Choline, a gut microbiome metabolite, is associated with cardiovascular risk and other chronic illnesses. The aim was to develop a high-throughput nuclear magnetic resonance (NMR)–based assay to measure choline on the Vantera® Clinical Analyzer.

A non-negative deconvolution algorithm was developed to quantify choline. Assay performance was evaluated using CLSI guidelines.

Deming regression analysis comparing choline concentrations by NMR and mass spectrometry (n = 28) exhibited a correlation coefficient of 0.998 (intercept = –9.216, slope = 1.057). The LOQ were determined to be 7.1 µmol/L in serum and 5.9 µmol/L in plasma. The coefficients of variation (%CV) for intra- and inter-assay precision ranged from 6.2 to 14.8% (serum) and 5.4–11.3% (plasma). Choline concentrations were lower in EDTA plasma by as much as 38% compared to serum, however, choline was less stable in serum compared to plasma. In a population of apparently healthy adults, the reference interval was <7.1–20.0 µmol/L (serum) and <5.9–13.1 µmol/L (plasma). Linearity was demonstrated well beyond these intervals. No interference was observed for a number of substances tested.

The newly developed, high-throughput NMR-based assay exhibited good performance characteristics enabling quantification of choline in serum and plasma for clinical use.