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Measurement of methylated metabolites using liquid chromatography-mass spectrometry and its biological application

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Abstract

Methylation aberrations play an important role in many metabolic disorders including cancer. Methylated metabolites are direct indicators of metabolic aberrations, and currently, there is no liquid chromatography-mass spectrometry (LC-MS) based method available to cover all classes of methylated metabolites at low detection limits. In this study, we have developed a method for the detection of methylated metabolites, and its biological application. In this approach, we used a HILIC based HPLC method combined with MS to measure methylated organic acids, amino acids, and nucleotides. These metabolites were separated from each other by their hydrophobic interactions and analyzed using a targeted metabolomics approach by single reaction monitoring in positive and negative modes of electrospray ionization. These metabolites were quantified, and the interday reproducibility was <10% relative standard deviation. Furthermore, by applying this method, we identified high levels of methylated metabolites in bladder cancer cell lines compared to benign cells. In vitro treatment of cancer cells with a methylation inhibitor, 5-aza-2′-deoxycytidine, showed a decrease in these methylated metabolites. These data indicate that HPLC analysis using this HILIC based method could be a powerful tool for measuring methylated metabolites in biological specimens. This method is rapid, sensitive, selective, and precise to measure methylated metabolites.

Graphical abstract: Measurement of methylated metabolites using liquid chromatography-mass spectrometry and its biological application

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Publication details

The article was received on 03 Oct 2018, accepted on 14 Nov 2018 and first published on 15 Nov 2018


Article type: Paper
DOI: 10.1039/C8AY02168F
Citation: Anal. Methods, 2019, Advance Article
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    Measurement of methylated metabolites using liquid chromatography-mass spectrometry and its biological application

    C. R. Ambati, V. Vantaku, S. R. Donepudi, C. S. Amara, S. S. Ravi, A. Mandalapu, M. Perla, V. Putluri, A. Sreekumar and N. Putluri, Anal. Methods, 2019, Advance Article , DOI: 10.1039/C8AY02168F

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