高效液相色谱—二极管阵列检测法测定原料乳中的舒巴坦

Determination of sulbactam in raw milk by the high-performance liquid chromatography-photodiode array detection

建立了利用高效液相色谱-二极管阵列法(high-performance liquid chromatography-photodiode array,HPLC-PDA)测定原料乳中舒巴坦含量的方法。采用C18色谱柱,以5.44g/L磷酸二氢钾溶液(pH5.0)-乙腈(95∶5)为流动相,流速为lmL/min,检测波长220nm,柱温30℃。样品经乙腈提取、乙酸锌沉淀蛋白,HPLC-PDA检测。舒巴坦浓度在10～200μg/mL范围内,线性良好(R2=0.9998),添加回收率为98.82%～101.42%,检出限为0.25mg/kg,定量限为0.75mg/kg。该方法操作简单,结果准确、可靠,适用于原料乳中舒巴坦的快速检测。

To develop a method for the determination of sulbactam in raw milk using high performance liquid chromatography-photodiode array detector（HPLC-PDA）. C18 column was used,the mobile phase consisted of 5.44g/L potassium dihydrogen phosphate solution （pH5.0） -acetonitrile （95：5） at the flow rate of lmL/min, the detection wavelength was 220nm.The column temperature was 30℃. Acetonitrile was used for extraction,zinc acetate was used for protein precipitation ,the filtered fluid was determined by high performance liquid chromatography-photodiode array detector. The linear calibration curve was obtained in the concentration range of 1-200μg/mL（R2=0.9998） with the lower limit of detection of 0.25mg/kg and lower limit quantification of 0.75mg/kg The average recovery of sulbactam was 98.82%-101.42%. The method was simple,accurate,reliable and could be used for rapid determination of sulbactam in raw milk.