CS224726B1 - Ergosterol isolation method - Google Patents

Description

The invention relates to a process for the isolation of ergosterol from a mixture of sterols obtained after

of nucleic acids and alkaline hydrolysis by non-polar solvent extraction.

The isolation of ergosterol from a mixture of sterols and by-products produced by yeast, and so far, is usually accomplished by extraction with benzene, hexane or another organic solvent and crystallizing the product either directly or after addition of lower alcohols (methanol, ethanol). The disadvantage of these processes is to obtain either relatively little pure product requiring recrystallization or low ergoetherol yield. The most commonly used solvent is benzene, which, while still providing a product of satisfactory quality without recrystallization, gives a low crystallization yield of about 75%. Another disadvantage is the long crystallization time;

The new method is used for the extraction of n-butenol, the authors perform the isolation by pouring the concentrated extract into water (Japan Kokai No. 75 142 787). However, this method requires, on the one hand, recrystallization of the product, and, on the other hand, the stratification of the hopping system often results in considerable problems due to the negative action of the salts of higher fatty acids formed by alkaline hydrolysis.

These disadvantages are overcome by a process for isolating ergosterol from a sterol mixture obtained from hydrolysed yeast by extraction with an organic solvent and either direct distillation of the solvent, or crystallization after partial concentration of the extract, or distillation of the solvent from the mother liquors after this crystallization. SUMMARY OF THE INVENTION The invention comprises adding from 2 to 20 times the amount of alcohol to G, preferably C, or acetone and water to the sterol mixture in an amount ranging from 1 to 20% by weight, after which the mixture is heated to 50 to 90%. And ergoaterol is isolated from the resulting solution after cooling to +5 to -5 ° C. The pH of the solution may be adjusted to 5 to 9 at any stage of the isolation. Weak acids such as acetic or acidic salts such as ammonium chloride, hydrogen and sodium dihydrogen phosphate, sodium hydrogen sulfate can be used to adjust the pH of the ergoaterol solution.

In the case of processing the extract according to the invention, i.e. after distilling off the solvent used for yeast extraction, ergosterol with a purity of about 90% and a crystalline water content of 5 to 6% is obtained, which is suitable for further treatment by irradiation to vitamin Dg. If P ° is first crystallized and only the mother liquors are processed, a product of about 75% purity is obtained which, by recrystallization, gives a pure substance. The mother liquors after isolation of ergoaterol from the alcohol-water mixtures containing yet another quantity of ergoaterol, which can be obtained after partial concentration and replenishment of water in the form of less pure fractions, but which can be purified.

For recrystallization, either the repetition of the aqueous alcohol crystallization process according to the process of the invention or the crystallization itself from commonly used solvents such as benzene, benzene-ethanol, ethanol, isopropyl alcohol, etc. can be used. Crystallization of the crude product by addition to the original sterol-containing extract in an organic solvent and crystallization or crystallization under mother liquors after isolation of the first product fractions in the case of using two-stage crystallization. The mother liquors after recrystallization of ergosterol can be treated together with 9 mother liquors after crystallization of the first fractions of ergoaterol. For extraction of the mixture. The preferred yeast sterols after hydrolysis are the most preferred aromatics such as benzene, toluene and optionally saturated hydrocarbons such as hexane and cyclohexane. To reduce the oxidation of the product in an alkaline solution, a small amount of antioxidants such as butylhydroxytoluene may be added.

The advantage of the process according to the invention is to increase the ergoaterol yield by 5 to 15% while maintaining the quality of the minimum cost, to reduce the crystallization time and to reduce the amount of waste.

In order that the invention may be more fully understood, the following examples are provided.

Example 1

From the mother liquors (benzene) after isolation of ergoaterol, benzene was distilled off up to a flask temperature of 130 ° C and the benzene residues were distilled off under reduced pressure so that the distillation residue contained less than 2% by weight. benzene.

