alexa Heterogeneous Ganglioside Standards in LC-MS/MS: Sensitive Method for Quantifying the Major Molecular Components in Mono-Sialo Ganglioside Standards | Abstract
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
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Research Article

Heterogeneous Ganglioside Standards in LC-MS/MS: Sensitive Method for Quantifying the Major Molecular Components in Mono-Sialo Ganglioside Standards

Ashta Lakshmi Prasad Gobburi1, Renliang Zhang2, Belinda Willard2, Denise Inman3 and David J Anderson1*

1Department of Chemistry, Cleveland State University, Cleveland, Ohio, USA

2Research Core Services, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA

3Department of Pharmaceutical Sciences, Northeast Ohio Medical University, Rootstown, Ohio, USA

*Corresponding Author:
David J Anderson
Department of Chemistry
Cleveland State University
2121 Euclid Avenue, Cleveland
Ohio 44115, USA
Tel: 001-216-687-2453
Fax: 001-216-687-9298
E-mail: [email protected]

Received date: November 04, 2015 Accepted date: November 21, 2015 Published date: November 28, 2015

Citation: Gobburi ALP, Zhang R, Willard B, Inman D, Anderson DJ (2015) Heterogeneous Ganglioside Standards in LC-MS/MS: Sensitive Method for Quantifying the Major Molecular Components in Mono-Sialo Ganglioside Standards. J Anal Bioanal Tech S13:009. doi:10.4172/2155-9872.S13-009

Copyright: © 2015 Gobburi ALP, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

A commercially-available mono-sialo (GM1) ganglioside standard consists of three major components with different ceramide structures: C18:0 fatty acid/C18-sphingosine, C18:0 fatty acid/C20-sphingosine and C20:0 fatty acid/C18-sphingosine. The usual multiple reaction monitoring (MRM) liquid chromatography-tandem mass spectrometry (LC-MS/MS) of gangliosides, which monitors the dehydrated sialic acid fragment (m/z 290), cannot differentiate between the individual iso-molecular weight components C18:0 fatty acid/C20-sphingosine and C20:0 fatty acid/C18-sphingosine. Present characterization of ganglioside standards quantifies only the fatty acid content by gas chromatography-flame ionization detection (GC-FID) analysis of the ganglioside mixture and does not parse out the percentages of the individual mono-sialo ganglioside components. In the present work analyzing a heterogeneous GM1 standard, results from a dehydrated sialic acid daughter ion MRM LC-MS/MS determination employing hydrophilic interaction liquid chromatography were combined with results of the fatty acid content determined by GC-FID analysis to sensitively quantify the three predominant individual molecular GM1 components in standards at concentrations as low as 50 ng/mL (Method 1). These dehydrated sialic acid MRM results (Method 1) were confirmed by a less sensitive fatty acid daughter ion MRM LC-MS/MS technique (Method 2) which could only determine molecular GM1 components in standards at high concentrations (1 μg/mL-10 μg/mL). Method 2, however, has the advantage of directly quantifying the three predominant individual molecular GM1 components for comparison with Method 1 results. Equations are derived which incorporate the combined data (Method 1) to calculate percentages of individual mono-sialo gangliosides in the standard. Percentages for the individual mono-sialo gangliosides in the standard differed by at most 2% (absolute difference in the percentages) in comparing the results obtained by the two methods.

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