An extension of current mobility resolution equations as they apply to high-field ion mobility spectrometry is presented. The new resolution expression is applied to arrival time distributions for ions having a large range of ion mobilities and mass-to-charge ratios (m/z). The results indicate that the new equation can be utilized to predict the mobility resolution over a broader range of applied electric fields than previous ion mobility resolution expressions. (J Am Soc Mass Spectrom 2004, 15, 1320 -1324) © 2004 American Society for Mass Spectrometry S eparation based on the mobility of an ion through a neutral buffer gas (i.e., ion mobility spectrometry (IMS)) [1,2] is an important technology for studying long-lived electronic states of gas-phase ions and a sensitive method for detection of air-borne species [3, 4]. Recent work, which demonstrates the utility of IMS for analyzing gas-phase ionized biopolymers (i.e., peptides, proteins, DNA, etc.), has renewed interest in the analytical applications of IMS and led to the development of several high-resolution drift tubes in combination with mass spectrometry [5][6][7][8][9][10][11]. In order to perform a gas-phase separation of near thermal, structurally isomeric gas-phase ion populations, most IMS separations are performed near the low-field limit. Low-field conditions are important for studies of gas-phase peptide/protein structure because collisional activation can, in some cases, lead to structural rearrangement and/or fragmentation.In addition to the distinctions drawn for IMS operation in terms of separation field strength, the use of IMS can be divided into two pressure regimes, viz. high pressure (typically low-field) and low pressure (either low or high applied fields), and there are distinct advantages associated with both pressure regimes. At high pressure, the number of ion-neutral collisions is high, i.e., C 60 ϩ⅐ undergoes 10 10 collisions/s at 760 torr He compared with about 10 7 collisions/s for the same ion at 1 torr He. It is also important to consider that collisions with low-level impurities may significantly influence the separation and total drift time of an analyte ion. For example, in a 30 cm drift cell maintained at 760 torr He, C 60 ϩ⅐ will undergo 10 5 -10 6 collisions with a of 0.01% impurity, compared with ca. 10 collisions with the same level of impurity at 1 torr He. Another advantage of low-pressure drift tubes is the ease of coupling ion mobility with high vacuum mass spectrometers. Lastly, low-pressure (or high-field) mobility decreases ion separation times, leading to higher throughput analysis (approximately 3 orders of magnitude faster for analysis times at 1 torr over 760 torr).Low-field mobility conditions are defined in terms of the kinetic energy acquired by the ion in the presence of an applied field (E o ). That is, the translational energy gained by the ion between collisions should be less than the thermal energy of the collision gas [12, 13],where m is the mass of the ion, M is the mass of the neutral drift gas, ...