, determine after loading the water wash of first using 1 times of column volume, use again 10%～15% methyl alcohol drip washing of 1 times of column volume, discard leacheate, elutriant is through follow-up optimization, and the pure methanol-based of 1.4～1.5 times of column volumes originally can be complete by the arbutin wash-out adsorbed on resin.Meoh eluate is concentrated into dry, obtains the about 3.0g of arbutin crude extract, wherein arbutin content approximately 34%.
Step 3, use ENV polymerization filling purify the arbutin crude extract.
Through a large amount of preliminary experiments, select the ENV adsorption stuffing further to purify the arbutin crude extract.The ENV filler is a kind of polystyrene-divinylbenzene copolymer material, arbutin in the aqueous solution is had to adsorption preferably, and the large polar materials such as carbohydrate are not had to absorption substantially, and the arbutin be adsorbed on ENV is easily eluted by 10% methanol solution, other impurity that are adsorbed on the ENV filler need the methyl alcohol of higher concentration just can elute, thereby reach good decontamination effect improving.
Concrete experimental procedure: get 24g ENV filler dress post (the about 64mL of column volume), use successively the methyl alcohol of 1～1.5 times of column volume, the water activation pillar of 1～2 times of column volume, get the 3.0g arbutin crude extract that rough purification obtains through LSA-900B, use the 60mL water dissolution, filter, filtrate is crossed column purification, collect effluent liquid, water wash with 2 times of column volumes, in the loading effluent liquid, arbutin content is all lower than 0.005mg/mL, discard effluent liquid, use again 10% methanol-eluted fractions of 10 times of column volumes, each elutriant (being numbered 1～10) of collecting 1 times of column volume, whole process flow velocity keeps 2mL/min, in the elutriant of first three time wash-out gained, arbutin content is the highest, be respectively 2.9mg/mL, 4.9mg/mL, 1.8mg/mL, in the elutriant of the 4th wash-out gained, arbutin content is lower, be only 0.05mg/mL, from the 5th gained elutriant, in elutriant, be mainly other composition.So according to detection case, collect first three time gained elutriant, concentrated, obtain the arbutin crude product that about 0.84g purity is higher (for solid state), after testing, arbutin content is about 85%.
Step 4, recrystallization obtain the arbutin monomer
Get the arbutin crude product that about 0.84g is purer, add the 50mL ethyl acetate, reflux 1h, filtered while hot, hold over night, separate out the white powder solid, filter, drying, obtain the about 0.4g of white powder solid, and in this pulverulent solids, arbutin content is greater than 95%, if need the higher arbutin of preparation purity, can repeat recrystallization, recrystallization 3 times, the arbutin purity is more than 99%.
The conclusive evidence of step 5, arbutin
To the arbutin that separation obtains, adopt high performance liquid chromatography, mass spectrum, infrared spectra and nucleus magnetic resonance H spectrum and C spectrum to prove conclusively, all can be corresponding with the standard spectrum storehouse.
Shaanxi Province is the large province of Production of fruit, and wherein the cultivated area of pear tree is nearly 1,000,000 mu, and leaf of pear tree, as the by product of pear tree plantation, is originated very abundant, and cost is low.
Claims (8)
1. from leaf of pear tree, extracting the method for arbutin, it is characterized in that, from leaf of pear tree, extracting the leaf juice that contains arbutin, obtain arbutin through concentrated, purification and recrystallization.
2. the method from extraction arbutin leaf of pear tree as claimed in claim 1, is characterized in that, specifically comprises the following steps:
Step 1, after leaf of pear tree is dried, pulverized, with the methyl alcohol dipping, filter, by filtrate simmer down to medicinal extract;
Step 2, get resin dress post, by step 1 gained medicinal extract water dissolution, filters, and in filtrate, arbutin content is 2mg/mL, and column purification on filtrate, obtain scavenging solution, concentrates to obtain the arbutin crude extract;
Step 3, get ENV filler dress post, and step 2 gained arbutin crude extract upper prop is purified again, obtains the arbutin crude product;
Step 4, carry out crystallization to step 3 gained arbutin crude product, obtains arbutin.
