Rapid and sensitive HILIC-ESI-MS/MS quantitation of polar metabolites of acrylamide in human urine using column switching with an online trap column

The carcinogen acrylamide (AA) is formed during the processing of food. AA is metabolized to mercapturic acids, which are excreted with urine. A hydrophilic interaction liquid chromatography tandem mass spectrometry method (HILIC-MS/MS) using a zwitterionic stationary phase (Zic-HILIC) was developed and validated to quantitate the mercapturic acids of AA (AAMA) and glycidamide (GAMA), and AAMA-sulfoxide in human urine. In contrast to reversed phases, the application of Zic-HILIC resulted in efficient retention and separation of these highly polar compounds. Off-line sample workup was avoided by application of column switching with a Stability BS-C17 trap column prior to the analytical column, thus minimizing interferences with the urinary matrix. Limit of quantification values (LOQs) were 0.5 microg/L (AAMA), 2.0 microg/L (AAMA-sulfoxide), and 1.0 microg/L (GAMA) in human urine. Median concentrations in urine samples ( n = 54) of six nonsmoking human subjects were 24.0 microg/L (AAMA, 7.8-79.8 microg/L), 16.7 microg/L (AAMA-sulfoxide, 6.8-70.1 microg/L), and 3.82 microg/L (GAMA, 1.0-23.6 microg/L).