A possible origin of [M - n H + m X] ( m-n ) + ions (X = alkali metal ions) in electrospray mass spectrometry of peptides Christopher F. Rodriquez, Rene ´ Fournier, Ivan K. Chu, Alan C. Hopkinson, K.W. Michael Siu* Department of Chemistry, York University, 4700 Keele Street, Toronto, Ontario M3J 1P3, Canada Received 26 January 1999; accepted 10 May 1999 Abstract The [M - nH + mX] ( m-n) + (X = alkali metal ion) are common ions in the mass spectrum of a peptide that is electrosprayed in the presence of an alkali metal salt or hydroxide. The feasibility of forming [M - nH + mX] ( m-n) + ions in the gas phase including those in the lens region of the mass spectrometer via ion–molecule reactions and/or reactions between components of collisionally activated adducts was investigated. The Li + ion was selected for examination since its salts are computationally the least expensive among alkali metal salts. The lithium ion affinities of the [M - H] - ions of N-methylacetamide, acetic acid, and 1-propanamine were calculated by means of density functional theory (DFT) at various levels of theory, including B3LYP/6-311++G( d, p). These three compounds were selected as representatives of relevant functional groups on a peptide. The calculated lithium ion affinities, together with evaluated thermochemical data, were used to calculate the enthalpies of reactions between the model compounds and LiOH, LiCl, and Li(H 2 O) + that might lead to the formation of [M - nH + mX] ( m-n) + . A number of these reactions were found to be exothermic or slightly endothermic ( H°+20 kcal/mol). DFT calculations on the energetics of a model reaction revealed a relatively flat potential energy hypersurface containing a well of approximately 35 kcal/mol in depth and devoid of significant barriers. These results are used to postulate the formation of [M - nH + mX] ( m-n) + ions in the gas phase in the ion source and/or in the lens region via collisions between an ionic peptide and neutral lithium compounds or collisional activation of lithium–peptide adducts. (Int J Mass Spectrom 192 (1999) 303–317) © 1999 Elsevier Science B.V. Keywords: Adduct ions; Alkali metal; Peptide; Collisional activation 1. Introduction The sodium ion is an ubiquitous contaminant of samples in electrospray mass spectrometry. For pep- tide samples, the presence of high concentrations of Na + typically results in envelopes of ions of [M - nH + mNa] ( m-n) + in the positive ion detection mode, where n is the number of protons abstracted from the peptide M, and m is the number of sodium ions incorporated [1,2]. For a given charge state, i.e. constant ( m - n) +, the peptide containing peaks are readily identified by their separation on the m/z axis by (23 - 1)/( m - n) units; however, the presence of a large number of sodium-containing adducts of the peptide crowds the mass spectrum and decreases sensitivity as the peptide signal is distributed among many peaks, and is generally considered to be a * Corresponding author. E-mail: kwmsiu@yorku.ca 1387-3806/99/$20.00 © 1999 Elsevier Science B.V. All rights reserved PII S1387-3806(99)00116-5 International Journal of Mass Spectrometry 192 (1999) 303–317