Plant hormones (phytohormones) are long time known as important components of signaling cascades in plant development and plant responses to various abiotic and biotic challenges. Chromatographic methods have been used increasingly to exclude interfering compounds, separate different hormones from each other, and increase the precision of hormones quantification. The combination of GC or LC with MS has been a major advance in the analysis of hormones. Here Creative Proteomics established a HPLC-MS/MS-based platform for quantitative analysis of plant hormones. This platform is sensitive and reliable for simultaneous quantification of multiple analytes in a small amount of tissue.