The internal energy deposition of a Venturi-assisted array of micromachined ultrasonic electrosprays (AMUSE), with and without the application of a DC charging potential, is compared with equivalent experiments for Venturi-assisted electrospray ionization (ESI) using the "survival yield" method on a series of para-substituted benzylpyridinium salts. Under conditions previously shown to provide maximum ion yields for standard compounds, the observed mean internal energies were nearly identical (1.93-2.01 eV). Operation of AMUSE without nitrogen flow to sustain the air amplifier focusing effect generated energetically colder ions with mean internal energies that were up to 39% lower than those for ESI. A balance between improved ion transfer, adequate desolvation, and favorable ion energetics was achieved by selection of optimum operational ranges for the parameters that most strongly influence the ion population: the air amplifier gas flow rate and API capillary temperature. Examination of the energy landscapes obtained for combinations of these parameters showed that a low internal energy region (<or=1.0 eV) was present at nitrogen flow rates between 2 and 4 L min(-1) and capillary temperatures up to 250 degrees C using ESI (9% of all parameter combinations tested). Using AMUSE, this region was present at nitrogen flow rates up to 2.5 L min(-1) and all capillary temperatures (13% of combinations tested). The signal-to-noise (S/N) ratio of the intact p-methylbenzylpyridinium ion obtained from a 5 microM mixture of thermometer compounds using AMUSE at the extremes of the studied temperature range was at least fivefold higher than that of ESI, demonstrating the potential of AMUSE ionization as a soft method for the characterization of labile species by mass spectrometry.