CN103525892A - Method for quickly preparing rare panaxoside Rd - Google Patents
Method for quickly preparing rare panaxoside Rd Download PDFInfo
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- CN103525892A CN103525892A CN201310429027.8A CN201310429027A CN103525892A CN 103525892 A CN103525892 A CN 103525892A CN 201310429027 A CN201310429027 A CN 201310429027A CN 103525892 A CN103525892 A CN 103525892A
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- ginsenoside
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Abstract
The invention relates to a method for quickly preparing rare panaxoside Rd, and in particular relates to a method for degrading panaxoside Rb1 to generate panaxoside Rd. The method comprises the following steps: uniformly mixing panaxoside Rb1 liquor and an enzymic preparation under a normal temperature condition; directly introducing the mixture to a high-voltage pulsed electric field for treatment, wherein the electric field intensity is 5-20kV/cm, the number of treatment pulses is 1-10, the addition amount of the enzymic preparation is 1-4% and the adopted enzyme is commercial cellulase and beta-glucosidase which are low in price. According to the method, the high-voltage pulsed electric field is introduced in a panaxoside Rb1 enzymolytic process, so that the enzyme activity is improved, the reaction speed of degrading panaxoside Rb1 is increased, and the panaxoside Rb1 is quickly and efficiently promoted to be converted into rare panaxoside Rd. Compared with conventional conversion methods, the method is free from heating, short in preparation time, high in yield and less in energy consumption.
Description
Technical field
The present invention relates to a kind of method of preparing fast scarce ginsenoside Rd, particularly relate to pulsed electric field coupled enzymatic preparation method.
Background technology
Ginseng (Panax ginseng C.A.Meyer) is araliaceae ginseng plant, is widely used in medicine and pharmacology field.Ginsenoside, as ginseng metabolism significant secondary product, has multiple pharmacologically active.Wherein scarce ginsenoside Rd belongs to former ginseng dibasic alcohol, has neural system provide protection, anti-cancer metastasis, promotes cell proliferation of nerve cord, prevents chemicals to cause injury of the kidney and stop the effects such as vasoconstriction.
But scarce ginsenoside Rd content in ginseng is lower, and content only has some thousandths of, from ginseng, directly extraction does not obviously meet reality.Current chemical synthesising technology step is complicated, productive rate is very low, is also difficult to suitability for industrialized production.
The chemical structure of scarce ginsenoside as Rb1 compares, has just lacked one or several glycosyl with the ginsenoside that in ginseng, some content is high, therefore can obtain scarce ginsenoside by being hydrolyzed the glycosyl of some high-content ginsenoside.At present, the method for ginsenoside glycosyl hydrolase mainly contains chemical transformation and biotransformation method.Wherein chemical transformation reaction comprises acid hydrolysis and basic hydrolysis, but exists specificity poor, the shortcoming that by product is many, productive rate is low; Biotransformation method comprises microbial transformation and enzymatic conversion method, but reaction time is long, complex process.The inventive method has mild condition, does not destroy saponin structure, operation steps is simple, selectivity is high, yield advantages of higher, and the enzyme adopting is cheap commercialization enzyme.
Summary of the invention
The object of the invention is to propose a kind of method that pulsed electric field and enzyme coupling degraded ginsenoside Rb1 prepare scarce ginsenoside Rd, to overcome current chemical method and biological process degraded ginsenoside Rb1's shortcoming.
The present invention relates to a kind of method that ginsenoside Rb1 of degraded prepares scarce ginsenoside Rd fast, utilize high-voltage pulse electric field technology coupling enzyme preparation degrades ginsenoside Rb1 to prepare scarce ginsenoside Rd, at normal temperatures by ginsenoside Rb1's sample dissolution in buffered soln, add zymin to mix, directly passing into pulsed electric field processes, its strength of electric field is 5-20kV/cm, the umber of pulse of processing is 1-10, material after pulsed electric field is processed adds organic solvent extraction, centrifugation, get supernatant liquor, solvent is removed in volatilization, obtain ginsenoside Rd.
Described zymin is cellulase or beta-glucosidase.
Described zymin addition is 1-4%.
Described buffered soln is used Acetate-acetate buffer solution.
Water saturated solution when described organic solvent is used propyl carbinol.
Present method, owing to introducing pulsed electric field in enzyme digestion reaction process, improves enzymic activity, accelerates enzyme digestion reaction speed, promotes fast and efficiently ginsenoside Rb1 to transform and generates scarce ginsenoside Rd.
