Research On Separation And Purification Of Tobramycin

The purity of tobramycin in crude extracts from fermentation broth can only achieve to 50?-60% in industrial production by strongly-acid resin, which can hardly meet the standards recorded in Chinese Pharmacopoeia.Adsorption performance of tobramycin with different macroporous weakly-acid resins was determined by static adsorption and dynamic adsorption experiments and HZ-3B resin was the best choice of absorbent. The optimal conditions for purification of tobramycin were as follows:concentration of tobramycin was 5mg/mL and pH range of tobramycin solution was 6.8-7.2 at sample flow velocity of 9BV/h; and then 0.125mol/L of ammonium hydroxide was used as desorption solution with flow velocity of 6BV/h. HPLC results showed that the purity of tobramycin increased from 50?60% to 97.15% with yield of 89.52% after purification, and no other aminoglycoside antibiotics were found. The purity of tobramycin can meet the standards of Chinese Pharmacopoeia through technology improvement.D301 resin was studied as the best anion-exchange resin for purifying Tobramycin. The best condition for anion-exchange resin were as follows:concentration of tobramycin was 5mg/mL and pH ranged from 8.5-9.5 at sample flow velocity of 9BV/h;then 0.6% of ammonium hydroxide was used as desorption solution with flow velocity of 6BV/h.HPLC results showed that the purity of tobramycin increased from 50?60% to 93.15% with yield of 85.41% after purification and compared the result with HZ-3B and finally chose HZ-3B as the best resin to purify fermentation solution.At last this paper also studied the process to remove impurities in fermentation solution.The best technology were as follows:make fermentation flow through 711 resin at the velocity of 5BV/h,then adjust ethyl alcohol rate to 30%,load 10BV sample through HZ-806 resin by 5BV/h.The solution from resins were mainly aminoglycoside antibiotics.The purity of Tobramycin solution by this technology can reach to at least 97.35% and other impurities were less than 0.5%.The stability of resin was good,RSD was less than 2%.This technology provided a new and economical way to purify Tobramycin for industry.