高效液相色谱-荧光检测法(HPLC-FLD)测定水体中孔雀石绿

Determination of malachite green content by HPLC-fluorescence detection method in water

实验旨在建立水样中孔雀石绿测定的液相色谱-荧光检测方法.水样中孔雀石绿采用二氯甲烷为溶剂超声波辅助提取,提取液离心分离后经MCX柱净化,并用5%氨水甲醇溶液洗脱,以Develosil C18色谱柱为分析柱,0.05 mol/L的乙酸铵(pH 4.5)-乙腈(20:80,V/V)为流动相,采用荧光检测器分析,外标法定量.结果表明:孔雀石绿在0.05μg^1.0μg/mL范围内线性良好,相关系数为0.9993,检出限为0.5μg/L.空白水样在加标孔雀石绿含量分别为1.0μg、2.5μg、5.0μg时,孔雀石绿的平均加标回收率为72.0%~86.4%,相对标准偏差为5.4%~7.5%.该方法准确度高、精密度良好,适用于水产养殖用水中孔雀石绿的测定.

A gradient high performance liquid chromatography （HPLC） coupled with fluorescence detection was developed for the detection of malachite green from water. Methylene chloride was used as extracting solvent assisted with ultrasonic method, the malachite green extract was purificated by MCX column after centrifuged, then eluted by 5 % ammonia solution in methanol elution, Develosil C 18 column was used as analytical column, 0.05 mol/L ammonium acetate （ pH 4.5） - acetonitrile （20 ： 80, v/v） as mobile phase, analyzed by fluores- cence detector, quantified by external standard method. The results showed that the linear range of malachite green was 0.05 μg/mL ~ 1.0 μg/mL （ r2 = 0.9993 ）, and the detection limit was 0.5 μg/L. When the added quality of malachite green in blank water was 1.0 μg, 2.5 μg, 5.0 μg, respectively, the average recovery ranged from72.0% to 86.4%, and the relative standard deviation was 5.4% ~7.5%. This method was precise and accurate, and suitable for determination of malachite green in aquaculture water.