SU1525167A1 - Method of obtaining mixture of ribonucleoside-2ъ and 3ъ monophosphates - Google Patents

Abstract

The invention relates to nucleosides, in particular to the preparation of a mixture of ribonucleoside-2 ¹ , and 3 ¹ -monophosphates, which can be used in biochemistry, medicine and pharmaceuticals. The goal is to simplify the process, increase its safety and protect the environment. Obtaining the target mixture is treated with an aqueous suspension of the raw material containing ribonucleic acid / RNA / calcium hydroxide. The process is carried out in a molar ratio of calcium hydroxide and organic phosphorus RNA: 2-2.5: 1. The method allows, in simple technological conditions, to obtain a product of high yield and purity with the exception of sorption on coal and evaporation, as well as work with toxic methanol. 1 tab.

Description

This invention relates to an improved process for preparing a mixture of ribonucleoside-δ. and 3-monophosphates by alkaline hydrolysis of natural RNA and can be used in biochemistry, medicine and pharmaceuticals.

The purpose of the invention is to simplify the process, increase its safety and protect the environment using calcium hydroxide as a reagent for hydrolysis in the established molar ratio.

The invention is illustrated by the following examples.

Example 1. A mixture of ribonucleoside-2 and 3-monophosphates.

100 g of RNA (content of organic phosphorus 7.5%) are suspended in 250 ml of water and a suspension of 35.8 g (2 mol) is added with stirring

calcium hydroxide in 50 ml of water. The reaction mixture is kept for 1 h at. The mixture is cooled to 5-10 ° C, the precipitate is filtered off and washed on the filter with 30 ml of a saturated solution of calcium hydroxide. Output 8A, OA g (the content of the basic substance, as determined by the method of A.S. Spirin,).

Analysis of a mixture of ribonucleoside-2 and 3-monophosphates for the content of adenosine-2 (3) -monophosphate, guanosin- -2 (3) -monophosphate, cytidine-2 (3) -mon-phosphate and uridine-2 (3) -monophosphate was carried out on a Varian 8500 liquid chromatograph on a Micro Pak AX-5 anion-exchange column; eluent was phosphate buffer / acetonitrile (2S nm) by calculating the components by area

SP

to

ate

about:

registered peaks (compared to standards).

The precipitate contains, mmol: guanosin- -2 (3) -monophosphate, b5 mmol, adenrzine-2 (3) -monophosphate 60, uridine-2 (3) -monophosphate, 67 cytidin-2 (3) - -monophosphate 50

Examples 2-4, in which the influence of the temperature of time and the amount of the reagent on the yield and quality of the target product were clarified, were carried out under similar conditions (see table).

Example 5. A Simax 50-liter glass reactor was charged with 30 liters of demineralized water and 10 kg of RNA was metered in with stirring (the content of organic phosphorus in RNA was 7.5). 1.1 kg (2.29 mol) were added to the suspension in portions calcium hydroxide, incubated with stirring for 1b h at 20 ° C. The reaction mixture is fed to a suction filter and the precipitate formed is filtered off. The precipitate is washed on the filter with 2.5 l of water and dried at. Yield 8.32 kg (basic substance content 95%).

Example 6. Suspended 100 g of RNA-containing material obtained by extracting yeast biomass of the protein-vitamin complex (containing

five

about

five

0

organic phosphorus 1.3, RNA 13.8%) in 250 ml of water. With stirring, 6.58 g (2 mol) of calcium hydroxide suspended in 20 ml of water are added with stirring. The reaction mixture is kept for 1 h at 1 h, cooled to 5-10 ° C, kept at this temperature for 6 h, the precipitate is filtered off. Yield 1b, 15 g (content of the main substance is 90%).

Claims (1)

Thus, this method allows, under simple technological conditions, to achieve a high yield and purity of the target product, while excluding the operations of sorption on coal and evaporation, as well as work with toxic methanol. Invention Formula The method of obtaining a mixture of ribonucleoside-2 and 3-monophosphates by treating an aqueous suspension of the raw material containing ribonucleic acid (RNA) with an alkaline agent, which is characterized by the fact that, in order to simplify the process, increase its safety and protect the environment , as an alkaline agent, calcium hydroxide is used in a molar ratio of calcium hydroxide to organic RNA phosphorus of 2-2.5: 1.