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@ARTICLE{Reiter:891762,
      author       = {Reiter, Alexander and Herbst, Laura and Wiechert, Wolfgang
                      and Oldiges, Marco},
      title        = {{N}eed for speed: evaluation of dilute and shoot-mass
                      spectrometry for accelerated metabolic phenotyping in
                      bioprocess development},
      journal      = {Analytical and bioanalytical chemistry},
      volume       = {413},
      issn         = {0016-1152},
      address      = {Heidelberg},
      publisher    = {Springer},
      reportid     = {FZJ-2021-01722},
      pages        = {3253-3268},
      year         = {2021},
      abstract     = {With the utilization of small-scale and highly parallelized
                      cultivation platforms embedded in laboratory robotics,
                      microbial phenotyping and bioprocess development have been
                      substantially accelerated, thus generating a bottleneck in
                      bioanalytical bioprocess sample analytics. While microscale
                      cultivation platforms allow the monitoring of typical
                      process parameters, only limited information about product
                      and by-product formation is provided without comprehensive
                      analytics. The use of liquid chromatography mass
                      spectrometry can provide such a comprehensive and
                      quantitative insight, but is often limited by analysis
                      runtime and throughput. In this study, we developed and
                      evaluated six methods for amino acid quantification based on
                      two strong cation exchanger columns and a dilute and shoot
                      approach in hyphenation with either a triple-quadrupole or a
                      quadrupole time-of-flight mass spectrometer. Isotope
                      dilution mass spectrometry with 13C15N labeled amino acids
                      was used to correct for matrix effects. The versatility of
                      the methods for metabolite profiling studies of microbial
                      cultivation supernatants is confirmed by a detailed method
                      validation study. The methods using chromatography columns
                      showed a linear range of approx. 4 orders of magnitude,
                      sufficient response factors, and low quantification limits
                      (7–443 nM) for single analytes. Overall, relative standard
                      deviation was comparable for all analytes, with < $8\%$ and
                      < $11\%$ for unbuffered and buffered media, respectively.
                      The dilute and shoot methods with an analysis time of 1 min
                      provided similar performance but showed a factor of up to 35
                      times higher throughput. The performance and applicability
                      of the dilute and shoot method are demonstrated using a
                      library of Corynebacterium glutamicum strains producing
                      l-histidine, obtained from random mutagenesis, which were
                      cultivated in a microscale cultivation platform.},
      cin          = {IBG-1},
      ddc          = {540},
      cid          = {I:(DE-Juel1)IBG-1-20101118},
      pnm          = {2171 - Biological and environmental resources for
                      sustainable use (POF4-217)},
      pid          = {G:(DE-HGF)POF4-2171},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {33791825},
      UT           = {WOS:000635497000002},
      doi          = {10.1007/s00216-021-03261-3},
      url          = {https://juser.fz-juelich.de/record/891762},
}