Multiply-charged noncovalent cluster anions of adenosine-5=-monophosphate (AMP) were formed by electrospray ionization (ESI). Ions in higher charge states were observed when the ions were accumulated in an ion trap with helium buffer gas before detection. We determined the smallest size (n a ) or appearance size as a function of charge state (q), i.e., n a ϭ 4 for q ϭ 2, n a ϭ 8 for q ϭ 3, and n a ϭ 13 for q ϭ 4. The relation between n a and q can be described by a charged droplet model. When the size is larger than n a for a given q, the fragmentation pathway of an anion cluster is dominated by loss of neutral fragments. In contrast, when the size approaches the appearance size, only charged fragments are formed. Cluster cations such as metal clusters have been extensively studied during the last two decades [3,4]. Recently, the study of biomolecular cluster cations has become an active research field, and much work has dealt with the formation, dissociation, and charge partitioning among fragments of protein cluster cations [5][6][7][8][9][10]. Magic numbers for amino acid cationic clusters [11][12][13] and preference for homochirality [14,15] have been reported. Negatively charged clusters of amino acids have also been produced [13].The formation and energetics of gas-phase multiplycharged anions (MCA) is a subject that has attracted much attention [16 -18]. Even though several multiplycharged molecular anions have been measured, observations of multiply-charged cluster anions are few. Multiply-charged metal cluster anions were observed by electron attachment to monoanions in a Penning trap [19 -22] . Here we report, to our knowledge, the first observation of negatively, multiply-charged noncovalent clusters made of adenosine 5=-monophosphate (AMP). The dependence of cluster size on charge state was measured, and the results, are modeled by a charged droplet model. The fragmentation pathways of multiply-charged negative clusters are also presented.
ExperimentalTwo setups were applied in the experiments: an electrostatic ion storage ring (ELISA) and an accelerator mass spectrometer (AMS). In both setups, negative clusters of AMP were produced by ESI. A saturated solution of AMP in methanol was sprayed.Detailed information about the ESI source (Figure 1a) is described in reference [32]. Briefly, the source is mounted on the high-voltage platform of the accelerator. A stainless steel hypodermic needle is, via a fused silica capillary, connected to a syringe containing a solution of the analyte. A syringe pump (Harvard apparatus) delivers a constant flow through the needle. A typical flow rate is 1ϳ2 l/min. A voltage of 3 to 4 kV relative to the heated capillary is put on the needle. The capillary is normally heated for desolvation of solvent molecules from analyte molecules. At the exit of the capillary, ions emerge into the first vacuum zone where a pressure of around 1 mbar is maintained. The ions focused by a tube lens go through a skimmer into a second region. The voltage on the tube lens varies from 50 to...