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Abstract: We applied a novel approach to respiratory waveform analysis—Monotone Signal Segments Analysis (MSSA) on 6-h recordings of respiratory signals in rats. To validate MSSA as a respiratory signal analysis tool we tested it by detecting: breaths and breath-to-breath intervals; respiratory timing and volume modes; and changes in respiratory pattern caused by lesions of monoaminergic systems in rats. MSSA differentiated three respiratory timing (tachypneic, eupneic, bradypneic–apneic), and three volume (artifacts, normovolemic, hypervolemic-sighs) modes. Lesion-induced respiratory pattern modulation was visible as shifts in the distributions of monotone signal segment amplitudes, and of breath-to-breath intervals. Specifically, noradrenergic lesion induced an increase in mean volume (p ≤0.03), with no change of the mean breath-to-breath interval duration (p ≥0.06). MSSA of timing modes detected noradrenergic lesion-induced interdependent changes in the balance of eupneic (decrease; p ≤0.02), and tachypneic (an increase; p ≤0.02) breath intervals with respect to control. In terms of breath durations within each timing mode, there was a tendency toward prolongation of the eupneic (p ≤0.08) and bradypneic–apneic (p ≤0.06) intervals. These results demonstrate that MSSA is sensitive to subtle shifts in respiratory rhythmogenesis not detectable by simple respiratory pattern descriptive statistics. MSSA represents a potentially valuable new tool for investigations of respiratory pattern control. [Copyright &y& Elsevier]