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The Scientific World Journal
Volume 2012 (2012), Article ID 359721, 8 pages
Research Article

A Novel HPLC Method for the Concurrent Analysis and Quantitation of Seven Water-Soluble Vitamins in Biological Fluids (Plasma and Urine): A Validation Study and Application

1Centre for Health Services Studies, University of Kent, Canterbury, Kent CT2 7NF, UK
2Department of Biosciences, University of Kent, Canterbury, Kent CT2 7PD, UK
3School of Health, Nursing and Midwifery, University of the West of Scotland, Ayr Campus, Ayr KA8 0SX, Scotland, UK

Received 10 October 2011; Accepted 30 November 2011

Academic Editors: D. Agbaba and M. Mahmoud

Copyright © 2012 Margherita Grotzkyj Giorgi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


An HPLC method was developed and validated for the concurrent detection and quantitation of seven water-soluble vitamins (C, B1, B2, B5, B6, B9, B12) in biological matrices (plasma and urine). Separation was achieved at 30°C on a reversed-phase C18-A column using combined isocratic and linear gradient elution with a mobile phase consisting of 0.01% TFA aqueous and 100% methanol. Total run time was 35 minutes. Detection was performed with diode array set at 280 nm. Each vitamin was quantitatively determined at its maximum wavelength. Spectral comparison was used for peak identification in real samples (24 plasma and urine samples from abstinent alcohol-dependent males). Interday and intraday precision were <4% and <7%, respectively, for all vitamins. Recovery percentages ranged from 93% to 100%.