Helena Hruskova

• Samkumpim, Thidarat; Dowlatshah, Samira; Alahmad, Waleed; Varanusupakul, Pakorn; Hruskova, Helena & Hansen, Frederik Andre [Show all 7 contributors for this article] (2025). 96-Well Agarose-Gel Electromembrane Extraction. Analytical Chemistry. ISSN 0003-2700. 98(1), p. 1090–1097. doi: 10.1021/acs.analchem.5c06783. Full text in Research Archive Show summary

  For the first time, gel electromembrane extraction was demonstrated in a 96-well system. Gel membranes of 3% w/v agarose and with a thickness of 3.5 mm were immobilized in hydrophilic polyvinylidene fluoride (PVDF) filters in a 96-well filter plate. The filters provided mechanical support for the gel membranes, and were important for the stability and robustness of the system. A selection of 90 basic pharmaceuticals in the polarity range −4.2 < log P < 8.1 was used as model analytes (compounds). The compounds were extracted from 200 μL sample (water or human plasma) adjusted to pH 4.0 with dilute formic acid, through the gel membrane, and into 200 μL of 100 mM formic acid as acceptor. The extraction potential was 25 V, and the extraction time was 20 min for careful operation to limit Joule heating and electroendosmosis. The majority of the compounds in the polarity range −4.0 < log P < 3.0 were extracted with high recovery (40–100%). Compounds with log P > 3.0 were discriminated due to interactions with the gel membrane. Proteins and phospholipids were not extracted, and the system therefore provided efficient cleanup from human plasma samples. The 96-well agarose-gel electromembrane extraction (EME) system showed great potential. Selectivity was controlled by interactions with the aqueous gel membrane. This is fundamentally very different from traditional EME with oil membranes, where selectivity is controlled by electro-assisted partition in and out of the oil membrane. 96-Well electromembrane extraction with gel membranes of agarose is favorable in terms of greenness and efficiency for polar analytes, as compared with systems based on oil membranes.

• Kogler, Stian; Skottvoll, Frøydis Sved; Hruskova, Helena; Rise, Frode; Aizenshtadt, Aleksandra & Krauss, Stefan Johannes Karl [Show all 9 contributors for this article] (2025). Electromembrane Extraction Provides Unprecedented Selectivity for Drugs in Cell Culture Media Used in Organoid and Organ-on-Chip Systems. Analytical Chemistry. ISSN 0003-2700. 97(9), p. 4923–4931. doi: 10.1021/acs.analchem.4c04994. Full text in Research Archive Show summary

  The use of organoids and organ-on-chip technologies as nonanimal methodologies in drug discovery and personalized medicine is rapidly expanding. However, the complexity and small volumes of organoid culture samples present significant analytical challenges, e.g., in drug analysis using liquid chromatography–mass spectrometry (LC–MS). Essentially an electrophoresis across an oil membrane, electromembrane extraction (EME) offers a promising approach for measuring drugs, as it is, for example, compatible with small samples such as organoid and organ-on-chip formats. Given the potential of the technology, there is a need to assess the extraction purity of EME extracts to ensure EME’s compatibility with high-throughput, downstream analysis. This study evaluates the effectiveness of EME for sample cleanup in various common cell culture media used for organoids and organs-on-chips. The media were spiked with 90 small-molecule drugs. Using gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis), high-resolution nuclear magnetic resonance spectroscopy, and LC–MS, we demonstrate that EME provides exhaustive removal of unwanted medium components (proteins, polar molecules, and apolar/neutral molecules) while selectively extracting the spiked small-molecule drugs. The approach was demonstrated with human stem-cell-derived liver organoids, allowing simple detection and monitoring of telltale cytochrome P450 metabolism. Taken together, our observations highlight an unprecedented ability of EME to provide sample cleanup for drug analysis in matrixes compatible with organoids and organ-on-chip technology.

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