کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1177423 1491413 2007 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Determination of carbon labeling distribution of intracellular metabolites from single fragment ions by ion chromatography tandem mass spectrometry
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Determination of carbon labeling distribution of intracellular metabolites from single fragment ions by ion chromatography tandem mass spectrometry
چکیده انگلیسی

Liquid chromatography tandem mass spectrometry coupling is a highly sensitive and specific technique allowing molecule detection in the femtomolar range. This article introduces a straightforward approach to apply this technique in 13C metabolic flux analysis. Based on a theoretical analysis of the correlation between molecule ions and corresponding fragments, a method was developed to determine the carbon labeling of intracellular metabolites without increasing the number of measurements per metabolite compared with direct molecule ion analysis. The method was applied to phosphorylated metabolites because their fragmentation results in high yields of [PO3]− and/or [H2PO4]− ions. Comparing the accuracy of the carbon labeling determination of phosphorylated metabolites between direct analysis of the molecule ions with that of corresponding phosphate fragment ions, it could be demonstrated that the introduced approach resulted in significantly higher accuracy and sensitivity for all tested metabolites. When applying the techniques to Escherichia coli cell extracts, 2 μg cell dry weight per injection was sufficient to determine the natural abundances of the carbon fractions m and m + 1 from six phosphorylated metabolites with high accuracy, predestining the approach for very small cultivation volumes in the microliter range.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 360, Issue 2, 15 January 2007, Pages 182–188
نویسندگان
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