title: A liquid chromatography-tandem mass spectrometry assay for the detection and quantification of trehalose in biological samples.
abstract: Trehalose is an important disaccharide that is used as a cellular protectant by many different organisms, helping these organisms better survive extreme conditions, such as dehydration, oxidative stress, and freezing temperatures. Methods to detect and accurately measure trehalose from different organisms will help us gain a better understanding of the mechanisms behind trehalose's ability to act as a cellular protectant. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay using selected reaction monitoring mode for the detection and quantification of trehalose using maltose as an internal standard has been developed. This assay uses a commercially available LC column for trehalose separation and a standard triple quadrupole mass spectrometer, thus allowing many scientists to take advantage of this simple assay. The calibration curve from 3 to 100μM trehalose was fit best by a single polynomial. This LC-MS/MS assay directly detects and accurately quantifies trehalose, with an instrument limit of detection (LOD) that is 2-1000 times more sensitive than the most commonly-used assays for trehalose detection and quantification. Furthermore, this assay was used to detect and quantify endogenous trehalose produced by Escherichia coli (E. coli) cells, which were found to have an intracellular concentration of 8.5±0.9mM trehalose. This method thus shows promise for the reliable detection and quantification of trehalose from different biological sources.
authors: Departments of Chemistry, Union College, 807 Union St. Schenectady, NY 12308, USA.
authors: Philip M
authors: PM
authors: Kretschmer
authors: Departments of Chemistry, Union College, 807 Union St. Schenectady, NY 12308, USA.
authors: Austin M
authors: AM
authors: Bannister
authors: Departments of Biology, Union College, 807 Union St. Schenectady, NY 12308, USA.
authors: Molly K
authors: MK
authors: O Brien
authors: Departments of Chemistry, Union College, 807 Union St. Schenectady, NY 12308, USA.
authors: Laura A
authors: LA
authors: MacManus-Spencer
authors: Departments of Chemistry, Union College, 807 Union St. Schenectady, NY 12308, USA. Electronic address: paulickm@union.edu.
authors: Margot G
authors: MG
authors: Paulick
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D057230:Limit of Detection; D016014:Linear Models; D020011:Protective Agents / Q000032:analysis*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014199:Trehalose / Q000032:analysis*
chemical_list: D020011:Protective Agents; D014199:Trehalose
keywords: Biological samples; Cellular protectant; LC–MS/MS; Quantification; Trehalose