A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept
Abstract
:1. Introduction
2. Experimental Section
2.1. Fluorophore
2.2. SL Standards and Plasma Samples
2.3. Sample Treatment
2.4. Plates for HPTLC Separation
2.5. AMD-FDIC-MS Hyphenated System: Description and Conditions for SL Analysis
2.5.1. Sample Application
2.5.2. HPTLC Using AMD
MeOH (vol %) | DCM (vol %) | Migration (mm) | |
---|---|---|---|
Step 1 | 50 | 50 | 30 |
Step 2 | 50 | 50 | 30 |
Step 3 | 50 | 50 | 30 |
Step 4 | 50 | 50 | 30 |
Step 5 | 50 | 50 | 30 |
Step 6 | 30 | 70 | 60 |
Step 7 | 10 | 90 | 90 |
MeOH (vol %) | DCM (vol %) | Migration (mm) | |
---|---|---|---|
Step 1 | 80 | 20 | 20 |
Step 2 | 50 | 50 | 50 |
2.5.3. FDIC Using Fluorescence Scanning Densitometry
2.5.4. Coupling AMD-FDIC to API-MS
2.6. Repeatability of Sample Treatment
2.7. Calibration and Quantification of SM in Plasma Using FDIC
3. Results and Discussion
3.1. Primuline as an FDIC Fluorophore
3.2. AMD-Gradient Separation of SLs
3.3. FDIC of Separated SLs Using Primuline
3.4. Online AMD-FDIC-MS Coupling
3.5. Identification of SLs Using AMD-FDIC-(API)-MS
3.6. Quantitative Analysis of SLs by FDIC
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References and Notes
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Domínguez, A.; Jarne, C.; Cebolla, V.L.; Galbán, J.; Savirón, M.; Orduna, J.; Membrado, L.; Lapieza, M.-P.; Romero, E.; Sanz Vicente, I.; et al. A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept. Chromatography 2015, 2, 167-187. https://doi.org/10.3390/chromatography2020167
Domínguez A, Jarne C, Cebolla VL, Galbán J, Savirón M, Orduna J, Membrado L, Lapieza M-P, Romero E, Sanz Vicente I, et al. A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept. Chromatography. 2015; 2(2):167-187. https://doi.org/10.3390/chromatography2020167
Chicago/Turabian StyleDomínguez, Andrés, Carmen Jarne, Vicente L. Cebolla, Javier Galbán, María Savirón, Jesús Orduna, Luis Membrado, María-Pilar Lapieza, Elena Romero, Isabel Sanz Vicente, and et al. 2015. "A Hyphenated Technique based on High-Performance Thin Layer Chromatography for Determining Neutral Sphingolipids: A Proof of Concept" Chromatography 2, no. 2: 167-187. https://doi.org/10.3390/chromatography2020167