CN102260304A - Method for purifying rebaudioside A - Google Patents
Method for purifying rebaudioside A Download PDFInfo
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- CN102260304A CN102260304A CN2010101865284A CN201010186528A CN102260304A CN 102260304 A CN102260304 A CN 102260304A CN 2010101865284 A CN2010101865284 A CN 2010101865284A CN 201010186528 A CN201010186528 A CN 201010186528A CN 102260304 A CN102260304 A CN 102260304A
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Abstract
The invention provides a method for purifying rebaudioside A, comprising the following steps: dispersing the raw material containing rebaudioside A in a solvent system consisting of absolute ethyl alcohol, acetone and water, filtering and collecting undissolved substance to obtain a crude product of rebaudioside A; dispersing the crude product of rebaudioside A in a solvent system consisting of absolute ethyl alcohol and water, filtering and recrystallizing the filtrate, collecting crystals, and drying to obtain the purified rebaudioside A. The method has simple steps, and has the advantages of low cost, high conversion rate, environmental protection and the like, and is suitable for industrialization production.
Description
Technical field
The application relates to the pharmacognosy field, is specifically related to the crude drug extracting and purifying method, particularly the method for refining purifying content rebaudioside-A from stevioside.
Background technology
Stevioside is a kind of high sugariness that extracts from Folium Chrysanthemi. the natural sweeteners of non-nutritive low in calories.At present, from stevioside, found eight kinds of glucosides, i.e. stevioside (slevioside), rebaudioside (Rebaudioside A, B, C, D, E), but (Dul～ide and this are for dimension uncle glucoside (steviobi～siti) for Dole's glucoside A.Wherein the sugariness of content rebaudioside-A (being designated hereinafter simply as RA) is the highest, and greater than 300 times of sucrose, and flavor matter is best, does not contain any bad pleasant impression, is that a kind of ideal crude sweet is hidden agent.
Extracting the high stevioside product of preparation content rebaudioside-A content is the research focus of domestic and international stevioside manufacture.
In the existing patented technology, the method for existing disclosed acquisition high purity RA is as follows:
The method of publication number CN101270138A is from sweet Stevia, by steps such as extraction, resin isolation, decolouring, crystallization, recrystallization, washings, obtains the RA crystal of purity greater than 90%-98%.
The method one of publication number CN101200480A is from the rebaudioside, by silica gel column chromatography, contain the RA that cut merge to concentrate, crystallization gets purity 〉=99% of RA.
Method two is from sweet Stevia, gets the RA of purity 〉=99% by extraction, ultrafiltration, resin isolation, crystallization.
The method of publication number CN101100477A is from containing the stevioside of RA30%, getting the RA of purity＞90% by adopting methyl alcohol and Virahol (volume ratio 99: 1) crystallization and recrystallization, membrane sepn.
Part also comes with some shortcomings in the aforesaid method.These methods or step are more, are unfavorable for suitability for industrialized production; The purity that perhaps obtains RA is not high, only is＞90%, and the solvent of Shi Yonging is not the solvent of conventional suitability for industrialized production simultaneously, and for example methyl alcohol is toxic, Virahol price height, all non-common solvent.
Summary of the invention
In order to improve the production technique of RA, make its step simple, have advantages such as lower production cost, high conversion and environmental protection simultaneously, and propose the present invention.Method of the present invention is suitable for suitability for industrialized production.
According to technical scheme provided by the invention, the method for refining purifying content rebaudioside-A comprises the following steps: from stevioside
-the raw material that will contain content rebaudioside-A is scattered in dehydrated alcohol, in the solvent system that acetone and water constitute, filters and collects insolubles, to obtain the content rebaudioside-A crude product;
-described content rebaudioside-A crude product is scattered in the solvent system of dehydrated alcohol and water formation, make the filtrate recrystallization after the filtration, collect crystal, oven dry obtains the content rebaudioside-A of purifying.
Described dehydrated alcohol, the volume ratio of all kinds of SOLVENTS is in the solvent system that acetone and water constitute: 3～8 parts of dehydrated alcohols: 5～10 parts of acetone: 0.4～0.8 part of water; Preferably, the volume ratio of all kinds of SOLVENTS is in the described solvent system: 4～6 parts of dehydrated alcohols: 6～8 parts of acetone: 0.4～0.6 part of water.Under described preferred proportion, can make product have higher transformation efficiency and purity.As non-special instruction, theoretical content * 100% of product in transformation efficiency of the present invention=final finished quality/raw material, purity=straight product quality/final finished quality * 100%.
According to preferred version of the present invention, describedly contain the raw material of content rebaudioside-A and there is the fixed proportionlity in the consumption of above-mentioned solvent system, in proportion relation, raw material is calculated in mass, the solvent by volume is calculated, and mass unit mg is suitable with volume unit ml, for example, when 1 part of raw material referred to the 1mg raw material, 1 part of solvent referred to the 1ml solvent.The concrete proportioning of this preferred version is: 1 part of raw material: 3～8 parts of dehydrated alcohols: 5～10 parts of acetone: 0.4～0.8 part of water; Preferably, the volume ratio of all kinds of SOLVENTS is in the described solvent system: 1 part of raw material: 4～6 parts of dehydrated alcohols: 6～8 parts of acetone: 0.4～0.6 part of water, promptly aforementioned solvents system consumption and 1 part of raw material that contains content rebaudioside-A are complementary.Under described preferred proportion, can make product have higher transformation efficiency and purity.
