Chemical composition of propolis samples from north Algeria was characterized by chromatographic and spectroscopic analyses. High-performance liquid chromatorgaphy with diode-array detection (HPLC-DAD) fingerprint of the methanol extracts allowed the definition of two main types of Algerian propolis (AP) directly related to their secondary metabolite composition. Investigation of two representative types of AP by preparative chromatographic procedure and mass spectrometric (MS) and NMR techniques led to the identification of their main constituents: caffeate esters and flavonoids from an AP type rich in phenolic compounds (PAP) and labdane and clerodane diterpenes, together with a polymethoxyflavonol, from an AP type containing mainly diterpenes (DAP). Subsequently, two specific HPLC-MS/MS methods for detection of PAP and DAP markers were developed to study the chemical composition of propolis samples of different north Algerian regions. Antioxidant activity of AP samples was evaluated by 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay, and a significant free-radical scavenging effect was observed for propolis of the PAP series rich in polyphenols.