CN104710414A - Preparation method of 2,3-cis-silybin B - Google Patents
Preparation method of 2,3-cis-silybin B Download PDFInfo
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- CN104710414A CN104710414A CN201310689466.2A CN201310689466A CN104710414A CN 104710414 A CN104710414 A CN 104710414A CN 201310689466 A CN201310689466 A CN 201310689466A CN 104710414 A CN104710414 A CN 104710414A
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
- C07D407/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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Abstract
The invention relates to a preparation method of 2,3-cis-silybin B. With silybin B as a raw material, and under an alkaline condition, silybin B is subjected to catalytic conversion into low-content 2,3-cis-silybin B, industrial chromatography enrichment is adopted and combined with semi-preparative chromatographic purification and separation, and thus high-purity 2,3-cis-silybin B is obtained. The 2,3-cis-silybin B is a flavone lignin compound which naturally exists but has low content, a conventional chemical separation method is unable to prepare the high-purity compound, and the method can be used for effective separation to obtain the high-purity 2,3-cis-silybin B.
Description
Technical field
The present invention relates to the method that one prepares the cis-Silybin B of 2,3-.
Background technology
Silymarin is the medicinal plants of Compositae Silymarin seed, treatment liver and gall diseases has the history of more than 2000 year, the damage that can prevent chemical toxicant, sitotoxin and medicine etc. from causing liver, promotes hepatocellular regeneration and reparation, is called as " natural hepatoprotective "; In addition, Silymarin, as powerful antioxidant, can be removed the free radical in human body, delay senility.In recent years, its anticancer and tumor prevention effect has demonstrated good potentiality to be exploited.
Silymarin (Silmyarin) is from the seed coat of Silymarin seed, extract the flavone Lignanoids compounds obtained, it is the main active ingredient of Silymarin, comprise 10 kinds of isomeric compounds such as silibinin (Silybin), Isosilybin (Isosilybin), Silychristin (Silychristin) and Silydianin (Silydianin), modern pharmacological research proves that these compounds all have very strong pharmacologically active.The cis-Silybin B of 2,3-(2,3-cis-Silybin B) is a kind of compound in silymarin, and structure is as follows:
This compound content in plant is low, disturb by other isomeric compounds in sepn process, conventional chemical separation method cannot prepare high-purity compound, proves that it is present in Silymarin, be not separated acquisition high-purity compound so far by means of only chromatography-mass spectroscopy method.
Summary of the invention
The present invention relates to the method that one prepares the cis-Silybin B of 2,3-.Take Silybin B as raw material, after carrying out being catalytically conveted to the cis-Silybin B of low levels 2,3-in the basic conditions, adopt industrial chromatography enrichment, in conjunction with half preparative chromatography purifies and separates, obtain the cis-Silybin B of high purity 2,3-.
Concrete steps are as follows:
1) base catalysis of Silybin B transforms
Silybin B raw material mixes with alkali and carries out catalyzed reaction, and reaction terminates rear HCl neutralization reaction liquid, filters 2, the 3-cis-Silybin B conversion products that acquisition content is 1-3%.
2) richness of 2,3-cis-Silybin Bs is amassed
The cis-Silybin B conversion product of 2,3-, through industrial chromatography enrichment, obtains 2, the 3-cis-Silybin B enriched substance that content is 30-70%.
3) purifying of 2,3-cis-Silybin Bs
The cis-Silybin B enriched substance of 2,3-, through half preparative chromatography purifies and separates, obtains 2,3-cis-Silybin B products of more than 95% purity.
Described Silybin B raw material choose Silybin B monomeric compound or mixture.
Base catalysis process alkali used comprises Na
2cO
3, NaHCO
3, ammoniacal liquor, K
2cO
3, KHCO
3one or more.
When using catalyzed by solid base, Ji guest raw material and alkali mass ratio are between 1:0.5 to 1:10, and when using ammonia-catalyzed, silibinin raw material and ammoniacal liquor ratio are between 1:5 to 1:20 (g:ml).
Catalyzed reaction reflux in 40-100% methyl alcohol or ethanolic soln is carried out.
Between HCl neutralization reaction liquid to PH4-7.
Industrial chromatography uses the filling of 10-45um particle diameter C18 filler.
Half preparative chromatography uses 5-10um particle diameter C18 filler.
Industrial chromatography and half preparative chromatography all adopt methanol solution wash-out, and methanol concentration scope is between 50-70%.
Employing the present invention have the following advantages:
The cis-Silybin B of 2,3-is a kind of low levels flavones lignin compound, disturbs in sepn process by other high-content isomeric compounds, is difficult to adopt conventional separation methods to be separated.In the basic conditions, Silybin B can be converted into the cis-Silybin B of 2,3-, and content improves.The reaction conversion ratio that Silybin B alkali is converted into the cis-Silybin B of 2,3-is low, and applicable industry chromatogram carries out Fu Jihou, then through half preparative chromatography purifying, obtains the cis-Silybin B of high purity 2,3-.Compared with being separated with direct post isolation technique, after transforming, the method for preparative separation reduces sample and solvent consumption, shortens process, and method is effective fast, can be separated and obtain this material of high purity.
