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Materials and methods

Milk samples (214 lame; 211 control) from dairy cows were collected from 10 commercial dairy farms across Derbyshire after cows were mobility scored by visual inspection using 0-3 mobility scoring (AHDB, 2024). Samples (n= 425) were prepared using biphasic extraction for untargeted metabolomics and lipidomics. For this study, only the organic phases were used. LC-MS was performed in 4 batches and pooled quality control (QC) samples were prepared. Pre-processing of the generated LC-MS datasets including lipid identification was performed using Compound Discoverer 3.3 SP2 software (Thermo Fisher Scientific, Hemel Hempstead, UK). Statistical analysis comprised multivariate analysis performed on all the detected peaks in the milk samples using SIMCA 16 software (Umetrics, Sweden). Principal component analysis was used to check the performance of the analytical run and …