By a News Reporter-Staff News Editor at Life Science Weekly -- Current study results on Gram-Positive Bacteria have been published. According to news originating from Lyngby, Denmark, by NewsRx correspondents, research stated, "Analysis of intracellular metabolites in bacteria is of utmost importance for systems biology and at the same time analytically challenging due to the large difference in concentrations, multiple negative charges, and high polarity of these compounds. To challenge this, a method based on dispersive solid phase extraction with charcoal and subsequent analysis with ion-pair liquid chromatography coupled with electrospray ionization tandem mass spectrometry was established for quantification of intracellular pools of the 28 most important nucleotides."
Our news journalists obtained a quote from the research from the Technical University of Denmark, "The method can handle extracts where cells leak during the quenching. Using a Phenyl-Hexyl column and tributylamine as volatile ion-pair reagent, sufficient retention and separation was achieved for mono-, di-, and triphosphorylated nucleotides. Stable isotope labeled nucleotides were used as internal standards for some analytes. The method was validated by determination of the recovery, matrix effects, accuracy, linearity, and limit of detection based on spiking of medium blank as well as standard addition to quenched Lactococcus lactis samples. For standard addition experiments, the isotope-labeled standards needed to be added in similar or higher concentrations as the analytes."
According to the news editors, the research concluded: "L lactis samples had an energy charge of 0.97 +/- 0.001 which was consistent with literature, whereas some differences were observed compared with legacy data based on P-33 labeling."
For more information on this research see: Dispersive solid phase extraction combined with ion-pair ultra high-performance liquid chromatography tandem mass spectrometry for quantification of nucleotides in Lactococcus lactis. Analytical Biochemistry, 2013;440(2):166-177. Analytical Biochemistry can be contacted at: Academic Press Inc Elsevier Science, 525 B St, Ste 1900, San Diego, CA 92101-4495, USA. (Elsevier - www.elsevier.com; Analytical Biochemistry - www.elsevier.com/wps/product/cws_home/622781)
The news correspondents report that additional information may be obtained from O. Magdenoska, Technical University of Denmark, Dept. of Syst Biol, Fungal Physiol & Biotechnol Grp, DK-2800 Lyngby, Denmark. Additional authors for this research include J. Martinussen, J. Thykaer and K.F. Nielsen (see also Gram-Positive Bacteria).
Keywords for this news article include: Lyngby, Europe, Denmark, Streptococcaceae, Imaging Technology, Lactococcus lactis, Gram-Positive Bacteria, High-Performance Liquid Chromatography
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