Atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) was used to study the forced unbinding of quadruple self-complementary hydrogen-bonded urea-aminotriazine (UAT) complexes in hexadecane (HD). To elucidate the bond strength of individual linkages the unbinding forces of UAT supramolecular polymers were investigated for the first time. The bond rupture was probed at three different, fixed piezo retraction rates in far from equilibrium conditions. The number of supramolecular bonds (N) between AFM tip and the surface was determined by independent knowledge of the linker length. The observed rupture force of urea aminotriazine (UAT)-based supramolecular polymer chains was found to decrease with increasing rupture length. The dependence of the most probable rupture force on N was in quantitative agreement with the theory of uncooperative bond rupture for supramolecular linkages switched in series. Experiments with three different, fixed loading rates provided identical values (within the experimental error) for the characteristic bond length x(beta) and the off-rate constant in the absence of force k(off)(f = 0). The value of x(beta) was found to agree with literature data on the hydrogen-bond distance obtained via crystallographic data of the hydrogen-bonded dimer. This work broadens the scope of our previous report showing that relevant parameters of the bond energy landscape can be derived from a single data set of rupture events at a fixed loading rate for supramolecular linkages switched in series.