HPLC-DAD法测定兽药恩诺沙星注射液中非法添加呋塞米

Determination of Furosemide Added illegally in Enrofloxacin Injection by HPLC-DAD

关键词:  HPLC-DAD法  兽药  恩诺沙星注射液  非法添加  呋塞米

Key words:  HPLC-DAD  veterianry drugs  Enrofloxacin Injection  illegal addition  Furosemide

摘要:

建立了兽药恩诺沙星注射液中非法添加呋塞米的HPLC－DAD检查方法。以十八烷基硅烷键合硅胶为填充剂，以水-四氢呋喃-冰醋酸（70:30:1）为流动相；柱温30℃；流速1.0 mL/min；进样量20 μL；二极管阵列检测器检测，采集波长范围为190~400 nm，提取波长为272 nm。通过液相色谱保留时间、紫外吸收光谱和峰纯度分析对非法添加物进行确认。结果表明，呋塞米与其他组分在色谱条件下分离情况良好。呋塞米在2.0~100.0 μg/mL范围内线性关系良好，R2=0.9999。回归方程为：Y=1.0629X+0.0685，R2=1.0000。按高、中、低三个浓度水平添加呋塞米，每个浓度水平各平行配制3份阳性添加样品溶液进行测定，每个平行进样2次，测得平均回收率为100.8%，RSD=1.0%。方法的检测限为1.0 μg/mL。方法准确，可靠，可以用于检查兽药恩诺沙星注射液中非法添加呋塞米。

Abstract:

A method has been established for determining furosemide added illegally in Enrofloxacin Injection. The method is carried out for high performance liquid chromatography with photo-diode array detector (HPLC-DAD)，using a column packed with octadecylsilane bonded silica gel and a mixture of water-tetrahydrofuran-glacial acetic (70:30:1) as the mobile phase. The column temperature is 30℃; The flow rate was 1.0 mL/min; The detection wavelength is 272 nm, while acquiring wavelength are between 190 nm and 400 nm; The furosemide is detected and verified with similar retention time and spectrum, and good peak purity as well. Results show the furosemide is separated finely with other compounds under the chromatographic condition. When concentration of reference furosemide solutions are in the range of 2.0~100.0 μg/mL, the linear relationship is good, with R2=0.9999. The average recovery of furosemide is 100.8% at 3 spiked levels of 3.0, 4.0 and 5.0 g/L, with RSD of 1.0%, and the detection limit is 1.0 μg/mL. The method is accurate and reliable，and can be used for determining furosemide added illegally in Enrofloxacin Injection.