2024-03-28T19:31:13Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1226932022-03-03T14:05:05Zcom_10261_35com_10261_5com_10261_31com_10261_3com_10261_52707col_10261_288col_10261_284col_10261_52708
A hyphenated technique based on high-performance thin layer chromatography for determining neutral sphingolipids: A proof of concept
Domínguez Carrasco, Andrés
Jarne, Carmen
Cebolla, Vicente L.
Galbán, Javier
Savirón, María
Orduna, Jesús
Membrado, Luis
Lapieza Remón, Mª Pilar
Romero Giménez, Elena
Sanz Vicente, Isabel
Marcos, Susana de
Garriga, Rosa
Consejo Superior de Investigaciones Científicas (España)
Ministerio de Economía y Competitividad (España)
Diputación General de Aragón
European Science Foundation
European Commission
CSIC - Unidad de Recursos de Información Científica para la Investigación (URICI)
TLC-MS
Induced fluorescence
Automated multiple development
Primuline
Sphingolipids
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license.-- This article belongs to the Special Issue "New Trends in Thin-Layer Chromatography".
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.
The authors thank the Spanish Ministry of Economy and Competitiveness (MINECO) and FEDER (UE) (Plan Nacional de I+D+I, projects CTQ2012-035535 and CTQ2012-34774), as well as DGA-ESF, for financial support. Carmen Jarne thanks CSIC and ESF for a JAE-doc grant.
We acknowledge the support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).
Peer reviewed
2015-09-28T06:32:14Z
2015-09-28T06:32:14Z
2015
artículo
http://purl.org/coar/resource_type/c_6501
Chromatography 2(2): 167-187 (2015)
http://hdl.handle.net/10261/122693
10.3390/chromatography2020167
2227-9075
http://dx.doi.org/10.13039/501100003339
http://dx.doi.org/10.13039/501100003329
http://dx.doi.org/10.13039/501100000782
http://dx.doi.org/10.13039/501100000780
Publisher's version
http://dx.doi.org/10.3390/chromatography2020167
Sí
open
Multidisciplinary Digital Publishing Institute