Honeybee-collected pollen is promoted as a health food with a wide range of nutritional and therapeutic properties. A high-performance capillary electrophoresis with amperometric detection method has been developed for the simultaneous determination of bioactive ingredients in 10 samples of honeybee-collected pollen in this work. Under the optimum conditions, 13 phenolic components can be well-separated or nearly baseline-separated (apigenin and vanillic acid peaks) within 29 min at the separation voltage of 14 kV in a 50 mM borax running buffer (pH 9.0), and adequate extraction was obtained with ethanol for the determination of the above 13 compounds. Recovery (94.1-104.0%), repeatability of the peak current (<5.4%), and detection limits (6.9 x 10(-7)-6.4 x 10(-9) g mL(-1)) for the method were evaluated. This procedure was successfully used for the analysis and comparison of the phenolic content of honeybee-collected pollen samples originating from different floral origins based on their electropherograms or "phenolic profiles".