Dai Ming, Xie Ping, Liang Gaodao, Chen Jun, Lei Hehua
Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology of China, Institute of Hydrobiology, Chinese Academy of Sciences, Donghu South Road 7, Wuhan 430072, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Feb 1;862(1-2):43-50. doi: 10.1016/j.jchromb.2007.10.030.
A sensitive and selective liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantitative determination of microcystin-LR (MC-LR) and its glutathione conjugate (MC-LR-GSH) in fish tissues. The analytes were extracted from fish liver and kidney using 0.01M EDTA-Na(2)-5% acetic acid, followed by a solid-phase extraction (SPE) on Oasis HLB and silica cartridges. High-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry, operating in selected reaction monitoring (SRM) mode, was used to quantify MC-LR and its glutathione conjugate in fish liver and kidney. Recoveries of analytes were assessed at three concentrations (0.2, 1.0, and 5microg g(-1) dry weight [DW]) and ranged from 91 to 103% for MC-LR, and from 65.0 to 75.7% for MC-LR-GSH. The assay was linear within the range from 0.02 to 5.0microg g(-1) DW, with a limit of quantification (LOQ) of 0.02microg g(-1) DW. The limit of detection (LOD) of the method was 0.007microg g(-1) DW in both fish liver and kidney. The overall precision was determined on three different days. The values for within- and between-day precision in liver and kidney were within 15%. This method was applied to the identification and quantification of MC-LR and its glutathione conjugate in liver and kidney of fish with acute exposure of MC-LR.
建立了一种灵敏且具选择性的液相色谱 - 串联质谱法，并进行了验证，用于同时定量测定鱼组织中的微囊藻毒素 - LR（MC - LR）及其谷胱甘肽共轭物（MC - LR - GSH）。使用0.01M乙二胺四乙酸二钠 - 5%乙酸从鱼肝和肾中提取分析物，随后在Oasis HLB和硅胶柱上进行固相萃取（SPE）。采用电喷雾电离质谱的高效液相色谱（HPLC），在选择反应监测（SRM）模式下运行，用于定量鱼肝和肾中的MC - LR及其谷胱甘肽共轭物。在三个浓度（0.2、1.0和5μg g⁻¹干重[DW]）下评估分析物的回收率，MC - LR的回收率为91%至103%，MC - LR - GSH的回收率为65.0%至75.7%。该测定法在0.02至5.0μg g⁻¹ DW范围内呈线性，定量限（LOQ）为0.02μg g⁻¹ DW。该方法在鱼肝和肾中的检测限（LOD）均为0.007μg g⁻¹ DW。在三个不同日期测定了总体精密度。肝和肾中日内和日间精密度值均在15%以内。该方法应用于急性暴露于MC - LR的鱼的肝和肾中MC - LR及其谷胱甘肽共轭物的鉴定和定量。
