How to extract both polar and lipophilic compounds from a single tumor specimen?


Szymon Macioszek, Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gdansk, Gdansk, Poland (macioszek@gumed.edu.pl)


Cancer tissue metabolomics is a powerful tool to acquire knowledge about molecular background and current state of the tumor. Along with better understanding of tumor pathophysiology, it is a way to find new therapeutic targets. However, tissue metabolomics has many challenges, starting from proper sampling through normalization process to reproducible extraction. Here, we tried to choose a proper method for sample preparation of gastrointestinal stromal tumor (GIST) samples. Most often GIST develops as a consequence of mutations in KIT gene; however, metabolic patterns during GIST development have not yet been investigated and elucidated.



To track changes in biochemical pathways in the cancer tissue, we chose to apply untargeted metabolomics with the use of LC-MS. The first stage of the study was focused on selecting the most appropriate sample preparation method. We aimed at developing a method that provides possibility of extracting both polar and lipophilic metabolites from one small GIST specimen. Tissue samples were homogenized with a 50:50 mixture (v/v) of methanol and waterin ratio 1:10 (w/v) and obtained homogenate was subjected to four different methods of extraction. They included monophasic extraction with methyl tert-butyl ether (MTBE), methanol and water, biphasic extraction with different ratios of MTBE, methanol and water as well as with dichloromethane, methanol and water. Organic extracts were analyzed with reversed phase LC-TOF-MS, while HILIC separation was used for polar extracts. Criteria for sample preparation method selection were based on the number of features detected in the extracts and reproducibility of extraction procedure.



The selected method will be further used in metabolomic fingerprinting of GIST tissue specimens, which can complement genomic studies on GIST or propose potential new therapeutic targets.



Abstract Reference & Short Personal Biography of Presenting Author


The presenting author, Szymon Macioszek graduated in Pharmaceutical Sciences at the Medical University of Gdansk in 2016. The same year, he started PhD studies under the supervision of Prof. Michal Markuszewski. His research interests focus on untargeted metabolomics with the use of liquid and gas chromatography coupled to mass spectrometry, as well as chemometric analysis of MS data.


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