Record number :
Title of article :
Measurement of 2′,3′-cyclic nucleotides by liquid chromatography–tandem mass spectrometry in cells
Author/Authors :
Bنhre، نويسنده , , Heike and Kaever، نويسنده , , Volkhard، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2014
Pages :
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Abstract :
Recently, the occurrence of 2′,3′-cyclic nucleoside monophosphates (2′,3′-cNMPs) in addition to 3′,5′-cNMPs in mammalian tissues was reported. We developed a liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for the measurement of four 2′,3′-cyclic nucleotides, i.e., 2′,3′-cAMP, 2′,3′-cCMP, 2′,3′-cGMP, 2′,3′-cUMP, in cell samples. Chromatographic separation was achieved using a Zorbax eclipse XCB-C18 (50 mm × 4.6 mm; 1.8 μm column; Agilent) connected to a QTRAP5500 system (AB Sciex) operating in positive ionization mode. Calibration curves were constructed in the range 0.41 fmol/μL to 1666.6 fmol/μL for 2′,3′-cAMP, 2′,3′-cCMP, and 2′,3′-cGMP, and 3.3–1666.6 fmol/μL for 2′,3′-cUMP, respectively, showing squared correlation coefficients >0.9992. Accuracy and inter- and intra-day precision lay within the required ranges of <20% for LLOQ and <15% for higher concentration levels. The method was applied to the analysis of nucleotides in two different cell lines (Hek293T and HuT-78).
Keywords :
cells , 2? , Liquid chromatography–tandem mass spectrometry , 3?-Cyclic nucleotide monophosphates
Journal title :
Journal of Chromatography B
Journal title :
Journal of Chromatography B
Serial Year :
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