Abstract
Although detergents are often essential in protocols, they are usually incompatible with further biochemical analysis. There are several methods for detergent removal, but the procedures are complicated or suffer from sample loss. Here, we describe a convenient and rapid method for detergent removal from sialic acid-containing glycosphingolipids (gangliosides) and neutral glycolipids in detergent-resistant membrane (DRM) microdomain. It is based on selective detergent extraction, in which the sample is dried on a glass tube, followed by washing with organic solvent. We investigated 18 organic solvents and used high performance thin-layer chromatography (HPTLC) and matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS) to confirm that dichloroethane (DCE) was the most suitable solvent and completely removed the nonionic detergent Triton X-100. Furthermore, DCE extraction effectively removed interference caused by other nonionic, zwitterionic, or ionic detergents in MALDI-QIT-TOF MS analysis.
Highlights
Detergents are often essential in protocols, they are usually incompatible with further biochemical analysis
These results indicated that Triton X-100 needs to be removed at a concentration range of 1–10 g for ganglioside analysis by MALDI-QITTOF MS
Half of the quantity was applied to high performance thin-layer chromatography (HPTLC); the remainder was separated into gangliosides or other lipids via Svennerholm’s partition, and the resulting lower phase was applied to MALDI-QIT-TOF MS for analysis (Fig. 6)
Summary
Detergents are often essential in protocols, they are usually incompatible with further biochemical analysis. DCE extraction effectively removed interference caused by other nonionic, zwitterionic, or ionic detergents in MALDI-QIT-TOF MS analysis.—Suzuki, Y., and K. To confirm the detergent extraction ability from the ganglioside of the organic solvent, GM3 (4 g) and Triton X-100 (4 mg for MS, 30 g for HPTLC) were mixed and dried in Pyrex glass tubes. Confirmation of detergent interference for MALDI-QIT-TOF MS analysis of gangliosides
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