title: Simultaneous determination of three dipeptides (JBP485, Gly-Sar and JBP923) in the cell lysates by liquid chromatography-tandem mass spectrometry: application to identify the function of the PEPT1 transfected cell.
abstract: A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of JBP485, Gly-Sar and JBP923 in the cell lysates using methanol as a deproteinization solvent was developed and validated. Detection was performed by turbo ionspray ionization in multiple reaction monitoring mode using the transitions of m/z 147.1 → m/z 90.1 for Gly-Sar, m/z 201.1 → m/z 86.1 for JBP485, m/z 219.1 → m/z 86.1 for JBP923 and m/z 152.0 → m/z 110.0 for paracetamol (internal standard). The analytes were separated on a Hypersil ODS C18 HPLC column using isocratic elution mode with a mobile phase containing 0.1% formic acid in water-methanol (97:3, v/v) at a flow rate of 0.2 mL/min. The calibration curves were demonstrated to be linear over the concentration range of 5.00-5000 nm with coefficient of 0.9968 for Gly-Sar, 0.9975 for JBP485 and 0.9952 for JBP923. The intra- and inter-day precisions were <10.2% for each quality contro; level, and the accuracy was within ±5.6% for each analyte. The matrix effect, the extraction recovery and stabilities of LC-MS/MS analysis were also investigated. This validated method was successfully applied to the simultaneous determination of JBP485, Gly-Sar and JBP923 in the cell lysates for identification of stably transfected HeLa cells with human PEPT1.
authors.affiliation: Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, China.
authors.forename: Xinjin
authors.initials: X
authors.lastname: Guo
authors.forename: Qiang
authors.initials: Q
authors.lastname: Meng
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.forename: Changyuan
authors.initials: C
authors.lastname: Wang
authors.forename: Xiaokui
authors.initials: X
authors.lastname: Huo
authors.forename: Zhe
authors.initials: Z
authors.lastname: Zhang
authors.forename: Taiichi
authors.initials: T
authors.lastname: Kaku
authors.forename: Kexin
authors.initials: K
authors.lastname: Liu
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004151:Dipeptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004355:Drug Stability; D006367:HeLa Cells; D006801:Humans; D000074011:Peptide Transporter 1; D010456:Peptides, Cyclic / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D027981:Symporters / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D004151:Dipeptides; C412308:JBP 485; C412307:JBP 923; D000074011:Peptide Transporter 1; D010456:Peptides, Cyclic; C082186:SLC15A1 protein, human; D027981:Symporters; C004194:glycylsarcosine
keywords: Gly-Sar; JBP485; JBP923; LC-MS/MS; cell lysates