The volume ratio of two kinds of solvents is in the solvent system that described dehydrated alcohol and water constitute: 12～16 parts of dehydrated alcohols: 1.5～1.6 parts of water; Preferably, the volume ratio of all kinds of SOLVENTS is in the described solvent system: 14 parts of dehydrated alcohols: 1.6 parts of water.Under described preferred proportion, can make product have higher transformation efficiency and purity.
According to preferred version of the present invention, there is the fixed proportionlity in the consumption of described content rebaudioside-A crude product and above-mentioned solvent system, in proportion relation, the content rebaudioside-A crude product is calculated in mass, the solvent by volume is calculated, and mass unit mg is suitable with volume unit ml, for example, when 1 part of content rebaudioside-A crude product referred to 1mg content rebaudioside-A crude product, 1 part of solvent referred to the 1ml solvent.The concrete proportioning of this preferred version is: 1 part of crude product: 12～16 parts of dehydrated alcohols: 1.5～1.6 parts of water; Preferably, the volume ratio of all kinds of SOLVENTS is in the described solvent system: 1 part of crude product: 14 parts of dehydrated alcohols: 1.6 parts of water, promptly aforementioned solvents system consumption and 1 part of content rebaudioside-A crude product are complementary.Under described preferred proportion, can make product have higher transformation efficiency and purity.
It is 20%～80% that the raw material that contains content rebaudioside-A of the present invention adopts RA content, preferred 40%～60% stevioside.
Dispersion of the present invention is that the particle with a kind of material is distributed in the another kind of material, is about to disperse phase and is distributed in the dispersion medium, to form the process of dispersion system.Those skilled in the art can select various means according to actual needs, for example, can adopt the stirring jolting, ultra-sonic dispersion, and one or more means such as reflux are disperseed.Preferably, technical scheme of the present invention adopts the mode of reflux.
The invention has the advantages that:
1, technology is simple, is fit to suitability for industrialized production.This technology only need reflux, recrystallization gets final product, and the arts demand of other patent applied for is crossed silica gel column chromatography or macroporous adsorptive resins recrystallization again, and technology is comparatively loaded down with trivial details.
2, the production time weak point can save production cost.
3, transformation efficiency height.It is that 40%～60% Steviosides is a raw material that this technology can adopt RA content, produces 〉=97%RA, and transformation efficiency is greater than 50%.
4, environmental protection.This technology is only used 2 kinds of organic solvents, low price and most of recyclable utilization again.Do not adopt poisonous and the high organic solvent of price.
Description of drawings
Fig. 1 tests the 3. HPLC collection of illustrative plates of prepared RA in the comparative example;
Fig. 2 is for containing the HPLC collection of illustrative plates of 50%RA raw material among the embodiment 2;
Fig. 3 is the HPLC collection of illustrative plates of RA crude product among the embodiment 2;
Fig. 4 is the HPLC collection of illustrative plates of embodiment 2 final finished.
Embodiment
Comparative example
Carry out following experiment according to condition well known in the prior art
1, investigates of the influence of different multiples 90% ethanol to refining 50%RA
1. precision takes by weighing 5g 50%RA, adds 50mL90% ethanol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
2. precision takes by weighing 5g 50%RA, adds 40mL90% ethanol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
3. precision takes by weighing 5g 50%RA, adds 30mL90% ethanol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
4. precision takes by weighing 5g 50%RA, adds 20mL90% ethanol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
When above-mentioned solution is placed 6 days, 2. 3. there is small amount of crystalline to separate out in 4., when placing 15 days, 1. do not have crystallization to separate out yet, will 2. 3. 4. filter, collect crystallization, dry, weigh, the results are shown in Table 2.
The experiment 3. prepared RA the HPLC collection of illustrative plates as shown in Figure 1.As shown in Figure 1, directly with ethanol water recrystallization, RA purity is lower, does not reach 97% according to the method for prior art.
By above experiment, checking can not get 97% RA by the method for patent, and transformation efficiency is not high yet simultaneously.
2, investigate the influence of different concentration ethanol to refining 50%RA
1. precision takes by weighing 5g 50%RA, adds the 30mL dehydrated alcohol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
2. precision takes by weighing 5g 50%RA, adds 30mL96% ethanol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
3. precision takes by weighing 5g 50%RA, adds 30mL92% ethanol, is back to dissolving fully, filters, and filtrate is placed to be crystallized separating out.
When above-mentioned solution is placed 6 days, only 3. there is small amount of crystalline to separate out.
By above experiment, checking is by the method for patent, and no crystallization is separated out, and can not get 97% RA.
