Method ergot alkaloid extraction from ergot

FIELD: chemistry.

SUBSTANCE: ergot alkaloid is extracted from ergot with high yield and purity level using method including extraction of Claviceps purpurea, i.e. ergot, mix of solvents toluene/ethanol that leads to production of primary extract. The primary extract can be additionally treated within two stages of liquid-liquid extraction to provide alkaloid purification that gives purified toluene extract. Toluene extract can be additionally partially softened by stream, and crystalline product is produced by toluene crystallisation or mixture of toluene and aliphatic hydrocarbon.

EFFECT: development of method of ergot extraction with high yield and purity level.

23 cl, 3 ex

 

The prior art TO WHICH the INVENTION RELATES.

The present invention relates to a method of extraction and purification of alkaloids of ergot, in particular extraction and purification of peptide alkaloids of ergot fungus Claviceps purpurea.

RATIONALE INVENTIONS

Peptide ergot alkaloids, also called ergopeptine are natural products used to obtain drugs. It is known that ergopeptine have a healing themselves (eg, ergotamine) or hydrated form, such as dihydrobromide, for example digidroergotamin, dihydroergocristine and so on. In addition, these ergopeptine known as starting compounds for the partial synthesis of some semi-synthetic drugs such as enicillin, pergolid etc.

The alkaloids of ergot peptide of the type produced by the fungus Claviceps purpurea, which can be grown in parasitic conditions (growing in the fields using rye as the host plant) or saprophytic conditions (i.e. fermentation). Although the methods used to allocate ergopeptine field of LPV and fermentative broth, have common characteristics (i.e. similar solvents and cleaning methods), these methods differ widely depending on the nature of the source material.

Some with the person extraction of ergot have been described previously. Separate ways, mainly used for separation of ergot alkaloids on an industrial scale, used by different solvents. For example, in these known methods used aqueous ethanol or methanol (see DP 47315 and DP 697760), more recent methods have used chlorinated hydrocarbons (see DE 2113281, DE 2637764 and DD programs 10059), diethyl ether (see CS 264880 and CS 264881), acetone (see DE 1695986), methylisobutylketone (see BE 891421) or ethyl acetate (see DE 2949593 and EP 22418). Some of these solvents, however, is currently not acceptable for large-scale method for security reasons, and ecology (e.g., diethyl ether and chlorinated hydrocarbons). In addition, some of these solvents are not sufficiently selective for the synthesis of alkaloids of the ergot of a purity suitable for practical realization of production, such as aqueous methanol, aqueous ethanol and acetone.

The LPV also contains up to 30% oil and other lipids. Previous methods that used solvents, extracting other components of ergot, mainly oil, require surgery for the separation of alkaloids from the surrounding lipids. However, not all known solvents suitable for the immediate removal of lipids by liquid-liquid extraction, and therefore the extraction procedure, including to the ncentratio primary extracts by evaporation and dissolution of the residue in another solvent, additionally complicated way. In addition, evaporation of the primary extract containing oil and other ballast components, dangerous for the extracted alkaloids.

Natural ergopeptine are derivatives of lysergic acid diethylamide and easily isomerized to derivatives isolysergic acid, the so-called ergopeptine. This circumstance usually complicates the selection ergopeptine because the primary extracts derived from ergot, always contain a mixture of ergopeptine and ergopeptine, making it difficult to obtain a crystalline product, thereby eliminating the techniques of crystallization, which in other cases are effective ways of cleaning. Thus, there remains a need for methods of large-scale allocation of ergot alkaloids, based on the use of safe and environmentally friendly solvents and simple and effective cleanup operations (e.g., crystallization).

The INVENTION

The present invention relates to a new method of extraction of ergot alkaloids from ergot.

The presented method is a new method of purification of the extract of ergot, leading to the obtaining of ergot alkaloids with high yield and quality.

These and other advantages will become apparent from the following detailed described what I were achieved thanks to made by the applicants discovery that the LPV can be extracted with a high yield of a mixture of toluene and ethanol. Additional advantages include, but are not limited to, the ability to purify the extract of ergot using a relatively safe solvent and this relatively simple method according to the present invention can be brought to an industrial scale.

DETAILED description of the INVENTION

The present invention concerns the allocation of ergot alkaloids from ergot, i.e. Claviceps purpura, by a simple efficient method for the extraction with the use of relatively safe and environmentally solvents. One of the embodiments of the present invention the LPV extracted with a mixture of toluene/ethanol, getting the primary extract. This primary extract may be further subjected to liquid-liquid extraction for further separation and purification of ergot alkaloids obtained in the primary extract.

