We evaluated short echo time two-dimensional 1H magnetic resonance spectroscopy (2D-1HMRS) with the point-resolved spatial selection (PRESS) protocol in temporal lobe epilepsy (TLE) patients with negative magnetic resonance imaging (MRI) findings and described the characteristics of 2D-(1)HMRS in the epileptic focus. Thirty-eight TLE patients with negative conventional MRI findings and 10 healthy controls were studied by 2D-1HMRS. A 128-channel prolonged video-electroencephalographic (EEG) method was used as the standard for epileptogenic focus localization to evaluate N-acetyl aspartate/(choline + creatine + phosphocreatine) (NAA/[Cho + Cr-PCr]), glutamate + glutamine/creatine (Glx/Cr-PCr) and myo-inositol/Cr-PCr ratios in patients with negative MRI findings. The 2D-1HMRS showed that 32/38 patients and all healthy controls had stable baselines and good signal-to-noise ratios. Compared with the healthy controls, 32 patients showed abnormal NAA/(Cho + Cr-PCr) ratios in the hippocampus and, in 25 of these patients, focus lateralization agreed with the EEG. It is concluded that short echo time 2D-1HMRS with the PRESS protocol can help find abnormalities in lateralization of temporal epilepsy in patients with negative MRI findings.