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Simple separation of glycosphingolipids in the lower phase of a Folch’s partition from crude lipid fractions using zirconium dioxide

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Abstract

A simple method was developed for the separation of glycosphingolipids (GSLs) from lipid mixtures, including phospholipids and cholesterol, using zirconium dioxide (zirconia, ZrO2). Although this procedure does not incorporate a mild alkali treatment, which is commonly used for eliminating glycerophospholipids, it can be used to remove both alkali-resistant sphingomyelin and glycerophospholipids possessing ether bonds. Importantly, when GSLs were dissolved in organic solvent together with cholesterol (Chol) and phospholipids, and loaded onto ZrO2, Chol did not bind to the ZrO2 but both the GSLs and phospholipids did. When eluted with 5 mg/mL of 2,5-dihydroxybenzoic acid in methanol, GSLs but not phospholipids were recovered, leaving the phospholipids bound to the ZrO2 particles. This method is particularly applicable for GSLs such as triglycosylceramides, tetraglycosylceramides and some pentaglycosylceramides, sulfatide and GM3 located in the lower phase of a Folch’s partition, where significant amounts of phospholipids, Chol and neutral lipids reside along with GSLs. This method was successfully used to easily isolate GSLs from biological materials for their subsequent analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry with high resolution.

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Acknowledgements

We thank Renee Mosi, PHD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

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  1. Division of Microbiology and Molecular Cell Biology, Nihon Pharmaceutical University, 10281 Komuro, Inamachi, Saitama, 362-0806, Japan

    Hideharu Nagasawa & Mamoru Kyogashima

  2. GL Sciences Inc., 237-2 Sayamagahara, Saitama, 358-0032, Japan

    Shota Miyazaki

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  1. Hideharu Nagasawa
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Correspondence to Mamoru Kyogashima.

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Nagasawa, H., Miyazaki, S. & Kyogashima, M. Simple separation of glycosphingolipids in the lower phase of a Folch’s partition from crude lipid fractions using zirconium dioxide. Glycoconj J 39, 789–795 (2022). https://doi.org/10.1007/s10719-022-10080-w

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