| Full text | |
| Author(s): |
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]
Total Authors: 6
|
| Affiliation: | [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
Total Affiliations: 5
|
| Document type: | Journal article |
| Source: | SPRINGERPLUS; v. 5, FEB 25 2016. |
| Web of Science Citations: | 6 |
| Abstract | |
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) | |
| FAPESP's process: | 13/23920-8 - Development of new mass spectrometry techniques and their general applications in science: chemistry, biochemistry, material sciences, forensics, medicine, food science, pharmaceutical and veterinary medicine |
| Grantee: | Lygia de Azevedo Marques |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 10/50114-4 - The role of saccharopine pathway in diverse biological models |
| Grantee: | Paulo Arruda |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 12/00235-5 - Mechanisms of saccharopine pathway induction in human cells |
| Grantee: | Izabella Agostinho Pena |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |