Circulating pyridoxal 5′-phosphate in serum and whole blood: implications for assessment of vitamin B6 status

Abstract Objectives Concentrations of pyridoxal 5′-phosphate (PLP) in serum and whole blood are routinely measured. The suitability of these markers in capturing vitamin B6 insufficiency is not well studied. Methods In 212 subjects, concentrations of PLP and thiamine pyrophosphate (TPP) were simultaneously measured in EDTA-whole blood using Chromsystems® (52052) method on HPLC devices. The whole blood PLP concentrations were compared to serum PLP concentrations measured using reagents from Immundiagnostik® (KC 2100). The whole blood TPP concentrations measured with the Chromsystems® (52052) were compared to those measured by the Chromsystems® (35000) method. Concentrations of homocysteine (tHcy) and cystathionine (Cys) were measured and used to judge the PLP methods. Results Serum PLP concentrations were on average 41% lower than whole blood PLP [mean (SD)=55.4 (83.0) vs. 131 (217) nmol/L]. Serum and whole blood PLP showed a strong correlation [Pearson correlation coefficient=0.724, p<0.001, n=204]. Eighty-five samples showed discrepant results for PLP status (serum PLP ≤30 nmol/L, but whole blood PLP >51 nmol/L) while 102 samples showed coherent results (reference group). The discrepancy group showed higher odds ratio for elevated tHcy >12.0 μmol/L compared to the reference group [OR (95% confidence intervals, CI)=2.1 (1.2–4.0)]. The OR (95% CI) of elevated Cys >300 nmol/L was 1.9 (1.0–3.5) in the discrepancy group compared to the reference group. TPP concentrations were 6% lower when using the Chromsystems®, 52052 compared to levels measured with Chromsystems®, 35000. Conclusions Serum and whole blood PLP concentrations disagree in a substantial number of samples. Serum PLP was better in reflecting elevated tHcy and Cys compared to whole blood PLP. Whole blood PLP underestimates the prevalence of vitamin B6 insufficiency. Methods of measuring TPP concentrations in whole blood were exchangeable.