Analysis of Heterocyclic Amines in Human Urine Using Multiple Solid-Phase Extraction by Liquid Chromatography/Mass Spectrometry

摘要

A multiple solid-phase extraction (SPE) method was used with liquid chromatography, coupled with mass spectrometry (LC/MS), for the analysis of heterocyclic amines (HCAs) in human urine. Separation efficiencies based on the pH of the mobile phase and the types of columns were compared. An amide column showed better baseline separation and narrower HCApeak widths at pH 5.0 for the mobile phase than a C_8 column. Each SPE step, HLB, MCX, and Hy-bridSPE, was optimized by controlling the pH conditions. The combined method with the three SPEs effectively removed interfering species that cause ion-suppression during HCA detection. Validation of the method, performed with SIM and SRM detection, showed correlation coefficients above 0.991 in the range 0.3 -16.7 ng/mL. Recovery rates were 45.4 - 97.3% on the C_8 column and 71.8-101.4% on the amide column, and method detection limits were 0.11 - 0.65 ng/mL on the C_8 column and 0.12 - 0.48 ng/mL on the amide column. This method using multiple SPEs offers significant benefits for high-throughput determination of HCAs in urine.