Method of extraction of alkaloids

 

(57) Abstract:

The invention relates to a method by which you can lower cost and with greater purity to obtain alkaloids from plants. Describes the way in which the alkaloid extract or fraction of this extract, he still does not have the form of an aqueous solution, dissolved in water. Set the pH of the aqueous alkaloid extract to alkaline. Then carry out the extraction liquid in a liquid with simple ether and separate the organic phase from the aqueous phase. In the organic phase is almost completely alkaloid-lupain, while in the aqueous phase are present in the original extract, alkaloids. 13 C.p. f-crystals.

The invention relates to a method of extraction of alkaloids, especially lupinine of alkaloids.steroid plant restrictive part of paragraph 1 of the claims.

The prior art is still known to produce alkaloids extraction, for example, from flour obtained from the seeds of the plants with an organic solvent. While in the literature as extractants offer simple diethyl ethers or chlorinated hydrocarbons, such as chloroform or dichloromethane. Podobna, that simple ethyl esters have the tendency to form peroxides, which required large maintenance costs and equipment costs. To prevent such explosive peroxides add stabilizers simple ether. But they pollute the extracts, which is considered a disadvantage. Therefore, the purity of the obtained alkaloids is poor.

Further, it is known, especially to reduce the bitterness of bitter Lipunov, obtaining the so-called water alkaloid extracts. Such water alkaloid extracts can be obtained in various ways. Example to obtain such water alkaloid extract disclosed in EP 0084547. In this patent the seeds of the lupine grind to flour with a specific grain size and then the flour in various technological operations is mixed with water and again separated.

Proceeding from the aforementioned prior art, the present invention is to propose ways by which you can simply, with less cost and high purity to obtain alkaloids from plants.

This problem is solved thanks to the distinctive characteristics of paragraph 1. The result is given in the dependent claims operations docno invention lies an unexpected discovery that lupain in pure form can be distinguished from simple esters from the aqueous alkaloid extract by the method of extraction liquid in the liquid.

Therefore alkaloid extract according to the invention, since it is not presented as an aqueous solution, dissolved in water. Water alkaloid extract set at the alkaline pH value. If alkaloid extract has previously been dissolved in another way, you can also use hence the resulting fraction of this alkaloid extract. Then spend named the extraction fluid in the fluid with simple ether and separate the organic phase from the aqueous phase. Now in the organic phase is almost completely alkaloid lupain, while in the aqueous phase contains other available in the original extract alkaloids.

From now presents the organic phase can be obtained respectively lupain, while the water phase can be used to obtain at least another, different from lupinine alkaloid.

A particularly large output lupinine or particularly good separation lupinine from the aqueous phase is possible at pH values in the range of 10 to 13.5 during extraction. Very good results were achieved in the woods should not be used acid-resistant materials.

As a simple ether used is preferably a simple tert.-butyl methyl ether (tBME). Special preference tBME before the usual simple diethyl esters is that this simple ether has no tendency to the formation of peroxide and as a result not a cause for concern with the technical application. Thanks to this process easier and, consequently, also more economical.

Preferably to highlight lupinine containing loanin the organic phase to evaporate at least partially, for example, in a vacuum extractor, so that the organic extract at least concentrate.

In a particularly pure form, you can get lupain from the organic phase, in which loanin or preferably narrowed organic extract is cooled until, until crystallizes loanin. In addition to the described high purity, this method provides the advantage that in addition to water and simple ether as the extracting agent does not use any other solvents to highlight lupinine. The fact that lupain can be obtained in crystalline, molecular form without the other operations directly from the extractant, is the following neozed is stylizacji lupinine recommended the addition of crystallization centers.

Already remarkably high degree of purity obtained in this way lupinine increases, and the resulting crystals are recrystallized, preferably again in tM. Obtained in this way lupain has a purity of > 98%.

As already mentioned, the aqueous phase remaining when conducted according to the method of the invention the extraction liquid in the liquid, can also be mined to get at least another lepinoy alkaloid. Here take into account, in particular, alkaloids sparteine, 13-oxylipin, 3-oxylipin, 17-oxoguanine, n-medillin, angustifolii, tetrahydrobiopterin, lupinine, multiflori, Albin, ammodendron and alkaloids of ester formed from 13 - and 3-oxylipin.

Now for the further identification of such alkaloids aqueous phase from the extraction according to the method of the invention, the liquid in the liquid set to a pH value of more than 12.5. Especially good results were achieved in the range of pH-values between 13 and 13.5. Now again conduct the extraction liquid in a liquid organic solvent. For use as solvents are particularly suitable was chlorinated hydrocarbons and particularly dichloromethane or chloroform, is here the organic phase can also separate other alkaloids, for example, chromatography, and should encourage the preceding contraction, for example, in a vacuum. For the separation we should mention especially the use of so-called instant chromatography, the application of which in this regard still was not known.

Obtaining different from lupinine alkaloid can be made through distillation. This method can be applied, in particular, directly to the aqueous phase. First of all you can select by distillation alkaloid sparteine.

As starting material for the present method is suitable all alkalotolerant plants, especially plants with alkaloids finalizada and especially their seeds. These plants are mainly in the field pea, among which should again highlight lupine, for example, Lupinus albus (white Lupin), Lupinus luteus (yellow Lupin), Lupinus mutabilis, Lupinus angustifolius (narrow-leafed Lupin), Lupinus polyphyllus (overlapping lupine), Lupinus varius, Lupinus micrantus or Luinus cosentinii. The high number lupinine contain Lupinus albus, Lupinus angustif olius and Lupinus mutabilis. Next, Lupinus luteus should mention great content sparteine. Other alkaloids found in various combinations in all lupine, primarily in the so-called bitter it must however be below 0.05 wt.%.

