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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Potential diagnostic of Branched-Chain Ketoaciduria by HPLC-DAD

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Author(s):
Trintinalia, Maira Magalhaes [1] ; Lopes Alves, Atecla Nunciata [1] ; Fernandes, Liliam [1] ; Henriques Bechara, Etelvino Jose [1, 2] ; Assuncao, Nilson Antonio [1]
Total Authors: 5
Affiliation:
[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
Total Affiliations: 2
Document type: Journal article
Source: Journal of the Brazilian Chemical Society; v. 25, n. 10, p. 1857-1863, OCT 2014.
Web of Science Citations: 1
Abstract

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)

FAPESP's process: 12/02514-9 - Use of proteomics techniques to study radical acetylation of proteins triggered by products of the reaction biacetyl/peroxinitrite
Grantee:Nilson Antonio de Assunção
Support Opportunities: Regular Research Grants