高效液相色谱法测定猪排泄物中帕托珠利  

Determination of ponazuril in pig excrement by high performance liquid chromatography

作　　者：聂晶晶 王文湘[1] 华成云 梁立杰 黄显会[1] NIE Jingjing;WANG Wenxiang;HUA Chengyun;LIANG Lijie;HUANG Xianhui(College of Veterinary Medicine,South China Agricultural University/Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院/广东省兽药研制与安全评价重点实验室,广东广州510642

出　　处：《华南农业大学学报》2021年第1期34-41,共8页Journal of South China Agricultural University

基　　金：国家科技支撑计划子课题(2015BAD11B03-06)。

摘　　要：【目的】建立高效液相色谱(HPLC)结合二极管阵列检测器(PDA)检测猪排泄物中帕托珠利的方法。【方法】尿液样品用0.2%(φ)乙酸酸化乙腈和二氯甲烷进行2次提取;粪便样品经乙腈涡旋提取,亲水−亲油平衡(HLB)固相萃取柱净化。流动相为0.005 mol/L磷酸二氢钾溶液(A)−乙腈(B),尿液和粪便样品的流动相比例V(A)∶V(B)分别为55∶45和56∶44。检测波长为255 nm,柱温为35℃,进样量为30μL。【结果】猪尿液中帕托珠利的检测限和定量限分别为0.02和0.05μg/mL,在0.05~5.00μg/mL范围内呈良好的线性关系,决定系数(R2)为0.9998;在3个添加剂量(0.05、1.00和5.00μg/mL)下,帕托珠利在猪尿液中的平均回收率为93.49%~99.16%,批内和批间相对标准偏差(RSD)为0.97%~7.62%。猪粪便中帕托珠利的检测限和定量限分别为0.10和0.25μg/g,在0.25~100.00μg/g范围内呈良好的线性关系,R2为0.9995;在3个添加剂量(0.25、25.00、100.00μg/g)下,帕托珠利在猪粪便中的平均回收率为89.55%~95.88%,批内和批间RSD为1.76%~3.63%。帕托珠利在尿液和粪便样品中的提取回收率均大于89.50%,批内和批间RSD均小于8%。【结论】本研究方法对样品的前处理操作简单、灵敏度高,可用于猪排泄物中帕托珠利的检测分析。【Objective】To establish a method of high performance liquid chromatography(HPLC)with a photodiode array detector(PDA)for the determination of ponazuril in pig feces and urine.【Method】Urine samples were extracted twice with 0.2%(φ)acetic acetonitrile and dichloromethane.Feces samples were vortexextracted by acetonitrile and purified by hydrophile-lipophile balance(HLB)solid phase extraction column.The mobile phase was 0.005 mol/L potassium dihydrogen phosphate solution(A)-acetonitrile(B),the mobile phase ratios of V(A)∶V(B)for urine and feces samples were 55∶45 and 56∶44 respectively.The detection wavelength was 255 nm,the column temperature was 35℃and the injection volume was 30μL.【Result】The detection limit and quantitative limit of ponazuril in urine were 0.02 and 0.05μg/mL,respectively,which showed a good linear relationship in the range of 0.05−5.00μg/mL,and the determination coefficients(R2)was 0.9998.The average recovery rates ranged from 93.49%to 99.16%at three spiked levels of 0.05,1.00 and 5.00μg/mL,and the intra-batch and inter-batch relative standard deviations(RSDs)ranged from 0.97%to 7.62%.The detection limit and quantitative limit of ponazuril in feces were 0.10 and 0.25μg/g,respectively,which showed a good linear relationship within the range of 0.25−100.00μg/g,and R2 was 0.9995.The average recovery rates ranged from 89.55%to 95.88%at three spiked levels of 0.25,25.00 and 100.00μg/g,and the intra-batch and inter-batch RSDs ranged from 1.76%to 3.63%.The recovery rates of ponazuril in feces and urine were both higher than 89.50%,and the intra-batch and inter-batch RSDs were both lower than 8%.【Conclusion】This method has simple sample pretreatment and sensitive detection,and is suitable for the determination of ponazuril in pig excrement.

关 键 词：帕托珠利 高效液相色谱 猪 排泄物 回收率 精密度 稳定性 

分 类 号：S859.7[农业科学—临床兽医学]