3. as claimed in claim 2 from leaf of pear tree, extracting the method for arbutin, it is characterized in that: also comprise step 5, step 5 is for to carry out recrystallization to step 4 gained arbutin.
4. as claimed in claim 3 it is characterized in that: recrystallization can repeat repeatedly from leaf of pear tree, extracting the method for arbutin, and arbutin purity reaches more than 99%.
5. the method from extraction arbutin leaf of pear tree as described as claim 2-4 any one, is characterized in that, in step 1, and yield of extract 7%～10%; In medicinal extract, arbutin content approximately 6%～10%.
6. the method from extraction arbutin leaf of pear tree as described as claim 2-4 any one, is characterized in that, the resin in step 2 is the LSA-900B resin; Column purification on filtrate, the drip washing of different concns methyl alcohol, keep 0.4～0.5 times of column volume of flow velocity/hour, obtain the arbutin crude extract, wherein arbutin content approximately 30%～34%.
7. as described as claim 2-4 any one from leaf of pear tree, extracting the method for arbutin, it is characterized in that, in step 3, after column purification, collect effluent liquid on filtrate, water wash with 2 times of column volumes, in the loading effluent liquid, arbutin content, all lower than 0.005mg/mL, discards effluent liquid, then uses 10% methanol-eluted fractions of 10 times of column volumes, each elutriant of collecting 1 times of column volume, whole process flow velocity keeps 2～3mL/min; In first three time wash-out gained elutriant, arbutin content is the highest, is respectively 3.0mg/mL, 4.8mg/mL, 1.6mg/mL, collects the elutriant of first three time wash-out gained, concentrated, obtains solid state arbutin crude product, and its arbutin content is greater than 82%.
8. as described as claim 2-4 any one from leaf of pear tree, extracting the method for arbutin, it is characterized in that, in step 4, the arbutin crude product adds ethyl acetate, reflux 0.5～1h, filtered while hot, hold over night, separate out the white powder solid, filter, drying, obtain the white powder solid, and in this pulverulent solids, arbutin content is greater than 95%.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310320988.5A CN103408610B (en) | 2013-07-26 | 2013-07-26 | The method of arbutin is extracted from leaf of pear tree |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107163093A (en) * | 2017-06-16 | 2017-09-15 | 江苏天晟药业股份有限公司 | A kind of preparation method of ursin |
| CN107722080A (en) * | 2017-11-08 | 2018-02-23 | 长沙爱扬医药科技有限公司 | A kind of method that ursin is extracted in the leaf from purple bergenia herb |
| CN108558964A (en) * | 2018-04-18 | 2018-09-21 | 山东众山生物科技有限公司 | A kind of purification process of alpha-arbutin |
| CN110551160A (en) * | 2019-09-29 | 2019-12-10 | 李世� | extraction and purification method of arbutin and freckle removing and whitening application |
| CN110684061A (en) * | 2018-07-05 | 2020-01-14 | 大湾汉唯(广州)中药研究开发有限公司 | Extract containing alpha-arbutin and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107163093A (en) * | 2017-06-16 | 2017-09-15 | 江苏天晟药业股份有限公司 | A kind of preparation method of ursin |
| CN107722080A (en) * | 2017-11-08 | 2018-02-23 | 长沙爱扬医药科技有限公司 | A kind of method that ursin is extracted in the leaf from purple bergenia herb |
| CN108558964A (en) * | 2018-04-18 | 2018-09-21 | 山东众山生物科技有限公司 | A kind of purification process of alpha-arbutin |
| CN110684061A (en) * | 2018-07-05 | 2020-01-14 | 大湾汉唯(广州)中药研究开发有限公司 | Extract containing alpha-arbutin and preparation method thereof |
| CN110684061B (en) * | 2018-07-05 | 2021-09-03 | 大湾汉唯(广州)中药研究开发有限公司 | Extract containing alpha-arbutin and preparation method thereof |
| CN110551160A (en) * | 2019-09-29 | 2019-12-10 | 李世� | extraction and purification method of arbutin and freckle removing and whitening application |
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| CN103408610B (en) | 2016-06-01 |
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