UPLC-MS/MS测定人源Caco-2细胞中人参炔醇的含量

Abstract

Objective To develop an ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) method to quantify the uptake of panaxynol in cultured Caco-2 cells.MethodsThe chromatographic separation was performed on Acquity BEH C18 column(2.1 mm×100 mm,1.7 μm) with aqueous methanol as the mobile phase,using gradient elution at 0.3 mL/min.A triple-quadruple mass spectrometer employing electrospray ionization in positive ion mode was developed,and panaxynol in Caco-2 cells was determined using multiple reaction monitoring of precursor→product ion transitions at m/z 227→129 for quantification and m/z 227→143 for confirmation.ResultsThe established method was validated by determining the linearity(r20.99),precision(≤6.2%) and accuracy(-6.7% to 2.1%).The limit of detection for panaxynol was 4 ng/mL.When incubated for 2 h at 37 ℃,the uptake was(20.7±1.8) nmol/mg protein for 50 μmol/L panaxynol and(21.8±1.7) nmol/mg protein for 100 μmol/L panaxynol,and there was no significant difference between them(P0.05).ConclusionThis method is sensitive,reliable and specific,and can be used in determination of panaxynol in Caco-2 cells.