CS269105B1 - A method of separating agroclavin from a mixture of keyboard alkaloids - Google Patents

Abstract

The solution concerns the isolation or removal of agroclavine from a mixture of clavine alkaloids contained in fermentation fluids, aqueous solutions or biological media. The separation is achieved by adsorption on modified silica gel at an optimum pH. The procedure can achieve high specificity of separation and thus perfect removal of agroclavine. The solution can be used in the pharmaceutical industry for industrial and analytical purposes.

Description

The invention relates to a process for the separation of agroclavine from a mixture of ergot alkaloids contained in fermentation fluids, aqueous solutions, biological fluids such as blood plasma, etc.

Ergot alkaloids are produced in saprophytic cultures of Claviceps purpurea and Claviceps paspall, Aspergilus sp., Pennicillium sizoa, Pennicillium roqueforti, etc. (M. Abe et al., Jap. Pat. 178 336, M. Abe. U.S. Pat. No. 2,835,675 , D. Grbger and VE Tylar Iv. Lhoydia 26, 174 (1963).

Piano wave alkaloids form the raw material base for the production of various semi-synthetic, therapeutically used derivatives. (Eich E., Pharmacia 30, 8, 1975, Tung-Chung Choong and H. R. Shough, Tetrahedron Lett. 19, 1627, 1977; J. Křepelka et al. Coll. Czech. Chem., Comm 42 1412 (1977).

Several inventions, for example EP 22 418, DD 146 - 620 and DT 2840 - 670, describe a process for the isolation of peptide ergot alkaloids from fermentation fluids by extraction or sorption on bentonite with activated carbon. 199 986 (Řeháček Z. et al., 1980) and author's certificate No. 329 441 (Stuchlík et al. 1984).

The first MS. The author's certificate deals with the isolation of alkaloids by extraction with an organic solvent and their separation by column chromatography on silica gel. The disadvantage of the solution is the formation of difficult-to-separate emulsions during extraction.

The second MS. The author's certificate mainly deals with the operational processing of fermentation fluids and the separation of agroclavine from elymoclavine by the extraction method. This procedure achieves a maximum alkaloid purity of about 95%.

The present invention relates in particular to the separation of agroclavine from a mixture of other more polar ergot alkaloids, in particular from mixtures in which agroclavine is present in a maximum concentration of 20% of the total alkaloid content. For the separation, specific sorption of agroclavine onto modified silica gel is used - modified by attachment of a non-polar substituent such as an alkane group containing two to eighteen carbons, a phenyl group, preferably an octadecyl group. The specific adsorption and Freundlich and Langmuir isotherm values, respectively, are given in the table.

Freundlichova isotherm Langmuirova isotherm TO N TO KS Elymoclavine 8.28635E-28 10.2111 -2.3O2O2E-O5 -218,114 Chano-aldehyde 3.1833E-31 10.5972 -3.Θ5419Ε-Ο5 -393,662 Agroclavine 4.O6993E-O4 .35359 3.16814E-O3 47.3833 Chanoclavine 3.94185E-28 10.33333 4.23O9E-O5 -892,372 Alk. total 1.65763E-13 3.17546 -1.24211E-03 -2408.48

pH of separations - 5.2

As can be seen from Table 1, the adsorption of agroclavine is many times higher than the adsorption of other more polar alkaloids. The optimum for the specific sorption of agroclavine lies in the range of pH 5 to 7. In the range of higher pH (8 to 11) the values of the coefficients of individual alkaloids of adsorption are equalized, and thus the specificity of the sorpoe of agroclavine is suppressed.

The described procedure can advantageously remove lower concentrations of agroclavine from various preparations, especially elymoclavine, preferably purification of crystalline preparations, for analytical purposes or to isolate agroclavine from highly dilute biological materials, for example when testing the effect of agroclavine and its derivatives.

Example

1 filtered fermentation broth containing 2.1 g of elymoclavin, 0.15 g of agrocla

EN 269 105 B1 wine and 0.05 g of chanoclavine were adjusted to pH 6.8 with aqueous ammonia solution. To this solution, 2 x 35 g of allicagel e bound by an octadecyl group with a particle size of 50 μm (Separon SGX C 18 Teesek Prague) was added and stirred for 15 minutes on a magnetic stirrer.

The mixture was filtered, the filter cake washed with 100 ml of distilled water and filtered off with suction. The filter cake was extracted with 3 x 150 mL of methanol containing 5% aqueous ammonia. The extracts were combined and concentrated to a residue.

The following alkaloid contents were found in the remaining fermentation fluid by high performance liquid chromatography: Elymoclavine 2.05 g / l, chanoclavine 0.05 g / l, agroclavine traces. HPLC analysis of the residue: A residue with a total weight of 0.30 g and an aspen content of alkaloids of 70%, containing 0.14 g of agroclavine, 0.05 g of elymoclavine.

Claims (1)

OBJECT OF THE INVENTION Process for the separation of agroclavine from a mixture of clavin alkaloids contained in fermentation broth, aqueous solutions, biological fluids such as blood plasma, characterized in that agroclavine is selectively adsorbed from a mixture of alkaloids on silica gel modified with phenyl, alkane group containing 2 to 18 carbons, s preferably an octadecyl group at pH 5 to 7 and released from the sorbent by desorption.