SU111196A1 - The method of isolation and purification of antibiotic albicin - Google Patents

Description

A known industrial method for the isolation and purification of the antibiotic albumin, obtained by desorption from activated carbon, on which it was sorbed from the culture fluid, makes it possible to obtain a preparation containing about 80% of the ballast substances, which therefore has undesirable pyrogenic properties.

The proposed method allows, in the process of industrial production of albicin from an acetone eluate, to obtain a pyrogen-free preparation with high activity and a significant yield.

A distinctive feature of the method is that albicin contained in the acetone eluate, in order to separate it from the ballast impurities, is subsequently sorbed on the sulfonic cation resin SDW-3, taken in a mixed hydrogen-ammonium form, then on the same resin in the hydrogen form and.

finally, on the KFU resin in the hydrogen form, after which the resulting purified concentrate of Albicin is treated by known methods, and the ion exchange resins are regenerated as usual.

The method is carried out as follows.

The acetone eluate is acidified to a pH of 2.3-2.6 with 2.5% sulfuric acid, filtered on a filter and fed to a chromatographic column with SDK-3 sulfonic resin and in a mixed ammonium-hydrogen form at a rate of 100-120 ml 1 cm 1 hour . The amount of eluate is based on 10–25 billion units per 1 kg of air-dry resin.

The chromatographic separation of albicin and related impurities is carried out using an elution method. The albicin zone moves through the column while passing 0.05 p acetic ammonium buffer solution pH 5.1-5.3 through the column at a rate of 40

60 ml / cm / hour. At the exit from the column, little-colored impurities appear first, and then a colored solution of albicin (a colorimetric method for iron analysis) comes out. A solution of albicin with an iron concentration of more than 20–30 gamma1 ml is passed through the column to a subsequent concentration, and a solution with a lower iron concentration is added to the original acetone eluate for repeated purification.

The yield at this stage is 70-85%, with 60-65% being concentrated, and 10-20% being recycled.

The solution of albicin after chromatographic purification is fed to the column with the SDV-3 resin in the hydrogen form in the amount of 8-10 liters of solution per 0.1 kg of air-dry resin. The flow rate is 100-300 ml / cm hour. Albicin is sorbed completely, the elution is carried out with 1N ammonia solution with a speed of 30-35 ml / cm / hour.

The concentrated ammoniacal solution of albicin (dark red) discharged from the column is passed through a column containing KFU resin in hydrogen form 60-100 ml 1 cm 1 hour in the amount of one liter of ammonia concentrate per 0.2 kg of resin. Next, the column is washed with distilled water at a rate of 40-

50 ml1cm 1 hour for complete crowding out of albicin.

The resulting albicin concentrate is characterized by a specific activity of 550,000-700,000 units of 1 mg, is not pyrogenic and non-toxic.

The yield at this stage is 75-80%.

Using the freeze drying method, the finished product albicin is obtained from the concentrate.

The yield from (k) a ltural liquid, taking into account losses on sorption on coal, is 40-45%.

Subject invention

The method of isolation and purification of the antibiotic Albicin from acetone zljuat during its industrial production, characterized in that, in order to obtain a pyrogen-free drug with high activity, albicin in the acetone eluate to separate it from the ballast impurities is sequentially sorbed on the sulfonic cation exchanger SDN-3 resin, taken in the mixed hydrogen-ammonium form, then on the same resin in the hydrogen form and, finally, on the KFU resin in the hydrogen form, after which the resulting purified concentrate of albicin is treated with known techniques, and ion-exchange resins are regenerated as usual.