Preferably, the extraction mixture of toluene/ethanol contained about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 up to 30% ethanol (about/on). If the ethanol concentration is below 5%, the extraction of ergot alkaloids from ergot is not as effective. The upper limit of the amount of ethane is l based on a given selectivity of extraction. For example, the more ethanol is present, the more polar the ballast component is extracted, and the more difficult the further processing of the primary extract. More preferably, in the extraction mixture was attended by 10-20% ethanol (o/o).

The temperature of the extraction process preferably is not limited, since it is usually undesirable isomerization ergopeptine, which may occur at high temperatures, does not interfere with the crystallization of the desired product. However, temperatures above about 50°C is not feasible. In a preferred embodiment, the LPV extracted with a mixture of toluene/ethanol in the range of temperatures from about 20, 25, 30, 35, 40, 45 up to 50°C, while the ambient temperature is preferred.

In another variant implementation of the primary extract is subjected to liquid-liquid extraction using an aqueous acid solution. Such liquid-liquid extraction of the primary extract allows you to separate the generally more polar and hydrophilic alkaloids from the less polar hydrophobic oils and lipids. Thus, liquid-liquid extraction of the primary extract with an aqueous solution containing the acid, it leads to the transfer of ergot alkaloids in aqueous solution, leaving the oil in the primary extract. Polucen is th at this aqueous extract, containing alkaloids, can then be easily separated from the primary extract and further facilitates the isolation and purification of alkaloids. Any acid may be used with a water solution. Hydrochloric acid is preferred because of the high solubility of hydrochloride ergopeptine in aqueous solutions of hydrochloric acid.

According to another variant implementation of the water solution can be added alcohol to prevent or reduce the formation of emulsions. It is established that under certain conditions an aqueous solution can form an emulsion, which reduces the possibility of the release of alkaloids. Thus, inhibiting the formation of emulsions amounts of alcohol or similarly acting solvent can be added to the aqueous solution to improve the results. For example, ethanol may be added to aqueous solution containing hydrochloric acid used for extraction of alkaloids from the primary extract. The concentration of acid in aqueous solution is not critical, but preferably the solution contains at least one equivalent of acid for quantitative extraction of alkaloids from the primary extract. For example, an aqueous acid solution preferably contains from about 30% to 60% (o/o) water, from about 70% is about 40% (about/on) ethanol and about from 0.05% to 1.0% (m/m) acid. The aqueous acid solution preferably contains about 40-50% (o/o) of water, about 60-50% (o/o) of ethanol and approximately 0.1% to 0.3% (m/m) acid. Even more preferably the aqueous acid solution contains about 50% (V/o) water, about 50% (V/o) ethanol and about 0.2% (m/m) acid or 40% (V/o) water, about 60% (o/o) ethanol and about 0.2% (m/m) acid.

In another variant implementation of the resulting aqueous extract is alkalinized (e.g., pH>7,0)that facilitates the transition of the alkaloid in an organic solvent while the other liquid-liquid extraction. This can be accomplished using any aqueous solution of alkali, as, for example, using an aqueous solution of sodium hydroxide, more preferably 5% aqueous sodium hydroxide (m/m).

In another variant implementation of the aqueous extract, after increasing alkalinity over 7,0, is subjected to liquid-liquid extraction with toluene. Toluene extract obtained at this stage liquid-liquid extraction, contains in fact only ergot alkaloids and therefore can be called purified toluene extract.

In another variant implementation of purified toluene extract is partially evaporated. It is assumed that such partial evaporation causes the formation of a crystalline product. Found that some of the alkaloids, and what about the ergotamine and ergocristine, can be obtained as crystalline products simply by evaporation of toluene extract. The fact that the organic solvent may contain a mixture of ergopeptine and ergopeptine no negative impact on the crystallization of these products because it is made of a crystalline mixture of the corresponding ergopeptine and ergopeptine, for example a mixture of ergotamine and ergotamine or a mixture ergokristina and ergocristine. Without going into theory, it is believed that the methods of extraction and purification of the present invention lead to the selection of ergot alkaloids, essentially, in pure form and without inhibiting the crystallization of impurities. Thus, even when there is a mixture of the extracted alkaloids, crystalline product can be obtained after partial evaporation of the solvent. In addition, the possible transformation of ergot alkaloids, such as isomerization of ergot alkaloids, which may be caused by high temperature, apparently, does not affect the isolation and crystallization of the products obtained after evaporation of the solvent. Therefore, extraction at high temperatures does not adversely affect this method. Crystallization of alkaloids from toluene unexpectedly gave products of high purity, as illustrated in examples 1 and 2 by composition Alka is oidov first and second charges.

