2015
DOI: 10.3390/chromatography2020167
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A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept

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 ioniza… Show more

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Cited by 14 publications
(20 citation statements)
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“…Standard addition approach is commonly used to quantify biomarkers that contain high endogenous level in the human samples [8] , [9] , [10] in order to minimize matrix effect difference between standard curve and patient samples. However, it is technically challenging to develop a method with an LLOQ less than the normal reference range of the endogenous analyte present in the blank plasma (standard zero).…”
Section: Resultsmentioning
confidence: 99%
“…Standard addition approach is commonly used to quantify biomarkers that contain high endogenous level in the human samples [8] , [9] , [10] in order to minimize matrix effect difference between standard curve and patient samples. However, it is technically challenging to develop a method with an LLOQ less than the normal reference range of the endogenous analyte present in the blank plasma (standard zero).…”
Section: Resultsmentioning
confidence: 99%
“…The use of primuline as a fluorophore for quantitative purposes was first described by Domínguez et al . 53 as FDIC, i.e. fluorescence detection by intensity changes.…”
Section: Methodsmentioning
confidence: 99%
“…Images from stained plates were acquired using Ettan DIGE Imager (GE Healthcare, Chicago, IL, USA). Bands corresponding to Cers (C20-C24), Cers (C14-C18), SMs (C20-C24) and SMs (C14-C18) were compared among NW, -vitDNW and –vitDO, considering primuline as a FDIC (fluorescence detection by intensity changes) fluorophore 53,58 .…”
Section: Methodsmentioning
confidence: 99%
“…For this reason, primuline-induced fluorescence was used to quantify saturated PC. Primuline experiences increases in emission in the presence of long hydrocarbon chains, including saturated ones due to dipole-induced dipole interactions, as it was described elsewhere [20,21].…”
Section: Quantitative Densitometry For Pl Classesmentioning
confidence: 72%