SU878769A1 - Method of separating mixture of xanthosin and guanosin - Google Patents

Description

The invention relates to a method for separating a mixture of nucleosides, namely xanthosine and guanosine, which are used in biochemical studies. Xanthosine is obtained by desamynir0; va, by the use of guanosine and incomplete deamination, it is necessary to separate the mixture of xanthosine and guaiosin.

The known method for selective separation of a mixture of xanthosine and guanosine using ion exchange chromatography I.

Such a method is as follows. A mixture of xanthosine l guanosine in an aqueous solution after adjusting the pH to 6.0-8.5 is passed through a strongly basic anionnt in the form of a salt. The column is washed with water and get guaios. In with the content of the main substance of 96.7% (yield. Guanosine 90.88%). Then the ion exchange material is eluted with 1N. hydrochloric acid yielded 94 g of xacosin with a content of the main substance of 95% (xanthosine yield 89.3%).

The known method provides for the separation of xanthosine and guanosine by means of xantosine sorption on strongly basic aniovite, but it is laborious, as preparation, regeneration of the ion exchange material and evaporation of eluant-TOiB are required.

The disadvantages of this method are also relatively low yield (yield of xanthosine 89.3%, guanozine 90.88%) and relatively low purity of the target products (xanthosine 95%, guanosine 96.7%).

The aim of the invention is to increase the yield and purity of the slit products and simplify the process of separating the components.

The goal is achieved by the fact that when the separation method of a mixture of xanthosine and guanosine in an aqueous medium is carried out, the initial mixture is acidified to a pH of 0.6-1.0 and stirred for 30- 35 minutes at 15-25 ° C to a complete solution of 15 no guanosine, then the xanthosine in the precipitate is separated, and the pH is adjusted to 3,. 0, the guanosine dihydrate is precipitated, which is converted to guanosine by known methods.

Guanosin and xanthosia in cold water

20 practically "they are dissolved, and in the crystallization process, when the dissolution is carried out at elevated temperature, they precipitate together. The separation of the mixture reaches 25%; using the differing solubilities of various forms of nucleosides. Different p / Ca values for guanosine (1.6-1.8) and xanthosine () will allow a pH of 0.6-1.0 to be established, at which time 30 all guanosine will practically form a protonated, more soluble form even at room temperature, it can dissolve into solution. Xanthosis in such conditions remains in the precipitate "can be separated by filtration from the dissolved; guanosine. After separation of xanthosine, an alkali solution is added to the solution and ipH is stabilized in the range at which guanosine forms a neutral little soluble in water form, namely pH 3.0-6.0. With such a pH, the olichesta venio separates guanosine from the uterine. Both nucleosides by the proposed method are obtained by chromatographic chromatography without additional recrystallization.

The separation is carried out in two stages: Stage I — isolation of chromatographically pure xanthosine; Stage II - isolation of chromic. Mathematically pure guaiosia.

Fractionated precipitation of xanthosine and guanosine provides high yield and purity of the target and products.

Example 1.2 g of xanthosine and 0.44 g of guaiosin are suspended in 40: ml of distilled water, the pH is adjusted to 0.8 1N. at 30 ° C, the salt is stirred for 30 minutes. The precipitate is filtered off, PRO: WASHED with hydrochloric acid at pH 1.0, then with water and ethyl alcohol. Wash solutions are not attached to the filtrate. 1.9 g (94%) of chromatographically pure xanthosine are obtained, the content of basic vesch, 99%

1n. sodium hydroxide solution set: pH of the filtrate 6.0 and precipitated guaiosin dihydrate. Get 0.45 g of the latter. Guanosine dihydrate is converted to .3 guanoscine by heating for 3 hours at 60 ° C. 0.4 g of guanosine is obtained, yield 90%. The content of the main substance is 100%.

Example 2. 1.5 g of xanthosis ina and 0.53 g of guanosine are suspended in 50 ml of distilled water, the pH is adjusted to 1.0 with 1N. salt KI.SLOTOY at 15 ° C and mix for 30 minutes. Gssanthosine is filtered off, washed with hydrochloric acid at a pp of 1.0, then with water and ethyl alcohol. Washing waters are not attached to the filtrate. 1.44 g (95%) of chromatographically physically pure xanthosine are obtained. Content of the main substance is 99%.

