Hydrophilic interaction LC combined with electrospray MS for highly sensitive analysis of underivatized amino acids in rhizosphere research

Analysis of underivatized amino acids is challenging regarding the separation as well as the detection of these small polar analytes. Hydrophilic interaction LC using a 2.1 x 150 mm ZIC (R) (ZIC, zwitterionic)-hydrophilic interaction LC from SeQuant as stationary phase with 1% v/v formic acid in water and ACN as eluents was combined with MS/MS in multiple reaction monitoring mode for the separation and the detection of 16 underivatized amino acids. Regression coefficients of eight or seven point calibrations varied from 0.9454 to 0.9993. Absolute LODs and LOQ (on column) were in the fmol range (0.1-12 and 0.4-41 fmol, respectively). A fast screening method of 19 min total runtime has been developed offering applicability to samples from rhizosphere studies - characterized by low analyte concentrations and complex matrices. A successful application to the analysis of tyrosine in samples from soil adsorption experiments is presented as well as an evaluated enrichment procedure for amino acids derived from plant culture in nutrient solution.