›› 2011, Vol. 31 ›› Issue (10): 1370-.doi: 10.3969/j.issn.1674-8115.2011.10.003

• Original article (Basic research) • Previous Articles     Next Articles

UPLC-MS/MS method for determination of panaxynol in cultured Caco-2 cells

YAN Zhong-hong1, LI Lin2, WANG Guo-quan3, JIN Yu-e3, YANG Ruo-lin1, LU Yang1   

  1. 1.Department of Pharmacy, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China；2.Unilever R&D Shanghai, Shanghai 200233, China；3.Shanghai Municipal Center for Disease Control and Prevention, Shanghai 200336, China
  • Online:2011-10-28 Published:2011-10-27
  • Supported by:

    Natural Science Foundation of Shanghai, 06ZR14059, 11ZR1419000；Shanghai Municipal Health Bureau Foundation, 2006J006A

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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. Methods The 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. Results The established method was validated by determining the linearity (r2>0.99), 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 （P>0.05）. Conclusion This method is sensitive, reliable and specific, and can be used in determination of panaxynol in Caco-2 cells.

Key words: panaxynol, ultra-performance liquid chromatography tandem mass spectrometry, Caco-2