Simultaneous determination of ammonia nitrogen and L-glutamine in bioreactor media using flow injection

A novel split stream flow injection (FI) system suitable for the simultaneous determination of L-glutamine and ammonia nitrogen (ammonia-N) in cell culture media is described. Potentiometric detection of ammonia-N in one portion of the manifold is achieved using a commercial ammonia gas-sensing elec...

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Bibliographic Details
Published inAnalyst (London) Vol. 118; no. 11; pp. 1361 - 1365
Main Authors PALSSON, B. O, SHEN, B. Q, MEYERHOFF, M. E, TROJANOWICZ, M
Format Journal Article
LanguageEnglish
Published Cambridge Royal Society of Chemistry 1993
Subjects
Ammonia - analysis
Biological and medical sciences
Biosensing Techniques
Biosensors
Biotechnology
Culture Media
Electrodes
Flow Injection Analysis - methods
Fundamental and applied biological sciences. Psychology
Glutamine - analysis
Methods. Procedures. Technologies
Nitrogen - analysis
Potentiometry
Various methods and equipments
Culture medium
Ammonia
Ammonium
Bioreactor
Biosensor
Method
Quantitative analysis
Glutamine
Flow injection
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Summary:A novel split stream flow injection (FI) system suitable for the simultaneous determination of L-glutamine and ammonia nitrogen (ammonia-N) in cell culture media is described. Potentiometric detection of ammonia-N in one portion of the manifold is achieved using a commercial ammonia gas-sensing electrode fitted with a wall-jet cap. L-Glutamine is quantified in the other part of the split sample by potentiometric detection of ammonium ions (by an ammonium-selective polymer membrane electrode), liberated from the hydrolysis of glutamine after the sample flows through a glass bead reactor containing immobilized glutaminase. Endogenous ammonia-N and potassium ions that would normally interfere with the glutamine measurement are removed upstream using a unique tubular cation-exchange unit. Using 50 microliters sample volumes and mixed solutions of ammonium chloride and L-glutamine in Iscove's Modified Dulbecco's Medium to calibrate the FI measuring system, values for ammonia-N and L-glutamine determined for 22 media samples obtained from a bioreactor growing retroviral producer cells correlate well with those measured with commercial, manual enzymic-spectrophotometric assay kits.
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ISSN:0003-2654
1364-5528
DOI:10.1039/an9931801361