Dynamic calibration and dissolved gas analysis using membrane inlet mass spectrometry for the quantification of cell respiration.
A membrane inlet mass spectrometer connected to a miniaturized reactor was applied for dynamic dissolved gas analysis. Cell samples were taken from 7 mL shake flask cultures of Corynebacterium glutamicum ATCC 13032, and transferred to the 12 mL miniaturized reactor. There, oxygen uptake and carbon dioxide and its mass isotopomer production rates were determined using a new experimental procedure and applying nonlinear model equations. A novel dynamic method for the calibration of the membrane inlet mass spectrometer using first-order dynamics was developed. To derive total dissolved concentration of all carbon dioxide species (C(T)) from dissolved carbon dioxide concentration ([CO(2)](aq)), the ratio of C(T) to [CO(2)](aq) was determined by nonlinear parameter estimation, whereas the mass transfer coefficient of CO(2) was determined by the Wilke-Chang correlation. Subsequently, the suitability of the model equations for respiration measurements was examined using residual analysis and the Jarque-Bera hypothesis test. The resulting residuals were found to be random with normal distribution, which proved the adequacy of the application of the model for cell respiration analysis. Hence, dynamic changes in respiration activities could be accurately analyzed using membrane inlet mass spectrometry with the novel calibration method.
Yang
Tae Hoon
TH
Technische Biochemie, Saarland University, Im Stadtwald, Bldg. 2, D-66123 Saarbruecken, Germany.
Wittmann
Christoph
C
Heinzle
Elmar
E
D000465:Algorithms; D019149:Bioreactors / Q000382:microbiology; D002138:Calibration / Q000592:standards; D002245:Carbon Dioxide / Q000032:analysis*; D019069:Cell Respiration / Q000502:physiology*; D003198:Computer Simulation; D003352:Corynebacterium / Q000737:chemistry* / Q000502:physiology*; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods / Q000592:standards*; D008567:Membranes, Artificial; D008954:Models, Biological; D017711:Nonlinear Dynamics; D010100:Oxygen / Q000032:analysis* / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008567:Membranes, Artificial; D002245:Carbon Dioxide; D010100:Oxygen