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From the first issue of 2016, Chromatography has changed its name to Separations.

Open AccessArticle
Chromatography 2015, 2(2), 167-187; doi:10.3390/chromatography2020167

A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept

1
ICB-CSIC, C/ Miguel Luesma, 4, 50018 Zaragoza, Spain
2
Departamento de Química Analítica, Universidad de Zaragoza, 50009 Zaragoza, Spain
3
CEQMA-CSIC, Facultad de Ciencias, Universidad de Zaragoza, 50009 Zaragoza, Spain
4
ICMA-CSIC, Facultad de Ciencias, Universidad de Zaragoza, 50009 Zaragoza, Spain
5
Departamento de Química-Física, Universidad de Zaragoza, 50009 Zaragoza, Spain
*
Author to whom correspondence should be addressed.
Academic Editor: Mark Devlin Maloney
Received: 21 January 2015 / Revised: 16 March 2015 / Accepted: 26 March 2015 / Published: 8 April 2015
(This article belongs to the Special Issue New Trends in Thin-Layer Chromatography)
View Full-Text   |   Download PDF [700 KB, uploaded 9 April 2015]   |  

Abstract

Hyphenated HPTLC has been used to analyze several neutral sphingolipids acting as lysosomal storage disease (LSD) biomarkers. Automated multiple development (AMD) provides separation of lipid peaks, which are detected and quantified using fluorescence detection by intensity changes (FDIC) after primuline post-impregnation. A final online transfer to a mass spectrometer by means of an elution-based interface allows their identification using electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI).Given that the increases in fluorescent emission detected by FDIC are produced by non-specific, electrostatic interactions between the primuline and hydrocarbon chains in the ceramide backbones of sphingolipids, it is a non-destructive detection technique, allowing the precise location and transfer of biomarker peaks to a mass spectrometer using an elution interface. By using primuline as a fluorophore, the technique is also compatible with ESI-APCI and does not interfere with the MS of sphingolipids. APCI provides useful and complementary structural information to the ESI for sphingolipid identification. Moreover, FDIC emission can be used for quantitative purposes. Results include the determination of sphingomyelin (SM) in human-plasma samples (RSD < 6%) by means of a standard addition method with non-linear calibration, and the identification of globotriaosylceramide (Gb3) in the plasma of a Fabry patient. Only one HPTLC plate is needed to perform the analysis. View Full-Text
Keywords: TLC-MS; induced fluorescence; automated multiple development; primuline; sphingolipids TLC-MS; induced fluorescence; automated multiple development; primuline; sphingolipids
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Domínguez, A.; Jarne, C.; Cebolla, V.L.; Galbán, J.; Savirón, M.; Orduna, J.; Membrado, L.; Lapieza, M.-P.; Romero, E.; Sanz Vicente, I.; de Marcos, S.; Garriga, R. A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept. Chromatography 2015, 2, 167-187.

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