Roldán-Pijuán Mercedes, Pedersen-Bjergaard Stig, Gjelstad Astrid
Department of Analytical Chemistry, Institute of Fine Chemistry and Nanochemistry, University of Córdoba, Marie Curie Building, Campus de Rabanales, 14071, Córdoba, Spain.
Anal Bioanal Chem. 2015 Apr;407(10):2811-9. doi: 10.1007/s00216-015-8505-9. Epub 2015 Feb 15.
The new sample preparation concept "Parallel artificial liquid membrane extraction (PALME)" was evaluated for extraction of the acidic drugs ketoprofen, fenoprofen, diclofenac, flurbiprofen, ibuprofen, and gemfibrozil from human plasma samples. Plasma samples (250 μL) were loaded into individual wells in a 96-well donor plate and diluted with HCl to protonate the acidic drugs. The acidic drugs were extracted as protonated species from the individual plasma samples, through corresponding artificial liquid membranes each comprising 2 μL of dihexyl ether, and into corresponding acceptor solutions each comprising 50 μL of 25 mM ammonia solution (pH 10). The liquid membranes and the acceptor solutions were located in a 96-well filter plate, which was sandwiched with the 96-well donor plate during extraction. Parallel extraction of several samples was performed for 15 to 60 min, followed by high-performance liquid chromatography-ultraviolet detection of the individual acceptor solutions. Important PALME parameters including the chemical composition of the liquid membrane, extraction time, and sample pH were optimized, and the extraction performance was evaluated. Except for flurbiprofen, exhaustive extraction was accomplished from plasma. Linearity was obtained for all six drugs in the range 0.025-10 μg/mL, with r (2) values ranging between 0.998 and 1.000. Precision data were in the range 3-22% RSD, and accuracy data were within 72-130% with spiked plasma samples. Based on the current experiences, PALME showed substantial potential for future high-throughput bioanalysis of non-polar acidic drugs.
新的样品制备概念“平行人工液膜萃取（PALME）”用于从人血浆样品中萃取酸性药物酮洛芬、非诺洛芬、双氯芬酸、氟比洛芬、布洛芬和吉非贝齐。将血浆样品（250μL）加入96孔供体板的各个孔中，并用盐酸稀释以使酸性药物质子化。酸性药物以质子化形式从各个血浆样品中，通过各自包含2μL二己醚的相应人工液膜，萃取到各自包含50μL 25 mM氨溶液（pH 10）的相应接受溶液中。液膜和接受溶液位于96孔滤板中，在萃取过程中该滤板与96孔供体板夹在一起。对多个样品进行15至60分钟的平行萃取，随后对各个接受溶液进行高效液相色谱 - 紫外检测。对包括液膜化学成分、萃取时间和样品pH在内的重要PALME参数进行了优化，并评估了萃取性能。除氟比洛芬外，从血浆中实现了完全萃取。所有六种药物在0.025 - 10μg/mL范围内均获得线性，r²值在0.998至1.000之间。加标血浆样品的精密度数据在3 - 22% RSD范围内，准确度数据在72 - 130%以内。基于目前的经验，PALME在未来非极性酸性药物的高通量生物分析中显示出巨大潜力。