1 n. the pH of the filtrate is adjusted to 3.0 with a solution of 1 hydro. sodium. and the guranosium dilidrate is precipitated. Get 0.55 g of the latter. Guanosine dihydrate is drilled 13 guanosine by heating for 3 hours at 60 ° C. 0.48 g (91%) of chromatographically pure guanosine is obtained. Soda, the basic substance neighing 100%.

Example 3. 4.0 g of “Santozin” and 1.0 g of guanosine are suspended in 100 ml of distilled water, the pH is adjusted to 0, .6 1 n. hydrochloric acid at 28 ° C and stirred for 3.0 min. Isantozine is filtered off, washed on the filter with dilute hydrochloric acid at pH 1.0, then with water and ethyl alcohol. Wash solutions are not attached to the filtrate. 3.80 g (96%) of chromatographically pure xanthosine are obtained. Content of the main substance is 99%.

1 n. A solution of sodium hydroxide is used. A RP of the filtrate is set at 4.5 and guanosine dihydrate is precipitated. Osado.k.pro.wash on

filter with water and ethyl opirt. 1.03 g of chromantotraphic guanosine dihydrate is obtained. The latter is converted into guanosine by heating for 3 hours at 60 ° C. 0.9 g are obtained chromatographically.

pure guanosine. Yield 91%. Content of the main substance of 1009%.

Example 4. A mixture of 1 g of xanthosine and 3 g of guanosine is suspended in 50 ml. 1, of the inhaled water, and a RP of 1.0 is established.

at 20 ° C and stirred for 30 minutes. Then filtered, nro.washed with dilute hydrochloric acid (pH 1.0), water, ethyl alcohol. Wash solutions to the filtrate are not used. Get

0.79 g (78%) of chromatographic of physically pure xanthosine. Content of the main substance is 99%.

The pH of the filtrate is adjusted to 5.0 and guanosine is filtered off. Get 3.09 g

(91%) chromatographically pure guanosine dihydrate. Content of the main substance is 100%.

Example 5. A mixture of 2.0 g of xanthozine and 1.0 g of guanidine is suspended with IB with 100 ml of distilled water, adjusted to pH 0.6 at 20 ° C and stirred for 35 minutes, then filtered, the precipitate washed with dilute hydrochloric acid (pH 1.0), water, ethyl alcohol. Washing

solutions are not attached to the filtrate. 1.86 g (94%) of chromatographically pure xanthosine are obtained. Content of the main substance. And 99%.

The pH of the filtrate is adjusted to 5.0 and the guanosine is separated. 1.02 g (90.5%) of chromatographically pure guanosine dihydrate is obtained. Content of the main substance is 100%.

Ex. 6. A mixture of 10 g of Xanthosine and 1 g of guanosine is suspended / simulated. in 100 ml of distilled water, the pH is adjusted to 10 at 18 ° C with dilute hydrochloric acid and transferred within 30 minutes,

then filtered, washed with dilute hydrochloric acid, water, ethyl alcohol. Wash water is not added to the filtrate. 9.6 g (95%) of chromatographically pure xanthosine are obtained. Co. main substance 99%.

The pH of the filtrate is adjusted to 3.0. Gua.nozin is separated. 1.01 g (90%) of chromatographic guanosis dihydrate is obtained. The content of the main substance

100%.

Claims (1)

1. A method of separating a mixture of xantosis a and guanosine in an aqueous medium, characterized in that, in order to increase the yield and quality of the target product and simplify the process, the initial mixture is acidified to a pH of 0.6-1.0 and stirred within 30-35 minutes at 15-25 ° C, then the xaitosin in the precipitate is separated and the pH of the solution is adjusted to 3.0-6.0 with guanosine dihydrate precipitated, which is converted into guanosine. The source of information taken into account in the examination: 1. Japan patent No. 73/21960, cl. 16 E 33, published. 1973 (prototype).