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Food chain length is a fundamental ecosystem property, and plays a central role in determining ecosystem functioning. Recent advances in the field of stable isotope ecology allow the estimation of food chain length (FCL) from stable nitrogen isotope (δ15N) data. We conducted a global literature synthesis and estimated FCL for 219 lake, stream, and marine ecosystems. Streams had shorter food chains (∼3.5 trophic levels) than marine and lake ecosystems (∼4.0 trophic levels). In marine systems, inclusion of marine mammals increased FCL by 2/3 of a trophic level. For each ecosystem type, estimates of FCL were normally distributed and spanned two full trophic levels. Comparison with published connectance food webs revealed similar mean FCL values, though stable isotope-derived FCL estimates were less variable. At the global scale, FCL showed weak or no relationships with ecosystem size, mean annual air temperature, or latitude. Our study highlights the utility of stable isotopes for quantifying among-system food web variability, and the application of this approach for assessing global-scale patterns of food chain length.