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Abstract
Macromolecular impurities in penicillin G sodium (PGS) may cause allergic reactions. Separation of these impurities and PGS by gel filtration chromatography was investigated in this study. The effects of gel, mobile phase, and temperature on separation were examined. The penamaldate method was applied to detect penicilloylated impurities separated on gel columns. Three gels, Sephadex G-10, G-25, and Toyopearl HW-40F were examined with water, acetate, phosphate, citrate buffers as mobile phases. Separation on HW-40F was better than on G-10 and G-25. Mobile phase significantly affected both the separation of macromolecular impurities and the chromatographic behavior of PGS. PGS was over excluded in water. The distribution coefficient of PGS was largest in citrate buffer, and smallest in acetate buffer. Ion strength influenced the chromatographic behavior of PGS greatly. Higher ion strength resulted in a greater distribution coefficient. Temperature hardly affected the chromatographic behavior of macromolecular impurities and PGS, but it greatly influenced the retention of small impurities. The suitable conditions for separation were using gel HW-40F as stationary phase, 5 mM citrate buffer as mobile phase, and temperature below 15°C.