CS240315B1 - The method of isolation of D-volemitol from primrose spring - Google Patents

Abstract

The solution concerns a method of isolating D-volemitol from primrose, Primula officinalis. The essence of the invention lies in the extraction of the above-ground part of primrose 3 to 7% by weight with an aqueous solution of formic acid at a temperature of 75 to 95 °C for 15 to 25 hours and from the extract the protein part is precipitated with 50% by volume, ethanol, fermentable carbohydrates are fermented by fermenting the solution with baker's yeast and from the residue D-volemitol is crystallized from a methanol-aqueous solution. D-volemitol is used as a standard in studies of carbohydrates in natural material and as a starting compound in biochemical syntheses.

Description

The present invention relates to a method of isolating D-volemitol from Primula officinalis. SUMMARY OF THE INVENTION The invention consists in extracting the above-ground part of the primrose spring 3 to 7% by weight. aqueous solution of formic acid at 75-95 ° ^C for 15-25 hours and the protein portion is precipitated with 50% by volume, ethanol, quaternary carbohydrates are fermented by fermentation of the solution with baker's yeast and crystallized from the residue D-volemitol from methanol-aqueous solution. D-volemitol is used as a standard in carbohydrate studies in natural materials and as a starting compound in biochemical syntheses.

The invention relates to a method for the isolation of D vole mitol from spring primrose (Prinmhis offieinalis).

In the natural material D-volemitol (D-glycero-D-taloheptitol, D glycero-D-iuanoheptitol) has been found and isolated in many cases. It was found in all parts of the plant in 90 species of primrose [B. Kremer: Z. Pflanzenphysiol. 86, 453 (1978)] and isolated from the above-ground part of the primrose by spring extraction with water in an amount of 1.7 wt. [R. Begbie, N.K. Richtmyer: Carbohydr. Res. 2, 272 (1966)]. Also found in various lichen species (Dermatocarpon lachneum, D. miniatum, D. fluviatile, Endacarpon adscendens) [B. Lindberg, A. Misiorny, C.A. Wachtmeister: Acta Chem. Scand. 7, 591 (1955)] and isolated from D. miniatum in an amount of 4 wt. [B. Lindberg, Acta Chem. Scand. 9, 917 (1955)], from D. moulinsii in an amount of 0.01 wt. [J. S. Rao, K.G. Sarma, T. R. Seshari: Indian J. Chem. 5, 177 (1967)]. D-volemitol is found in the three-lobed liverwort (Plagiochila asplenioides) [A. Suleiman A.A. Bacon J., Christie A., Lewis D. H. New. Phytologist. 82, 439 (1979); A. A. A. Suleiman, D. H. Lewis: New Phytol. 84 45 (1980)], in stonecrop (Sedum) [N. K. Richtmyer, Carbohyd. Res. 12, 139 (1970)] and in avocado seeds [N. K. Richtmyer, Carbohyd. Res. 12, 135 (1970)]. D-volemitol was isolated from 0.01% by weight of red algae (Pophyra ummbilicalis). [B. Lindberg: Acta Chem. Scand. 9, 1097 (1955)] and from algae (Pelvetia canaliculata) in an amount of 0.7 wt. and simultaneously mono- and di-O-D-glucosyl derivatives of D-volemitol with D-glucose binding + glycosidic bond to the primary hydroxyl groups of D-volemitol [B. Lindberg, J. Paju: Acta Chem. Scand. 8, 817 (1954)]. When cultivated with yeast Torulopsis versatilis on lactose, D-volemitol is produced in an amount of 1.1-3.3% by weight. [H. Onichi, Μ. B. Lips: Can. J. Microbiol. 18, 925 (1972)], D-volemitol is obtained from natural material by extraction of individual plant parts with methanol, optionally ethanol, water, or acetone.

SUMMARY OF THE INVENTION The invention is based on the extraction of the above-ground part of the primrose 3 to 7% by weight. aqueous solution of glacial acid at 75-95 ° C for 15-25 hours and the protein part is removed from the extract by precipitation of 50% by volume, ethanol, fermentable carbohydrates by fermentation with baker's yeast solution and crystalline D-volemitol is obtained from the residue. .

