Highlights 17 Proteins are the dominant compound of zooplankton 18 Biochemical and energy content differ between zooplankton groups and size 19 classes 20 Copepods, decapod larvae and chaetognaths represent the most energy-rich 21 groups 22 Biochemical and energy content vary seasonally with environmental 23 parameters 24 Zooplankton energy peaks in 200-1000 µm size classes in all seasons 25 2 Abstract 26 Zooplankton plays a prominent role in marine pelagic food webs, but its contribution 27 to organic matter transfer from phytoplankton to upper level consumers is modulated 28 by size and specific composition. The biochemical composition (protein, carbohydrate 29 and lipid concentrations) and energy content of size-and group-fractionated 30 zooplankton were analyzed in the NW Mediterranean Sea (Marseille) over 18 months. 31 Proteins were the most abundant biochemical compounds in all size classes and 32 zooplankton groups, and provided the largest part of the energy content. The medium 33 (200-1000 μm) size classes, mainly composed of copepods, crustacean larvae and 34 eggs, presented a higher protein and energy content than both the smallest (80-200 µm) 35 size class, mainly composed of phytoplankton, and the largest (>1000 μm) size 36 classes, composed of siphonophores, salps and chaetognaths. Strong and similar 37 seasonal variations in zooplankton biochemical composition were observed in all size 38 classes, with higher energy content associated with cold oxygen-and nutrient-rich 39 waters in spring 2017 and winter 2018. This study evidenced the importance of size-40 and group-fractionating methods to achieve a better insight into energy and nutrient 41 transfer up into pelagic food webs, and understanding of zooplanktivorous fish 42 feeding preferences, depending on the variable quality of zooplankton prey. 43 44 45 46 47 48