Practice of solid-phase extraction and protein precipitation in the 96-well format combined with high-performance liquid chromatography-ultraviolet detection for the analysis of drugs in plasma and brain

Journal of Chromatography. B, Biomedical Sciences and Applications
M C RouanG Maurer

Abstract

C18 Empore 96-well extraction disc plates have been employed for the analysis of three drugs with different polarities in plasma in conjunction with HPLC-UV, rufinamide, ICL670 and an anticonvulsant agent (AA1) in an early stage of development. With the most polar compound (AA1), ion-pair extraction at pH 12 was applied. The method developed for the assay of AA1 in plasma was applied to its determination in brain using an Oasis HLB plate following homogenisation in a pH 7.4 buffer and protein precipitation with NaOH-ZnSO4, thereby saving time for method development. Protein precipitation in the 96-well format with filtration of the precipitate was applied to the determination of ICL670, a highly protein-bound compound (>99.5%), with a good recovery (78%). Reversed-phase chromatography was applied using a short 5 cm column packed with 3 microm particles for the determination of ICL670 and AA1 and two parallel columns (15 cm long) for the determination of rufinamide. The methods were used routinely, one plate per analysis day being processed, resulting in increase in sample throughput and saving in solvents.

Citations

Oct 6, 2012·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ramakrishna NirogiHimabindu Vurimindi
Mar 23, 2012·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Helen PligoropoulouIrene Panderi
Aug 11, 2011·Luminescence : the Journal of Biological and Chemical Luminescence·Jamshid L ManzooriJalil Vaez-Gharamaleki
Apr 12, 2016·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Jafar AbolhasaniJavad Hassanzadeh

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