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Clinical Chemistry 45: 862-868, 1999;
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(Clinical Chemistry. 1999;45:862-868.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Quantification of Riboflavin, Flavin Mononucleotide, and Flavin Adenine Dinucleotide in Human Plasma by Capillary Electrophoresis and Laser-induced Fluorescence Detection

Steinar Hustada, Per Magne Ueland and Jørn Schneede

Department of Pharmacology, University of Bergen, Armauer Hansens Hus, 5021 Bergen, Norway.
a Author for correspondence. Fax 47-55-974605; e-mail steinar.hustad{at}farm.uib.no

Background: Riboflavin is the precursor of flavin mononucleotide (FMN) and FAD, which serve as cofactors for several redox enzymes. We have developed a capillary electrophoresis method for the determination of riboflavin and its two coenzyme forms in human plasma.

Methods: Trichloroacetic acid-treated plasma was subjected to solid-phase extraction on reversed-phase columns. The analytes were separated by micellar electrokinetic capillary chromatography in uncoated fused- silica capillaries filled with borate buffer containing 50 mmol/L sodium dodecyl sulfate, methanol, and N-methylformamide. Native fluorescence was monitored at 530 nm, using an argon laser operating at 488 nm as excitation source.

Results: The assay was linear over a concentration range of two orders of magnitude, and the limit of detection was far below physiological concentrations for all vitamers. The within-day and between-day coefficients of variation were 4–9% and 6–12%, respectively. The reference values (median, 5–95 percentiles) obtained by analyzing plasma from 63 healthy subjects were 8.6 nmol/L (2.7–42.5 nmol/L) for riboflavin, 7.0 nmol/L (3.5–13.3 nmol/L) for FMN, and 57.9 nmol/L (44.5–78.1 nmol/L) for FAD.

Conclusions: Capillary electrophoresis with laser-induced fluorescence detection allows determination of all riboflavin vitamers far below physiological concentrations. The method may become a useful tool for the assessment of riboflavin status in humans.© 1999 American Association for Clinical Chemistry




The following articles in journals at HighWire Press have cited this article:


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J.-Y. Hautem, C. Morel, R. Couderc, and F. Moussa
Liquid chromatographic determination of b2 vitamers in human plasma and whole blood.
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O. Midttun, S. Hustad, E. Solheim, J. Schneede, and P. M. Ueland
Multianalyte Quantification of Vitamin B6 and B2 Species in the Nanomolar Range in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry
Clin. Chem., July 1, 2005; 51(7): 1206 - 1216.
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S. Hustad, B. G Nedrebo, P. M. Ueland, J. Schneede, S. E. Vollset, A. Ulvik, and E. A Lien
Phenotypic expression of the methylenetetrahydrofolate reductase 677C->T polymorphism and flavin cofactor availability in thyroid dysfunction
Am. J. Clinical Nutrition, October 1, 2004; 80(4): 1050 - 1057.
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S. Hustad, M. C. McKinley, H. McNulty, J. Schneede, J.J. Strain, J. M. Scott, and P. M. Ueland
Riboflavin, Flavin Mononucleotide, and Flavin Adenine Dinucleotide in Human Plasma and Erythrocytes at Baseline and after Low-Dose Riboflavin Supplementation
Clin. Chem., September 1, 2002; 48(9): 1571 - 1577.
[Abstract] [Full Text] [PDF]

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S. Hustad, P. M. Ueland, S. E. Vollset, Y. Zhang, A. L. Bjorke-Monsen, and J. Schneede
Riboflavin as a Determinant of Plasma Total Homocysteine: Effect Modification by the Methylenetetrahydrofolate Reductase C677T Polymorphism
Clin. Chem., August 1, 2000; 46(8): 1065 - 1071.
[Abstract] [Full Text] [PDF]


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Copyright © 1999 by the American Association for Clinical Chemistry.