The presence of natural ingredients as bioactive components found in essential oils has been a major field of research due to their wide applicability. Essential oils are analyzed using advanced analytical methods to determine secondary substances or minor components. The analytical challenges that arise during analysis are mainly the separation of analytes and preparation of samples. The preparation of samples is considered the most crucial step in developing analytical methods for the detection of bioactive compounds in essential oils. This chapter describes several extraction methods and analytical techniques for extracting bioactive compounds from essential oils.