| Texto completo | |
| Autor(es): |
Pena, Izabella A.
[1]
;
Marques, Lygia A.
[2]
;
Laranjeira, Angelo B. A.
[3]
;
Yunes, Jose A.
[3, 4]
;
Eberlin, Marcos N.
[2]
;
Arruda, Paulo
[1, 5]
Número total de Autores: 6
|
| Afiliação do(s) autor(es): | [1] Univ Estadual Campinas UNICAMP, Ctr Biol Mol & Engn Genet, BR-13083875 Campinas, SP - Brazil
[2] Univ Estadual Campinas UNICAMP, Thomson Mass Spectrometry Lab, BR-13083861 Campinas, SP - Brazil
[3] Univ Estadual Campinas UNICAMP, Ctr Infantil Boldrini, BR-13083210 Campinas, SP - Brazil
[4] Univ Estadual Campinas UNICAMP, Dept Genet Med, Fac Ciencias Med, BR-13083887 Campinas, SP - Brazil
[5] Univ Estadual Campinas UNICAMP, Dept Genet & Evolucao, Inst Biol, BR-13081970 Campinas, SP - Brazil
Número total de Afiliações: 5
|
| Tipo de documento: | Artigo Científico |
| Fonte: | SPRINGERPLUS; v. 5, FEB 25 2016. |
| Citações Web of Science: | 6 |
| Resumo | |
Detection and quantification of lysine degradation metabolites in plasma is necessary for the diagnosis and follow-up of diseases such as pyridoxine-dependent epilepsy. The principal metabolites involved in the disease are related to the first steps of lysine oxidation, either through the saccharopine or the pipecolate pathways. Currently, there are three different analytical methods used to assess the content of these metabolites in urine and plasma, but they require different sample preparations and analytical equipment. Here, we describe a protocol that calls for a simple sample preparation and uses liquid chromatography tandem mass spectrometry (LC-MS/MS) that allows simultaneous detection and quantification of underivatized L-saccharopine, L-aminoadipic acid, L-pipecolic acid, piperideine-6-carboxylate, L-glutamic acid, and pyridoxal-5-phosphate in plasma samples. To validate the method we analyzed the time course degradation after intraperitoneal injection of L-lysine in C57BL/6/J mice. We observed that the degradation of lysine through the saccharopine pathway reached a maximum within the first 2 h. At this time point there was an increase in the levels of the metabolites saccharopine, aminoadipic acid, and pipecolic acid by 3-, 24- and 3.4-fold, respectively, compared to time zero levels. These metabolites returned to basal levels after 4-6 h. In conclusion, we have developed a LC-MS/MS approach, which allows simultaneous analysis of lysine degradation metabolites without the need for derivatization. (AU) | |
| Processo FAPESP: | 13/23920-8 - Técnicas modernas em espectrometria de massas e novas aplicações em ciências: química, bioquímica, materiais, forense, medicina, alimentos, farmácia e veterinária |
| Beneficiário: | Lygia de Azevedo Marques |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |
| Processo FAPESP: | 10/50114-4 - O papel da via da sacaropina em diversos modelos biológicos |
| Beneficiário: | Paulo Arruda |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 12/00235-5 - Mecanismos de indução da via da sacaropina em células humanas |
| Beneficiário: | Izabella Agostinho Pena |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado Direto |