The relationship between DM intake (DMI) and enteric methane emission is well established in ruminant animals but may depend on measurement technique (e.g. spot v. continuous gas sampling) and rumen environment (e.g. use of fermentation modifiers). A previous meta-analysis has shown a poor overall (i.e. 24 h) relationship of DMI with enteric methane emission in lactating dairy cows when measured using the GreenFeed system (GF; Symposium review: uncertainties in enteric methane inventories, measurement techniques, and prediction models. Journal of Dairy Science 101, 6655 to 6674). Therefore, we examined this relationship in a 15-week experiment with lactating dairy cows receiving a control diet or a diet containing the investigational product 3-nitrooxypropanol (3-NOP), an enteric methane inhibitor, applied at 60 mg/kg feed DM. Daily methane emission, measured using GF, and DMI were clustered into 12 feed-intake timeslots of 2 h each. Methane emission and DMI were the lowest 2 h before feeding and the highest within 6 h after feed provision. The overall (24 h) relationship between methane emission and DMI was poor (R2 = 0.01). The relationship for the control (but not 3-NOP) cows was improved (R2 = 0.31; P < 0.001) when DMI was allocated to timeslots and was strongest (R2 = 0.51; P < 0.001) 8 to 10 h after feed provision. Analysis of the 3-NOP emission data showed marked differences in the mitigation effect over time. There was a lack of effect in the 2-h timeslot before feeding, the mitigation effect was highest (45%) immediately after feed provision, persisted at around 32% to 39% within 10 h after feed provision, and decreased to 13%, 4 h before feeding. These trends were clearly related to DMI (i.e. 3-NOP intake) by the cows. The current analysis showed that the relationship of enteric methane emission, as measured using GF, and DMI in dairy cows depends on the time of measurement relative to time of feeding. The implication of this finding is that a sufficient number of observations, covering the entire 24-h feeding cycle, have to be collected to have representative emission estimates using the GF system. This analysis also revealed that the methane mitigation effect of 3-NOP is highest immediately after feed provision and lowest before feeding.