One‐carbon metabolite ratios as functional B‐vitamin markers and in relation to colorectal cancer risk

One‐carbon metabolism biomarkers are easily measured in plasma, but analyzing them one at a time in relation to disease does not take into account the interdependence of the many factors involved. The relative dynamics of major one‐carbon metabolism branches can be assessed by relating the functional B‐vitamin marker total homocysteine (tHcy) to transsulfuration (total cysteine) and methylation (creatinine) outputs. We validated the ratios of tHcy to total cysteine (Hcy:Cys), tHcy to creatinine (Hcy:Cre) and tHcy to cysteine to creatinine (Hcy:Cys:Cre) as functional markers of B‐vitamin status. We also calculated the associations of these ratios to colorectal cancer (CRC) risk. Furthermore, the relative contribution of potential confounders to the variance of the ratio‐based B‐vitamin markers was calculated by linear regression in a nested case–control study of 613 CRC cases and 1,190 matched controls. Total B‐vitamin status was represented by a summary score comprising Z‐standardized plasma concentrations of folate, cobalamin, betaine, pyridoxal 5′‐phosphate and riboflavin. Associations with CRC risk were estimated using conditional logistic regression. We found that the ratio‐based B‐vitamin markers all outperformed tHcy as markers of total B‐vitamin status, in both CRC cases and controls. In addition, associations with CRC risk were similar for the ratio‐based B‐vitamin markers and total B‐vitamin status (approximately 25% lower risk for high vs. low B‐vitamin status). In conclusion, ratio‐based B‐vitamin markers were good predictors of total B‐vitamin status and displayed similar associations as total B‐vitamin status with CRC risk. Since tHcy and creatinine are routinely clinically analyzed, Hcy:Cre could be easily implemented in clinical practice.