Accompanying drawing explanation
Fig. 1 is sample thing analysis chart
Wherein A: raw material; B: conversion product; C: enriched substance; D: product; The cis-Silybin B of 1:2,3-.
Embodiment
Now be described in further details the present invention with accompanying drawing, embodiment is only limitted to the present invention is described in conjunction with the embodiments, but not limitation of the invention.
Embodiment 1
10g silibinin mixture (Figure 1A is shown in analysis) and 5g Na2CO3 mixed dissolution are in 40% methyl alcohol, reflux is after 12 hours, and cooling, with HCl neutralization reaction liquid to pH7, filter and obtain 2, the 3-cis-Silybin B conversion products (Figure 1B is shown in analysis) that purity is 3%.Conversion product is separated through industrial chromatography post (190 × 80mm, 15um), 55% methanol-eluted fractions, collect the flow point containing target compound, concentrating under reduced pressure obtains 2, the 3-cis-Silybin B enriched substance (Fig. 1 C is shown in analysis) that purity is 70%, and enriched substance is through half preparative chromatography post (250 × 20mm, 5um) be separated, 60% methanol-eluted fractions, collects the flow point containing target compound, after concentrating under reduced pressure, obtain 2, the 3-cis-Silybin B products (Fig. 1 D is shown in analysis) that purity is 98.4%.
Embodiment 2
10g silibinin mixture and 100g NaHCO3 mixed dissolution are in 100% methyl alcohol, and reflux is after 16 hours, and cooling, with HCl neutralization reaction liquid to pH7, filters and obtain 2, the 3-cis-Silybin B conversion products that purity is 3%.Conversion product is separated through industrial chromatography post (190 × 80mm, 15um), 55% methanol-eluted fractions, collect the flow point containing target compound, concentrating under reduced pressure obtains 2, the 3-cis-Silybin B enriched substance that purity is 30%, and enriched substance is through half preparative chromatography post (250 × 20mm, 10um) be separated, 60% methanol-eluted fractions, collects the flow point containing target compound, after concentrating under reduced pressure, obtain 2, the 3-cis-Silybin B products that purity is 95.5%.
Embodiment 3
0.1g Silybin B monomeric compound and 0.5ml ammoniacal liquor mixed dissolution are in 40% ethanol, and reflux is after 24 hours, and cooling, with HCl neutralization reaction liquid to pH7, filters and obtain 2, the 3-cis-Silybin B conversion products that purity is 1%.Conversion product is separated through industrial chromatography post (240 × 100mm, 45um), 55% methanol-eluted fractions, collect the flow point containing target compound, concentrating under reduced pressure obtains 2, the 3-cis-Silybin B enriched substance that purity is 50%, and enriched substance is through half preparative chromatography post (250 × 10mm, 5um) be separated, 55% methanol-eluted fractions, collects the flow point containing target compound, after concentrating under reduced pressure, obtain 2, the 3-cis-Silybin B products that purity is 97.3%.
Embodiment 4
0.1g Silybin B monomeric compound and 2ml ammoniacal liquor mixed dissolution are in 100% methyl alcohol, and reflux is after 24 hours, and cooling, with HCl neutralization reaction liquid to pH7, filters and obtain 2, the 3-cis-Silybin B conversion products that purity is 3%.Conversion product is separated through industrial chromatography post (200 × 80mm, 45um), 50% methanol-eluted fractions, collect the flow point containing target compound, concentrating under reduced pressure obtains 2, the 3-cis-Silybin B enriched substance that purity is 70%, and enriched substance is through half preparative chromatography post (250 × 20mm, 10um) be separated, 70% methanol-eluted fractions, collects the flow point containing target compound, after concentrating under reduced pressure, obtain 2, the 3-cis-Silybin B products that purity is 98.3%.
Embodiment 5
10g silibinin mixture and 10g K
2cO
3mixed dissolution is in 40% methyl alcohol, and reflux is after 12 hours, and cooling, with HCl neutralization reaction liquid to pH6, filters and obtain 2, the 3-cis-Silybin B conversion products that purity is 3%.Conversion product is separated through industrial chromatography post (190 × 80mm, 15um), 55% methanol-eluted fractions, collect the flow point containing target compound, concentrating under reduced pressure purity is 2, the 3-cis-Silybin B enriched substance of 30%, and enriched substance is through half preparative chromatography post (250 × 20mm, 10um) be separated, 60% methanol-eluted fractions, collects the flow point containing target compound, after concentrating under reduced pressure, obtain 2, the 3-cis-Silybin B products that purity is 98.4%.
