Kadar Ali, Peyre Ludovic, de Souza Georges, Wortham Henri, Doumenq Pierre, Rahmani Roger
Aix Marseille Univ, CNRS, LCE, Marseille, France; INRA, UMR 1331 TOXALIM, Laboratoire de Toxicologie Cellulaire et Moléculaire des Xénobiotiques, BP 167, 400 Route des Chappes, 06903, Sophia Antipolis Cedex, France.
INRA, UMR 1331 TOXALIM, Laboratoire de Toxicologie Cellulaire et Moléculaire des Xénobiotiques, BP 167, 400 Route des Chappes, 06903, Sophia Antipolis Cedex, France.
Chemosphere. 2017 Oct;184:20-26. doi: 10.1016/j.chemosphere.2017.05.157. Epub 2017 May 29.
A method for the determination of chlorfenvinphos, ethion and linuron in liver samples by LC-MS/MS is described. Sample treatment was performed by using Sola™ polymeric reverse phase SPE cartridges after protein precipitation. Gradient elution using 10 mM ammonium formate in methanol (A) and 10 mM ammonium formate in water (B) was used for chromatographic separation of analytes on a Hypersil™ end-capped Gold PFP reverse phase column (100 mm × 2.1 mm, 3 μm). All analytes were quantified without interference, in positive ionization mode using multiple reaction monitoring (MRM) with chlorfenvinphos-d10 as internal standard. The whole procedure was validated according to the FDA guidelines for bioanalytical methods. The calibration curves for chlorfenvinphos, linuron and ethion compounds were linear over the concentration range of 0.005-2 μM (i.e. 0.0018-0.720 μg/mL, 0.0019-0.770 μg/mL and 0.0012-0.500 μg/mL respectively) with coefficients of determination higher than 0.998. A Lower limit of quantification of 0.005 μM was achieved for all analytes, i.e. 5.76, 6.08 and 3.84 μg/kg of liver for chlorfenvinphos, ethion and linuron respectively. Compounds extraction recovery rates ranged from 92.9 to 99.5% with a RSD of 2.3%. Intra- and inter-day accuracies were within 90.9 and 100%, and imprecision varied from 0.8 to 8.2%. Stability tests proved all analytes were stable in liver extracts during instrumental analysis (+12 °C in autosampler tray for 72 h) at the end of three successive freeze-thaw cycles and at -20 °C for up to 9 months. This accurate and robust analytical method is therefore suitable for contamination or metabolism studies.
描述了一种通过液相色谱-串联质谱法（LC-MS/MS）测定肝脏样本中毒虫畏、乙硫磷和利谷隆的方法。在蛋白质沉淀后，使用Sola™聚合物反相固相萃取柱进行样品处理。在Hypersil™封端金五氟苯基反相柱（100 mm×2.1 mm，3μm）上，使用甲醇中10 mM甲酸铵（A）和水中10 mM甲酸铵（B）进行梯度洗脱，以对分析物进行色谱分离。所有分析物均在正离子模式下，以毒虫畏-d10作为内标，采用多反应监测（MRM）进行定量分析，无干扰。整个程序根据美国食品药品监督管理局（FDA）生物分析方法指南进行了验证。毒虫畏、利谷隆和乙硫磷化合物的校准曲线在0.005 - 2μM浓度范围内呈线性（即分别为0.0018 - 0.720μg/mL、0.0019 - 0.770μg/mL和0.0012 - 0.500μg/mL），测定系数高于0.998。所有分析物的定量下限均为0.005μM，即毒虫畏、乙硫磷和利谷隆在肝脏中的定量下限分别为5.76、6.08和3.84μg/kg。化合物的萃取回收率在92.9%至99.5%之间，相对标准偏差（RSD）为2.3%。日内和日间准确度在90.9%至100%之间，不精密度在0.8%至8.2%之间。稳定性测试证明，在连续三个冻融循环结束后，以及在-20°C下长达9个月的时间里，所有分析物在仪器分析期间（自动进样器托盘在+12°C下放置72小时）在肝脏提取物中均稳定。因此，这种准确且稳健的分析方法适用于污染或代谢研究。
