SU110650A1 - The method of direct extraction of vitamin B12 from the waste of the antibiotic industry and other natural liquids - Google Patents

Description

The subject of the invention is a method for the direct extraction of vitamin B12 from wastes of the antibiotic industry and other natural liquids using cyanides.

The proposed method, in comparison with the known ones, allows increasing the yield of the target product and simplifying the process by eliminating the most difficult operations — adsorption of vitamin with activated carbon and its subsequent desorption.

The peculiarity of the method is that cyanide is added before extraction, which is maintained by a mixture consisting of tricresol, kerosene and butanol, followed by processing the resulting liquid in a known manner by converting vitamin Bi2 into water, purifying it and crystallizing it.

In the extraction mixture, tricresol, kerosene and butanol are contained in a 1: 1: 0.1 ratio.

The method is carried out as follows.

Prior to extraction, 0.03-0.05% of sodium cyanide or potassium cyanide is added to the culture liquid, the pH is adjusted to 10 and maintained for 1 hour. The vitamin is extracted with a mixture containing tricresol, kerosene, butanol in a ratio of 1: 1: 0.1 taken in the amount of b-8% of the total volume of the extracted liquid.

After separation at the separator or by settling, isopropyl alcohol (50% of the volume of the liquid) is added to the organic liquid containing vitamin Bia, and the vitamin is extracted with small portions of water. Further, the vitamin enters the purification and crystallization, which is carried out by one of the known methods.

An example of obtaining vitamin BIO from the culture fluid in the production of streptomycin.

The input flow of the mother liquor is significant; therefore, the primary purification of the culture liquid, which is received with heating up to 80 ° C, is carried out at separation of the mycelium at a pH of about 4.5 (blue - violet and bromophenolblau). This does not interfere with the preparation of streptomycin and facilitates the extraction of the vitamin. Big.

The mother liquor is neutralized, 0.05% sodium cyanide is added to it and the pH is adjusted to 10 (blue for thymolphthalein), after which it is aged for 1 hour. 8% tricresolkerosine-butanol 1: 1: 0.1 mixture is added to the mother liquor, the liquids mix well and are separated into a separator to separate the organic mixture with vitamin BI. The mixture yield 75% of the specified amount. The mother liquor goes to the garbage. In the absence of a separator, the liquids are separated by settling for 3 hours.

The output of the mixture is 65-70%.

The resulting mixture is washed several times with small portions of water {2-3 times 5% of the volume of the mixture), then to it a volume of isyropy alcohol and butyl alcohol and vitamin Bi2 is extracted 4-5 times with small portions of water. The total volume of the aqueous extract is about 2/3 of the volume of the added alcohol (or 20 liters of liquor). The aqueous extract is filtered from the suspension through a suction filter, on the canvas of which a small layer of AO3 is applied, and the organic mixture is fed to the regeneration (distillation of alcohol) and returned to production.

Phenol - butanol cleaning. 0.25% sodium cyanide is added to the filtered aqueous extract and the extract is aged for 1 hour. After that, it is saturated with sodium chloride (food grade) and vitamin B | 2 is extracted in small portions of the phenol-butanol mixture 1: 3 until the mixture ceases to be colored. The total mixture is consumed 25% of the volume of the aqueous extract (5 l /. And mother liquor). The mixture is washed with a saturated solution of common salt until the staining of the wash water is stopped. Then

1 volume of chloroform is added to the mixture and the vitamin Bi2 is converted to water.

The output of the aqueous extract is about 50% of the phenol-butanol mixture (2.5 liters per 1 m of mother liquor).

Phenol-chloroform and cleaned. The phenol-chloroform mixture 1: 6 is added to the aqueous extract in small portions. The first 2–3 servings do not entrap the vitamin, remove impurities (turn yellow without pink hues) and discard, and the subsequent portions extract vitamin Bi2 and collected. The output of the mixture is about 60% of the aqueous extract (1.5-2 per 1 m of the queen cell).

The conversion of vitamin Bi2 to water is made after adding 2.5 volumes of chloroform to the mixture. The output of the aqueous extract is 45-50% of the initial mixture (0.7 l per 1 m of mother liquor).

Tricresol-CC1 | cleaning up The vitamin is extracted from the aqueous solution in small portions of the trnresol-CCB 1: 1 mixture, and then transferred to water when added to the organic mixture, 3 volumes of CCl.i and 0.8 volumes of butiol. The final volume of aqueous extract of 0.4 l per 1 m of the queen cell.

After the final aqueous extract of the residue of organic solvents, it is washed 2-3 times with small portions of sulfuric ether (anesthetic), the remnants of which are in turn removed by gently heating the extract in a water bath.

The resulting final extract is concentrated under vacuum to a content of about 10 mg of vitamin Bi2 in 1 ml (up to 40 ml per 1 mother liquor) and is fed for chromatographic purification on an AO3 column and crystallization.

The yield of crystals, the trial of 6 semi-production loads, amounted to 350-400 mg per 1 m of the queen cell.

Subject invention

1. The method of direct extraction of vitamin B | 2 from antibiotic waste