Accumulation of 3‐aminopropylphosphonate in the ex vivo brain observed by phosphorus‐31 nuclear magnetic resonance

3‐aminopropylphosphonate (3‐APP) is known for its use as an exogenous indicator of extracellular volume and pH in phosphorus‐31 nuclear magnetic resonance ((31)P NMR) studies. We used 3‐APP for estimating the extracellular volume in NMR studies of several ex vivo preparations including retrograde perfused mouse heart (n = 4), mouse liver slices (n = 2), xenograft breast cancer tumors (n = 7, MCF7), and rat brain slices (n = 4). In the former three preparations, the 3‐APP signal was stable in lineshape and intensity for hours and the chemical shift of the signal in the presence of the biological sample was the same as in the perfusion medium without the biological sample. However, in studies of brain slices, the 3‐APP signal appeared split into two, with an upfield component (0.7 ± 0.1 ppm to the left) increasing with time and showing a wider linewidth (66.7 ± 12.6 vs. 39.1 ± 7.6 Hz, the latter is of the perfusion medium signal). This finding suggests that 3‐APP inadvertently accumulated in brain slices, most likely as a membrane bound form. This observation limits the use of 3‐APP as an inert biochemical indicator in brain preparations and should be taken into account when using 3‐APP in vivo.