Quantification of F(2)-isoprostane isomers in cultured human lung epithelial cells after silica oxide and metal oxide nanoparticle treatment by liquid chromatography/tandem mass spectrometry.
F(2)-isoprostanes are lipid peroxidation products of arachidonic acid in cell membrane and are reliable biomarkers for oxidative stress and cell membrane damage. Nanomaterials are widely used as raw materials in many industries and will have high potentials to be used in life science and medical fields. However, the human health impact of nanoparticles has caused people's great concern. Unfortunately, the mechanisms of cytotoxicity of many nanoparticles are not well defined. By measuring the levels of F(2)-isoprostane isomers in cultured cells after nanoparticle exposure, the information can be used to explain whether the cytotoxicity of nanoparticles is caused by lipid peroxidation and to investigate the biological pathways. In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to separate and quantify F(2)-isoprostane isomers in nanoparticle-treated human lung cancer cells. Silica oxide (15nm) and other four metal oxide nanoparticles including Fe(2)O(3) (30nm), Al(2)O(3) (13nm), TiO(2) (40nm) and ZnO (70nm) are chosen in this study. The isotope forms of F(2)-isoprostane isomers, 8-iso-PGF(2alpha)-d(4) and PGF(2alpha)-d(4), were used as internal standard (IS). After human lung epithelial cells were exposed to different nanoparticles for 24h, F(2)-isoprostanes were extracted by a single step solid phase extraction with Oasis HLB cartridge. For the first time, six F(2)-isoprostane isomers were tentatively identified and quantified in human lung epithelial cells. The levels of F(2)-isoprostane isomers in the cells increased after the treatment with nanoparticles. For SiO(2), Fe(2)O(3), and ZnO nanoparticles, F(2)-isoprostane isomers increasing are consistent with nanoparticles' cytotoxicity data. For Al(2)O(3) and TiO(2) nanoparticles, F(2)-isoprostane isomers levels increased even before nanoparticles showed significant cytotoxicity at 100microg/mL concentration in 24h. Based on our best knowledge, this is the first study on the F(2)-isoprostane isomers corresponding to nanoparticles' exposure in vitro. Our study demonstrates that SiO(2) (15nm) nanoparticle showed the highest degree of lipid peroxidation and cell membrane damage among the studied nanoparticles.
Liu
Xiaoqian
X
Department of Chemistry and Environmental Research Center, Missouri University of Science and Technology, Rolla, MO 65409, USA. xlvw9@mst.edu
Whitefield
Philip D
PD
Ma
Yinfa
Y
D002462:Cell Membrane / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D004847:Epithelial Cells / Q000166:cytology*; D028441:F2-Isoprostanes / Q000737:chemistry*; D006801:Humans; D008168:Lung / Q000166:cytology*; D013058:Mass Spectrometry / Q000379:methods; D008670:Metals / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods; D018384:Oxidative Stress; D010087:Oxides / Q000737:chemistry*; D015203:Reproducibility of Results; D012822:Silicon Dioxide / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
D028441:F2-Isoprostanes; D008670:Metals; D010087:Oxides; D012822:Silicon Dioxide