Raw data from an untargeted ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) metabolomics study, in this case of sorghum plants responding dynamically to infection by a fungal pathogen, Colletotrichum sublineolum, was employed. Briefly, two groups of samples were used in this study: fungal-infected (treated) and control (non-infected) samples, labelled T and C, respectively. The control group consisted of nine samples, whereas the treated group consisted of 15 samples. Each sample was a methanol extract from 10 plants. Analytical data of methanol-based plant extracts were acquired in both positive and negative centroid ion mode; but for this paper, only the positive data were further processed. The m/z range was 100–1000 Da and the data were acquired by applying a Waters Acquity UHPLC system coupled in tandem to a Waters photodiode array detector and an electrospray SYNAPT G1 HDMS Q-TOF mass spectrometer (Waters, Milford, MA, USA), applying a method as previously described . A lock spray source was used allowing online mass correction to obtain high mass accuracy of analytes. Leucine encephalin, [M + H]+ = 556.2766 and [M − H]− = 554.2615, was used as the lock mass, being continuously sampled every 15 s, thus producing an average intensity of 350 counts scan−1 in centroid mode. By using a lock mass spray as a reference and continuously switching between sample and reference, the MassLynxTM software can automatically correct the centroid mass values in the sample for small deviations from the exact mass measurement.