兽用预混剂中非法添加杆菌肽锌UPLC-PDA检查方法的建立

Establishment of Detection Method for Bacitracin Zinc in Veterinary drug Premixes by UPLC-PDA

关键词:  超高效液相色谱法  地克珠利预混剂  盐霉素预混剂  莫能菌素预混剂  杆菌肽锌

Key words:  UPLC  diclazuril premix  salinomycin premix  monensin premix  bacitracin zinc

摘要:

建立了兽用地克珠利预混剂、盐霉素预混剂、莫能菌素预混剂中非法添加杆菌肽锌的超高效液相色谱-二极管阵列检查方法。采用Waters ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7 μm),以0.05 mol/L甲酸铵溶液(甲酸调节pH值至4.0)-乙腈为流动相,梯度洗脱,流速0.5 ml/min,二极管阵列检测器(PDA),波长采集范围为200～400 nm,记录色谱图波长217 nm,柱温30 ℃,进样量5 μl。采用峰纯度检查和光谱相似度检查辅助标准品比对方法,对非法添加杆菌肽锌进行确证,并对建立的方法进行方法学验证。结果显示,杆菌肽锌质量浓度在0.4～2 mg/ml内峰面积呈良好线性关系(R_²=0.9996),在地克珠利预混剂、盐霉素预混剂、莫能菌素预混剂中杆菌肽锌的平均回收率分别为100.34%、99.22%和95.92%,RSD分别为1.40%、1.21%和1.26%,主成分杆菌肽A与其他成分分离度较好,峰纯度检查和光谱相似度检查均符合要求。本方法操作简便、准确、可靠,适用于以上三种兽用预混剂中非法添加杆菌肽锌的检查。

Abstract:

A detection method of bacitracin zinc in diclazuril premix, salinomycin premix and monensin premix was established by UPLC-PDA. The Waters ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7 μm)was used, the mobile phase consisted of 0.05 mol/L ammonium formate solution (adjust pH value to 4.0 with formic acid)- acetonitrile by gradient elution. The flow rate was 0.5 ml/min, the column temperature had remained 30 ℃, photodiode array detector wavelength range was 200 to 400 nm, the recorded wavelength was 217 nm, the injection volume was 5 μl. Peak purity test and spectrum similar test were helped to identify bacitracin zinc. The linear range of bacitracin zinc was 0.4～2 mg/ml(R²=0.9996). The mean recovery for bacitracin zinc in diclazuril premix, salinomycin premix and monensin premix were 100.34%, 99.22% and 95.92%,RSD were 1.40%, 1.21% and 1.26%, respectively. Main component bacitracin A was separated from other components well. The peak purity test and spectra matching test were applied and the method was verified. This method was accurate, easy and reliable, it was suitable for detection of illegally added bacitracin zinc in the above three veterinary drug premixes.