Effect of liquid chromatography separation of complex matrices on liquid chromatography–tandem mass spectrometry signal suppression

The effect of liquid chromatography separation on liquid chromatography–tandem mass spectrometry (LC–MS–MS) signal response for the characterization of low-molecular-mass compounds in a complex matrix was investigated. Matrix induced signal suppression appears throughout the entire LC–MS–MS analysis of wheat forage extract, with greatest suppression occurring at early retention times. Experimental results show that co-elution of matrix components and analytes from the LC column may be most strongly attributed to column overloading rather than similar analyte and matrix retention behavior. As a result, two-dimensional (LC–LC) separation can be a highly effective approach to address signal suppression effects for the quantitative LC–MS–MS analysis of complex matrix samples.