Department of Pharmacy and Biotechnology, University of Bologna, Via Belmeloro 6, Bologna 40126, Italy.
Department of Chemistry "Giacomo Ciamician", University of Bologna, Via F. Selmi 2, Bologna 40126, Italy.
J Chromatogr A. 2022 Oct 11;1681:463452. doi: 10.1016/j.chroma.2022.463452. Epub 2022 Aug 28.
Glyphosate or N-(phosphonomethyl)glycine, widely used as herbicide in agriculture to control weeds and to facilitate harvesting, has been included in Group 2A pollutants (probably carcinogenic to humans) by the International Agency for Research on Cancer (IARC). In intensive agricultural areas, runoff and soil leaching are likely to drive glyphosate to surface waters, where the compound is often detected together with its main microbial metabolite, aminomethylphosphonic acid (AMPA). In the present study a method based on capillary electrophoresis coupled with light-emitting diode-induced fluorescence detection has been developed and validated for the determination of the two compounds in whole soft mass of marine mussels (Mytilus galloprovincialis). The method is based on the acidic hydrolysis of lyophilized tissue using 6 M HCl (oven at 110 °C for 22 h) to release the target analytes; their subsequent derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole, was found to be suitable for the sensitive fluorescence detection. To achieve optimum separation of the analytes from the matrix and degradation reagent interferences, the background electrolyte constituted by borate buffer (pH 9.2, 30 mM) was supplemented with 10 mM heptakis(2,6-di-O-methyl)-β-cyclodextrin. The method was validated for linearity, precision, accuracy, robustness and sensitivity showing LOQ of 0.2 and 1.0 µg/g in fresh tissues, for AMPA and glyphosate, respectively; the recovery values ranged within 88.5 - 94.6% for glyphosate and 70.4 - 76.6% for AMPA. Experimental samples of Mediterranean mussels M. galloprovincialis treated with 100 µg/L or 500 µg/L of both glyphosate and AMPA, showed a dose dependent bioaccumulation of the compounds reaching maximum level of 77.0 µg/g and 11.3 µg/g of AMPA and glyphosate, respectively. The study demonstrates for the first time M. galloprovincialis as potential sentinel organisms for the environmental occurrence of these small amphoteric pollutants.
草甘膦或 N-(膦酸甲基)甘氨酸，作为农业中广泛使用的除草剂，用于控制杂草并促进收获，已被国际癌症研究机构 (IARC) 归类为 2A 类污染物（可能对人类致癌）。在集约化农业区，径流和土壤淋滤可能会将草甘膦带到地表水中，而该化合物通常与主要的微生物代谢物氨甲基膦酸 (AMPA) 一起被检出。在本研究中，开发并验证了一种基于毛细管电泳与发光二极管诱导荧光检测相结合的方法，用于测定海洋贻贝（贻贝）全软体组织中的两种化合物。该方法基于使用 6 M HCl（在 110°C 下放置 22 小时）对冻干组织进行酸性水解，以释放目标分析物；随后使用 4-氟-7-硝基-2,1,3-苯并恶二唑对其进行衍生化，发现该方法适合于灵敏的荧光检测。为了实现从基质和降解试剂干扰中最佳分离分析物，背景电解质由硼酸盐缓冲液（pH 9.2，30 mM）组成，并用 10 mM 七（2,6-二-O-甲基）-β-环糊精补充。该方法对线性、精密度、准确度、稳健性和灵敏度进行了验证，结果表明 AMPA 和草甘膦的新鲜组织的 LOQ 分别为 0.2 和 1.0 μg/g；草甘膦和 AMPA 的回收率范围分别为 88.5-94.6%和 70.4-76.6%。用 100 μg/L 或 500 μg/L 的草甘膦和 AMPA 处理的地中海贻贝 M. galloprovincialis 的实验样品显示出化合物的剂量依赖性生物积累，达到 AMPA 和草甘膦的最大水平分别为 77.0 μg/g 和 11.3 μg/g。该研究首次证明贻贝是这些小两性污染物环境发生的潜在指示生物。
