Quantification of the B6 vitamers in human plasma and urine in a study with pyridoxamine as an oral supplement; pyridoxamine as an alternative for pyridoxine

Mathias D. G. Van den Eynde, Jean L. J. M. Scheijen, Coen D. A. Stehouwer, Toshio Miyata, Casper G. Schalkwijk*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


Background and aims: Vitamin B6 is involved in a large spectrum of physiological processes and comprises of the vitamers pyridoxamine (PM), pyridoxal (PL), pyridoxine (PN), and their phosphorylated derivatives including the biological active pyridoxal 50-phosphate (PLP). While PN toxicity is known to complicate several treatments, PM has shown promise in relation to the treatment of metabolic and age-related diseases by blocking oxidative degradation and scavenging toxic dicarbonyl compounds and reactive oxygen species. We aimed to assess the metabolization of oral PM supplements in a single and three daily dose.

Materials and methods: We optimized and validated a method for the quantification of the B6 vitamers in plasma and urine using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Five healthy volunteers were recruited to study PM metabolization after a single oral dose of 200 mg PM or a three daily dose of 67 mg PM. A third protocol was implemented as control for dietary intake. Venous blood samples, 24 h urine and fasted second void urine samples were collected.

Results: After a single oral dose of 200 mg PM, plasma PM increased in the first 3 h to a maximum of 2324 +/- 266 nmol/L. While plasma PM levels returned to baseline after similar to 10 h of PM intake, PLP increased to a maximum of 2787 +/- 329 nmol/L and reached a plateau. We found a small increase of PN to a maximum of 13.5 +/- 2.1 nmol/L; it was nearly undetectable after similar to 12 h. With a three daily dose of 67 mg PM we observed an increase and decline of plasma PM, PL, and PN concentrations after each PM intake. PLP showed a similar increase as in the single dose protocol and accumulated over time.

Conclusion: In this study we showed high plasma levels of PM after oral PM supplementation. We found steadily increasing levels of the biologically active PLP, with minimal formation of PN. The B6 vitamer PM is an interesting supplement as an inhibitor of harmful processes in metabolic diseases and for the treatment of vitamin B6 deficiency. (C) 2021 The Authors. Published by Elsevier Ltd.

Original languageEnglish
Pages (from-to)4624-4632
Number of pages9
JournalClinical Nutrition
Issue number7
Publication statusPublished - Jul 2021


  • Vitamin B6
  • Pyridoxamine
  • Metabolization
  • Pharmacokinetics

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