Background: Trophic interactions are key processes, which determine the ecological function and performance of organisms. Many decapod crustaceans feed on plant material as a source for essential nutrients, e.g. polyunsaturated fatty acids. Strictly herbivorous feeding appears only occasionally in marine decapods but is common in land crabs. To verify food preferences and to establish trophic markers, we studied the lipid and fatty acid composition of the midgut glands of two marine crab species (Grapsus albolineatus and Percnon affine), one semi-terrestrial species (Orisarma intermedium, formerly Sesarmops intermedius), and one terrestrial species (Geothelphusa albogilva) from Taiwan. Results: All species showed a wide span of total lipid levels ranging from 4 to 42% of the dry mass (%DM) in the marine P. affine and from 3 to 25%DM in the terrestrial G. albogilva. Triacylglycerols (TAG) were the major storage lipid compound. The fatty acids 16:0, 18:1(n-9), and 20:4(n-6) prevailed in all species. Essential fatty acids such as 20:4(n-6) originated from the diet. Terrestrial species also showed relatively high amounts of 18:2(n-6), which is a trophic marker for vascular plants. The fatty acid compositions of the four species allow to clearly distinguish between marine and terrestrial herbivorous feeding due to significantly different amounts of 16:0, 18:1(n-9), and 18:2(n-6). Conclusions: Based on the fatty acid composition, marine/terrestrial herbivory indices were defined and compared with regard to their resolution and differentiating capacity. These indices can help to reveal trophic preferences of unexplored species, particularly in habitats of border regions like mangrove intertidal flats and estuaries.