Bioanalytical validation and clinical application of a liquid chromatography-tandem mass spectrometry method for the quantification of 3-orthomethyldopa, 5-hydroxytryptophan, 5-hydroxyindolacetic acid and homovanillic acid in human cerebrospinal fluid.

Inherited disorders of monoamine neurotransmitters are a subset of inborn errors of metabolism affecting biochemical pathways of catecholamines, serotonin or their enzymatic cofactors. Usually, their clinical presentation is similar to those of other common neurological syndromes. For this reason, they are frequently under-recognized and misdiagnosed. Because cerebrospinal fluid concentration of catecholamine metabolites (3-orthomethyldopa and homovanillic acid) and serotonin metabolites (5-hydroxytryptophan and 5-hydroxyindolacetic acid) presents a direct correlation with their brain levels, analysis of this group of compounds is critical to reach an accurate diagnosis. Although there are several published liquid chromatography-based bioanalytical methods for the quantification of these compounds, most of them present disadvantages, making their application difficult to implement in routine clinical practice. In this study, a rapid and simple UHPLC-MS/MS method for simultaneous quantification of 3-orthomethyldopa, 5-hydroxytryptophan, 5-hydroxyindolacetic acid and homovanillic acid in human cerebrospinal fluid was validated. All the evaluated performance parameters, including linearity, carryover, accuracy and precision (within and between-day), lower limit of quantitation, recovery, matrix effect and stability under different conditions met the acceptance criteria from international guidelines. Additionally, 10 human cerebrospinal fluid samples collected via lumbar puncture from 10 pediatric patients were quantified using the validated method to assess its clinical application and diagnostic utility for inherited monoamine neurotransmitter metabolism.

The level of exposure affects the arsenic urinary methylation profile of a population of children.

Differences in the As methylation capacity of Argentine children, exposed to different levels of As in drinking water were evaluated, considering the gender and the presence of the As3MT T860C gene polymorphism. Inorganic As (%IAs), monomethylated As (%MMA) and dimethylated As (%DMA), primary methylation index (PMI) and secondary methylation index (SMI) were evaluated and represented the As methylation capacity. Urinary As ranged from 18 to 5106 µg/g creatinine. Comparisons were performed between lowest and highest quartiles of urinary As. The level of exposure was positively related to urinary %MMA and negatively to %DMA and to SMI. Considering the presence of the As3MT T860C polymorphism, the level of exposure increased %MMA, and decreased %DMA and the SMI in carriers of the T/T genotype. SMI OR for T/T carriers was 10.61 (95% CI: 2.16-52.16, p: 0.0036). Regarding the gender, the level of exposure increased %MMA, and decreased %DMA and the SMI in girls and boys. SMI OR for girls was 8.71 (95% CI: 1.48-51.08, p: 0.0165) and for boys, OR: 18.15 (95% CI: 2.03-162.35, p: 0.0095). It was possible to identify the level of exposure as a factor that can modify the influence that other factors have on the methylation of As.