LC/MS Assay for Quantification of the Carbon Sources in Proliferating Cells. Applications in Anticancer Drug Discovery Research

Dimitri Kovalerchik, Chemistry, Metabomed Ltd., Yavne, Israel (dimitri.kovalerchik@metabomed.com)

Proliferating cells (including tumor cells) need to source carbon for their growth. The most common source of carbon in these cells are glucose and glutamine.  However, sometimes cells can use other sugars, amino acids, as well as fatty acids, acetate and other nutrients as carbon source. Acetyl CoA, is the common biosynthetic intermediate of all aforementioned carbon sources, where the Acetyl group is the actual 2 carbon unit offered.

When cells are grown in media where one of these carbon sources is labeled with ¹³C, it is possible to quantify the utilization of this source by determining the ratio of labeled/non-labeled Acetyl CoA (for example use of ¹³C 6-glucose to determine glucose usage).

LC/MS quantification of Acetyl CoA isotopes directly is usually challenging, due to its poor ionization and low concentration in the cells, however it can be measured indirectly.  We show here two easy and robust LC/MS methods for carbon source quantification.  The first measures fatty acids isotopes using RP C18 chromatography and the second Acetyl CoA polar metabolites using HILIC chromatography. These two methods are used in the quantification of inhibition of various biosynthetic pathways by bioactive compounds in cells. 

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Short Biography of Presenting Author

Dr. Kovalerchik Joined Metabomed as Senior Scientist in May 2016, with more than 7 years of analytical chemistry research. His professional interests focus on cancer cell metabolomics using Orbitrap LCMS system and drug discovery. Dr. Kovalerchik earned a Ph.D. in organic and analytical chemistry in Tel Aviv University.