The advantage of the proposed method of isolation of D-volemitol from spring primrose is that the technical implementation is simple and its yield is substantially higher. Another advantage is that the isolation technique can be applied to isolate D-volemitol from other plant species, respectively. parts of plants.

Ex

The air - dried aerial part of primrose harvested at flowering time (400 g) is macerated in a 7% by weight aqueous formic acid solution (4 L) for 10 hours at 75 ° C, then the drug is filtered off and gently heated for 7 hours. The combined filtrates were washed with charcoal and the solution was concentrated to dryness. The distillation residue was dissolved in water (1 L) and after the addition of 96% by volume of ethanol (11) the solution was filtered and concentrated. the residue is dissolved in drinking water (2 L) and after addition of baker's yeast (10 g) the solution is fermented until complete removal of D-glucose (2 days) .The solution is filtered, concentrated slightly (to ca 300 ml) and deionized on columns (with a diameter of 3.2 cm and a length of 40 cm) respectively an ion exchanger with functional sulphate groups (Ostion KS 0210) and an ion exchanger with quaternary groups (Ostion AT 0209) in the carbonate cycle. 70 ml) ap at 60 ° C, methanol (210 mL) was added to the solution, and then the solution was allowed to crystallize at 20-25 ° C (2 days) to give a first crystalline fraction of D-volemitol (8.2, i.e. 2.1). % wt. calculated on dry drug). The mother liquor is concentrated to dryness, the residue is dissolved in water (20 ml) and, after addition of methanol (80 ml), a second crop of D-volemitol (4.6 g, i.e. 1.1% by weight) is obtained by crystallization. From the mother liquor, chromatographic fractionation on an ion exchanger (3.5 cm diameter and 120 cm length, Dowex 50 W, X-8, 0.07-0.15 mm particle size) in a barium cycle eluted with water (30 mL flow rate). h ^-1 ) from the fraction containing D-volemitol (elution volume 540-700 ml) by crystallization a third crop (1.2 g, m.p. 0.3% by weight) of D-volemitol was obtained by crystallization. The total yield of crystalline D-volemitol is 3.5% by weight. calculated on the dry mass of spring primrose.

Recrystallized D-volemitol from methanol-water has a melting point of 151-151.5 ° C (Kofler) and / a / _D ²⁰ + 1.5 + 0.2 ° (c 2, water), w / d ²⁰ + 54 + ¹⁰ (c 2, 4% by weight aqueous ammonium molybdate solution, pH 5.9), [α] ²⁰ d + 100 + 1 ° (c 2, 4% acidic solution by weight ammonium molybdate, pH- 1.9). The literature gives for D-volemitol a melting point of 153 ° C, [α] D ²⁰ + 2.07 ° (c 3, water) [V. Ettel Collection Czechoslov. Chem. Commun. 4, 505 (1932)], and / '/ ²⁰ D + 55 ° (c 0.4, 5 wt.% Aqueous solution of ammonium molybdate), / a / ²⁰ _D + 109 ° (c 0.4, 4% by weight. acidic ammonium molybdate solution) [NK Rychtmyer: J. Amer. Chem. Soc. 73, 2249 (1951)]. Monitoring the purity of D-volemitol, respectively. its content in carbohydrate mixtures is carried out on paper chromatography (Whatman No 1), eluting with an acetone: 1-butanol: water 7: 2: 1 (20 h) flow rate, respectively. 1-butanol: ethanol: water 5: 1: 4 by volume (flow rate 40-50 h) wherein the mobility of D-volemitol relative to the mobility of D-glucose is 0.75 and 0.75, respectively. 0.95.

31S β

Example 2

The procedure is as in Example 1 except that the extraction of the drug is carried out with 3% by weight. aqueous formic acid solution at 95 ° C for 15 and 10 hours. The yield of D-volemitol is the same as in Example 1.

Claims (1)

SUBJECT Method for the isolation of D-volemitol from Primula officinalis, characterized in that the above-ground part of the plant is extracted with 3-7% by weight. aqueous formic acid solution at 75 to 95 ° C for 15 hours OF THE INVENTION up to 25 hours and from the extract the protein portion is precipitated with 50% volume, with ethanol, the quaternary carbohydrates are fermented by fermentation of the solution with baker's yeast and the residue is crystallized in methanol-aqueous solution.