Many direct immunoassays in use today for testosterone analysis are little better than a guess at the low levels that are sometimes required. This has led many clinical societies to advocate for research into mass spectrometry-based testing. Given this recommendation, and the often high volume of testing, many clinical laboratories are considering the development of their own in-house mass spectrometry-based testosterone analytical methods. This consideration requires selecting an instrument and workflow that fits laboratory operations and the requisite analytic specifications of the assay. The trade-off in these considerations is typically between more advanced, and more expensive, instruments or more involved and time consuming sample preparation techniques. This webinar will present research on different liquid chromatography tandem mass spectrometry testosterone analytical methods that can be performed on low-, mid- and high-sensitivity mass spectrometers. With each of these analytical methods, the increasing cost of the instrument comes with the advantage of decreasing sample preparation. The improvements in mass spectrometers will be discussed to provide a better understanding of what makes for more sensitive instruments; the operational challenges and costs of various sample preparation strategies will also be discussed. The goal is for clinical researchers to be aware of these tradeoffs when evaluating instruments in their research so they can select the instrument and workflow that best suits their laboratory’s needs.