AbstractPurposeNeurochemicals of interest quantified by MRS are often composites of overlapping signals. At higher field strengths (i.e., 7T), there is better separation of these signals. As the availability of higher field strengths is increasing, it is important to re‐evaluate the separability of overlapping metabolite signals.MethodsThis study compares the ability of stimulated echo acquisition mode (STEAM‐8; TE = 8 ms), short‐TE semi‐LASER (sLASER‐34; TE = 34 ms), and long‐TE semi‐LASER (sLASER‐105; TE = 105 ms) acquisitions to separate the commonly acquired neurochemicals at 7T (Glx, consisting of glutamate and glutamine; total N‐acetyl aspartate, consisting of N‐acetyl aspartate and N‐acetylaspartylglutamate; total creatine, consisting of creatine and phosphocreatine; and total choline, consisting of choline, phosphocholine, and glycerophosphocholine).ResultssLASER‐34 produced the lowest fit errors for most neurochemicals; however, STEAM‐8 had better within‐subject reproducibility and required fewer subjects to detect a change between groups. However, this is dependent on the neurochemical of interest.ConclusionWe recommend short‐TE STEAM for separation of most standard neurochemicals at 7T over short‐TE or long‐TE sLASER.