A trimethylamine (TMA) moiety is present in carnitine and acetylcarnitine, and both molecules play critical roles in muscle metabolism. At 7 T, the chemical shift dispersion was sufficient to routinely resolve the TMA signals from carnitine at 3.20 and from acetylcarnitine at 3.17 ppm in the ¹H‐MRS (Magnetic Resonance Spectroscopy) of human soleus muscle with a temporal resolution of about 2 min. In healthy, sedentary adults, the concentration of acetylcarnitine increased nearly 10‐fold, to 4.1 ± 1.0 mmol/kg, in soleus muscle after 5 min of calf‐raise exercise and recovered to a baseline concentration of 0.5 ± 0.3 mmol/kg. While the half‐time for decay of acetylcarnitine was the same whether measured from the TMA signal (18.8 ± 5.6 min) or measured from the methyl signal (19.4 ± 6.1 min), the detection of acetylcarnitine by its TMA signal in soleus has the advantage of higher sensitivity and without overlapping from lipid signals. Although the activity of carnitine acetyltransferase is sufficient to allow equilibrium between carnitine and coenzyme‐A pools, the exchange in TMA signal between carnitine and acetylcarnitine is slow in soleus following exercise on 7T ¹H‐NMR time scale. The TMA signal provides a simple and direct measure of the relative amounts of carnitine and acetylcarnitine. Magn Reson Med, 2013. © 2012 Wiley Periodicals, Inc.