Analytical method for urinary homovanillic acid and 5-hydroxyindoleacetic acid levels using HPLC with electrochemical detection applied to evaluate children environmentally exposed to manganese

Homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) are the urinary metabolites of dopamine (DA) and serotonin (5-HA), respectively. We aimed to develop an extraction method for the determination of HVA and 5-HIAA, using strong anionic exchange cartridges combined with HPLC with electrochemical detection, and apply it to measure the levels of HVA and 5-HIAA in children living near a ferro-manganese alloy plant in Simoes Filho, Brazil. The validated method showed good selectivity, sensitivity, precision, and accuracy. The limits of detection (LOD) were 4 and 8 & mu;mol/L for 5-HIAA and HVA, respectively, in urine. Recoveries ranged from 85.8 to 94%. The coefficients of determination (R-2) of the calibration curves were greater than 0.99. Spot urine samples of 30 exposed children and 20 nonexposed ones were processed accordingly. The metabolite levels in exposed and reference children were within the physiological ranges. The medians (range) for 5-HIAA and HVA of the exposed ones were 36.4 & mu;mol/L (18.4-58.0) and 32.9 & mu;mol/L (<LOD-91.9), respectively. There was no significant difference between the values presented by children in the reference group: 25.7 & mu;mol/L (19.9-81.4) and 35.2 & mu;mol/L (<LOD-67.6) for 5-HIAA and HVA, respectively. These results suggest that the quantification of the urinary metabolites possibly does not reflect the interference of manganese in the metabolism of DA and 5-HA in the central nervous system.