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Reverse isotope dilution analysis of 13 CO 2 using gas chromatography/isotope ratio mass spectrometry: Application to the quantitative determination of 13 CO 2 released by human polymorphonuclear leukocytes
Author(s) -
Lamrini Rachid,
Lacan Philippe,
Francina Alain,
Guilluy Roger,
Brazier Jean Louis
Publication year - 1995
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1290091109
Subject(s) - chemistry , isotope dilution , isotope ratio mass spectrometry , mass spectrometry , isotope , carbon dioxide , isotopes of carbon , carbon 13 , chromatography , analytical chemistry (journal) , stable isotope ratio , gas chromatography , oxygen 18 , deuterium , hydrogen , isotope analysis , radiochemistry , environmental chemistry , nuclear chemistry , isotopes of oxygen , organic chemistry , total organic carbon , physics , quantum mechanics , ecology , biology
We propose a new method for the quantitative determination of carbon dioxide released by a biological microgenerator: a suspension of human polymorphonuclear leukocytes (PMNL). This method is based on the reverse isotope dilution analysis by gas chromatography/isotope ratio mass spectrometry of 13 CO 2 released by PMNL in a controlled isotope abundance atmosphere containing 3% CO 2 . 13 CO 2 release is effective after PMNL stimulation in the presence of [ 13 C]glucose, labeled on positions 1, 2 or 6. The validation of this method is carried out by the measurement of the isotope ratio 13 CO 2 / 12 CO 2 using known amounts of [ 13 C]sodium hydrogen carbonate and the comparison with theoretical isotope abundances derived from various CO 2 equilibria. Complete release of CO 2 is achieved by the acidification of the medium. This method requires only few cells, displays high sensitivity and specificity and can be applied to the analysis of large series of samples using an automatic sample injector. In addition, this method can also be applied to other types of biological microgenerators of carbon dioxide.