Determination Of Enramycin By The Method Of HPLC

Enramycin, one of polypeptide antibiotic, was obtained by the fermentation of Streptomyces fungicidicus. Because of its unique molecular structure, Enramycin was active to Gram-positive bacteria, especially to the harmful Clostridium of intestinal tract.Enramycin can promote the growth of pigs, chickens, and so on. Thus, as a new feed additive, enramycin was widely used in the production of animal husbandry. Currently, the content of Enramycin was detected by the method of Microbiological assay in accordance with Quality standards for imported veterinary drugs(1999), issued by the Ministry of Agriculture. This study established the method of HPLC to detect the content of enramycin according to the structural characteristics of Enramycin. The optimum chromatographic conditions was: the Chromatography Column: Thermo C18, 5?m, 4.6mm×250mm; the mobile phase: acetonitrile: 0.05mol/L sodium dihydrogen phosphate solution=30:70;detection wavelength: 267nm; column temperature: 30?; flow rate: 1.0mL/min; injection volume: 20?L. Results: The linear relation of Enramycin A and B was good within the range from 12.5 to 200?g/mL. The regression equation of Enramycin A and B were A1=11587x+18926(R2 =0.998) and A2=35969x-46745(R2=0.996), respectively. The average recovery of enramycin A and enramycin B were 98.9% and 98.7%, respectively.The relative standard deviation(RSD) was 0.27%. The results showed that the method of high-performance liquid chromatography(HPLC) detecting the content of Enramycin has the advantage of wide linear range, short analysis time, simple sample preparation,accurate, reproducible, and high stability and recovery, and so on. Thus, the method of HPLC can be used as quality control method in enramycin content detection. Comparing the same sample in the two methods, HPLC and Microbiological assay, we found that the method of HPLC is simple, fast, accurate, which provided reliable basis for quality control.