Novel Three-Dimensional MALDI Plate for Interfacing High-Capacity LC Separations with MALDI-TOF

Novel matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) sample plates employing collimated-hole structures have been developed that allow capture and concentration of samples while simultaneously acting as a sink for carrier solvents. These plates were designed to provide an efficient interface between higher-capacity liquid chromatography (LC) separations and MALDI-TOF mass spectrometry (MS). LC-MALDI using conventional plates can accommodate the low-flow (< 1 mu L/min) separation protocols typically used in on-line LC-MS methods, and can also be used, with higher flow rate, larger columns, but are ultimately limited by the capacity of the two-dimensional surface onto which the sample is deposited. Typically, about 1 mu L of chromatographic effluent plus 1 mu L of matrix solution can be deposited and dried on a ca. 3 mm diameter spot Deposition rates (spot dwell time) are determined by the chromatographic resolution and the flow rate. To overcome this limitation, a new three-dimensional MALDI sample plate has been developed using collimated-hole structures (CHS) with monolithic chromatography media filling the holes in the collimated-hole structures. These new plates retain all of the required features of conventional sample plates, commonly formed from stainless steel, but provide additional capacity for capturing and concentrating samples. Results are presented from reversed-phase separation of peptides on a 1 mm W. column operating at flow rate of 50 mu L/min. Typically, 10 mu L of effluent can be collected on a single spot, and sample and matrix dried on a 1 mm diameter spot, to yield about 30-fold enrichment of sample concentration in matrix crystals on the surface compared to the conventional plate. Sample loadings ranging from 1 fmol to 10 pmol/spot were investigated.