The Paracas Necropolis on the south coast of Peru is renowned for the elaborate funerary bundles recovered from that dry sandy region. These bundles contained the remains of male leaders within the Paracas communities surrounded by multiple layers of plain cloth and garments with embroidered designs. The methods and materials used in dyeing the yarns used to embellish these ancient fabrics are still not well understood, and the research presented herein seeks to add to that body of knowledge. To investigate the sources of dye colourants in samples from Paracas Necropolis textiles, we applied direct analysis in real time-time-of-flight mass spectrometry (DART-MS). This new methodology has both advantages (analyses are rapid and require little or no sample preparation) and disadvantages (e.g. inability to identify intact carminic acid) compared to existing ones used in the analysis of dye colourants. Direct analysis in real time mass spectra were collected on chemical standards, botanical materials and comparative reference samples prepared with Relbunium roots and cochineal insects (Dactylopius sp.), and the results are compared to red fibres obtained from several different textiles within two different funerary bundles from the Museo Nacional de Arqueología Antropología e Historia in Lima, Peru. The results from the DART-MS analysis of these red fibres show that the compounds present are consistent with the presence of dye obtained from Relbunium species, and cochineal insects were not used to colour these fibres. Statement of significance DART-MS is an efficient and effective method to identify red colourant composition in small samples of fibres removed from archaeological textiles. Of significant note is the ability to differentiate isomers present in Relbunium and other anthraquinone dyes. Colourants in Relbunium are readily distinguished. Although a component of carminic acid from cochineal is less reliably positively identified by DART-MS, it is possible to use a simple extraction and ionization method to confirm or exclude the presence of carminic acid. Relbunium and cochineal are known to have been used in Paracas textiles. Not only does DART-MS aid in identification of colourants using small amounts of material and without significant sample preparation, the information can be used to distinguish fibres that were prepared differently thus indicating past knowledge of dyeing technology. The information can also aid in separating periods of artefact manufacture, and guide development of a conservation strategy.