Abstract
To aid in the detection of trace quantities of neuropeptides in a biological matrix (as saliva), the influence of different electrolytes and a supercharging reagent on the positive electrospray ionisation mass spectrometry (ESI-MS) response was investigated. Ammonium acetate, sodium chloride and sodium bicarbonate (10(-7) M to 10(-3) M) and the supercharging reagent m-nitrobenzyl alcohol (m-NBA) was added to the mobile phase and the effect on the ESI response and charge-states distribution (CSD) was studied in a group of peptides (molecular weight range 2.2kDa to 3.5kDa; CGRP, VIP, GLP1, CRF and PrRP). As expected, the result indicates that the ESI response is affected by the presence of additives: ammonium acetate shifted the observed charge states ratio whereas the addition of m-NBA resulted in the appearance of higher maximum charge state ions. This increase in higher charge state for all the peptide ions, [M + nH]n+ to [M + (n + 1)H](n+1), was atttributed to protonation of the C-terminal. However, when the composition of MeCN in a mobile phase containing m-NBA was increased, an enhancement of the total ion signal was observed for non-polar peptide samples. This is a very interesting observation as this is not observed in samples without m-NBA and could be a result of how these peptide ions are solubilised and positioned relative to the droplet surface/air interface.
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