Wikipedia is the largest free online encyclopedia and the seventh most visited website worldwide, containing >45,000 freely accessible English-language medical articles accessed nearly 1.6 billion times annually. Concerns have been expressed about the balance of content related to biological sex on Wikipedia. This study aims to categorize the top 1000 most-read (most popular) English-language Wikipedia health articles for June 2019 according to the relevance of the article topic to each sex and quality. In the first step, Wikipedia articles were identified using WikiProject Medicine Popular Pages. These were analyzed on 13 factors, including total views, article quality, and total number of references. In the second step, 2 general medical textbooks were used as comparators to assess whether Wikipedia's spread of articles was typical compared to the general medical coverage. According to the article's content, we proposed criteria with 5 categories: 1="exclusively female," 2="predominantly female but can also affect male individuals," 3="not sex specific or neutral," 4=predominantly male but can affect female individuals," and 5="exclusively male." Of the 1000 Wikipedia health articles, 993 (93.3%) were not sex specific and 67 (6.7%) were sex specific. There was no statistically significant difference in the number of reads per month between the sex-specific and non-sex-specific articles (P=.29). Coverage of female topics was higher (50/1000, 5%) than male topics (17/1000, 1.7%; this difference was also observed for the 2 medical textbooks, in which 90.2% (2330/2584) of content was not sex specific, female topics accounted for 8.1% (209/2584), and male topics for accounted for 1.7% (45/2584; statistically significant difference; Fisher exact test P=.03). Female-category articles were ranked higher on the Wikipedia medical topic importance list (top, high, or mid importance) than male-category articles (borderline statistical significance; Fisher exact test P=.05). Female articles had a higher number of total and unique references; a slightly higher number of page watchers, pictures, and available languages; and lower number of edits than male articles (all were statistically nonsignificant). Across several metrics, a sample of popular Wikipedia health-related articles for both sexes had comparable quality. Wikipedia had a lower number of female articles and a higher number of neutral articles relative to the 2 medical textbooks. These differences were small, but statistically significant. Higher exclusively female coverage, compared to exclusively male coverage, in Wikipedia articles was similar to the 2 medical textbooks and can be explained by inclusion of sections on obstetrics and gynecology. This is unlike the imbalance seen among biographies of living people, in which approximately 77.6% pertain to male individuals. Although this study included a small sample of articles, the spread of Wikipedia articles may reflect the readership and the population's content consumption at a given time. Further study of a larger sample of Wikipedia articles would be valuable.