Analytical Sciences

Untargeted LC-MS Metabolomics for the Analysis of Micro-scaled Extracellular Metabolites from Hepatocytes

Rodi ABDALKADER,*1 Romanas CHALECKIS,*2,*3 Isabel MEISTER,*2,*3 Pei ZHANG,*2,*3 Craig E. WHEELOCK,*2,*3 and Ken-ichiro KAMEI*1,*4

*1 Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-Ushinomiya-cho, Sakyo, Kyoto 606-8501, Japan
*2 Gunma University Initiative for Advanced Research (GIAR), Gunma University, Gunma 371-8511, Japan
*3 Division of Physiological Chemistry II, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
*4 Wuya College of Innovation, Shenyang Pharmaceutical University, Liaoning 110016, China

Metabolome analysis in micro physiological models is a challenge due to the low volume of the cell culture medium (CCM). Here, we report a LC-MS-based untargeted metabolomics protocol for the detection of hepatocyte extracellular metabolites from micro-scale samples of CCM. Using a single LC-MS method we have detected 57 metabolites of which 27 showed >2-fold shifts after 72-hour incubation. We demonstrate that micro-scale CCM samples can be used for modelling micro-physiological temporal dynamics in metabolite intensities.

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