Development of a fast, online three-phase electroextraction hyphenated to fast liquid chromatography-mass spectrometry for analysis of trace-level acid pharmaceuticals in plasma

Yupeng He; Nicolas Drouin; Bert Wouters; Paul Miggiels; Thomas Hankemeier; Peter W Lindenburg

doi:10.1016/j.aca.2021.339364

Development of a fast, online three-phase electroextraction hyphenated to fast liquid chromatography-mass spectrometry for analysis of trace-level acid pharmaceuticals in plasma

Anal Chim Acta. 2022 Feb 1:1192:339364. doi: 10.1016/j.aca.2021.339364. Epub 2021 Dec 11.

Authors

Yupeng He¹, Nicolas Drouin¹, Bert Wouters¹, Paul Miggiels¹, Thomas Hankemeier², Peter W Lindenburg³

Affiliations

¹ Analytical Biosciences and Metabolomics, Systems Biomedicine and Pharmacology, Leiden Academic Centre for Drug Research, Leiden University, the Netherlands.
² Analytical Biosciences and Metabolomics, Systems Biomedicine and Pharmacology, Leiden Academic Centre for Drug Research, Leiden University, the Netherlands. Electronic address: hankemeier@lacdr.leidenuniv.nl.
³ Analytical Biosciences and Metabolomics, Systems Biomedicine and Pharmacology, Leiden Academic Centre for Drug Research, Leiden University, the Netherlands; Research Group Metabolomics, Leiden Center for Applied Bioscience, University of Applied Sciences Leiden, the Netherlands.

PMID: 35057963
DOI: 10.1016/j.aca.2021.339364

Abstract

Sample preparation is a challenge for high-throughput analysis, especially for volume-limited samples with low-abundant analytes. Ideally, sample preparation enriches the analytes of interest while removing the interferents to reduce the matrix effect and improve both sensitivity and quantification. In this study, a three-phase electroextraction (EE) method hyphenated to fast online liquid chromatography-mass spectrometry (LC-MS) was developed. Four model acidic drugs of relevance for drug monitoring in plasma, i.e. naproxen, fenoprofen, flurbiprofen, and ibuprofen, were utilized for the optimization and evaluation of the method. A Design of Experiment approach (Box-Behnken design) was used to optimize the critical parameters of the method, i.e., the type of organic solvent, pH of the sample and acceptor phase, and the extraction voltage and time. Good fitness (P < 0.02, R² > 0.95) was observed for the developed quadratic model. Extraction could be achieved in less than 2 min (115 s) with enrichment factors (EF) up to 190 and extraction recoveries (ER) up to 38% for academic samples. Additionally, the optimized three-phase EE method was successfully applied to spiked plasma samples with low-abundant (50 ng mL^-1) analytes and a low sample volume of 15 μL plasma in 10-fold diluted samples. Finally, two crucial contributors to the matrix effect of three-phase EE application on plasma samples were determined. Specifically, the ion-suppression effect in the MS source was reduced by the fast LC separation, and the matrix effect during extraction was negligible for the diluted protein-precipitated plasma samples. The developed three-phase EE method is easy to operate and provides fast and online extraction of trace-level acidic analytes from volume-limited biological samples. Therefore, this method can provide a potential solution for sample-preparation bottlenecks in high-throughput bioanalysis workflows.

Keywords: Bioanalysis; Electroextraction; Human plasma; Online fast LC-MS; Sample preparation.

MeSH terms

Acids*
Chromatography, Liquid
Mass Spectrometry
Pharmaceutical Preparations*
Proteins

Substances

Acids
Pharmaceutical Preparations
Proteins