As FFQ are subject to measurement error, associations between self-reported intake by FFQ and outcome measures should be adjusted by correction factors obtained from a validation study. Whether the correction is adequate depends on the characteristics of the reference method used in the validation study. Preferably, reference methods should (1) be unbiased and (2) have uncorrelated errors with those in the FFQ. The aim of the present study was to assess the validity of the duplicate portion (DP) technique as a reference method and compare its validity with that of a commonly used reference method, the 24 h recall (24hR), for protein, K and Na using urinary markers as the unbiased reference method. For 198 subjects, two DP, two FFQ, two urinary biomarkers and between one and fifteen 24hR (web based and/or telephone based) were collected within 1·5 years. Multivariate measurement error models were used to estimate bias, error correlations between FFQ and DP or 24hR, and attenuation factors of these methods. The DP was less influenced by proportional scaling bias (0·58 for protein, 0·72 for K and 0·52 for Na), and correlated errors between DP and FFQ were lowest (protein 0·28, K 0·17 and Na 0·19) compared with the 24hR. Attenuation factors (protein 0·74, K 0·54 and Na 0·43) also indicated that the DP performed better than the 24hR. Therefore, the DP is probably the best available reference method for FFQ validation for nutrients that currently have no generally accepted recovery biomarker.