Direct Electromembrane Extraction‐Based Mass Spectrometry: A Tool for Studying Drug Metabolism Properties of Liver Organoids

Abstract

This work introduces a strategy for organoid analysis -direct Electromembrane Extraction based Mass Spectrometry (dEME-MS) -for coupling liver organoids with mass spectrometry (MS). dEME-MS comprises electrophoresis of selected small molecules from a culture chamber across an oil membrane, and to a MS compatible solution. This enables clean micro-extraction of drugs and their metabolites as produced in the liver organoids to capillary liquid chromatography-mass spectrometry. Applying dEME-MS, proof-of-concept of directly measuring methadone metabolism is demonstrated on adult liver organoids. With 50 liver organoids and 1 μM methadone, methadone metabolism was monitored from 0 to 24 hours (11 time points). All analytes had < 0.4 % variance in retention times with > 100 measurements. dEME-MS is capable of automated and selective monitoring of drug metabolism in liver organoids, and could serve as a valuable tool for automated drug discovery efforts.

On‐line reduction of insulin disulfide bonds with photoinduced radical reactions, upstream to nano liquid chromatography‐mass spectrometry

“…These biomaterials/microphysiological systems, which may be grown from, for example, induced pluripotent stem (iPS) cells or patient cells, are emerging as powerful tools for, for example, drug discovery and development biology, serving as alternatives to animal models and conventional cell cultures [2]. Recently, we have studied drug metabolism traits of liver organoids, using 96‐well electromembrane extraction (EME) [3] and on‐chip EME [4]. In the latter study, we coupled the on‐chip format directly with mass spectrometry, allowing a high degree of automation and specificity, leading us to conclude that organoids/chip systems and mass spectrometry is a well‐suited pairing.…”

Determination of insulin secretion from stem cell-derived islet organoids with liquid chromatography-tandem mass spectrometry

“…Our research group focuses on applying liquid chromatography-mass spectrometry (LC-MS) to study the traits of various organoids [11][12][13][14], and we here describe the first MS-based method for highly selective monitoring of insulin excreted from islet organoids. Main ingredients of the method were:…”

“Organ-in-a-column” coupled on-line with liquid chromatography-mass spectrometry

“…However, EME requires an electrical current driven transfer of metabolites through an oil membrane into an MS-compatible solution which can potentially limit the spectrum of analytes that can be analyzed [14]. Additionally, a key challenge for coupling chips with MS is ensuring practical and robust connections and standardization.…”

“…Verpoorte and co-workers have previously combined organs/organ models, chip microfluidics and separation science, for studying metabolism of liver slices [11] and pharmacology in a gut-on-chip [12]. We have recently coupled liver organoids with sample preparation techniques such as electromembrane extraction (EME) [13], which we also found to be compatible with on-line coupling of organoid-containing chips to LC-MS for studying organoid drug metabolism [14]. However, EME requires an electrical current driven transfer of metabolites through an oil membrane into an MS-compatible solution which can potentially limit the spectrum of analytes that can be analyzed [14].…”

“…We have recently coupled liver organoids with sample preparation techniques such as electromembrane extraction (EME) [13], which we also found to be compatible with on-line coupling of organoid-containing chips to LC-MS for studying organoid drug metabolism [14]. However, EME requires an electrical current driven transfer of metabolites through an oil membrane into an MS-compatible solution which can potentially limit the spectrum of analytes that can be analyzed [14]. Additionally, a key challenge for coupling chips with MS is ensuring practical and robust connections and standardization.…”