Madridge Journal of Pharmaceutical Research

ISSN: 2638-1591

International Conference on Medicinal and Pharmaceutical Chemistry
December 5-7, 2016 | Dubai, UAE

Characterization of nucleotide pool in bacterial cells

Eva Zborníková1, 2*, Dominik Rejman1, Libor Krásný3, Vallo Varik4,5,6, Yuzuru Tozawa7 and Vasili Hauryliuk4,5,6

1Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences v.v.i., Czech Republic
2Charles University, Faculty of Science, Department of Analytical Chemistry, Czech Republic
4University of Tartu, Institute of Technology, Estonia
5Department of Molecular Biology, Umeå University, Sweden
6Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Sweden
7Graduate School of Science and Engineering, Saitama University, Japan

DOI: 10.18689/2638-1591.a1.004

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Various bacterial responses to environmental stimuli lead into changes in intracellular concentration of small molecules (nucleotides, nucleosides and their derivatives). Robust, sensitive and simple method for characterizationof thesechanges is therefore necessary. E.g. in the case of stringent response –a potential novel drug target - intracellular levels of alarmone nucleotides guanosine tetraphosphate (ppGpp) and pentaphosphate (pppGpp), need to be determined.

There are two key steps in the process of determining nucleotide levels in bacterial biomass (complex samples with strong matrice effect):1) extraction method with quantitative and reproducible yield, and 2) good analytical method capable todistinguish and quantify individual nucleotides.

To characterize nucleotide pools in bacteriawe have chosen LC-MS in HILIC mode of separation that exhibits many advantages over commonly used ion pair (IP) LC coupled with UV-VIS detector. Moreover, the ballast mass from bacteria are well separated from majority of analytes and do not disturb the analysis.

Biography:
After the period of pesticide analysis in certified laboratories, I switched to medicinal chemistry and continue working with liquid chromatography and mass spectrometry techniques. After several years, I have started PhD studies. Modern analytical chemistry is my hobby and could be often quite challenging.
2003-2008: Faculty of Food and Biochemical Technology, UCT, Prague
2008: National Veterinary Institute, Prague
2009-2010: Institute of Organic Chemistry and Biochemistry, CAS
2010-now: Institute of Organic Chemistry and Biochemistry, CAS; Charles University, Department of Analytical Chemistry, Prague; Ph.D.study

Acknowledgments
This work was supported by the Czech Science Foundation grant number 15-11711S (DR) and internal IOCB Pragueproject 9330/33.

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