The pollen spectrum and the physicochemical characteristics were analyzed in 13 samples of honeys produced by Apis mellifera in two areas with different degree of human disturbance in the Region of Biobio, Province of Nuble, Chile: a) Central Valley and b) Preandean foothills of the Cordillera de los Andes. Principal component analysis was carried out for pollen and physicochemical data and pollen diversity was calculated with the Shannon-Weaver diversity index (H'). Some native species such as Gevuina avellana, Lomatia hirsuta, Lithraea caustica and Luma apiculata were found only in honeys from San Fabian de Alico (Preandean foothills) but introduced species dominated the pollen spectra in both areas. Ash content, pH, acidity and water content were the most important physico-chemical factors to distinguish the honeys from both areas. All honeys showed characteristics that met the standards of the Chilean food health regulations. Melissopalynology allow us to detect one bifloral honey from the Central Valley and two monofloral Lotus honeys from the Preandean foothills of Cordillera de los Andes.