The liquid impingement samplers, represented by BioSampler and AGI-30, are widely employed in the field of public health and epidemic control due to their simple operation and suitability for multi-component analysis. However, the quantitative assessment of their performance and differences between these two types of samplers is often overlooked, with both typically used as standard methods. This study investigated the physical collection efficiency of the two samplers using the aerodynamic particle sizer spectrometer. The colony cultivation assays and real-time quantitative PCR (qPCR) were employed to assess the biological viability of the microbial sample. Results revealed that for 1 μm particles, BioSampler achieved a superior collection efficiency of 76.7%, compared to AGI-30. The physical collection efficiency of both samplers increased with the rise in flow rate. It was discovered that liquid impingement samplers inflicted less damage on microorganisms and retrieved a higher number of cultivable microorganisms compared to filter-based samplers. At higher concentrations, the impingement samplers notably outperformed the filter-based sampler in terms of sampling effect. The trends observed in qPCR results echoed those from the aerodynamic particle sizer, indicating while the filter-based sampler had the highest physical collection efficiency, its biological viability rate was significantly lower than the impingement samplers. Consequently, differing sampling environments and analytical approaches yielded varying evaluations of sampler performance, depending on their operational principles. This research offers crucial insights for the performance evaluation and selection of liquid impingement samplers.