Liquid chromatography has been widely employed in the analysis of materials in Heritage Science, due to its ease of use and relatively low-cost, starting from thin layer chromatography of organic binders in paintings, of archaeological waxes and resins, and finally of natural dyes. High performance systems employing analytical columns containing packed stationary phases gradually supplanted thin layer chromatography (TLC) in the field, since the separation, detection and quantitation of specific species contained in a sample in the field of Cultural Heritage requires selective, sensitive and reliable methods, allowing for analysing a wide range of samples, in terms of analyte types and concentration range. Today, the main applications of High-Performance Liquid Chromatography in this field are related to the separation and detection of dyestuffs in archaeological materials and paint samples by reversed-phase liquid chromatography with suitable detectors. Proteomics and lipidomics are also gaining momentum in the last decade, thanks to the increased availability of instrumentation and procedures. In this chapter, principles and theory of liquid chromatography will be presented. A short review of the instrumentation needed to perform an analysis will be provided and some general principles of sample preparation revised. More details on the detection systems, the chromatographic setups and specific sample treatment strategieswill be provided in the individual sections dedicated to the applications to Heritage Science of the main types of liquid chromatographic techniques. In particular, the applications of thin layer chromatography will be shortly described in paragraph 4.1. The applications of Reverse Phase High Performance Liquid Chromatography (RP-HPLC) will be discussed in detail in paragraph 4.2, including the analysis of natural and synthetic dyes and pigments and the profiling of lipid materials. The possibility to perform proteomic analysis will be presented and a link to the relevant Chapter in this book provided. The most important and promising applications of ion exchange chromatography (IC) will be discussed in paragraph 4.3. Finally, size exclusion and gel permeation chromatography (GPC) will be presented in paragraph 4.4, including applications to the study of polymeric network formation in paint binders, of the phenomena related to the depolymerisation of cellulose in paper and of cellulose and lignin in wood samples. The possibility to study synthetic polymers as artists’ materials and restorers’ tools by size exclusion (SEC) or gel permeation (GPC)will also be introduced. In the conclusions, future perspectives of liquid chromatography in Heritage Science will be briefly discussed.