Field-amplified sample injection and sweeping micellar electrokinetic chromatography in analysis of glyphosate and aminomethylphosphonic acid in wheat

Glyphosate, a widely used herbicide, has been classified as probably carcinogenic to humans by the International Agency for Research on Cancer (IARC). In the present study a method based on Field-Amplified Sample Injection and Sweeping Micellar Electrokinetic Chromatography (FASI sweep-MEKC) has been developed and validated for determination of glyphosate and its microbial metabolite aminomethylphosphonic acid (AMPA) in wheat flour. The method involved a preliminary solid phase extraction for cleanup of the aqueous extracts from wheat flour, based sequentially on C18 and strong anion exchange cartridges, followed by derivatization using 9-fluorenylmethylchloroformate. Optimization of sample cleanup and derivatization procedure was carried out by a HPLC-UV method, whereas FASI sweep-MEKC was applied for achieving the sensitivity necessary for analysis of real samples. To this regard, optimum conditions involved the use of an extended path fused-silica capillary (80 cm total length, 50 μm, i.d.) filled with a high concentration buffer (sodium phosphate 100 mM, pH 2.2). Electrokinetic sampling was carried out at −10 kV with injection time of 700 s and the separation of the loaded analytes was performed under MEKC conditions using sodium phosphate buffer 50 mM at pH 2.2, supplemented with sodium dodecyl sulfate, 100 mM. The method was validated for linearity, precision, accuracy and sensitivity, showing that using conventional UV detection (210 nm) the achieved limit of quantitation (LOQ) values for both the analytes were widely lower than those set by Authorities. In particular, LOQ for glyphosate and AMPA were found to be 5 and 2.5 ng/mL, respectively, corresponding to 0.1 and 0.05 mg/kg, in wheat flour. The method, applied to commercially available real samples (wheat flour from different manufacturers) and to an experimental sample obtained by cv. Svevo wheat, can be considered as a convenient alternative to the existing approaches in analysis of complex matrices.