The present invention is high-voltage pulse electric field technology and the commercialization zymin rare ginseng ginsenoside Rd of preparation that is coupled, have non-thermal, speed of response is fast, by product is few, low cost and other advantages, for ginsenoside structural modification provides novel method.
Accompanying drawing explanation
Fig. 1 ginsenoside Rb1 degrades and generates the reaction path of ginsenoside Rd
Embodiment
In conjunction with the embodiment providing below, the inventive method is described in further detail.
Embodiment 1
The present invention degrades ginsenoside Rb1 to generate the method for ginsenoside Rd, be to add 3% beta-glucosidase in 1mM ginsenoside Rb1 solution, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 15kV/cm, and umber of pulse is 8; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, through the yield of efficient liquid phase chromatographic analysis ginsenoside Rd, is 91.42%.
Embodiment 2
Ginsenoside Rb1 is degraded and generates the method for ginsenoside Rd, be to add 3% beta-glucosidase in 1mM ginsenoside Rb1 solution, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 10kV/cm, and umber of pulse is 10; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, through the yield of efficient liquid phase chromatographic analysis ginsenoside Rd, is 70.35%.
Embodiment 3
Ginsenoside Rb1 is degraded and generates the method for ginsenoside Rd, is to add 3% beta-glucosidase in 1mM ginsenoside Rb1 solution, mixes, and directly passes at normal temperatures pulsed electric field and processes, and its strength of electric field is 20kV/cm, and umber of pulse is 6; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, through the yield of efficient liquid phase chromatographic analysis ginsenoside Rd, is 44.52%.
Embodiment 4
Ginsenoside Rb1 is degraded and generates the method for ginsenoside Rd, is to add 4% beta-glucosidase in 1mM ginsenoside Rb1 solution, mixes, and directly passes at normal temperatures pulsed electric field and processes, and its strength of electric field is 20kV/cm, and umber of pulse is 8; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, through the yield of efficient liquid phase chromatographic analysis ginsenoside Rd, is 56.29%.
Embodiment 5
Ginsenoside Rb1 is degraded and generates the method for ginsenoside Rd, be in 1mM ginsenoside Rb1 solution, to add 2% cellulase Cellulase1.5L FG, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 15kV/cm, and umber of pulse is 10; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, through the yield of efficient liquid phase chromatographic analysis ginsenoside Rd, is 51.06%.
Embodiment 6
Ginsenoside Rb1 is degraded and generates the method for ginsenoside Rd, in 1mM ginsenoside Rb1 solution, to add 2% cellulase Cellulase(Aspergillus niger), mix, directly passing at normal temperatures pulsed electric field processes, its strength of electric field is 15kV/cm, and umber of pulse is 10; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, through the yield of efficient liquid phase chromatographic analysis ginsenoside Rd, is 46.81%.
Claims (10)
1. prepare fast a method of scarce ginsenoside Rd, utilize high-voltage pulse electric field technology coupling enzyme preparation degrades ginsenoside Rb1 to prepare scarce ginsenoside Rd, it is characterized in that:
At normal temperatures by ginsenoside Rb1's sample dissolution in buffered soln, add zymin to mix, directly passing into pulsed electric field processes, its strength of electric field is 5-20kV/cm, and the umber of pulse of processing is 1-10, and the material after pulsed electric field is processed adds organic solvent extraction, centrifugation, get supernatant liquor, solvent is removed in volatilization, obtains ginsenoside Rd.
2. a kind of method of preparing fast scarce ginsenoside Rd according to claim 1, is characterized in that:
Described zymin is cellulase or beta-glucosidase, and addition is 1-4%.
3. a kind of method of preparing fast scarce ginsenoside Rd according to claim 1, is characterized in that:
Described buffered soln is used Acetate-acetate buffer solution.
4. a kind of method of preparing fast scarce ginsenoside Rd according to claim 1, is characterized in that:
Water saturated solution when described organic solvent is used propyl carbinol.
5. according to a kind of method of preparing fast scarce ginsenoside Rd described in claim 1 to 4 any one, it is characterized in that:
In 1mM ginsenoside Rb1 solution, add 3% beta-glucosidase, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 15kV/cm, and umber of pulse is 8; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, and the yield of ginsenoside Rd is 91.42%.
