分散固相萃取/高效液相色谱法测定水产品中氯苯胍的残留量

摘要

A dispersive solid phase extraction coupled with high performance liquid chromatographic (HPLC) method was established for the determination of robenidine in fishery products. The sample was extracted with acidified ethyl acetate as extraction solvent and C18 as the sorbent, then purified by alumina-N. After evaporated to dryness, the residue was redissolved in acetonitrile water solution and defatted with n-hexane. The external standard method was used to quantify robenidine. The optimization chromatographic conditions were as follows; separation column; Agilent ODS-C,8 chromatographic column(250 mm x 4. 6 mm, 5 |xm) , mobile phase; pH 6. 8 acetonitrile - ammonium di-hydrogen phosphate buffer (65 : 35 , by volume), flow rate; 1.0 mlVmin, column temperature; 40 °C , sample volume; 20 (jlL, detection wavelength; 353 nm. Under the optimal conditions, the calibration curve showed a good linearity in the range of 0. 01 - 1. 00 mg/L with a correlation coefficient of 0. 999 9. Four common representative samples(tilapia, eel, shrimp and turtle) were chosen for the validation of the method by recovery and precision. The obtained recoveries were 76% -106% and the relative standard deviations were 0. 81% -8. 2% . The limit of detection was 10 |xg/ kg, and the limit of quantitation was 30 |xg/kg. Results of three inter-laboratory validation tests showed that the recoveries of robenidine in fish, shrimp and turtle samples at three spiked concentration levels were greater than 70% with relative standard deviations not more than 11% . With the ad-vantages of simple operation, good sensitivity, accuracy and reproducibility, the proposed method was successfully used in the determination of robenidine in real samples, which can be recommended as the related test method for application.%建立了水产品中氯苯胍残留的分散固相萃取/高效液相色谱检测法.采用酸化乙酸乙酯为提取溶剂、C18填料为基质分散剂提取水产品中残留的氯苯胍,经中性氧化铝柱净化,浓缩后用乙腈水溶液定容,经正己烷脱脂后上机检测,外标法定量.优化的色谱条件为:采用Agilent ODS-C18色谱柱(250 mm×4.6 mm,5μm)分离,以体积比为65:35的乙腈-磷酸二氢铵缓冲液(三乙胺调pH值至6.8)为流动相,流速1.0 mL/min,柱温40 ℃,进样量20μL,紫外检测器检测波长353 nm.在该色谱条件下,氯苯胍在质量浓度为0.01 ～1.00 mg/L范围内线性良好,相关系数为0.999 9.以罗非鱼、鳗鲡、对虾及甲鱼为基质进行3个水平的加标回收实验,回收率为76％～ 106％,相对标准偏差为0.81％ ～ 8.2％,检出限为10 μg/kg,定量下限为30 μg/kg.通过3家实验室间验证实验表明:在鱼、虾及甲鱼中3个加标水平的回收率均大于70％,相对标准偏差不高于11％.该方法操作简单,灵敏度、准确度与再现性良好,已成功用于实际样品检测,可作为相关检验方法进行推广应用.