We evaluated the performance of an FFQ in estimating phytosterol intake against multiple 24-h dietary recalls (24HDR) using data from 1011 participants of the calibration sub-study of the Adventist Health Study-2 (AHS-2) cohort. Dietary assessments of phytosterol intake included a self-administered FFQ and six 24HDR and plasma sterols. Plasma sterols were determined using the GLC flame ionisation method. Validation of energy-adjusted phytosterol intake from the FFQ with 24HDR was conducted by calculating crude, unadjusted, partial and de-attenuated correlation coefficients (r) and cross-classification by race. On average, total phytosterol intake from the FFQ was 439·6 mg/d in blacks and 417·9 mg/d in whites. From the 24HDR, these were 295·6 mg/d in blacks and 351·4 mg/d in whites. Intake estimates of β-sitosterol, stigmasterol, other plant sterols and total phytosterols from the FFQ had moderate to strong correlations with estimates from 24HDR (r 0·41–0·73). Correlations were slightly higher in whites (r 0·42–0·73) than in blacks (r 0·41–0·67). FFQ estimates were poorly correlated with plasma sterols as well as 24HDR v. plasma sterols. We conclude that the AHS-2 FFQ provided reasonable estimates of phytosterol intake and may be used in future studies relating phytosterol intake and disease outcomes.