6. according to a kind of method of preparing fast scarce ginsenoside Rd described in claim 1 to 4 any one, it is characterized in that:
In 1mM ginsenoside Rb1 solution, add 3% beta-glucosidase, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 10kV/cm, and umber of pulse is 10; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, and the yield of ginsenoside Rd is 70.35%.
7. according to a kind of method of preparing fast scarce ginsenoside Rd described in claim 1 to 4 any one, it is characterized in that:
In 1mM ginsenoside Rb1 solution, add 3% beta-glucosidase, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 20kV/cm, and umber of pulse is 6; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, and the yield of ginsenoside Rd is 44.52%.
8. according to a kind of method of preparing fast scarce ginsenoside Rd described in claim 1 to 4 any one, it is characterized in that:
In 1mM ginsenoside Rb1 solution, add 4% beta-glucosidase, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 20kV/cm, and umber of pulse is 8; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, and the yield of ginsenoside Rd is 56.29%.
9. according to a kind of method of preparing fast scarce ginsenoside Rd described in claim 1 to 4 any one, it is characterized in that:
In 1mM ginsenoside Rb1 solution, add 2% cellulase Cellulase1.5L FG, mix, directly pass at normal temperatures pulsed electric field and process, its strength of electric field is 15kV/cm, and umber of pulse is 10; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, and the yield of ginsenoside Rd is 51.06%.
10. according to a kind of method of preparing fast scarce ginsenoside Rd described in claim 1 to 4 any one, it is characterized in that:
In 1mM ginsenoside Rb1 solution, add 2% cellulase Cellulase, Aspergillus niger, mixes, and directly passes at normal temperatures pulsed electric field and processes, and its strength of electric field is 15kV/cm, and umber of pulse is 10; With water saturation n-butanol extraction reaction solution, centrifugation, supernatant liquor volatilization is except desolventizing, and the yield of ginsenoside Rd is 46.81%.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104398549A (en) * | 2014-11-11 | 2015-03-11 | 长春师范大学 | Method for preparation of total saponins of panax ginseng by high-voltage pulse electric field assisted enzyme hydrolysis |
| CN106900856A (en) * | 2017-03-07 | 2017-06-30 | 吉林大学 | The balloonflower root fermented ginseng Yoghourt and its production method of a kind of green |
| CN108138210A (en) * | 2015-10-22 | 2018-06-08 | 株式会社爱茉莉太平洋 | The method for selectively preparing ginsenoside Rd from the saponin of ginseng using enzyme process |
| CN111388491A (en) * | 2020-05-08 | 2020-07-10 | 何莹懿 | Rare monomer ginsenoside composition and its preparation method |
| CN113005157A (en) * | 2021-03-29 | 2021-06-22 | 吉林大学 | Preparation method of soybean isoflavone aglycone |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86105166A (en) * | 1985-07-22 | 1987-03-04 | 武田药品工业株式会社 | Produce a kind of method of ginsenoside-rd |
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2013
- 2013-09-20 CN CN201310429027.8A patent/CN103525892A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86105166A (en) * | 1985-07-22 | 1987-03-04 | 武田药品工业株式会社 | Produce a kind of method of ginsenoside-rd |
Non-Patent Citations (1)
| Title |
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| 吴建生,等: "高压电场对茶叶β-糖苷酶及细胞膜的影响", 《西南农业大学学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104398549A (en) * | 2014-11-11 | 2015-03-11 | 长春师范大学 | Method for preparation of total saponins of panax ginseng by high-voltage pulse electric field assisted enzyme hydrolysis |
| CN104398549B (en) * | 2014-11-11 | 2018-03-16 | 长春师范大学 | The method that high-pressure pulse electric auxiliary enzyme hydrolysis prepares general ginsenoside |
| CN108138210A (en) * | 2015-10-22 | 2018-06-08 | 株式会社爱茉莉太平洋 | The method for selectively preparing ginsenoside Rd from the saponin of ginseng using enzyme process |
| CN106900856A (en) * | 2017-03-07 | 2017-06-30 | 吉林大学 | The balloonflower root fermented ginseng Yoghourt and its production method of a kind of green |
| CN111388491A (en) * | 2020-05-08 | 2020-07-10 | 何莹懿 | Rare monomer ginsenoside composition and its preparation method |
| CN113005157A (en) * | 2021-03-29 | 2021-06-22 | 吉林大学 | Preparation method of soybean isoflavone aglycone |
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Application publication date: 20140122 |