The demand of safe and high-quality foods of plant origin is increasing by the consumers, thus having an impact in the economic and medical aspects of life. To meet quality and safety standards, rapid, reliable, and robust chemical analyses are being developed. Phytofood products are commonly used for human consumption, and their analysis is crucial to prevent possible risk or health issues. Phytofood analysis is an important sector of analytical chemistry providing information on the chemical composition, quality control, and eventual contamination ensuring compliance with food and trade laws. The development of analytical methods to study complex phytofood matrices is complicated by the large variety of the physicochemical properties of their chemical components. The development of advanced analytical and spectroscopic techniques has simplified the analysis allowing to get fast response both in term of food product composition and eventual adulteration. The techniques used include nuclear magnetic resonance (NMR) spectroscopy, gas chromatography (GC), liquid chromatography (LC), mass spectrometry (MS), and also the use of hyphenated methods: gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–mass spectrometry (LC–MS). Data analysis and molecular networking methods are also used to interpret huge amount of data obtained by the chemical and spectroscopic analyses. The choice of the technique depends on the chemical and physical nature of chemical compounds present in the matrices. Usually, an integrated approach combining different methods affords best results. This special issue of Phytochemical Analysis includes papers exploring the analytical methods applied to phytofood analysis for food safety and quality assessment. Particular attention is given to the following topics: (i) application of new methods for extraction, isolation, structure elucidation, and quality control of food phytochemicals; (ii) novel computational technologies, such as molecular networking analysis, associated with phytochemical analytical tools; (iii) application of targeted and untargeted omics technologies, such as metabolomics, to study phytofood composition and origin; and (iiii) analytical methods to predict and evaluate the toxicology and the bioactivity of food phytochemicals.