title: Fast quantification of endogenous carbohydrates in plasma using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry.
abstract: Endogenous carbohydrates in biosamples are frequently highlighted as the most differential metabolites in many metabolomics studies. A simple, fast, simultaneous quantitative method for 16 endogenous carbohydrates in plasma has been developed using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry. In order to quantify 16 endogenous carbohydrates in plasma, various conditions, including columns, chromatographic conditions, mass spectrometry conditions, and plasma preparation methods, were investigated. Different conditions in this quantified analysis were performed and optimized. The reproducibility, precision, recovery, matrix effect, and stability of the method were verified. The results indicated that a methanol/acetonitrile (50:50, v/v) mixture could effectively and reproducibly precipitate rat plasma proteins. Cold organic solvents coupled with vortex for 1 min and incubated at -20°C for 20 min were the most optimal conditions for protein precipitation and extraction. The results, according to the linearity, recovery, precision, matrix effect, and stability, showed that the method was satisfactory in the quantification of endogenous carbohydrates in rat plasma. The quantified analysis of endogenous carbohydrates in rat plasma performed excellently in terms of sensitivity, high throughput, and simple sample preparation, which met the requirement of quantification in specific expanded metabolomic studies after the global metabolic profiling research.
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Bangjie
authors.initials: B
authors.lastname: Zhu
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.forename: Xituo
authors.initials: X
authors.lastname: Li
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Xue
authors.initials: X
authors.lastname: Gu
authors.forename: Jieyu
authors.initials: J
authors.lastname: Dai
authors.forename: Guiming
authors.initials: G
authors.lastname: Wang
authors.forename: Yu
authors.initials: Y
authors.lastname: Cheng
authors.forename: Chao
authors.initials: C
authors.lastname: Yan
mesh_terms: D000818:Animals; D002241:Carbohydrates / Q000097:blood / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002241:Carbohydrates
keywords: Endogenous carbohydrates; Hydrophilic interaction chromatography; Plasma; Tandem mass spectrometry