In this study, we aimed to assess the feasibility of re-collecting breath samples using the Centri® (Markes International, Bridgend, UK) followed by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC×GC-TOFMS) analysis. The work was conducted in two main phases. In the first phase, we evaluated the re-collection performance by analyzing two sets of standards, including a Grob mix primary solution and a standard mixture of 20 selected volatile compounds (VCs) covering different classes of organic species commonly found in breath samples. The intra-day and inter-day precision (reported as relative standard deviation (RSD), %) for the re-collection of the Grob mix primary solution were in the range of 1% to14% and 3% to12%, respectively. The re-collection accuracy ranged from 78% to 97%. The intra-day RSD for the re-collection of the standard mixture of selected VCs was within 20% for all compounds, except for acetone and nonane. The precision was within 25% for all compounds, except for nonane, n-hexane, 1,4-dichlorobenzene, and decane, which exhibited less than 36% RSD. The re-collection accuracy was in the range of 67% to 129%. In the second phase of the study, the re-collection performance in breath analysis was evaluated via five repetitive splitting and re-collection of six breath samples obtained from healthy adults, realizing a total of 30 breath analyses. Initially, we evaluated the re-collection performance by considering all features obtained from breath analysis and then focused on the 20 VCs commonly found in breath samples. The re-collection accuracy for total breath features ranged from 86 to 103%, and the RSDs were in the range of 1.0% to 10.4%. For the selected VCs, the re-collection accuracy of all compounds, except for undecane and benzene, was in the range of 71% to 132%.