The regulation of pituitary function via the hypothalamus and via intra-pituitary connections represents a complex system. Though hormones secreted from the anterior and posterior pituitary have been well studied, overall information of proteins expressed in the anterior and posterior pituitary gland is very limited. Protein expression profiling of normal pituitary tissue may lead to discovery of novel proteins playing an important role in the physiology of pituitary glands and can lead to better understanding of pituitary diseases. We aimed to carry out systematic proteomic profiling of adenohypophysis from human pituitary glands using high-resolution Fourier transform mass spectrometer. A total of 2,051 proteins were identified in this study of which, 653 proteins were identified for the first time from human pituitary gland. The comprehensive list of proteins identified in this study, many for the first time, will facilitate the better understanding the role of this important gland in health and disease.