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Rapid identification of beta-lactamase-producing strains of <i>Haemophilus influenzae</i> plays key role in diagnostics in clinical microbiology. Therefore, the aim of this study was the rapid determination of beta-lactamase’s presence in <i>H. influenzae</i> isolates via indirect detection of degradation ampicillin products using MALDI-TOF MS. <i>H. influenzae</i> isolates were subjected to antibiotic resistance testing using disk diffusion and MIC methodologies. Beta-lactamase activity was tested using MALDI-TOF MS, and results were compared to spectral analysis of alkaline hydrolysis. Resistant and susceptible strains of <i>H. influenzae</i> were distinguished, and strains with a high MIC level were identified as beta-lactamase-producing. Results indicate that MALDI-TOF mass spectrometry is also suitable for the rapid identification of beta-lactamase-producing <i>H. influenzae</i>. This observation and confirmation can accelerate identification of beta-lactamase strains of <i>H. influenzae</i> in clinical microbiology, which can have an impact on health in general.