Rich luprinol extract as the source of the extract method according to the invention can be obtained through supercritical CO2extraction from plants, especially from the flour of the seeds of plants, which should be processed further, as described, by way of the extraction liquid in the liquid.

An example embodiment of the invention explained in detail on the basis of the following description.

As a starting point can be obtained by applying cited in the prior art EP 0084547 water alkaloid extract or above is obtained by CO2-extraction of the extract. However, the original extract can also be obtained through the following extraction flour.

Extraction flour

Flour Lupinus albus or Lupinus angustifolius is mixed with water and long mix. This establishes the equilibrium concentration of alkaloids in the flour and liquid. During the initial quantities of 10 kg of flour and 90 kg of water can be obtained after separation of the phases (for example, with the help of decanter or filtering) about 75 kg of an aqueous extract containing alkaloid about 2.5 g/l to get from here to a clear solution, the extract should is this loss solution extract or loss due to the separation of solids comprise between 15 and 25%. For extraction of the fluid in the fluid use about 60 kg of extracts containing alkaloids 2.5 g/l In the extract contains 150 g of alkaloids.

Selection lupinine

The extraction liquid in a liquid

This aqueous extract using sodium liquor set to a pH value of about 11 and extracted with simple tert.-butylmethylamine ether (tBME) in the ratio of 1:1 (volume/volume). When multistage extraction of loanin almost completely transformed into the organic phase. For a constant pH-value during the extraction of the required pH-regulation, so as to increase the extraction pH-value drops and leaves for optimal extraction lupinine range. Consequently 105 g lupinine and even the rest of 14-15 g other alkaloids can be extracted in the organic phase. The organic extract contains about 120 g of alkaloids with the number lupinine from 88 to 90%. Aqueous extract after extraction contains about 30 g of alkaloids with number 13-oxylipin above 50%.

The organic extract

The organic phase is concentrated to approximately 1/100 of its original volume (e.g., by rotary evaporation apparatus in a vacuum of about 100 mbar, T = 40oC), and obtained in this way is to -12o. At this time of the solution precipitates already white crystals lupinine. The process can be accelerated by the persecution of the cooled solution with crystal lupinine.

Obtained, for example, by decantation of the solvent, the crystals contain about 93% lupinine (94,5 g) and about 7% of impurities (about 7 g). In the organic supernatant are about 10% of the total lupinine and residual alkaloids and other substances (dyes).

Recrystallization

The separated crystals are dissolved with approximately 3 - to 5-fold amount of weight in tBME and again cooled. After a new crystallization, phase separation, simple ether and determine the purity of lupinine the latter has a purity > 98%. The total number of detachable lupinine is about 85 grams, which corresponds to a yield of about 80%.

Repetition of this procedure, you can achieve pureness of above 99%. By recrystallization or washing of the crystals losses lupinine be about 20%.

Selection 13-oxylipin

Aqueous extract after the extraction lupinine differentiates 13-oxylipin (contains about 15 g of 13-oxylipin). For this pH-value is set to >12.5 and extracted with dichloromethane in Rel is torono distilled off. The remaining oily concentrate is served on a column of silica gel.

Due to the varying means to elution with increasing polarity (toluene ---> tBME ---> CHCl3--> CH2Cl2---> CH2Cl2/MeOH) one after another wash out impurities and dyes from the column, so that, finally, with a mixture of CH2C2/methanol can be obtained relatively pure fraction 13-oxylipin.

After separation of the solvent in this way it is possible to allocate about 10 g of 13-hydroxy-lupinine as oily liquid (purity > 80%).

Accordingly it is possible to allocate the remaining alkaloids Lupino, and particularly suitable turned out to be an instant chromatography.

1. Method of extraction of alkaloids, in particular lupinine, alkaloids.steroid plants such as lupine, by simple extraction with ether, characterized in that the aqueous extract of alkaloids from alkaloids.steroid plants, parts of such plants, in particular of their seeds or flour, and bring to alkaline pH values, b) the extraction is carried out, the liquid in the liquid with a simple ester, and C) separating the organic phase from the aqueous phase.

2. The method according to p. 1, characterized in that lupain separated from the body of the 2, characterized in that the pH value under method (a) is in the region of 10 to 13.5.

4. The method according to one of paragraphs.1 to 3, characterized in that the pH value under method (a) is in the area of 10,5 - 11,5.

5. The method according to one of paragraphs.1 to 4, characterized in that a simple live on stage method b) represents a tert.-butyl methyl simple ether.

6. The method according to one of paragraphs.1 to 5, characterized in that the organic phase is at least partially evaporated.

7. The method according to one of paragraphs.1 - 6, characterized in that the organic phase is cooled at least as long until crystallizes loanin.

8. The method according to one of paragraphs.1 to 7, characterized in that the crystallization lupinine add the crystallization centers.

9. The method according to one of paragraphs.1 to 8, characterized in that lupain purified by recrystallization.

10. The method according to one of paragraphs.1 to 9, characterized in that the aqueous phase in an alkaline medium is subjected to further extraction liquid in a liquid organic solvent to highlight 13-hydroxydopamine away from the resulting organic phase.

11. The method according to p. 10, characterized in that the pH value of the extraction is boob on PP.10 - 11, wherein the organic solvent is a chlorinated hydrocarbon.

14. The method according to p. 12, characterized in that the organic solvent is dichloromethane.

 

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