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For the determination of relative metabolite abundances, the integrated signal of all ions for each metabolite fragment was normalized by the signal from norvaline and sample protein content isolated by resuspending the protein layer in 50 l NaOH 200 mM, vertexing for 15 min at 96 C, and centrifuging at 4 C 14,000 rpm for 15 min

This result may be explained by the presence of an unentrapped drug in the outer layer of prepared vehicles

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