The increasing environmental concerns surrounding plastic waste have intensified recycling efforts, particularly in the textile industry, where poly(ethylene terephthalate) (PET) is widely used for sustainable material production. The growing use of recycled PET (rPET) in textiles has prompted the need for reliable analytical methods to detect and quantify rPET content. This study differentiates between virgin and recycled PET by simulating mechanical recycling through five reprocessing cycles of three distinct PET grades, assessing changes in crystalline structure, intrinsic viscosity, molecular weight, and specific degradation markers. Differential Scanning Calorimetry revealed bimodal melting behaviour in reprocessed samples, while intrinsic viscosity and Gel Permeation Chromatography indicated molecular degradation. Notably, the release of dimethyl terephthalate (DMT) and dimethyl isophthalate (DMiP) was consistently observed as a function of degradation. These markers were identified and quantified using High-Performance Liquid Chromatography (HPLC) and Gas Chromatography (GC), with GC offering higher sensitivity and lower matrix interference. This study demonstrates that DMT and DMiP are robust chemical indicators of PET degradation and recycled content. This analytical approach, combining thermal, rheological, and chromatographic techniques, provides a scientifically sound and potentially cost-effective basis for traceability systems, certification protocols, and regulatory compliance in sustainable textile production.