An important function of the auditory nervous system is to analyze the frequency content of environmental sounds. The neural structures involved in determining psychophysical frequency resolution remain unclear. Using a two-noise masking paradigm, the present study investigates the spectral resolution of neural populations in primary auditory cortex (A1) of awake macaques and the degree to which it matches psychophysical frequency resolution. Neural ensemble responses (auditory evoked potentials, multiunit activity, and current source density) evoked by a pulsed 60 dB SPL pure tone signal fixed at the best frequency (BF) of the recorded neural populations were examined as a function of the frequency separation (F) between the tone and two symmetrically flanking continuous 80 dB SPL 50 Hz-wide bands of noise. Fs ranged from 0% to 50% of the BF, encompassing the range typically examined in psychoacoustic experiments. Responses to the signal were minimal for F = 0% and progressively increased with F, reaching a maximum at F = 50%. Rounded exponential functions, used to model auditory filter shapes in psychoacoustic studies of frequency resolution, provided excellent fits to neural masking functions. Goodness-of-fit was greatest for response components in lamina 4 and lower lamina 3 and least for components recorded in more superficial cortical laminae. Physiological equivalent rectangular bandwidths (ERBs) increased with BF, measuring approximately 15% of the BF. These findings parallel results of psychoacoustic studies in both monkeys and humans, and thus indicate that a representation of perceptual frequency resolution is available at the level of A1.