Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection
Kenji Okumura, Kazunori Hayashi, Itsuro Morishima, Kichiro Murase, Hideo Matsui, Yukio Toki, Takayuki Ito
Ceramides and 1,2-diacylglycerol have been demonstrated in intracellular signaling pathways. A method of simultaneous mass determination of ceramides and 1,2-diacylglycerol in tissues was developed using the latroscan which combines thin layer chromatography and flame ionization detection (TLC/FID) techniques. Because of relatively low amounts of these components in tissues, the fraction of nonpolar lipids, which included ceramides and glycerides, was eluted with chloroform/acetone mixture (3:1, vol/vol) through a silicic acid column to eliminate the polar phospholipids. Development of Chromarods was carried out using three solvent systems in a four-step development technique. The relationship of the peak area ratio to weight ratio compared with cholesteryl acetate added as an internal standard was linear. The amount of ceramides increased with incubation of rat heart homogenate and human erythrocyte membranes in the presence of sphingomyelinase (E.C. 3.1.4.12). The location TLC/FID system provided a quick and reliable assessment of ceramides and 1,2-diacylglycerol.
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Thin layer chromatography Chemical Compounds
Flame Ionization Medicine & Life Sciences
1,2-diacylglycerol Chemical Compounds
Ionization Chemical Compounds
Thin Layer Chromatography Medicine & Life Sciences
Ceramides Chemical Compounds
Tissue Chemical Compounds
cholesterol acetate Chemical Compounds
Okumura, K., Hayashi, K., Morishima, I., Murase, K., Matsui, H., Toki, Y., & Ito, T. (1998). Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection. Lipids, 33(5), 529-532. https://doi.org/10.1007/s11745-998-0237-3
Okumura, Kenji ; Hayashi, Kazunori ; Morishima, Itsuro ; Murase, Kichiro ; Matsui, Hideo ; Toki, Yukio ; Ito, Takayuki. / Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection. In: Lipids. 1998 ; Vol. 33, No. 5. pp. 529-532.
@article{e4aff5b7ed2c463b9560c6c8a4709ba4,
title = "Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection",
abstract = "Ceramides and 1,2-diacylglycerol have been demonstrated in intracellular signaling pathways. A method of simultaneous mass determination of ceramides and 1,2-diacylglycerol in tissues was developed using the latroscan which combines thin layer chromatography and flame ionization detection (TLC/FID) techniques. Because of relatively low amounts of these components in tissues, the fraction of nonpolar lipids, which included ceramides and glycerides, was eluted with chloroform/acetone mixture (3:1, vol/vol) through a silicic acid column to eliminate the polar phospholipids. Development of Chromarods was carried out using three solvent systems in a four-step development technique. The relationship of the peak area ratio to weight ratio compared with cholesteryl acetate added as an internal standard was linear. The amount of ceramides increased with incubation of rat heart homogenate and human erythrocyte membranes in the presence of sphingomyelinase (E.C. 3.1.4.12). The location TLC/FID system provided a quick and reliable assessment of ceramides and 1,2-diacylglycerol.",
author = "Kenji Okumura and Kazunori Hayashi and Itsuro Morishima and Kichiro Murase and Hideo Matsui and Yukio Toki and Takayuki Ito",
journal = "Lipids",
Okumura, K, Hayashi, K, Morishima, I, Murase, K, Matsui, H, Toki, Y & Ito, T 1998, 'Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection', Lipids, vol. 33, no. 5, pp. 529-532. https://doi.org/10.1007/s11745-998-0237-3
Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection. / Okumura, Kenji; Hayashi, Kazunori; Morishima, Itsuro; Murase, Kichiro; Matsui, Hideo; Toki, Yukio; Ito, Takayuki.
In: Lipids, Vol. 33, No. 5, 1998, p. 529-532.
T1 - Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection
AU - Okumura, Kenji
AU - Hayashi, Kazunori
AU - Morishima, Itsuro
AU - Murase, Kichiro
AU - Matsui, Hideo
AU - Toki, Yukio
AU - Ito, Takayuki
N2 - Ceramides and 1,2-diacylglycerol have been demonstrated in intracellular signaling pathways. A method of simultaneous mass determination of ceramides and 1,2-diacylglycerol in tissues was developed using the latroscan which combines thin layer chromatography and flame ionization detection (TLC/FID) techniques. Because of relatively low amounts of these components in tissues, the fraction of nonpolar lipids, which included ceramides and glycerides, was eluted with chloroform/acetone mixture (3:1, vol/vol) through a silicic acid column to eliminate the polar phospholipids. Development of Chromarods was carried out using three solvent systems in a four-step development technique. The relationship of the peak area ratio to weight ratio compared with cholesteryl acetate added as an internal standard was linear. The amount of ceramides increased with incubation of rat heart homogenate and human erythrocyte membranes in the presence of sphingomyelinase (E.C. 3.1.4.12). The location TLC/FID system provided a quick and reliable assessment of ceramides and 1,2-diacylglycerol.
AB - Ceramides and 1,2-diacylglycerol have been demonstrated in intracellular signaling pathways. A method of simultaneous mass determination of ceramides and 1,2-diacylglycerol in tissues was developed using the latroscan which combines thin layer chromatography and flame ionization detection (TLC/FID) techniques. Because of relatively low amounts of these components in tissues, the fraction of nonpolar lipids, which included ceramides and glycerides, was eluted with chloroform/acetone mixture (3:1, vol/vol) through a silicic acid column to eliminate the polar phospholipids. Development of Chromarods was carried out using three solvent systems in a four-step development technique. The relationship of the peak area ratio to weight ratio compared with cholesteryl acetate added as an internal standard was linear. The amount of ceramides increased with incubation of rat heart homogenate and human erythrocyte membranes in the presence of sphingomyelinase (E.C. 3.1.4.12). The location TLC/FID system provided a quick and reliable assessment of ceramides and 1,2-diacylglycerol.
JO - Lipids
JF - Lipids
Okumura K, Hayashi K, Morishima I, Murase K, Matsui H, Toki Y et al. Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection. Lipids. 1998;33(5):529-532. https://doi.org/10.1007/s11745-998-0237-3