反相高效液相色谱法检测鲫鱼肌肉中伊维菌素

Determination of Ivermectin Residues in Crucian Muscle by Reversed Phase High Performance Liquid Chromatography

以鲫鱼肌肉为材料,建立了一种简便、准确的反相高效液相色谱法,用于鱼肉中伊维菌素的药物残留检测和药代动力学研究。鲫鱼肌肉中的伊维菌素用乙酸乙酯提取,离心后的上清液于45℃下氮气吹干,残渣用甲醇-水(4∶1,V/V)混合液溶解,正己烷去脂,离心后取下清液,进样20μL进行HPLC分析。本方法考察了提取剂、柱温和流动相的影响,确定了以乙酸乙酯为提取剂,柱温25℃和甲醇-水(90∶10,V/V)为流动相的优化条件。方法线性范围为0.025～2.50μg/g,相关系数r=0.9997,检出限为11ng/g。批内RSD为1.0%～5.8%,日间RSD为1.0%～7.7%,相对回收率(99.8±4.1)%,绝对回收率为(95.9±3.9)%。本方法具有操作简便,结果准确可靠的特点。

A simple reversed phase high performance liquid chromatographic method has been developed for the determination of ivermectin residues in fish muscle. The samples were extracted with ethyl acetate, after centrifugation, the supernatants were evaporated to dryness at 45℃ with nitrogen. The residues were dissolved with the mixture of methanol-water （4： 1, V/V） and defatted with hexane. After centrifugation, 20 μL of the underlayer was detected by reversed phase high performance liquid chromatography. The influencing factors including extraction solvent, column temperature and mobile phase were investigated. The optimum experimental conditions were as follows： ethyl acetate as extraction solvent, column temperature set at 25 ℃ and methanol-water（90：10, V/V） as mobile phase. The calibration curve was good linear between peak area and concentration of ivermectin from 0. 025 - 2.50μg/g with the correlation coefficient of r = 0. 9997. The method detection limit was 11 ng/g. The within-batch and the between-day standard deviations of this method are 1. 0% -5.8% and 1. 0% -7.7% , with a relative and absolute recovery of（99.8 ±4. 1 ）% and （95.9 ± 3.9） %, respectively. The method has the advantages of reliable, simple and fast in operation.