Capillary electrophoresis tandem mass spectrometry determination of glutamic acid and homocysteine's metabolites: Potential biomarkers of amyotrophic lateral sclerosis

被引:30
作者
Cieslarova, Zuzana [1 ]
Lopes, Fernando Silva [2 ]
do Lago, Claudimir Lucio [2 ]
Franca, Marcondes Cavalcante, Jr. [3 ]
Colnaghi Simionato, Ana Valeria [1 ,4 ]
机构
[1] Univ Estadual Campinas, Inst Chem, Dept Analyt Chem, POB 6154, BR-13083970 Campinas, SP, Brazil
[2] Univ Sao Paulo, Inst Chem, Dept Fundamental Chem, Ave Prof Lineu Prestes,748 Cidade Univ, BR-05508000 Sao Paulo, SP, Brazil
[3] Univ Estadual Campinas, Dept Neurol, Fac Med Sci, POB 6111, BR-13083970 Campinas, SP, Brazil
[4] Univ Estadual Campinas, Natl Inst Sci & Technol Bioanalyt, Inst Chem, Dept Analyt Chem, POB 6154, BR-13083970 Campinas, SP, Brazil
基金
巴西圣保罗研究基金会;
关键词
Capillary electrophoresis-tandem mass spectrometry; Target metabolomics; Amino acids; Amyotrophic lateral sclerosis; Protein precipitation; Sample preparation; CONTACTLESS CONDUCTIVITY DETECTION; AMINO-ACIDS; HUMAN PLASMA; ELECTROMIGRATION METHODS; ZONE-ELECTROPHORESIS; SEPARATION SYSTEMS; BIOLOGICAL-FLUIDS; ORGANIC-SOLVENTS; PROTEINS; DERIVATIZATION;
D O I
10.1016/j.talanta.2017.03.103
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that affects both lower and upper motor neurons, leading to muscle atrophy, paralysis, and death caused by respiratory failure or infectious complications. Altered levels of homocysteine, cysteine, methionine, and glutamic acid have been observed in plasma of ALS patients. In this context, a method for determination of these potential biomarkers in plasma by capillary electrophoresis tandem mass spectrometry (CE-MS/MS) is proposed herein. Sample preparation was carefully investigated, since sulfur-containing amino acids may interact with plasma proteins. Owing to the non-thiol sulfur atom in methionine, it was necessary to split sample preparation into two methods: i) determination of homocysteine and cysteine as S-acetyl amino acids; ii) determination of glutamic acid and methionine. All amino acids were separated within 25 min by CE-MS/MS using 5 mol L-1 acetic acid as background electrolyte and 5 mmol L-1 acetic acid in 50% methanol/H2O (v/v) as sheath liquid. The proposed CE-MS/MS method was validated, presenting RSD values below 6% and 11% for intra- and inter-day precision, respectively, for the middle concentration level within the linear range. The limits of detection ranged from 35 (homocysteine) to 268 nmol L-1 (glutamic acid). The validated method was applied to the analysis of plasma samples from a group of healthy individuals and patients with ALS, showing the potential of glutamic acid and homocysteine metabolites as biomarkers of ALS.
引用
收藏
页码:63 / 68
页数:6
相关论文
共 35 条
[1]   A cost-effective method to get insight into the peritoneal dialysate effluent proteome [J].
Araujo, J. E. ;
Jorge, S. ;
Teixeira e Costa, F. ;
Ramos, A. ;
Lodeiro, C. ;
Santos, H. M. ;
Capelo, J. L. .
JOURNAL OF PROTEOMICS, 2016, 145 :207-213
[2]   The Glutamate Hypothesis in ALS: Pathophysiology and Drug Development [J].
Blasco, H. ;
Mavel, S. ;
Corcia, P. ;
Gordon, P. H. .
CURRENT MEDICINAL CHEMISTRY, 2014, 21 (31) :3551-3575
[4]   Determination of reduced and oxidized homocysteine and related thiols in plasma by thiol-specific pre-column derivatization and capillary electrophoresis with laser-induced fluorescence detection [J].
Chassaing, C ;
Gonin, J ;
Wilcox, CS ;
Wainer, IW .
JOURNAL OF CHROMATOGRAPHY B, 1999, 735 (02) :219-227
[5]   Determination of amino acids by capillary electrophoresis-electrospray ionization-mass spectrometry: An evaluation of different protein hydrolysis procedures [J].
Colnaghi Simionato, Ana V. ;
Moraes, Edgar Perin ;
Carrilho, Emanuel ;
Maggi Tavares, Marina Franco ;
Kenndler, Ernst .
ELECTROPHORESIS, 2008, 29 (10) :2051-2058
[6]   Separation of twenty underivatized essential amino acids by capillary zone electrophoresis with contactless conductivity detection [J].
Coufal, P ;
Zuska, J ;
van de Goor, T ;
Smith, V ;
Gas, B .
ELECTROPHORESIS, 2003, 24 (04) :671-677
[7]   Capillary electrophoresis-mass spectrometry for the analysis of amino acids [J].
Desiderio, Claudia ;
Iavarone, Federica ;
Rossetti, Diana Valeria ;
Messana, Irene ;
Castagnola, Massimo .
JOURNAL OF SEPARATION SCIENCE, 2010, 33 (16) :2385-2393
[8]   Liquid-phase based separation systems for depletion, prefractionation, and enrichment of proteins in biological fluids and matrices for in-depth proteomics analysis-An update covering the period 2014-2016 [J].
El Rassi, Ziad ;
Puangpila, Chanida .
ELECTROPHORESIS, 2017, 38 (01) :150-161
[9]   A comparison of depletion versus equalization for reducing high-abundance proteins in human serum [J].
Fernandez, Carolina ;
Santos, Hugo M. ;
Ruiz-Romero, Cristina ;
Blanco, Francisco J. ;
Capelo-Martinez, Jose-Luis .
ELECTROPHORESIS, 2011, 32 (21) :2966-2974
[10]   Analytical methods involving separation techniques for determination of low-molecular-weight biothiols in human plasma and blood [J].
Isokawa, Muneki ;
Kanamori, Takahiro ;
Funatsu, Takashi ;
Tsunoda, Makoto .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2014, 964 :103-115