Comparison of a Direct and Indirect Method for Measuring Flavins- Assessing Flavin Status in Patients Receiving Total Parenteral Nutrition

J.S. Hamilton1, J.V. Woodside2, W.A. McIlveen1, M.C. McKinley2, I.S. Young*, 1, 2
1 epartment of Clinical Biochemistry, Royal Victoria Hospital, Grosvenor Road, Belfast BT12 6BA, UK
2 Nutrition and Metabolism Group, Centre for Clinical and Population Science, Queen’s University Belfast, Grosvenor Road, Belfast BT12 6BJ, UK

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© 2009 Hamiltonet al;

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Nutrition and Metabolism Group, Centre for Clinical and Population, Queen’s University Belfast, Institute of Clinical Science, Grosvenor Road, Belfast BT12 6BJ, UK; E-mail:


Background: Riboflavin, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) are cofactors in oxidation-reduction reactions. The aim of this study was to compare a direct high performance liquid chromatography (HPLC) method with an indirect enzymatic method to assess flavin status and establish reference intervals. The HPLC method was used to assess prevalence of flavin deficiency in total parenteral nutrition (TPN) patients.

Method: Blood was obtained from 90 healthy volunteers and 51 TPN patients. Results for each method were compared using Spearmans Rank correlation. Results from reference groups and TPN patients were compared using Mann-Whitney U tests.

Results: Erythrocyte glutathione reductase activity coefficients (EGRAC) correlated with flavin levels measured by HPLC (p<0.001). HPLC was chosen for TPN samples analyses due to superior sensitivity. There was no significant difference between FAD and FMN levels in controls and TPN patients, although TPN patients had significantly higher levels of riboflavin than controls (p<0.001).

Conclusion: HPLC is superior to the EGRAC assay for assessing flavin status. Hyporiboflavinosis is not prevalent in TPN patients.