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Analysis of sphingosine 1-phosphate by capillary electrophoresis coupled to laser-induced fluorescence detection: use of a transparent fused-silica capillary.
J Capill Electrophor Microchip Technol. 2007; 10(1-2):25-31.JC

Abstract

C18-sphingosine 1-phosphate (C18-SPP) is a sphingolipid with important functional and structural roles in cells. In this paper we report a new capillary electrophoresis technique that is coupled to a laser-induced fluorescence detection (CE-LIF) method to quantify the level of C18-SPP in biological samples. The method utilizes a commercial standard C17-sphingosine 1-phosphate (C17-SPP) derivatized with naphthalene-2,3-dicarboxaldehyde (NDA), a fluorogenic dye used to label primary amines. The detection limit for the C17-SPP NDA was 0.54 fmol. We quantified the C18-SPP in leukemic human cells before and after irradiation by gamma rays. We demonstrated that the amounts of this sphingolipid decreased after the irradiation. In a second part of this work, we used the technique to evaluate the ability of a novel transparent fused-silica capillary, which allows the use of fused-silica capillaries without burning a window. Solarization, homogeneity, and sensitivity were studied using this capillary. The results demonstrate that this new durable capillary can provide a sensitive and reproducible quantitation procedure for CE-LIF studies.

Authors+Show Affiliations

Rammouz G
Université Paul Sabatier, IMRCP, Toulouse, France.
Usta J
No affiliation info available
Ribeiro SJ
No affiliation info available
Messaddeq Y
No affiliation info available
Couderc F
No affiliation info available
Poinsot V
No affiliation info available

MeSH

AcrylatesCell Line, TumorElectrophoresis, CapillaryHumansImidesKineticsLasersLysophospholipidsMolecular StructureSilicon DioxideSpectrometry, FluorescenceSpectrometry, Mass, Electrospray IonizationSphingosineTandem Mass Spectrometry

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17685239

Citation

Rammouz, Georges, et al. "Analysis of Sphingosine 1-phosphate By Capillary Electrophoresis Coupled to Laser-induced Fluorescence Detection: Use of a Transparent Fused-silica Capillary." Journal of Capillary Electrophoresis and Microchip Technology, vol. 10, no. 1-2, 2007, pp. 25-31.
Rammouz G, Usta J, Ribeiro SJ, et al. Analysis of sphingosine 1-phosphate by capillary electrophoresis coupled to laser-induced fluorescence detection: use of a transparent fused-silica capillary. J Capill Electrophor Microchip Technol. 2007;10(1-2):25-31.
Rammouz, G., Usta, J., Ribeiro, S. J., Messaddeq, Y., Couderc, F., & Poinsot, V. (2007). Analysis of sphingosine 1-phosphate by capillary electrophoresis coupled to laser-induced fluorescence detection: use of a transparent fused-silica capillary. Journal of Capillary Electrophoresis and Microchip Technology, 10(1-2), 25-31.
Rammouz G, et al. Analysis of Sphingosine 1-phosphate By Capillary Electrophoresis Coupled to Laser-induced Fluorescence Detection: Use of a Transparent Fused-silica Capillary. J Capill Electrophor Microchip Technol. 2007;10(1-2):25-31. PubMed PMID: 17685239.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of sphingosine 1-phosphate by capillary electrophoresis coupled to laser-induced fluorescence detection: use of a transparent fused-silica capillary. AU - Rammouz,Georges, AU - Usta,Julnar, AU - Ribeiro,Sidney J L, AU - Messaddeq,Younes, AU - Couderc,François, AU - Poinsot,Véréna, PY - 2007/8/10/pubmed PY - 2007/11/10/medline PY - 2007/8/10/entrez SP - 25 EP - 31 JF - Journal of capillary electrophoresis and microchip technology JO - J Capill Electrophor Microchip Technol VL - 10 IS - 1-2 N2 - C18-sphingosine 1-phosphate (C18-SPP) is a sphingolipid with important functional and structural roles in cells. In this paper we report a new capillary electrophoresis technique that is coupled to a laser-induced fluorescence detection (CE-LIF) method to quantify the level of C18-SPP in biological samples. The method utilizes a commercial standard C17-sphingosine 1-phosphate (C17-SPP) derivatized with naphthalene-2,3-dicarboxaldehyde (NDA), a fluorogenic dye used to label primary amines. The detection limit for the C17-SPP NDA was 0.54 fmol. We quantified the C18-SPP in leukemic human cells before and after irradiation by gamma rays. We demonstrated that the amounts of this sphingolipid decreased after the irradiation. In a second part of this work, we used the technique to evaluate the ability of a novel transparent fused-silica capillary, which allows the use of fused-silica capillaries without burning a window. Solarization, homogeneity, and sensitivity were studied using this capillary. The results demonstrate that this new durable capillary can provide a sensitive and reproducible quantitation procedure for CE-LIF studies. SN - 1946-4940 UR - https://www.unboundmedicine.com/medline/citation/17685239/Analysis_of_sphingosine_1_phosphate_by_capillary_electrophoresis_coupled_to_laser_induced_fluorescence_detection:_use_of_a_transparent_fused_silica_capillary_ DB - PRIME DP - Unbound Medicine ER -