title: Multi-analyte quantification in bioprocesses by Fourier-transform-infrared spectroscopy by partial least squares regression and multivariate curve resolution.
abstract: This paper presents the quantification of Penicillin V and phenoxyacetic acid, a precursor, inline during Pencillium chrysogenum fermentations by FTIR spectroscopy and partial least squares (PLS) regression and multivariate curve resolution - alternating least squares (MCR-ALS). First, the applicability of an attenuated total reflection FTIR fiber optic probe was assessed offline by measuring standards of the analytes of interest and investigating matrix effects of the fermentation broth. Then measurements were performed inline during four fed-batch fermentations with online HPLC for the determination of Penicillin V and phenoxyacetic acid as reference analysis. PLS and MCR-ALS models were built using these data and validated by comparison of single analyte spectra with the selectivity ratio of the PLS models and the extracted spectral traces of the MCR-ALS models, respectively. The achieved root mean square errors of cross-validation for the PLS regressions were 0.22 g L(-1) for Penicillin V and 0.32 g L(-1) for phenoxyacetic acid and the root mean square errors of prediction for MCR-ALS were 0.23 g L(-1) for Penicillin V and 0.15 g L(-1) for phenoxyacetic acid. A general work-flow for building and assessing chemometric regression models for the quantification of multiple analytes in bioprocesses by FTIR spectroscopy is given.
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164 AC, 1060 Vienna, Austria.
authors.forename: Cosima
authors.initials: C
authors.lastname: Koch
authors.affiliation: Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Research Area Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1a, 1060 Vienna, Austria.
authors.forename: Andreas E
authors.initials: AE
authors.lastname: Posch
authors.affiliation: Laboratorio de Desarrollo Analítico y Quimiometría-LADAQ, Universidad Nacional del Litoral-CONICET, Facultad de Bioquímica y Ciencias Biológicas, Ciudad Universitaria, 3000 Santa Fe, Argentina.
authors.forename: Héctor C
authors.initials: HC
authors.lastname: Goicoechea
authors.affiliation: Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses, Research Area Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Strasse 1a, 1060 Vienna, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Herwig
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164 AC, 1060 Vienna, Austria. Electronic address: bernhard.lendl@tuwien.ac.at.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Lendl
mesh_terms: D000085:Acetates / Q000032:analysis* / Q000592:standards; D002138:Calibration; D005285:Fermentation; D016018:Least-Squares Analysis; D010404:Penicillin V / Q000032:analysis* / Q000592:standards; D010408:Penicillium chrysogenum / Q000737:chemistry / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared / Q000592:standards
chemical_list: D000085:Acetates; C415326:phenoxyacetic acid; D010404:Penicillin V
keywords: Chemometrics; FTIR spectroscopy; Inline bioprocess monitoring; Multivariate curve resolution; P. chrysogenum; Partial least squares regression