Conclusion: adopt the method for high concentration ethanol direct crystallization, length consuming time, transformation efficiency is low, and the purity of RA does not reach 97% yet, is not suitable for suitability for industrialized production.
97%RA embodiment
Embodiment 1: get the raw material that 100g contains 40%RA, by 1: 5: 7: the ratio of 0.5 (raw material is calculated in mass, and the solvent by volume is calculated, and following table is shown m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, get insolubles, oven dry gets the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
The refining experimental result of table 1RA
Embodiment 2: get the raw material that 100g contains 50%RA, in 1: 5: 7: the ratio of 0.5 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.Fig. 2～Fig. 4 has shown the HPLC collection of illustrative plates that contains the 50%RA raw material respectively, the HPLC collection of illustrative plates of RA crude product, and the HPLC collection of illustrative plates of the finished product.Can see intuitively from the HPLC collection of illustrative plates of Fig. 2～Fig. 4, before handling, contain in the collection of illustrative plates of 50%RA raw material that 6 tangible impurity peaks are still arranged except that main peak without the method for present embodiment; After the processing of present embodiment first step, the visible impurity peaks becomes 2 in the HPLC collection of illustrative plates of the RA crude product of acquisition; After further handling, in the HPLC collection of illustrative plates of the finished product, except that main peak, no longer there is tangible impurity peaks through the present embodiment second step recrystallization.
Embodiment 3: get the raw material that 100g contains 60%RA, in 1: 5: 7: the ratio of 0.5 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 4: get the raw material that 100g contains 50%RA, in 1: 4: 6: the ratio of 0.4 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 5: get the raw material that 100g contains 50%RA, in 1: 5: 7: the ratio of 0.5 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 2 hours, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 6: get the raw material that 100g contains 50%RA, in 1: 6: 8: the ratio of 0.6 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 3 hours, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 7: get the raw material that 100g contains 50%RA, in 1: 3: 5: the ratio of 0.8 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 8: get the raw material that 100g contains 50%RA, in 1: 8: 10: the ratio of 0.4 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 14: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 9: get the raw material that 100g contains 50%RA, in 1: 4: 6: the ratio of 0.4 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 12: 1.5 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Embodiment 10: get the raw material that 100g contains 50%RA, in 1: 4: 6: the ratio of 0.4 (m/v) added dehydrated alcohol, acetone and water, 80 ℃ of reflux 1 hour are filtered, and get insolubles, oven dry, the RA crude product; The RA crude product is weighed, and ratio adding dehydrated alcohol and water in 1: 16: 1.6 (m/v) refluxed 1 hour, filtered, and filtrate is placed 72 hours recrystallizations, filters, and collects crystal, dries, promptly.Sampling is carried out purity detecting with the HPLC method, the results are shown in Table 1.
Claims (10)
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104548648A (en) * | 2014-10-08 | 2015-04-29 | 浙江大学 | Crystallizing device for preparing high-purity rebaudioside A by employing continuous purification, and control method of crystallizing device |
| CN104725444A (en) * | 2015-03-24 | 2015-06-24 | 杨健 | Method for efficiently preparing high-purity rebaudioside A |
| CN105061527A (en) * | 2015-08-31 | 2015-11-18 | 湖南威嘉生物科技有限公司 | Process for refined production of stevioside |
| CN109563117A (en) * | 2016-08-09 | 2019-04-02 | 帝斯曼知识产权资产管理有限公司 | The crystallization of steviol glycoside |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1192447A (en) * | 1998-02-18 | 1998-09-09 | 南开大学 | Adsorption resin method for conectrating and separating vegetable baudy glucoside from stevioside |
| CN101200480A (en) * | 2006-12-15 | 2008-06-18 | 成都华高药业有限公司 | Rebaudioside A and extraction method thereof |
| CN101662960A (en) * | 2007-03-14 | 2010-03-03 | 爱尔兰浓缩加工公司 | Natural beverage products |
-
2010
- 2010-05-27 CN CN201010186528.4A patent/CN102260304B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1192447A (en) * | 1998-02-18 | 1998-09-09 | 南开大学 | Adsorption resin method for conectrating and separating vegetable baudy glucoside from stevioside |
| CN101200480A (en) * | 2006-12-15 | 2008-06-18 | 成都华高药业有限公司 | Rebaudioside A and extraction method thereof |
| CN101662960A (en) * | 2007-03-14 | 2010-03-03 | 爱尔兰浓缩加工公司 | Natural beverage products |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104548648A (en) * | 2014-10-08 | 2015-04-29 | 浙江大学 | Crystallizing device for preparing high-purity rebaudioside A by employing continuous purification, and control method of crystallizing device |
| CN104725444A (en) * | 2015-03-24 | 2015-06-24 | 杨健 | Method for efficiently preparing high-purity rebaudioside A |
| CN105061527A (en) * | 2015-08-31 | 2015-11-18 | 湖南威嘉生物科技有限公司 | Process for refined production of stevioside |
| CN109563117A (en) * | 2016-08-09 | 2019-04-02 | 帝斯曼知识产权资产管理有限公司 | The crystallization of steviol glycoside |
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