饲料中苯巴比妥钠检测方法的建立

发布时间：2018-10-05 15:23

【摘要】：苯巴比妥钠(Phenobarbital Sodium)是农业部176号公告规定的禁止在饲料和动物饮用水中使用的药物品种。目前,尚没有针对饲料中苯巴比妥钠的检测方法和标准,这就为饲料质量监管和人民饮食健康埋下了隐患,制定饲料中苯巴比妥钠的检测方法刻不容缓。苯巴比妥钠是弱酸强碱盐,在酸性环境中会发生水解以苯巴比妥的形式存在。饲料样品经过正己烷预处理后,加盐酸甲醇溶液提取,其中的苯巴比妥钠被以苯巴比妥(Phenobarbital)的形式提取出来。提取液经Waters Oasis HLB固相萃取小柱(SPE)净化,经甲醇+水(2+98)溶液淋洗后,用5%氨化甲醇洗脱。洗脱液经氮吹浓缩后,用甲醇+水(2+8)复溶,上高效液相色谱仪(HPLC)检测。所用色谱柱是粒径5μm,250 mm×4.6 mm的C8柱,柱温箱控制温度为30℃,流动相流速为1 mL/mi,梯度洗脱,紫外检测器于215 nm波长检测。经过公式计算和转化,得到苯巴比妥钠的含量。分析表明,该液相方法的检测限(LOD)为0.5mg/kg,定量限(LOQ)为1mg/kg,该方法具有良好的准确度和精密度,且对配合饲料、预混合饲料、浓缩饲料以及精料补充料均有较好的适用性,回收率均在在80%-120%之间,相对偏差(RSD)均在10%以内。标准曲线取点0.5μg/mL、1.0μg/mL、2.0μg/mL、5.0μg/mL、10.0μg/mL、20.0μg/mL、50.0μg/mL,曲线相关系数达到0.999,具有良好的线性回归。在确定的检测条件下,地西泮、司可巴比妥等镇定类药物无干扰。液相方法取得了良好的实验效果。苯巴比妥钠的液相色谱-串联质谱法(LC-MS-MS)是在液相方法的基础上深入研究的。饲料样品中苯巴比妥钠的提取、净化、浓缩在已有液相方法的基础上进一步调整和优化。液相色谱分离柱选择100mm×2.1 mm的C18柱,在流速为0.2 mL/min,柱温为40℃时苯巴比妥被分离出来,然后经串联质谱仪检测信号。在质谱仪上,苯巴比妥的定性定量监测离子对为231.084.9,231.0188.1,毛细管出口端电压为90V,碰撞能分别为5eV和3eV。苯巴比妥标准系列为0.05μg/mL,0.1μg/mL,0.2μg/mL,0.5μg/mL,1.0μg/mL,2.0μg/mL,相关系数达到0.997,具有良好的线性相关。经过公式计算和转化,得到苯巴比妥钠的含量。分析表明,方法的检测限为0.1mg/kg,定量限为0.2mg/kg,测试方法具有良好的准确度和精密度,且对配合饲料、预混合饲料、浓缩饲料以及精料补充料均有较好的适用性,回收率均在60%-120%之间,相对偏差均在20%以内。本方法建立的饲料中苯巴比妥钠液相色谱-串联质谱测定方法,为饲料中苯巴比妥钠的定性定量检测提供了方法依据。
[Abstract]:Phenobarbital sodium (Phenobarbital Sodium) is a drug banned from feed and animal drinking water as stipulated by the Ministry of Agriculture's Proclamation No. 176. At present, there are no methods and standards for the determination of phenobarbital sodium in feed, which is a hidden danger for the quality supervision of feed and the healthy diet of people. It is urgent to establish a method for the determination of phenobarbital sodium in feed. Phenobarbital sodium is a weak acid strong base salt, and hydrolysis occurs in acid environment in the form of phenobarbital. The feed sample was pretreated with n-hexane and extracted with methanol hydrochloride solution. The sodium phenobarbital was extracted in the form of phenobarbital (Phenobarbital). The extract was purified by Waters Oasis HLB solid-phase extraction column (SPE) and eluted with 5% ammoniated methanol after elution with methanol water (298) solution. The eluent was condensed by nitrogen blowing, then redissolved with methanol and water (28), and detected by (HPLC) with high performance liquid chromatography (HPLC). The chromatographic column was C8 column with a diameter of 5 渭 m of 250 mm 脳 4.6 mm. The column temperature was controlled at 30 鈩，

本文编号：2253892