Background/Objectives: Dartmoor Estate Tea plantation in Devon, UK, is renowned for its unique microclimate and varied soil conditions, which contribute to the distinctive flavours and chemical profiles of tea. The chemical diversity of fresh leaf samples from different were assessed via samples collected from various garden locations within the plantation. Methods: Fresh leaf, which differed by location, cultivar, time of day, and variety, were analysed using Flow Infusion Electrospray Ionisation Mass Spectrometry (FIE-MS). Results: Random forest classification revealed no significant differences between Georgian N2 cultivar garden locations. However, a significant degree of variability was observed within four tri-clonal variants (Tocklai Variety) with TV9 exhibiting greater similarity to the Georgian N2 cultivar compared to TV8 and TV11, while TV11 was found to be most like TV1. The intraclass variability in leaf composition was similar between the varieties. We explored the metabolic changes over the day in one variant (Camellia assamica Masters), yielding a model with a significant R2 value of 0.617 (p < 0.01, 3000 permutations). Starch and sucrose metabolism was found to be significant where the abundance of these chemical features increased throughout the day and then began to decrease at night.Conclusion: This research highlights the complex interplay of cultivars, geographical location, and temporal factors on the chemical composition of tea. It provides insightful data on the metabolic pathways influencing tea cultivation and production and underscores the importance of these variables in determining the final chemical profile and organoleptic characteristics of tea products.