There is both historical and increased current interest in the use of phytochemicals extracted from plant material (e.g., leaf, bark, seeds, fruit, etc.) as dietary supplements and cosmeceuticals to prevent and minimize the effects of degenerative diseases and aging.1,2 Numerous compounds extracted from plant materials have medical and/or benecial effects on humans (i.e., bioactives which include cosmeceuticals) and are the subjects of other chapters in this volume. In addition, several articles reviewing approaches for the analysis and isolation of plant products and bioactives have previously been published.3-10 This chapter will emphasize chromatographic approaches for identifying and quantifying key bioactives in botanicals. These same approaches can also be used to conrm the authenticity, potency, and composition of botanicals, once the components and their mean concentrations have been established for a plant. While both gas chromatographic (GC) and high performance liquid chromatographic (HPLC) methods can be used for analysis of volatile and nonvolatile compounds, recent advances in hyphenated mass spectrometry (MS) detection have vastly improved our ability to quantify targeted known compounds and to identify unknown bioactives from plant extracts. Therefore, this chapter will emphasize these hyphenated GC-MS and HPLC-MS techniques, providing a brief background of their principles and reviewing a select number of applications. This chapter is not intended to give a comprehensive review of all methods published to date for analysis of botanicals. The goal is to provide the reader with an overview of contemporary chromatographic and mass spectrometric approaches used in the chemical characterization of botanical extracts.