液相色谱紫外/荧光检测法测定水产品中孔雀石绿和隐性孔雀石绿

Determination of Malachite Green and Leucomalachite Green Content by HPLC-ultraviolet/fluorescence Detection Method in Aquatic Product

试验旨在建立一种检测水产品中孔雀石绿和隐性孔雀石绿的液相色谱-紫外/荧光检测方法,并对样品前处理方法进行简化。在乙腈作为溶剂提取步骤中同时加入酸性氧化铝吸附,提取液离心分离后采用串联的丙磺酸阳离子固相萃取柱(PRS)和酸性氧化铝柱净化,0.05 mol/L的乙酸铵/乙腈(20∶80,V/V)为流动相,采用液相色谱-紫外/荧光检测法同时分析孔雀石绿和隐性孔雀石绿的含量,外标法定量。结果表明:孔雀石绿和隐性孔雀石绿在0.05~1.0 mg/L范围内线性良好,相关系数分别为为0.9999和0.9996,最低检出限分别为0.002 mg/L和0.5μg/L。当加标量分别为10、25、50μg时,空白鱼样的加标回收率在72.0%~91.4%之间,RSD均小于7.5%。该方法准确度高,适于水产品中孔雀石绿及其代谢残留产物—隐性孔雀石绿的分析检测。

A gradient high performance liquid chromatography（ HPLC） coupled with ultraviolet detector and fluorescence detector was developed for the detection of malachite green and leucomalachite green from aquatic product,and the sample pretreatment methods were simplified. Acetonitrile was used as extracting solvent assisted with acidic aluminium oxide adsorption,after centrifugal separation column and acidic alumina column purification series of PRS and acidic aluminium oxide solid phase extraction column,0. 05 mol/L ammonium acetate- acetonitrile（20 ∶ 80,v/v） as mobile phase,methylene chloride and leucomalachite green content were analyzed by ultraviolet detector and fluorescence detector simultaneously,quantified by external standard method. The results showed that the linear range of malachite green and leucomalachite green were0. 05- 1. 0 mg/m L（ r^2= 0. 9999 and r^2= 0. 9996 respectively）,and the detection limit was 0. 002 mg/L and0. 5 ug/L respectively. When the added quality of malachite green and leucomalachite green in blank fish sample were all 10,25,50 μg,respectively,the average recovery ranged from 72. 0% to 91. 4%,and the relative standard deviation was all lower than 7. 5%. This method was precise and suitable for determination of malachite green and its metabolic product residues leucomalachite green in aquatic products.