Wilson’s disease (WD) is an autosomal recessive disorder of copper (Cu) transport caused by mutations of the ATP7B gene. International guidelines on the management of WD recommend serum non-ceruloplasmin-bound Cu (NCC; free Cu index) for diagnosis and therapeutic monitoring. The current ‘Standard of care’ evaluation of NCC levels involves the incubation of serum with EDTA to chelate copper from proteins other than ceruloplasmin prior to ultracentrifugation. However, a high molecular weight artefact binding copper from albumin retained above the 30KDa filter may result in underestimation of true NCC. We previously developed a Novel Copper Protein Speciation Method for calculating serum non-ceruloplasmin-bound copper however, the injection time is 20 minutes, and the glass column integrity is often compromised when using the GE Mono Q 5/50 GL Ion exchange column. An improved method was developed by Quotient Sciences that uses a steel TOSOH TSKgel Q-STAT Anion exchange column which reduces the robustness problems and delivers enhanced chromatographic performance, improved assay sensitivity and a reduced run time. We aim to compare the chromatography, relative abundance and NCC values of six different batches of serum when analysed with the two columns and methods. We hypothesise that the relative abundance and NCC data produced from the two columns will not be statistically different, but the chromatography will be significantly better when using the TOSOH column. We hope that our findings may contribute to the development of a better diagnostic method for patients with WD, thereby improving the prognosis of these patients.