Liquid-phase microextraction based on carrier mediated transport combined with liquid chromatography-mass spectrometry: New concept for the determination of polar drugs in a single drop of human plasma

Tung Si Ho*, Jan Leon Egge Reubsaet, Hanne Sofie Anthonsen, Stig Pedersen-Bjergaard, Knut Einar Rasmussen

*Corresponding author for this work
    95 Citations (Scopus)

    Abstract

    Recently, we demonstrated for the first time liquid-phase microextraction (LPME) of polar drugs based on carrier mediated transport. In this new extraction technique, selected analytes were extracted as ion-pairs from small volumes of biological samples, through a thin layer of a water immiscible organic solvent immobilised in the pores of a porous hollow fibre (liquid membrane), and into a μl volume of an acidic aqueous acceptor solution placed inside the lumen of the hollow fibre. In the current paper, this new extraction technique was combined with liquid chromatography-mass spectrometry (LC-MS) for the first time. Carrier mediated LPME was evaluated for several new model drugs (0.01 < log P < 1.76), the sample clean-up aspects were investigated in detail, and this new extraction technique was fully validated for the first time. Extractions were performed from 50 μl of human plasma samples, which provided sufficient material in combination with LC-MS. Sodium octanoate (50 mM) was added to the sample as carrier, 1-octanol (≈15 μl) was used as the liquid membrane in the wall of the hollow fibre, and 50 mM HCl was utilized as acceptor solution in the lumen of the hollow fibre. The addition of carrier to the samples was found to significantly improve extraction recoveries for the polar drugs tested, providing recoveries in the range 16-78%. Validation was accomplished for atenolol and cimetidine. Limits of quantification (S/N = 5) from 50 μl of plasma were 25 and 50 ng/ml for atenolol and cimetidine, respectively. The intra-day precision (R.S.D.) ranged from 7.8 to 17.2% and from 9.5 to 14.1% for atenolol and cimetidine, respectively, and corresponding inter-day precisions (R.S.D.) were within 6.7-1.4% and 7.7-20.3%. The method was linear in the range 25-1500 ng/ml for atenolol (r = 0.992), and 50-3500 ng/ml for cimetidine (r = 0.976). The accuracy of the method was found to be in range 89.1-99.6% and 83.4-86% for atenolol and cimetidine, respectively. The sample clean-up obtained by carrier mediated LPME was excellent, providing a significantly lower back-ground level in total ion current chromatograms by LC-MS as compared to protein precipitation.

    Original languageEnglish
    JournalJournal of Chromatography A
    Volume1072
    Issue number1
    Pages (from-to)29-36
    Number of pages8
    ISSN0021-9673
    DOIs
    Publication statusPublished - 22 Apr 2005

    Keywords

    • Carrier mediated transport
    • Drugs
    • Liquid-phase microextraction
    • Polar

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