Borrowing the notion of instantaneous frequency that was developed in the context of time-frequency signal analysis, an instantaneous frequency amplitude spectrum (IFAS) is introduced for estimating fundamental frequency of speech signal in both noiseless and adverse environments. We define harmonicity measure as a quantity that indicates degree of periodical regularity in the IFAS and that shows substantial difference between periodic signal and noise-like waveform. The harmonicity measure is applied to estimate the existence of fundamental frequency. We provide experimental examples to demonstrate the general applicability of the harmonicity measure and apply the proposed procedure to Japanese continuous speech signals. The results show that the proposed method outperforms the conventional methods with or without the presence of noise.