It is established that under certain conditions toluene extract may contain a mixture of alkaloids, such as a mixture of alkaloids ergotoksina, which limits the ability of these alkaloids to crystallization. Alkaloid composition of toluene extract is responsible spectrum of alkaloids that occur when using a strain of ergot. When this mixture of alkaloids ergotoksina present in the toluene extract directly from toluene can be obtained a small amount of crystalline product, or cannot be obtained at all. In another variant implementation of the present invention overcome this limitation by adding to the solution one or more aliphatic hydrocarbons, such as C5-C8-hydrocarbon, such as hexane or heptane. Hexane is the preferred hydrocarbon. This technique of crystallization does not affect the alkaloid composition, but may still cause the crystalline product is free from ballast components and convenient for further use, as shown in example 3.

Other characteristic features of the invention will become apparent from the subsequent descriptions of typical embodiments shown in order to illustrate the invention and are not to be considered as limiting examples.

EXAMPLES

Example 1/p>

About 20,000 kg LPV (strain ergokristina 130 GAL) extracted under conditions of countercurrent using a mixture of toluene and ethanol 87:13 (o/o) on the extractor continuous rotary type. Get about 64 m3the primary extract. Then the primary extract is extracted by the extractor continuous Westfalia using a mixture of ethanol and water 1:1 (on/about)containing 0.2% (m/m) of hydrogen chloride. Formed about 28 m3aqueous extract. The aqueous extract is alkalinized to approximately pH 7.3 with 5% (m/m) aqueous solution of sodium hydroxide and extracted with toluene extractor continuous Westfalia. Get about 16 m3toluene extract. Toluene extract is evaporated to approximately 1000 kg, the resulting crystalline product is separated by filtration, washed with toluene and dried for 3 hours in a vacuum dryer at 60°C and 50 mbar, getting 152 kg first collection of raw ergokristina. The mother liquor evaporated to approximately 400 kg and add 1500 l technical hexane. The precipitated crystalline product is separated by filtration, washed with technical hexane and dried, receiving 89 kg second collection of raw ergokristina.

Analytical results

First sessionThe second collection
Analysis by titration93.1%of92,6%
Ergocristine30,9%20,0%
Ergocristine59,3%36,6%
α-ergocryptine2,4%10,5%
α-ergocryptine3,9%17,2%
The sum of the other alkaloids2,6%15,7%

Example 2

About 20,000 kg LPV (strain ergotamine GAL 404) extracted under conditions of countercurrent using a mixture of toluene and ethanol 84:16 (o/o) on the extractor continuous carousel and get 78 m3the primary extract. The primary extract is extracted by the extractor continuous Westfalia using a mixture of ethanol and water 6:4 (o/o), containing 0.2% (m/m) of hydrogen chloride, and get 18 m3aqueous extract. The aqueous extract is alkalinized, increasing pH to 7.3 by adding 5% (m/m) aqueous sodium hydroxide. Then extracted with toluene extractor continuous Westfalia. Get up to 30 m3toluene extract. Toluene extract is evaporated to approximately 1000 kg, the resulting crystalline product is separated by filtration, washed with toluene and dried for 3 hours in a vacuum dryer at 60°C and 50 mbar, getting about 181 kg PE the first collection of raw ergotamine. The mother liquor evaporated to approximately 100 kg, the crystalline product is separated by filtration, washed with toluene and dried, obtaining 19 kg of the second collection of raw ergotamine.

Analytical results

First sessionThe second collection
Analysis by titration95,0%90,6%
Ergotamine30,9%20,0%
Ergotamine59,3%36,6%
The sum of the other alkaloids3,6%17,0%

Example 3

About 20,000 kg LPV (strain ergotoksina GAL 310) extracted under conditions of countercurrent using a mixture of toluene and ethanol 87:13 (o/o) on the extractor continuous rotary type. Get about 69 m3the primary extract. The primary extract is extracted by the extractor continuous Westfalia using a mixture of ethanol and water 1:1 (on/about)containing 0.2% (m/m) of hydrogen chloride, receiving 27 m3aqueous extract. The aqueous extract is alkalinized, increasing the pH of an aqueous extract of approximately $ 7.3 with 5% (m/m) aqueous sodium hydroxide. Then extracted with toluene extractor continuous Westfalia, getting about 17 m3toluene extract. Toluols the second extract is evaporated to approximately 800 kg and add 1800 l technical hexane. The precipitated crystalline product is separated by filtration, washed with technical hexane and dried for 3 hours in a vacuum dryer at 60°C and 50 mbar, getting 151 kg of raw ergotoxine.

Analytical results

Analysis by titration94,4%
Ergocornine18,1%
Ergocornine29,2%
α-ergocryptine11,7%
α-ergocryptine18,5%
β-ergocryptine6,3%
β-ergocryptine11,6%
The sum of the other alkaloids4,6%

This description is only the preferred embodiments of the invention and are just a few examples that reflect the possible diversity. It is obvious that the invention is applicable to various other combinations and conditions and acceptable changes or modifications within the scope of the concept of the invention is given below.

1. Extraction of the ergot alkaloid of ergot, including extraction of ergot using a mixture containing toluene and ethanol, to obtain a primary extract.

