title: Simultaneous quantification of labeled (2)H5-glycerol, (13)C6-glucose, and endogenous D-glucose in mouse plasma using liquid chromatography tandem mass spectrometry.
abstract: Monitoring the level of glucose and glycerol or their labeled derivatives in biological fluid for kinetic studies has always been challenging, especially in mice, because of the limited volume in addition to the complexity of plasma. For such application, we developed a simple, fast, and sensitive method for the simultaneous measurement of absolute concentrations of labeled (2)H5-glycerol and (13)C6-glucose as well as endogenous D-glucose using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). In our study, 15.0 μL of mouse plasma was processed by a one-step protein precipitation, followed by LC-MS/MS analysis. The quantification of the analytes was carried out by monitoring the product ion scan of their corresponding deprotonated molecular ions and constructing the extracted ion fragmentogram by choosing a specific product ion for each analyte (equivalent to precursor ion to product ion transitions). The limit of detection (LOD) was evaluated to be 1.0 μM for both (2)H5-glycerol and (13)C6-glucose, and the limit of quantitation (LOQ) was observed to be 5.0 μM for both (2)H5-glycerol and (13)C6-glucose in diluted mice plasma that corresponds to 50 μM in plasma or 4.60 and 9.01 mg/dL of glycerol and glucose in plasma, respectively. The extraction recoveries are 81.9 % (CV = 8.1 %) for (2)H5-glycerol and 26.2 % (CV = 13.6 %) for (13)C6-glucose.
authors.affiliation: Icahn Institute and Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
authors.forename: Farid
authors.initials: F
authors.lastname: Jahouh
authors.affiliation: Icahn Institute and Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. rong.wang@mssm.edu.
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002138:Calibration; D002247:Carbon Isotopes; D002853:Chromatography, Liquid / Q000379:methods*; D003903:Deuterium; D005990:Glycerol / Q000097:blood* / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods*; D007700:Kinetics; D057230:Limit of Detection; D051379:Mice; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001786:Blood Glucose; D002247:Carbon Isotopes; D003903:Deuterium; D005990:Glycerol
keywords: Glucose; Glycerol; LC-MS/MS; Mouse; Plasma; Stable isotope tracer