10.0 distillation residue containing 23.1 wt. of sterol was dissolved in 100 ml isoamyl alcohol by heating to 80-90 ° C and 10 ml water was added. The emulsion was stirred and cooled to + 5 ° C, the precipitated crystals were aspirated and washed 3 times with 10 ml of ethanol. After drying at 50 [deg.] C., 3.1 g of the product were obtained and contained 37.3% ergoaterol, i.e. 50.2% of theory.

Example 2

100.0 g of the distillation residue, containing 22.3% of ergosterol obtained according to example 1, were dissolved in 1000 ml of isobutyl alcohol and 200 ml of water by heating to 70-80 ° C, 2.0 g of activated carbon added, hot filtered and filtrate gradually cooled to 0 ° C with stirring. The crystalline fraction was vigorously aspirated and washed with 3x100 ml ethanol cooled to 0 ° C.

After drying at 50 ° C, 20.5% of the product with 69.4% by weight ergosterol was obtained, i.e. 63.8% yield.

Example 3

300.0 g of a distillation residue containing 30.3% of ergosterol obtained according to Procedure 1 were dissolved in 2700 ml of n-butanol and 450 ml of water by heating to 80-90 ° C. The resulting solution after addition of 2 g of diatomaceous earth was filtered while hot, the pH of the solution was adjusted to 5-8 by the addition of ammonium chloride, and the filtrate was gradually cooled to 0 ° C with stirring. The crystalline fraction was vigorously aspirated and washed 3 times with 150 ml of methanol cooled to 0 ° C. After drying the cake, 87.2 g of a product containing 70.0 wt. ergosterol, i.e. and a yield of 67.1% of theory.

Example 4

From the sterol benzene extract after yeast hydrolysis, i.e. without partial isolation of ergosterol, benzene was distilled off up to a flask temperature of 130 ° C and the benzene residues were distilled off under reduced pressure to give a distillation residue of less than 2% by weight. benezenu.

218.0 g of a distillation residue containing 50.8 wt. ergosterol was dissolved in 1000 ml n-butanol and 163 ml water by heating to 80-85 ° C, after which the solution was gradually cooled to -2 ° C. The precipitated product was vigorously aspirated and washed 3 times with 200 ml of methanol. After drying, 110.8 g of the product was obtained with a content of 90.9% by weight. ergosterol, i.e. with a yield of 90.9% of theory.

Example 5

100 g of crude ergosterol containing 78.3 wt. was dissolved in 2000 ml butanol by heating to 80 ° C, 2 g carboraffin was added and filtered hot. 400 ml of hot water was added to the filtrate and cooled to 0 ° C with stirring. The precipitated crystal was vigorously aspirated and after drying 55 g of ergosterol were obtained with a content of 90.2% of the active compound, corresponding to a yield of 63.4% of theory.

Example 6

218 g of a distillation residue containing 50.8 wt. ergosterol was dissolved in 2000 ml acetone, 100 ml water was added, heated to 50 ° C and the solution gradually cooled to 0 ° C with stirring. The product was aspirated vigorously, washed 3 times with 200 ml of ethanol. 115.1 g of product was obtained with a content of 91.2% by weight. ergosterol, i.e. and a yield of 94.2% of theory.

Claims (2)

SUBJECT OF THE INVENTION A process for the isolation of ergosterol from a sterol mixture obtained from a hydrolysed yeast by extraction with an organic solvent and either by direct distillation of the solvent, or by crystallization after partial concentration from the extract, or by distilling off the solvent from the mother liquors after crystallization; 2 to 20 times the amount of alcohol to C C, preferably C ^, or acetone and water are added, ranging from 1 to 20% by weight, then the mixture is heated to 50-90 ° C and the ergosterol is recovered from the solution after cooling to +5 to -5 ° C. 2. The process according to claim 1, wherein the pH of the solution is adjusted to a value of 5 to 9 at any stage of the isolation.