A comprehensive reinvestigation of the rotational spectrum of pyrimidine was carried out by using several different spectrometers. All singly substituted 13C- and 15N-isotopic species of pyrimidine have been measured in natural abundance with millimeter-wave free jet and waveguide Fourier transform microwave techniques, and complete rs and r0 heavy atom geometries have been determined. The ground state rotational spectrum in the centimeter-wave region was measured at sub-Doppler resolution of the cavity Fourier transform spectrometer and all elements in the inertial and principal nuclear quadrupole-coupling tensors of the nitrogen nuclei in pyrimidine have been determined. The room-temperature spectrum was measured up to 337 GHz and J = 66 with BWO-based spectrometers and sextic level centrifugal distortion constants in the rotational Hamiltonian have been determined for the ground state and three lowest vibrational fundamentals of pyrimidine. Copyright 1999 Academic Press.