| Texto completo | |
| Autor(es): |
Rodrigues, Karina T.
;
Mekahli, Djalila
;
Tavares, Marina F. M.
;
Van Schepdael, Ann
Número total de Autores: 4
|
| Tipo de documento: | Artigo Científico |
| Fonte: | ELECTROPHORESIS; v. 37, n. 7-8, p. 9-pg., 2016-04-01. |
| Resumo | |
A CE-ESI-MS method was developed and validated for the separation and quantitative analysis of amino acids (AA) in urine. Experimental parameters related to the CE-MS interface, BGE, and mass spectrometer (MS) settings were optimized providing a good separation of 27 AA, including the isomers L-leucine, L-isoleucine, and L-alloisoleucine, in less than 30 min. The sheath liquid was composed by 0.50% formic acid in 60% (v, v) methanol-water delivered at a flow rate of 5 mu L/min. The BGE consisted of 0.80 mol/L formic acid at pH 1.96 and 15% methanol. A pH stacking procedure was implemented to enhance sensitivity (a 12.5% NH4OH solution was injected at 0.5 psi/9 s prior to samples injected at 0.6 psi/20 s). The proposed method was validated according to FDA and ICH protocols exhibiting acceptable parameters. Analytical curves presented coefficients of determination from 0.996 to 0.9997 (with large F statistics and low p-values). LODs and quantification ranged from 0.63 to 29 mu mol/L and from 1.9 to 86 mu mol/L, respectively. Practical repeatability was obtained for all AA with coefficients of variation better than 0.55% CV (migration time) and 1.7% CV (peak area ratios; methionine sulfone as internal standard). Recoveries of AA in spiked urine ranged from 92.0 to 123% with few exceptions. Moreover, a successful quantification ofAAin pooled control and test urine samples, which compose a vesicoureteral reflux cohort, was achieved showing the potential applicability of the proposed method for targeted metabolomics studies using CE-ESI-MS with an Ion Trap as mass analyzer. (AU) | |
| Processo FAPESP: | 12/09822-0 - Avaliação do perfil metabólico global urinário do refluxo vésico-ureteral por CE-MS |
| Beneficiário: | Karina Trevisan Rodrigues |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |
| Processo FAPESP: | 14/18789-2 - Avaliação da CE-MS visando para o estudo metabolômico alvo do refluxo vesico ureteral |
| Beneficiário: | Karina Trevisan Rodrigues |
| Modalidade de apoio: | Bolsas no Exterior - Estágio de Pesquisa - Doutorado |