Embodiment 6
10g silibinin mixture and 30g KHCO
3mixed dissolution is in 40% methyl alcohol, and reflux is after 36 hours, and cooling, with HCl neutralization reaction liquid to pH4, filters and obtain 2, the 3-cis-Silybin B conversion products that purity is 1%.Conversion product is separated through industrial chromatography post (190 × 75mm, 10um), 55% methanol-eluted fractions, collect the flow point containing target compound, concentrating under reduced pressure obtains 2, the 3-cis-Silybin B enriched substance that purity is 30%, and enriched substance is through half preparative chromatography post (250 × 20mm, 5um) be separated, 50% methanol-eluted fractions, collects the flow point containing target compound, after concentrating under reduced pressure, obtain 2, the 3-cis-Silybin B products that purity is 98.0%.
Claims (9)
1. prepare the method for the cis-Silybin B of 2,3-for one kind, it is characterized in that:
Take Silybin B as raw material, after carrying out being catalytically conveted to the cis-Silybin B of low levels 2,3-in the basic conditions, adopt industrial chromatography enrichment, in conjunction with half preparative chromatography purifies and separates, obtain the cis-Silybin B of high purity 2,3-; Concrete steps are as follows:
1) base catalysis of Silybin B transforms
Silybin B raw material mixes with alkali and carries out catalyzed reaction, and reaction terminates rear HCl neutralization reaction liquid, filters 2, the 3-cis-Silybin B conversion products that acquisition content is 1-3%;
2) richness of 2,3-cis-Silybin Bs is amassed
The cis-Silybin B conversion product of 2,3-, through industrial chromatography enrichment, obtains 2, the 3-cis-Silybin B enriched substance that content is 30-70%;
3) purifying of 2,3-cis-Silybin Bs
The cis-Silybin B enriched substance of 2,3-, through half preparative chromatography purifies and separates, obtains 2,3-cis-Silybin B products of more than 95% purity.
2. method according to claim 1, is characterized in that: described Silybin B raw material choose Silybin B monomeric compound or mixture.
3. method according to claim 1, is characterized in that: base catalysis process alkali used comprises Na
2cO
3, NaHCO
3, ammoniacal liquor, K
2cO
3, KHCO
3one or more.
4. method according to claim 3, is characterized in that: when using catalyzed by solid base, and Ji guest raw material and alkali mass ratio are between 1:0.5 to 1:10, and when using ammonia-catalyzed, silibinin raw material and ammoniacal liquor ratio are between 1:5 to 1:20 (g:ml).
5. method according to claim 1, is characterized in that: catalyzed reaction reflux in 40-100% methyl alcohol or ethanolic soln is carried out.
6. method according to claim 1, is characterized in that: between HCl neutralization reaction liquid to PH4-7.
7. method according to claim 1, is characterized in that: industrial chromatography uses the filling of 10-45um particle diameter C18 filler.
8. method according to claim 1, is characterized in that: half preparative chromatography uses 5-10um particle diameter C18 filler.
9. method according to claim 1, is characterized in that: industrial chromatography and half preparative chromatography all adopt methanol solution wash-out, and methanol concentration scope is between 50-70%.
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| CN201310689466.2A CN104710414A (en) | 2013-12-13 | 2013-12-13 | Preparation method of 2,3-cis-silybin B |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CZ302204B6 (en) * | 2009-10-21 | 2010-12-15 | Mikrobiologický ústav AV CR, v.v.i. | Process for preparing optically pure stereomers of silybin A and silybin B |
| CN102174041A (en) * | 2011-03-24 | 2011-09-07 | 江苏中兴药业有限公司 | High-purity silymarin and preparation method thereof |
| CN102478553A (en) * | 2010-11-23 | 2012-05-30 | 天津天士力制药股份有限公司 | Method for detecting silybin |
| CN102746283A (en) * | 2011-04-20 | 2012-10-24 | 江苏天士力帝益药业有限公司 | Preparation method of high purity silybin |
-
2013
- 2013-12-13 CN CN201310689466.2A patent/CN104710414A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CZ302204B6 (en) * | 2009-10-21 | 2010-12-15 | Mikrobiologický ústav AV CR, v.v.i. | Process for preparing optically pure stereomers of silybin A and silybin B |
| CN102478553A (en) * | 2010-11-23 | 2012-05-30 | 天津天士力制药股份有限公司 | Method for detecting silybin |
| CN102174041A (en) * | 2011-03-24 | 2011-09-07 | 江苏中兴药业有限公司 | High-purity silymarin and preparation method thereof |
| CN102746283A (en) * | 2011-04-20 | 2012-10-24 | 江苏天士力帝益药业有限公司 | Preparation method of high purity silybin |
Non-Patent Citations (1)
| Title |
|---|
| 付艳辉，等: "水飞蓟宾原料药中微量成分的分离及结构鉴定", 《中药与天然药高峰论坛暨第十二届全国中药和天然药物学术研讨会论文集》 * |
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