Fiber examination is frequently performed in forensics, and gel permeation chromatography (GPC) is one candidate method for discriminating polyester fibers. Here, the effects of machine washing on weight-average molecular weight (M w ), polydispersity index (PDI), and the percentage peak area of cyclic ethylene terephthalate trimer (PPAL) of commercial polyester shirts and manufactured poly(ethylene terephthalate) (PET) yarns were investigated using GPC. GPC was performed using a 1,1,1,3,3,3-hexafluoro-propan-2-ol polymer solubilizer, styrene−divinylbenzene copolymer GPC columns, a chloroform mobile phase, and a 254 nm absorbance monitor. The statistical change in the polyester fibers during machine washing was evaluated by comparing three GPC parameters of the same fiber samples before and after machine washing. Among the commercial polyester shirts examined, the GPC parameters changed significantly after machine washing with a considerable PPAL decrease. In contrast, the GPC parameters of manufactured PET yarns changed significantly with a moderate increase in M w . This work elucidates the change on GPC parameters of polyester fibers by machine washing.