The determination of sialic acids (SIAs) has recently gained interest because of their potential role as markers of inflammatory disorders or chronic diseases. Hydrolysis of conjugated derivatives, solid-phase extraction (SPE) and derivatization steps constitute sample preparation prior to insertion of the analytical sample into a mu-liquid chromatograph-laser induced fluorescence (mu-LC-LIF) detector in the present method for the determination of two representative SIAs of human metabolism. Ultrasound-accelerated hydrolysis released free SIAs, which were efficiently concentrated in a dynamic manner using a lab-on-valve (LOV) module that allows automation of SPE for preconcentration and cleanup. This step was on-line connected with DMB-labeling of SIAs (derivatization), which was shortened from 180 min required with the conventional heating method to 20 min with ultrasound assistance. Individual separation of the target analytes was achieved within 20 min by mu-LC, while LIF detection endowed the overall method with high sensitivity. The LODs and LOQs provided by the method ranged 0.1-0.8 ng mL(-1) and 0.4-1.0 ng mL(-1) (between 0.1-0.8 pg and 0.4-1.0 pg expressed as on-column amount), respectively. High efficiency for interferents removal by SPE enabled the application of the method to four different biofluids serum, urine, saliva and breast milk for the determination of the target metabolites. (C) 2013 Elsevier B.V. All rights reserved.