超高效液相色谱-串联高分辨率质谱法同时测定猪毛发中12种吩噻嗪类药物的残留量

Simultaneous Determination of Residues of 12 Phenothiazines in Pig Hair by Ultra-High Performance Liquid Chromatography-Tandem High Resolution Mass Spectrometry

考虑到毛发样品取样方便、前处理简单,可实现动物源性农产品中违禁药物的持续动态监测,因此进行了题示研究。猪毛发样品于-40℃冷冻,清洗,氮气吹干,剪成小段,置于液氮冷冻研磨器中,设置撞子振荡频率至12次·s^(-1),研磨时间至3.5 min,稳定时间至5 min,将样品研碎成末。分取80 mg,加入4μL 1000μg·L^(-1)混合内标溶液以及10 mL含0.1%(体积分数,下同)甲酸的乙腈溶液,超声提取5 min,于4℃离心3 min。上清液过0.45μm滤膜,将全部滤液过Oasis PRIME HLB固相萃取柱。收集流出液,分取5 mL,于60℃氮吹至干。加入200μL甲醇复溶,于4℃离心1 min,上清液按照优化的仪器工作条件测定。在色谱分析中,以Waters Acquity BEH C_(18)色谱柱为固定相,不同体积比的0.1%甲酸溶液和甲醇的混合溶液为流动性进行梯度洗脱分离。在串联质谱分析中,以加热电喷雾离子源正离子模式电离,以母离子提取峰面积进行内标法定量,子离子的精确质量数进行定性。结果显示:氯丙嗪、氯丙嗪亚砜、异丙嗪、丙酰丙嗪、硫利达嗪、三氟拉嗪的质量浓度在1~200μg·L^(-1)内,异丙嗪亚砜、甲苯噻嗪、乙酰丙嗪、奋乃静、氟奋乃静、癸氟奋乃静在2~200μg·L^(-1)内和对应的峰面积比呈线性关系,检出限(3S/N)为2~4μg·kg^(-1)。按照标准加入法进行回收试验,回收率为84.6%~107%,测定值的相对标准偏差(n=5)为1.2%~13%。

Considering the hair sample had advantages of convenient sampling and simple pre-treatment,which can achieve continuous dynamic monitoring of prohibited drugs in animal derived agricultural products,the study mentioned by the title was conducted.The pig hair sample was forzen at-40℃,cleaned,blown to dryness by nitrogen,cuted into small pieces,and placed into a liquid nitrogen freezer grinder,of which the frequency of collision oscillation was set to 12 times·s^(-1),milling time was set to 3.5 min,and stabilization time was set to 5 min.The sample was ground into powder,an aliquot(80 mg)was taken,and 4μL of 1000μg·L^(-1)mixed internal standard solution and 10 mL of acetonitrile solution containing 0.1%(volume fraction,the same below)formic acid were added.The mixture was ultrasonicated for 5 min,and centrifuged at 4℃for 3 min.The supernatant was filtered by a 0.45μm filter membrane,and all the filtrate was passed through the Oasis PRIME HLB solid phase extraction column.The effluent was collected,and an aliquot(5 mL)was taken,and blown to dyrness by nitrogen at 60℃.Then 200μL of methanol was added for dissolution,and the solution obtained was centrifuged at 4℃for 1 min.The supernatant was determined according to the optimized instrument working conditions.In chromatographic analysis,gradient elution separation was performed using Waters Acquity BEH C_(18) column as the stationary phase,and mixed solutions composed of 0.1%formic acid solution and methanol at different volume ratios as the mobile phase.In the tandem mass spectrometry analysis,the positive ion mode of the heated electrospray ion source was used for ionization,the peak area of the parent ion was used for internal standard quantification,and the precise mass number of the daughter ion was used for qualitative analysis.It was shown that linear relationships between values of the mass concentration and the peak area ratio were kept in the range of 1-200μg·L^(-1)for chlorpromazine,chlorpromazine sulfoxide,promethazine,propionyl promethazine,thiolidazine,trifluoperazine and 2-200μg·L^(-1)for promethazine sulfoxide,methylthiazide,acetylpromethazine,perphenazine,fluphenazine,decafluphenazine,with detection limits(3S/N)in the range of 2-4μg·kg^(-1).Test for recovery was made according to the standard addition method,giving recoveries in the range of 84.6%-107%,and RSDs(n=5)of the determined values ranged from 1.2%to 13%.