When we read, our eyes move through the text in a series of fixations and high-velocity saccades to extract visual information. This process allows the brain to obtain meaning, e.g., about sentiment, or the emotional valence, expressed in the written text. How exactly the brain extracts the sentiment of single words during naturalistic reading is largely unknown. This is due to the challenges of naturalistic imaging, which has previously led researchers to employ highly controlled, timed word-by-word presentations of custom reading materials that lack ecological validity. Here, we aimed to assess the electrical neural correlates of word sentiment processing during naturalistic reading of English sentences. We used a publicly available dataset of simultaneous electroencephalography (EEG), eye-tracking recordings, and word-level semantic annotations from 7129 words in 400 sentences (Zurich Cognitive Language Processing Corpus; Hollenstein et al., 2018). We computed fixation-related potentials (FRPs), which are evoked electrical responses time-locked to the onset of fixations. A general linear mixed model analysis of FRPs cleaned from visual- and motor-evoked activity showed a topographical difference between the positive and negative sentiment condition in the 224–304 ms interval after fixation onset in left-central and right-posterior electrode clusters. An additional analysis that included word-, phrase-, and sentence-level sentiment predictors showed the same FRP differences for the word-level sentiment, but no additional FRP differences for phrase- and sentence-level sentiment. Furthermore, decoding analysis that classified word sentiment (positive or negative) from sentiment-matched 40-trial average FRPs showed a 0.60 average accuracy (95% confidence interval: [0.58, 0.61]). Control analyses ruled out that these results were based on differences in eye movements or linguistic features other than word sentiment. Our results extend previous research by showing that the emotional valence of lexico-semantic stimuli evoke a fast electrical neural response upon word fixation during naturalistic reading. These results provide an important step to identify the neural processes of lexico-semantic processing in ecologically valid conditions and can serve to improve computer algorithms for natural language processing.