Potential diagnostic of Branched-Chain Ketoaciduria by HPLC-DAD

Autor(es)

Trintinalia, Maira Magalhaes ^[1] ; Lopes Alves, Atecla Nunciata ^[1] ; Fernandes, Liliam ^[1] ; Henriques Bechara, Etelvino Jose ^{[1, 2]} ; Assuncao, Nilson Antonio ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09972270 Diadema, SP - Brazil

^[2] Univ Sao Paulo, Inst Quim, Dept Quim Fundamental, BR-05508000 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of the Brazilian Chemical Society; v. 25, n. 10, p. 1857-1863, OCT 2014.
• Citações Web of Science: 1

Resumo

A system of high performance liquid chromatography (HPLC) was used for the development and validation of efficient method for quantitative determination of three aminoacids involved in the inherited metabolic disease Branched-Chain Ketoaciduria (BCK), also called maple syrup urine disease. The analytical conditions were selected in order to obtain baseline separation profiles of the amino acids known to be altered in blood plasma of BCK patients, namely L-valine, L-isoleucine, and L-leucine. Most accurate data were obtained using HPLC/diode detector. As the analytes do not have chromophore groups, they were pre-derivatized with o-phthaldialdehyde (OPA), yielding an unsaturated adduct, making thus possible the detection of amino acids. The validation was conducted according to National Health Surveillance Agency (ANVISA) and Guidance for Industry (Bioanalytical Method Validation) United States Food and Drug Administration (U.S. FDA). The results were satisfactory, with high sensitivity, good linearity, precision and accuracy, limit of detection and quantification, all within the established parameters for bioanalytical methods, showing its applicability and low cost compared to other existing techniques such as sequential mass spectrometry. For the three amino acids, L-valine, L-isoleucine and L-leucine, the detection limits (LOD) found were: 1.61, 1.84 and 1.88 mmol L- 1 and the quantification limits (LOQ) 4.37, 6.13 and 6.27 mmol L- 1, respectively. (AU)

• Processo FAPESP: 07/59039-2 - Participação de receptores de cininas em vias de sinalização ativadas por angiotensina: aspectos vasculares, bioquímicos e moleculares
• Beneficiário: Liliam Fernandes
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 06/60245-3 - Desenvolvimento de metodologias por eletroforese capilar-espectrometria de massas para análise de metabólitos relacionados a desordens do metabolismo humano
• Beneficiário: Nilson Antonio de Assunção
• Modalidade de apoio: Auxílio à Pesquisa - Programa Primeiros Projetos
• Processo FAPESP: 12/02514-9 - Uso de técnicas proteômicas para o estudo da acetilação de proteínas via radicalar desencadeada pelos produtos da reação diacetil/peroxinitrito
• Beneficiário: Nilson Antonio de Assunção
• Modalidade de apoio: Auxílio à Pesquisa - Regular