S-adenosylmethionine (SAMe) is the most common methyl donor found in living organisms. It is the second most common cellular enzyme substrate and is an essential molecule in the carbon metabolic cycle. SAMe is widely used in both a clinical setting and as a nutritional supplement to treat a variety of disorders ranging from liver disease to osteoarthritis in both humans and animals. Demand for SAMe is rapidly expanding and the yeast Saccharomyces cerevisiae is often used as an industrial production host. Increasing clinical research into SAMe, as well as increasing commercial demand, has led to the need for rapid, accurate measurement of SAMe levels from a variety of sources. We report here a novel hydrophilic interaction liquid chromatography (HILIC) UltraPerformance liquid chromatography – mass spectrometry (UPLC-MS) benchtop method to measure intracellular levels of SAMe from S. cerevisiae. Research is ongoing to understand the cellular impact of variations in SAMe levels as well as mutation screens to identify yeast strains with increased SAMe production. This UPLC method allows researchers to measure SAMe with increased resolution, speed, and sensitivity while decreasing both cost and environmental impact, and has the potential to be adapted for the measure of many other metabolites.