2. The method according to claim 1, where the mixture contains toluene and about 5-30% (o/o) of ethanol.

3. The method according to claim 2, where the mixture will win toluene and about 10-20% (o/o) of ethanol.

4. The method according to claim 2, where the extraction is carried out at a temperature of about 20-50°C.

5. The method according to claim 4, where the extraction is carried out at ambient temperature.

6. The method according to claim 2, where the extraction is carried out counter-flow method on the battery percolation or extractor continuous action.

7. The method according to claim 2, additionally including extraction of the primary extract in an aqueous solution of acid to transfer the ergot alkaloid from the primary extract in aqueous extract.

8. The method according to claim 7, where the aqueous acid solution is an aqueous solution of hydrochloric acid.

9. The method of claim 8, where an aqueous solution of hydrochloric acid contains about 30-60% (o/o) water, about 70 pass within 40% (o/o) ethanol, and about 0.05 to 1.0% (m/m)model HC1.

10. The method according to claim 9, where an aqueous solution of hydrochloric acid contains about 40-50% (o/o) water, about 60-50% (o/o) ethanol, and about 0.1 to 0.3%(m/m)HCl.

11. The method of claim 8, further comprising increasing the pH of an aqueous extract to a value of more than 7.0.

12. The method according to claim 11, where the increase is realized by the addition of an aqueous sodium hydroxide solution (m/m).

13. The method according to item 12, where the increase carried out by adding 5%aqueous sodium hydroxide solution (m/m).

14. The method according to claim 11, further comprising extraction of the aqueous extract with a pH above 7.0 and toluene to transfer alkaloid sproingies aqueous solution and the production of purified toluene extract.

15. The method according to 14, further comprising a partial evaporation of the solvent from the purified toluene extract to obtain a crystalline alkaloid of ergot.

16. The method according to clause 15, further comprising separating the crystalline alkaloid of ergot from residual toluene.

17. The method according to clause 15, further comprising adding one or more C5-C8-aliphatic hydrocarbons to concentrate after partial evaporation of the toluene, which promotes crystallization of the ergot alkaloid.

18. The method according to 17, where one or more aliphatic C5-C8-hydrocarbon is selected from hexane and heptane.

19. The method according to p, where one or more aliphatic C5-C8-hydrocarbons is a hexane.

20. The method according to 17, further comprising separating the crystalline alkaloid of ergot from a mixture of toluene/aliphatic hydrocarbon.

21. The method according to claim 20, including the allocation of crystalline alkaloid of ergot with a purity of over 90%.

22. The allocation method of the ergot alkaloid of ergot, including:

A. extraction of ergot a mixture containing toluene and ethanol, to obtain a primary extract, where the mixture comprises toluene and about 5-30% (o/o) ethanol;

b. extraction of the primary extract in an aqueous solution of acid to transfer the ergot alkaloid and is the primary extract in an aqueous extract;

C. increase the pH of an aqueous extract to a value of over 7.0;

d. extraction of the aqueous extract with a pH above 7.0 and toluene to transfer the ergot alkaloid from the aqueous solution and the production of purified toluene extract;

E. partial evaporation of the solvent from the purified toluene extract in order to obtain a crystalline alkaloid of ergot and

f. Department of the ergot alkaloid from residual toluene.

23. The allocation method of the ergot alkaloid of ergot, including:

A. extraction of ergot a mixture containing toluene and ethanol, to obtain a primary extract, where the mixture comprises toluene and about 5-30% (o/o) ethanol;

b. extraction of the primary extract in an aqueous solution of acid to transfer the ergot alkaloid from the primary extract in an aqueous extract;

C. increase the pH of an aqueous extract to a value of over 7.0;

d. extraction of the aqueous extract with a pH above 7.0 and toluene to transfer the ergot alkaloid from the aqueous solution and the production of purified toluene extract;

E. partial evaporation of the solvent from the purified toluene extract in order to obtain a crystalline alkaloid of ergot;

f. adding one or more C5-C8-aliphatic hydrocarbons to concentrate after partial evaporation of the toluene which promotes crystallization of the ergot alkaloid, and

g. the selection of crystalline alkaloid of ergot from a mixture of toluene/aliphatic hydrocarbon.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: ergot alkaloid is extracted from ergot with high yield and purity level using method including extraction of Claviceps purpurea, i.e. ergot, mix of solvents toluene/ethanol that leads to production of primary extract. The primary extract can be additionally treated within two stages of liquid-liquid extraction to provide alkaloid purification that gives purified toluene extract. Toluene extract can be additionally partially softened by stream, and crystalline product is produced by toluene crystallisation or mixture of toluene and aliphatic hydrocarbon.

EFFECT: development of method of ergot extraction with high yield and purity level.

23 cl, 3 ex

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