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Original Articles

DETERMINATION OF TOTAL AND UNBOUND CONCENTRATIONS OF VALPROIC ACID IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY

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Pages 1171-1183 | Published online: 09 May 2012
 

Abstract

An analytical method for the determination of valproic acid in human plasma and ultrafiltrated human plasma has been developed and validated. The analytical method lies in the solid phase extraction of valproic acid (VPA), followed by its quantification through an LC-MS method, using benzoic acid as the internal standard. The intra-assay and inter-assay precisions of this technique ranged between 4.29% and 9.91% and its accuracy ranged between 1.00% and 7.73%, for VPA spiked in plasma matrix. For VPA spiked in ultrafiltrated plasma, matrix precisions were from 6.29% to 9.84% and accuracies, from −1.17% to 5.38%. The limit of quantification was set at 1 µg/mL (precision 3.20%, accuracy 11.31%). The linear working range of concentrations was established between 10 and 250 µg/mL for the plasma matrix and between 1 and 100 µg/mL for the ultrafiltrated plasma matrix. The absolute recovery of valproic acid was over 82% in the studied range of concentrations in both matrixes. Valproic acid was found to be stable in both matrixes at the assayed conditions. This method has shown to be suitable for the analysis of free and total valproic acid concentrations in human plasma samples.

ACKNOWLEDGMENT

We thank Dr. Olga Jauregui and Dr. Andres Miguel Miguelez for their invaluable advice.

Notes

Note: Correlation coefficient was significant in all cases (p < 0.05).

SD, standard deviation.

SD, standard deviation; %CV, coefficient of variation; %RE, relative error.

SD, standard deviation; %CV, coefficient of variation; %RE, relative error.

*Ratio is expressed as percentages comparing mean area ratios of extracted samples under experimental conditions with respect to the freshly extracted samples analyzed at time zero.

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