The commonly used methods of estimating the 24-h urinary sodium (UNa) and urinary potassium (UK) from spot urine (SU) are the Kawasaki method (K-method), INTERSALT method (I-method), and Tanaka method (T-method), but the method that is suitable for use in the general Chinese population is still uncertain. We aimed to assess and validate these methods in estimating the 24-h UNa and UK using SU samples in Chinese adults. We studied 1428 individuals aged 18-69 years using SU and 24-h urine samples. For the K-method, I-method, and T-method, the Pearson correlation coefficients of the 24-h UNa were 0.35, 0.35, and 0.33 (all p < 0.01), and the intraclass correlation coefficients (ICC) were 0.34, 0.26, and 0.26 (all p < 0.01), respectively. The estimated 24-h UK using the K-method and T-method had correlation coefficients of 0.36 and 0.39 (all p < 0.01) and ICCs of 0.31 and 0.27 (all p < 0.01). The mean bias for the K-method in estimating the 24-h UNa and UK were the least biased among these methods. The bias between the 24-h urine Na/K ratio and the spot urinary Na/K ratio by the Bland-Altman method was −0.22. These methods for estimating the 24-h UNa and UK from SU were inadequate at the population level in Zhejiang Province, although the K-method showed the least bias among these methods. The spot urine Na/K ratio may be a useful and alternative method for 24-h urine collection for the estimation of the urinary Na/K ratio in the Chinese population.