Christa M. Snyder

Indiana University Bloomington

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Miloš V. Novotný

Indiana University Bloomington

Stephen C. Jacobson

Indiana University Bloomington

Thomas H. Linz

Wayne State University

Margit I. Campos

Indiana University Bloomington

Susan M. Lunte

University of Kansas

Xiaomei Zhou

Lianyungang Oriental Hospital

William R. Alley

Waters (United States)

Indranil Mitra

Roche Pharma AG (Germany)

Jonathan A. Karty

Indiana University Bloomington

John A. Goetz

Indiana University Bloomington

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Indiana University Bloomington

Indiana University

Bloomington Health Foundation

Indiana University – Purdue University Indianapolis

University of North Carolina at Chapel Hill

University of Kansas

University of Defence

Masaryk Memorial Cancer Institute

Národný Onkologický Ústav

Indiana University School of Medicine

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Analyzing Methylated Arginines Using Capillary Electrophoresis and Laser Induced Fluorescence Detection

Christa M. Snyder Thomas H. Linz Susan M. Lunte

The methylated arginines monomethylarginine (MMA), asymmetrical dimethylarginine (ADMA) and symmetrical dimethylarginine (SDMA) have been shown to be independent predictors of cardiovascular disease (CVD). As a result, an analytical method was developed to determine the concentrations of MAs in human plasma in order to track the onset and progression of CVD. Capillary electrophoresis (CE) with laser induced fluorescence detection (LIF) was used for the analysis of naphthalene-2,3-dicarboxaldehyde (NDA)derivatized MAs. The optimization of run buffer composition, focusing on sodium borate and sulfylbutylether b-cyclodextrin (SBE-b-CD) concentrations, was crucial in obtaining adequate resolution between the analytes of interest. Under optimal conditions, SDMA, ADMA, MMA and arginine were baseline resolved and the limits of detection for those species were 41 amol, 30 amol, 41 amol and 10 amol, respectively. Plasma analysis was performed using strong cation exchange solid phase extraction (SPE) tips to preconcentrate the analytes of interest prior to derivatization. Recovery of MAs from the SPE tips was determined to be 43%.

Solid phase extraction

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Optimization of the Separation of NDA-Derivatized Methylarginines by Capillary and Microchip Electrophoresis

SLAS TECHNOLOGY (2012)

Thomas H. Linz Christa M. Snyder Susan M. Lunte

The methylated arginines (MAs) monomethylarginine (MMA), asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA) have been shown to be independent predictors of cardiovascular disease. This article describes progress regarding the development of an analytical method capable of rapidly analyzing MAs using capillary electrophoresis (CE) and microchip electrophoresis (MCE) with laser-induced fluorescence (LIF) detection. Several parameters including buffer composition and separation voltage were optimized to achieve an ideal separation. The analytes of interest were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) to produce fluorescent 1-cyanobenz[f]isoindole (CBI) derivatives and then subjected to CE analysis. Baseline resolution of SDMA, ADMA, MMA, and arginine was achieved in less than 8 min. The limits of detection for SDMA, ADMA, MMA, and arginine were determined to be 15, 20, 25, and 5 nM, respectively, which are well below the expected plasma concentrations. The CE separation method was then transferred to a glass MCE device with LIF detection. MAs were baseline resolved in 3 min on-chip using a 14 cm separation channel with detection limits of approximately 10 nM for each species. To the best of the authors' knowledge, this is the first report of the separation of MAs by MCE.

10.1177/2211068211424551

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Structural Characterization of Serum N-Glycans by Methylamidation, Fluorescent Labeling, and Analysis by Microchip Electrophoresis

Analytical Chemistry (2016)

Indranil Mitra Christa M. Snyder Xiaomei Zhou Margit I. Campos William R. Alley

To characterize the structures of N-glycans derived from human serum, we report a strategy that combines microchip electrophoresis, standard addition, enzymatic digestion, and matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). We compared (i) electrophoretic mobilities of known N-glycans from well-characterized (standard) glycoproteins through standard addition, (ii) the electrophoretic mobilities of N-glycans with their molecular weights determined by MALDI-MS, and (iii) electrophoretic profiles of N-glycans enzymatically treated with fucosidase. The key step to identify the sialylated N-glycans was to quantitatively neutralize the negative charge on both α2,3- and α2,6-linked sialic acids by covalent derivatization with methylamine. Both neutralized and nonsialylated N-glycans from these samples were then reacted with 8-aminopyrene-1,3,6-trisulfonic acid (APTS) to provide a fluorescent label and a triple-negative charge, separated by microchip electrophoresis, and detected by laser-induced fluorescence. The methylamidation step leads to a 24% increase in the peak capacity of the separation and direct correlation of electrophoretic and MALDI-MS results. In total, 37 unique N-glycan structures were assigned to 52 different peaks recorded in the electropherograms of the serum samples. This strategy ensures the needed separation efficiency and detectability, easily resolves linkage and positional glycan isomers, and is highly reproducible.

Electropherogram

Linkage isomerism

10.1021/acs.analchem.6b00882

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Capillary electrophoresis–mass spectrometry for direct structural identification of serum N-glycans

Journal of Chromatography A (2017)

Christa M. Snyder Xiaomei Zhou Jonathan A. Karty Bryan R. Fonslow Miloš V. Novotný

Linkage isomerism

Capillary electrophoresis–mass spectrometry

N-Acetylneuraminic acid

10.1016/j.chroma.2017.09.009

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Complementary Glycomic Analyses of Sera Derived from Colorectal Cancer Patients by MALDI-TOF-MS and Microchip Electrophoresis

Analytical Chemistry (2016)

Christa M. Snyder William R. Alley Margit I. Campos Martin Svoboda John A. Goetz

Colorectal cancer is the fourth most prevalent cancer in the United States, yet there are no reliable noninvasive early screening methods available. Serum-based glycomic profiling has the necessary sensitivity and specificity to distinguish disease states and provide diagnostic potential for this deadly form of cancer. We applied microchip electrophoresis and MALDI-TOF-MS-based glycomic procedures to 20 control serum samples and 42 samples provided by patients diagnosed with colorectal cancer. Within the identified glycans, the position of fucose units was located to quantitate possible changes of fucosyl isomeric species associated with the pathological condition. MALDI-MS data revealed several fucosylated tri- and tetra-antennary glycans which were significantly elevated in their abundance levels in the cancer samples and distinguished the control samples from the colorectal cancer cohort in the comprehensive profiles. When compared to other cancers studied previously, some unique changes appear to be associated with colorectal cancer, being primarily associated with fucosyl isomers. Through MS and microchip electrophoresis-based glycomic methods, several potential biomarkers were identified to aid in the diagnosis and differentiation of colorectal cancer. With its unique capability to resolve isomers, microchip electrophoresis can yield complementary analytical information to MS-based profiling.

10.1021/acs.analchem.6b02310

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Citations (47)