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We evaluated the impact of standardizing the originally measured serum total 25-hydroxyvitamin D (25(OH)D) values from Third National Health and Nutrition Examination Survey (NHANES III, 1988-1994) on the association between 25(OH)D and rate of all-cause mortality. Values were standardized to the gold-standard laboratory method. Follow-up from 1990-2006 consisted of 15,099 participants aged at least 20 years at baseline, among whom there were 3,784 deaths. Relative risk of death was adjusted for age, sex, race/ethnicity, and season using Poisson regression. Results were obtained for eight 25(OH)D (nmol/L) categories: &lt;20 nmol/L, 20-29 nmol/L, 30-39 nmol/L, 40-49 nmol/L, 50-59 nmol/L, 60-74 nmol/L, 75-99 nmol/L (reference), and ≥100 nmol/L. Assay standardization dramatically shifted original 25(OH)D values toward zero. Accordingly, risk ≥120 nmol/L could not be evaluated (i.e., n = 7 and ndeaths = 2). Relative risk (95% confidence interval (CI)) &lt;40 nmol/L remained significant (30-39 nmol/L: relative risk (RR) = 1.4 (95% CI: 1.1, 1.6); 20-29 nmol/L: RR = 1.6 (95% CI: 1.3, 1.9), and &lt;20 nmol/L: RR = 2.1 (95% CI: 1.6, 2.7). However, adjusted relative risk estimates for 25(OH)D levels ≥40 nmol/L were no longer significant (40-49 nmol/L: RR = 1.2 (95% CI: 0.99, 1.4); 50-59 nmol/L: RR = 1.2 (95% CI: 1.04, 1.4); 60-74 nmol/L: RR = 1.1 (95% CI: 0.94, 1.2); 75-99 nmol/L: RR = 1.0 (referent), and ≥100 nmol/L: RR = 1.1 (95% CI: 0.6, 2.1). In summary, after standardization, risk of death from all causes increased with decreasing 25(OH)D &lt;40 nmol/L, while there was no association with values in categories between 40 nmol/L and 120 nmol/L.

The Reverse J-Shaped Association Between Serum Total 25-Hydroxyvitamin D Concentration and All-Cause Mortality: The Impact of Assay Standardization