牛乳中三氮脒残留高效液相色谱检测方法的建立

以乙腈沉淀牛乳中的蛋白质,上清液采用Oasis WCX固相萃取小柱净化浓缩,用乙酸-甲醇溶液洗脱药物,研究了牛乳中三氮脒残留的高效液相色谱检测方法。洗脱液上机检测,紫外检测波长372nm,色谱分析柱为氰基柱,流动相为V(乙腈)∶V(甲酸铵溶液)=10∶90,等度洗脱。当样品中三氮脒的含量为0.010～1.00μg/mL时,药物质量浓度与检测信号存在良好的相关性(r2>0.999);牛乳中三氮脒的检测限为0.50×10-2μg/mL,定量限为0.010μg/mL。牛乳按0.010、0.10和1.00μg/mL 3个添加水平做三氮脒的回收率试验,回收率≥82%,变异系数在1.3～4.2之间。结果表明,本研究建立的三氮脒残留高效液相色谱检测方法重复性好,灵敏度高,操作简便,可用于牛乳中三氮脒残留的检测。

Development of a high performance liquid chromatography method for the detection of diminazene diaceturate residue in milk

A high performance liquid chromatography(HPLC) method was developed for the determination of diminazene diaceturate in milk.Diminazene diaceturate was extracted with acetonitrile from milk.Oasis WCX cartridges was used to purify and concentrate the drug after centrifugation.The drug was then eluted by acetic acid-methanol.The sample was detected under the ultraviolet(UV) wavelength of 372 nm.Mobile phase consisted of acetonitrile and 0.02 mol/L ammonium formate solution(10∶90,V∶V) were used for isocratic elution and the flow rate was 1 mL/min.Within the range of 0.010 μg/mL to 1.00 μg/mL of diminazene diaceturate in milk,good linearity between drug concentration and detection signal was obtained(r2>0.999).The limit of detection and limit of quantification of the examined diminazene diaceturate in milk were 0.5×10-2 μg/mL and 0.010 μg/mL for milk,respectively.The recoveries from spiked milk at three levels(0.010 μg/mL,0.10 μg/mL,and 1.00 μg/mL) were more than 82%,and the coefficient of variation ranged from 1.3 to 4.2.The developed method showed good reproducibility,high sensitive,expedience and could be applied to the determination of diminazene diaceturate in milk.