TNT (2,4,6-trinitrotoluene) readily and predominantly transforms to 2ADNT (2-amino-4,6-dinitrotoluene) and 4ADNT (4-amino-2,6-dinitrotoluene) in environmental matrixes and tissues. Solid phase microextraction (SPME) was used to extract ADNTs (amino-dinitrotoluenes) from tissue as a potential method to investigate the recalcitrance of metabolically-generated ADNTs versus absorbed ADNTs. Tubifex tubifex was allowed to metabolize TNT into ADNTs in 24-hr static non-renewal exposure test followed by 24-hr depuration in clean reconstituted hard water. Polyacrylate-coated (PA) SPME fibers were then deployed and agitated in tissue homogenates containing metabolically-generated ADNTs for 48 hr to provide a measure of available ADNTs. Extractability of ADNTs from T. tubifex tissue containing metabolically-generated ADNTs was significantly less than extractability of ADNTs from T. tubifex tissue containing absorbed ADNTs: 50-60% and 81-90% of expected extractability based on fiber-water partition ratio. The lower SPME extractability of metabolically-generated ADNTs may stem from the unavailability of metabolically-generated ADNTs sequestered in tissue or bound to tissue macromolecules during metabolism of TNT to ADNT. Tissue extractions using SPMEs may be able to estimate such bound organic residues in tissue and serve as potential indicators of toxicological bioavailability and biomagnification potential of tissue-associated organic compounds.