Safflower (Carthamus tinctorius L.) is currently being developed as a platform for the production of novel proteins.  Methods for detecting and quantifying transgenic safflower are needed to ensure seed quality and to monitor for its adventitious presence.  We developed and compared three methods of assaying for transgenic safflower presence in conventional seedlots: field bioassays, enzyme-linked immunosorbent assays (ELISA), and quantitative polymerase chain reaction (Q-PCR).  Limits for reliable quantification for both ELISA and Q-PCR are ~0.1%, although levels at least as low as 0.02% can be detected by Q-PCR.  Levels of quantification for the field bioassay are limited only by space and resources available.  Multiple sampling methods to detect and quantify transgenic safflower presence at levels lower than 0.1% were used on field collected samples from a pollen outcrossing experiment to quantify the adventitious presence of transgenic safflower.  Taking into account the potential utility and relative advantages or disadvantages of each detection method, it is recommended that the initial testing for the adventitious presence of transgenic seed be carried out using an antibody-based test if available and that Q-PCR-based assays to quantify transgenic proportion be used when it is necessary to identify specific transgenic constructs or if antibody-based assays are not readily available.