The textile industry uses many raw materials (natural and synthetic dyes and fibers) and different dyeing techniques that can be considered important pollutants with a negative impact on the environment (toxic working conditions, discharged wastewater, and contamination). Although synthetic dyes are intensively used, offer a wide range of colors and hues and properties of adhesion, longevity, and resistance to sunshine and chemical processes, and are cost-effective, they have begun to be restricted by many textile producers because they are nonbiodegradable and have toxic, carcinogenic, and mutagenic effects that generate some imbalances in plant, animal, and human life. Natural dyes of plant and animal origin exhibit very good tolerance to washing, rubbing, and light and are biodegradable and nontoxic; these properties have led to a call for the renewed use of these dyes. Modern analytical techniques (solid-phase extraction, spectrophotometry, HPLC, HPTLC, capillary electrophoresis) with different spectroscopy (UV-Vis, diode-array detection, pulsed amperometric detection) and/or MS/tandem mass spectrometry detectors have an important role in the textile industry in obtaining essential information about dyeing techniques, material origin, historical trade routes of ancient textiles, and environmental pollution. For this purpose, isolation, separation, and quantification methods of natural and synthetic textile dyes from various matrices (ancient and modern fabrics, water, biota, etc.) are presented.