Aluminium (Al) is associated with many clinical disorders in renal patients. Al accumulation in brain has also been related to the neurodegenerative processes in Alzheimer's disease. In order to better understand Al transport in the human body, it is necessary to identify and quantify chemical species in which Al is present in body fluids and tissues. Among a variety of biological samples, Al speciation was the most frequently investigated in human serum. Improvements were made in the development of analytical techniques for the determination of the amount and composition of high molecular mass Al (HMM-Al) and low molecular mass Al (LMM-Al) species in human serum. However, due to the complex chemistry of Al in serum, its low total concentration and the high risk of contamination, speciation of Al in biological samples is still a difficult task for analytical chemists. In this work, problems related to speciation of Al in human serum are critically discussed. An overview of the progress that was made by the use of different analytical procedures, in order to propose analytical protocols for reliable speciation of Al in serum at low ng mL(-1) concentration range, is presented.