A limitation of any current approach using solvent-free MALDI mass spectrometry is that only one sample at a time can be prepared and transferred to the MALDI-plate. For this reason, multiple-sample preparation approaches for solvent-free MALDI MS analysis of synthetic polymers were developed that are simple and practical. One approach multiplexed sample preparation by simultaneously preparing multiple samples. With this approach, as many as 384 samples could be prepared by addition of analyte, matrix, salt, and 1-mm metal beads to each well of a 384-well disposable bacti plate, capping the plate with the lid and homogenizing all samples simultaneously using a common laboratory vortex device. Besides the time savings achieved by a single vortex step for multiple samples, an additional advantage of this method relative to previously reported solvent-free preparation methods is that the mixing volume per sample is reduced, which allows a reduction in the amount of analyte required. This method, however, still requires the transfer of each homogenized sample to the MALDI plate for subsequent analysis. Here we report a novel approach that combines multiple simultaneous solvent-free sample preparation with automatic sample transfer to the MALDI target plate. This approach reduces the possibility of cross-contamination, the amount of sample and matrix consumed for an analysis, and the time required for preparation of multiple samples. These methods were shown to provide high-quality mass spectra for various synthetic polymer standards with M n values to 10 kDa. The methods are efficient in that small sample amounts are required, the sample/salt/matrix ratio is not critical, and the time necessary to achieve sufficient homogenization of multiple samples is less than 5 min. . A recent study based on the ball-mill homogenization/loose powder transfer method revealed that solvent-free MALDI-MS simplifies the measurement and improves the result qualitatively and quantitatively [3]. In general, the solvent-free method gives more homogeneous analyte/matrix mixtures and higher shot-to-shot and sample-tosample reproducibility than does the solvent-based method [3,4]. Consequently, less laser power has to be applied [3,4], which yields milder MALDI conditions and thus less fragmentation [1,3]. This method, for the same reasons, also reduces background signals [3] and improves the resolution of the analyte signals [3,4].For synthetic polymer analyses, three general methods exist for homogenization. These are grinding by mortar and pestle [4,5], ball-mill [1][2][3][4], and vortexing [6]. Analyte, matrix, and salt are added to a vessel and, in the case of the ball-mill or vortex methods, suitably sized balls are used to assist in the homogenization process. For polymers having M n Յ 10 kDa ball-mill, vortex, and mortar-and-pestle approaches appear to work equally well [4,6]. Two general methods exist for the dry transfer of the homogenized powder from the vessel to the MALDI plate. In one method a pressed