Pollen is a source of nutrients for honeybees (Apis mellifera L.) and suitable for human consumption as well. In our research, a multiresidue method for pesticide determination was developed and validated for the bee pollen matrix. 247 components met the validation criteria for limit of detection, limit of quantification, linearity, and interday repeatability. Average recoveries varied between 70 and 120% except for 14 analytes, which were corrected during on-going validation. The matrix effect was strong for certain analytes, which required the use of matrix-matched calibration. The pesticide residue profiles of 21 pollen samples of different botanical origins were identified by the developed method. The most common active substances were chlorpyrifos, thiacloprid, and acetamiprid. Some products contained pesticides that are already banned. According to our estimates, the tested samples do not pose an acute risk on honeybees, although the combination of pesticides may cause synergistic toxicity.