An apparatus for and method of estimating a one-way delay time between two hosts and an apparatus and method of clock synchronization using the same. The two hosts are connected to a network and communicate a predetermined packet; k-th, (k+1)-th, and (k+2)-th transmission times are measured at the first host, and k-th, and (k+1)-th transmission times are measured at the second host; a time difference of the m-th transmission time is measured at the first and second host, where m is k or k+1, and by using the measured transmission times, an m-th one-way delay time is calculated. If the time difference is identical to the calculated one-way delay time, a value equal to or less than the calculated one-way delay time is determined as the estimated one-way delay time. Accordingly, the estimation error of a one-way delay time is reduced for both symmetrically and asymmetrically connected hosts.