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Potential errors in conventional DOT measurement techniques in shake flasks and verification using a rotating

Sven Hansen1, Frank Kensy, Andreas Käser

  • 1AVT, Biochemical Engineering, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany.

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Conventional dissolved oxygen tension (DOT) measurements in shake flasks using Clark-type electrodes or fixed optical sensor spots are unreliable. A new rotating flexitube optical sensor accurately verifies these measurement errors.

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Area of Science:

  • Biotechnology
  • Bioprocess Engineering
  • Analytical Chemistry

Background:

  • Dissolved oxygen tension (DOT) is critical for bioprocess monitoring.
  • Conventional DOT measurement methods in shake flasks, such as Clark-type electrodes and fixed optical sensor spots, introduce systemic errors.
  • These errors stem from altered hydrodynamics and headspace gas interference, particularly under low fill volumes and high oxygen demand.

Purpose of the Study:

  • To verify the systemic errors associated with Clark-type electrodes for DOT measurement in shake flasks.
  • To evaluate the accuracy of conventional optical DOT measurements using fixed sensor spots.
  • To introduce and validate a novel "flexitube optical sensor" for reliable DOT assessment.

Main Methods:

  • Comparative analysis of DOT measurements using Clark-type electrodes versus systems without electrodes.
  • Investigation of fixed optical sensor spot performance under oxygen-limited conditions.
  • Development and application of a novel rotating flexitube optical sensor.

Main Results:

  • Clark-type electrodes significantly altered shake flask hydrodynamics, increasing oxygen transfer capacity.
  • Fixed optical sensor spots yielded erroneous high DOT readings under oxygen-limited conditions due to headspace oxygen influence.
  • The flexitube optical sensor demonstrated utility in verifying these conventional measurement inaccuracies.

Conclusions:

  • Clark-type electrodes are unsuitable for accurate DOT measurement in shake flasks.
  • Fixed optical sensor spots are prone to errors caused by headspace oxygen.
  • The novel rotating flexitube optical sensor provides a reliable method for validating conventional optical DOT measurements.