Animal studies to determine the role of ABCG2 in modulating PPIX distribution, metabolism, and excretion

WT and Abcg2-null mice were treated with D2-ALA (50 mg/kg, intraperitoneally), a stable isotope-labeled precursor of PPIX. One hour after D2-ALA treatment, liver and bile samples were collected for metabolomic analysis. In brief, liver and bile samples were analyzed using ultra-performance liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer (UPLC-QTOFMS, Waters Corp, Milford, MA). Centroid and integrated mass chromatographic data were processed by MarkerLynx software (Waters Corp, Milford, MA) to generate a multivariate data matrix. These data were then exported to SIMCA-P+ software (Umetrics, Kinnelon, NJ) for multivariate data analysis. Principal components analysis and orthogonal projection to latent structures discriminant analysis were conducted to represent the major latent variables in the data matrix. The variables that substantially contributed to the discrimination between groups were subjected to structure identification.

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