Breast cancer is the most common serious disease that occurs in the human body. Trace elements have an important function in biological and metabolism processes including activation or inhibition of enzymatic reaction, reactive oxygen species (ROS), competition between trace elements and metal proteins for binding positions and modifications in the permeability of cellular membranes which influence carcinogenic processes. A significant association between the abnormal concentration of trace elements and breast cancer has been found in many studies using XRF techniques like energy dispersive X-ray fluorescence (EDXRF), particle induced X-ray emission (PIXE), total reflection X-ray fluorescence (TXRF), wavelength dispersive X-ray fluorescence (WDXRF) and synchrotron induced X-ray fluorescence (SRIXE). This chapter considers trace elements like Fe, Cu, Zn, Cr, Cl, Ca, P, S, K, Na, Mg, Se, As and Sr. from the standpoint of their role as either inhibitory or causative agents of breast cancer. XRF techniques and sample preparation methods for analysis of biological samples are also reviewed.