反相高效液相色谱法测定对虾组织中噁喹酸残留

Determination of Oxolinic Acid Residue in Shrimp,Litopenaeus Vannamei by Reversed-Phase High Performance Liquid Chromatography

建立了测定对虾血淋巴、肌肉和肝胰腺等组织中噁喹酸含量的反相高效液相色谱法。采用乙腈提取对虾血淋巴、肌肉、肝胰腺和鳃组织中噁喹酸,色谱柱为Aglient Zorbax SB-C18柱（150×4.6mm,5μm）,以乙腈-0.01mol·L-1四丁基溴化铵（磷酸调节pH为2.75）为流动相,流速1.0mL·min-1,荧光检测器检测。噁喹酸在0.002-10μg·mL-1范围内线性关系良好,相关系数R2=0.9996-0.9999。加标回收率为84.70%-90.60%,日内、日间精密度（RSD）分别为1.60%-4.02%、2.91%-4.73%,定量限分别为0.02μg·mL-1、0.01μg·g-1、0.02μg·g-1和0.02μg·g-1。该方法操作简单,重现性好,适用于对虾样品中噁喹酸含量的分析。

A method of reversed-phase high performance liquid chromatography（RP-HPLC）was developed to determine oxolinic acid residue in shrimp,Litopenaeus vannamei.The influence of different detection wavelengths and mobile phases on chromatographic behavior of oxolinic acid,and influence of different extraction methods on recovery of oxolinic acid were studied.The mobile phase was acetonitrile and 0.01 mol·L-1tetrabutyl ammonium bromide（pH was adjusted to 2.75 with phosphate solution）.Aglient Zorbax SB-C18column（150×4.6 mm,5μm）was used with the column temperature of 40 ℃.Fluorescence detector was used with excitation wavelength of 265 nm and emission wavelength of 380 nm.Sampling volume was 10 μL.The method was found to be linear and reproducible in the concentration range of 0.002-10μg·mL-1 with the correlation coefficient of 0.9996-0.9999.Oxolinic acid in shrimp tissues were extracted by acetonitrile,and the average recoveries of oxolinic acid from hemolymph,muscle,hepatopancreas and gill were 84.70% to 90.60%.The relative standard deviations for intra-day and inter-day precisions were 1.60%-4.02% and 2.91%-4.73%,respectively.The quantitative detection limits of oxolinic acid of hemolymph,muscle,hepatopancreas and gill were 0.02μg·mL-1,0.01μg·g-1,0.02μg·g-1 and 0.02μg·g-1,respectively.The method is simple,reproducible and suitable for determining oxolinic acid in shrimp tissues.