The spectrum of the non-self-adjoint Zakharov-Shabat operator with periodic potentials is studied, and its explicit dependence on the presence of a semiclassical parameter in the problem is also considered. Several new results are obtained. In particular: (i) it is proved that the resolvent set has two connected components, (ii) new bounds on the location of the Floquet and Dirichlet spectra are obtained, some of which depend explicitly on the value of the semiclassical parameter, (iii) it is proved that the spectrum localizes to a cross in the spectral plane in the semiclassical limit. The results are illustrated by discussing several examples in which the spectrum is computed analytically or numerically.