Separation of pristinamycin by silica gel column chromatography

The separation and purification of pristinamycin from the fermentation broth of Streptomyces pristinaespiralis was investigated by using silica gel column chromatography under isocratic elution.The results showed that qualitative analysis could be elementarily made by the method of thin layer chromatography (TLC) with silica GF254(developing distance of 5.0~6.0 cm and developing time of 6~7 min),under the developing agent composed either of methylene chloride and methanol with their volume proportions of 90 and 10,or of methylene chloride,ethyl acetate and ethanol with their volume proportions of 60,35 and 5.The column chromatography was preferably conducted under such conditions: 25 ℃,100～200 mesh silica gel as the stationary phase,solvent system of methylene chloride,ethyl acetate and ethanol with their volume proportions of 80,35 and 5 as the eluent,the eluent flow rate of 2.0 mL/min,pristinamycin feeding concentration of 2.26 g/L,and the total loading amount of silica gel of about 0.81 mg/g.Under these operational conditions,pristinamycin with purity of more than 95% was recovered,and the product recovery was over 95% simultaneously.