Department of Agricultural Chemistry, University of Córdoba, C-3 Building, Campus of Rabanales, 14071, Córdoba, Spain.
Anal Bioanal Chem. 2014 Jan;406(2):611-20. doi: 10.1007/s00216-013-7484-y. Epub 2013 Nov 20.
A simple method for the simultaneous determination of glufosinate and its metabolites in plants based on liquid chromatography-ultraviolet (LC-UV) absorption detection after derivatization with fluorenylmethoxycarbonyl chloride (FMOC-Cl) of some analytes to facilitate separation is reported here. Nonavailable standard metabolites were identified by LC-TOF/mass spectrometry (MS), which also confirmed all target analytes. Ultrasound-assisted extraction was used for sample preparation (power of 70 W and duty cycle of 0.7 s/s for 10 min) with subsequent evaporation of the extractant, reconstitution and filtration as the cleanup/concentration step prior to derivatization, and chromatographic separation and detection at 270 nm for underivatized analytes and 340 nm for those that were derivatized. The chromatographic analysis was completed in 40 min using a Luna® column (C18 phase). The analytical characteristics of the method were linear dynamic range of the calibration curves within 0.047-700 μg/mL with a regression coefficient (rc) of 0.999 for glufosinate, 0.077-700 μg/mL with a rc of 0.998 for N-acetyl-glufosinate, and 0.116-600 μg/mL with a rc of 0.998 for 3-(methylphosphinico)propanoic acid. The precision for the determination of glufosinate (studied at two levels, 0.1 and 5 μg/mL) was 2.7 and 6.0 % for repeatability and 4.7 and 7.2 % for within-laboratory reproducibility, respectively. Identification and confirmatory analysis of the presence of glufosinate and metabolites in the extracts from treated plants was carried out by LC-TOF/MS in high-resolution mode for the precursor ion. The method was validated by analyzing wheat (Triticum aestivum) samples (resistant and susceptible biotypes) treated with 300 g of glufosinate/ha following conventional agronomical practices.
本文报道了一种基于柱前衍生化的高效液相色谱-紫外（LC-UV）检测法，同时测定植物中草铵膦及其代谢物的方法。某些分析物用芴甲氧羰基氯（FMOC-Cl）衍生化后，通过液相色谱-飞行时间/质谱（LC-TOF/MS）鉴定出非标准代谢物，同时也确认了所有的目标分析物。超声辅助提取用于样品制备（功率为 70 W，占空比为 0.7 s/s，持续 10 min），然后提取剂蒸发，萃取物浓缩，衍生化前进行净化/浓缩，未衍生化分析物在 270 nm 处进行色谱分离和检测，衍生化分析物在 340 nm 处进行色谱分离和检测。使用 Luna®柱（C18 相）在 40 min 内完成色谱分析。方法的分析特性为草铵膦校准曲线的线性动态范围为 0.047-700 μg/mL，相关系数（rc）为 0.999；N-乙酰草铵膦的线性动态范围为 0.077-700 μg/mL，rc 为 0.998；3-（甲基膦酰基）丙酸的线性动态范围为 0.116-600 μg/mL，rc 为 0.998。草铵膦的测定精密度（在两个水平，0.1 和 5 μg/mL 下研究），重复性的精密度为 2.7%和 6.0%，实验室内再现性的精密度为 4.7%和 7.2%。用 LC-TOF/MS 在高分辨率模式下对提取液中处理过的植物中的草铵膦及其代谢物进行了存在分析物的鉴定和确证分析。该方法通过分析常规农艺实践中用 300 g/ha 草铵膦处理的小麦（Triticum aestivum）（抗性和敏感生物型）样品进行了验证。
