Despite concerns about their health risks, e‑cigarettes have gained popularity in recent years. Concurrent with the recent increase in e‑cigarette use, social media sites such as Twitter have become a common platform for sharing information about e-cigarettes and to promote marketing of e‑cigarettes. Monitoring the trends in e‑cigarette–related social media activity requires timely assessment of the content of posts and the types of users generating the content. However, little is known about the diversity of the types of users responsible for generating e‑cigarette–related content on Twitter. The aim of this study was to demonstrate a novel methodology for automatically classifying Twitter users who tweet about e‑cigarette–related topics into distinct categories. We collected approximately 11.5 million e‑cigarette–related tweets posted between November 2014 and October 2016 and obtained a random sample of Twitter users who tweeted about e‑cigarettes. Trained human coders examined the handles’ profiles and manually categorized each as one of the following user types: individual (n=2168), vaper enthusiast (n=334), informed agency (n=622), marketer (n=752), and spammer (n=1021). Next, the Twitter metadata as well as a sample of tweets for each labeled user were gathered, and features that reflect users’ metadata and tweeting behavior were analyzed. Finally, multiple machine learning algorithms were tested to identify a model with the best performance in classifying user types. Using a classification model that included metadata and features associated with tweeting behavior, we were able to predict with relatively high accuracy five different types of Twitter users that tweet about e‑cigarettes (average F1 score=83.3%). Accuracy varied by user type, with F1 scores of individuals, informed agencies, marketers, spammers, and vaper enthusiasts being 91.1%, 84.4%, 81.2%, 79.5%, and 47.1%, respectively. Vaper enthusiasts were the most challenging user type to predict accurately and were commonly misclassified as marketers. The inclusion of additional tweet-derived features that capture tweeting behavior was found to significantly improve the model performance—an overall F1 score gain of 10.6%—beyond metadata features alone. This study provides a method for classifying five different types of users who tweet about e‑cigarettes. Our model achieved high levels of classification performance for most groups, and examining the tweeting behavior was critical in improving the model performance. Results can help identify groups engaged in conversations about e‑cigarettes online to help inform public health surveillance, education, and regulatory efforts.