Qian Kun, Tang Tao, Shi Tianyu, Wang Fang, Li Jianqiang, Cao Yongsong
College of Agriculture and Biotechnology, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing 100193, China.
Anal Chim Acta. 2009 Mar 9;635(2):222-6. doi: 10.1016/j.aca.2009.01.022. Epub 2009 Jan 18.
A pre-column derivatization high-performance liquid chromatographic method for glyphosate analysis has been developed. Derivatization of glyphosate was performed with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF). In pH 9.5 H(3)BO(3)-Na(2)B(4)O(7) media, the reaction of glyphosate with CNBF completed at 60 degrees C for 30min. The labeled glyphosate was separated on a Kromasil C18 column (250mmx4.6mm, 5microm) at room temperature and UV detection was applied at 360nm. The separation of labeled glyphosate was achieved within 15min by gradient elution mode. Compared to other pre-column derivatization, this derivatization was performed more mildly, the derivative was more stable, and the detection limits of a few reagents were higher than CNBF, except 9-fluorenylmethyl chloroformate (FMOC-Cl) using fluorescence and mass spectrometry, however, this reagent avoid to be removed after derivatization like FMOC-Cl. The detection limit of glyphosate was 0.009mgL(-1) (S/N=3) without preconcentration and reach MRL, which is set at the level of 0.1mgL(-1) in China. The method linearity correlation coefficient was 0.9999, in concentrations ranging from 0.3 to 48.5mgL(-1). The proposed method has been applied to the quantitative determination of glyphosate in environmental water with recoveries of 91.80-100.20% and R.S.D. of 2.27-6.80, depending on the sample investigated.
建立了一种用于草甘膦分析的柱前衍生高效液相色谱法。草甘膦的衍生化反应采用4-氯-3,5-二硝基三氟甲苯(CNBF)进行。在pH 9.5的硼酸-硼砂介质中,草甘膦与CNBF的反应在60℃下30分钟完成。标记后的草甘膦在室温下于Kromasil C18柱(250mm×4.6mm,5μm)上分离,并于360nm处进行紫外检测。通过梯度洗脱模式在15分钟内实现了标记草甘膦的分离。与其他柱前衍生方法相比,该衍生化反应条件更温和,衍生物更稳定,除了使用荧光和质谱的9-芴基甲基氯甲酸酯(FMOC-Cl)外,几种试剂的检测限均高于CNBF,然而,该试剂不像FMOC-Cl那样在衍生化后需要去除。草甘膦的检测限为0.009mgL⁻¹(S/N = 3),无需预浓缩即可达到中国设定的0.1mgL⁻¹的最大残留限量(MRL)。该方法在0.3至48.5mgL⁻¹的浓度范围内线性相关系数为0.9999。所提出的方法已应用于环境水中草甘膦的定量测定,回收率为91.80 - 100.20%,相对标准偏差(R.S.D.)为2.27 - 6.80,具体取决于所研究的样品。