用高效液相色谱法检测底泥环境中孔雀石绿的方法

Determination of malachite green in the sediments from aquaculture environment by HPLC with fluorescence detector

研究了用高效液相荧光法检测养殖池塘底泥中孔雀石绿的方法，比较了不同提取液对孔雀石绿的提取效果，针对相应的提取液用正交试验建立最佳提取方案，采用硼氢化钾将孔雀石绿还原成隐色孔雀石绿后用高效液相色谱荧光检测器进行定量分析。结果表明：前处理中采用甲醇和二氯甲烷混合提取比较简单，最适宜提取方案是提取液用量为20mL，震荡提取5min，重复提取2次；硼氢化钾还原效果良好，用量2mg即能将该体系中的孔雀石绿完全还原。本方法在0．5—800ng／mL范围内具有很好的线性，孔雀石绿总量最低检出限为0．36μg/kg。孔雀石绿和隐色孔雀石绿的加标回收率分别为82．4％～84．8％、88．1％～92．3％，相对标准偏差为3．16％～4．74％、1．91％～4．11％。

A method to determine malachite green （MG） in sediments from aquaculture environment was studied by high performance liquid chromatography （HPLC） with fluorescence detector. Different extraction solvents were compared and the optimum extract condition was confirmed by orthogonal test. The results showed that the mixture of methanol and dichloromethane was comparably convenient in the extraction assay. The optimum extraction condition was as the following： extracting for 5 min with 20 mL extraction solvent and repeat once, addition of 2 mg KBH4to the system reducing the MG to leumcomalachite green （LMG） with the linear range from 0.5 ng/mL to 800 ng/mL, and the detection limit of 0.36 μg/kg for LMG. The average recovery of the spiked sample was found to be 82.4% -84.8% for MG, and 88.1% -92.3% for LMG. The relative standard deviation （RSD） was 3.16% - 4.74% for MG, and 1.91% -4.11% for LMG. The method was found to be suitable for the trace analysis of MG in sediments from aquaculture environment and offer a better reference for establishing the related standards.