Electrospray ionization mass spectrometry (ESI-MS) is widely used in drug development, therapeutic drug monitoring, and other fields. However, unstable mass spectral signals, especially during the initial stages of instrument operation, plague analysts. Generally, in quantitative experiments, the stability of response can be achieved by running the analytical system for some time. However, the equilibration time required for the responses of different compounds to stabilize has been elusive. To investigate the response stability of the ESI-MS system, 72 compounds with different physicochemical properties were employed on three systems, and flow injection analysis was performed in positive ion mode. With the use of 5.00% (response stable factor, RSF) as the stability limit, about 80% of the compounds were stable within 60 min. Under a 2.00% criterion, the stabilization time was significantly longer. The stabilization time varies with different instruments and physicochemical properties of the compounds. When positive ion detection is performed in an acidic mobile phase, the octanol-water partition coefficient (Log P), molecular weight, and molar volume can all affect the time required to stabilize the response. In general, it is necessary to balance the ESI-MS system for an appropriate time before sample detection, especially for the analysis of compounds with strong hydrophilicity, small molecular weight, or small molar volume under the conditions above.
Keywords: electrospray ionization mass spectrometry; equilibrium; physicochemical properties; response stability.