Determination of free and total valproic acid in human plasma by capillary electrophoresis with contactless conductivity detection

作者：Thi Thanh Thuy Pham; See Hong Heng^*; Morand Rejane; Kraehenbuehl Stephan; Hauser Peter C

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2012, 907: 74-78.

DOI：10.1016/j.jchromb.2012.08.037

• Dispersive liquid-liquid microextraction
• Capillary electrophoresis
• Contactless conductivity detection
• Valproic acid
• Human plasma

A new approach for the determination of free and total valproic acid in small samples of 140 mu L human plasma based on capillary electrophoresis with contactless conductivity detection is proposed. A dispersive liquid-liquid microextraction technique was employed in order to remove biological matrices prior to instrumental analysis. The free valproic acid was determined by isolating free valproic acid from protein-bound valproic acid by ultrafiltration under centrifugation of 100 mu L sample. The filtrate was acidified to turn valproic acid into its protonated neutral form and then extracted. The determination of total valproic acid was carried out by acidifying 40 mu L untreated plasma to release the protein-bound valproic acid prior to extraction. A solution consisting of 10 mM histidine, 10 mM 3-(N-morpholino)propanesulfonic acid and 10 mu M hexadecyltrimethylammonium bromide of pH 6.5 was used as background electrolyte for the electrophoretic separation. The method showed good linearity in the range of 0.4-300 mu g/mL with a correlation coefficient of 0.9996. The limit of detection was 0.08 mu g/mL, and the reproducibility of the peak area was excellent (RSD = 0.7-3.5%, n = 3, for the concentration range from 1 to 150 mu g/mL). The results for the free and total valproic acid concentration in human plasma were found to be comparable to those obtained with a standard immunoassay. The corresponding correlation coefficients were 0.9847 for free and 0.9521 for total valproic acid.

• 出版日期2012-10-15