Determination of sorbitol in the presence of high amount of mannitol from biological samples

Abstract

Preparation of intact mitochondria requires the use of an osmoticum in high concentration that preserves the mitochondrial structure and prevents physical swelling and rupture of membranes. For this purpose mannitol is widely accepted as a component of the homogenization medium. The formation of sorbitol in plant mitochondria can be an important element of plant response to salinity and drought. The large difference in the concentration of possibly formed sorbitol and mannitol of the homogenisation medium makes the determination of sorbitol difficult. Gas-chromatography with flame ionization detection (GC-FID), gas-chromatography coupled to quadrupole mass spectrometry (GC-QPMS) and liquid chromatography/tandem mass spectrometry (LC/MS-MS) using hydrophilic interaction chromatography (HILIC) were tested to quantify this highly polar sugar alcohol: Although all three methods offered satisfactory results, the HPLC/MS-MS technique proved to be the best.