Thomas Beuermann

On-line carbon balance of yeast fermentations using miniaturized optical sensors.

Monitoring of microbiological processes using optical sensors and spectrometers has gained in importance over the past few years due to its advantage in enabling non-invasive on-line analysis. Near-infrared (NIR) and mid-infrared (MIR) spectrometer set-ups in combination with multivariate calibrations have already been successfully employed for the simultaneous determination of different metabolites in microbiological processes. Photometric sensors, in addition to their low price compared to spectrometer set-ups, have the advantage of being compact and are easy to calibrate and operate. In this work, the detection of ethanol and CO(2) in the exhaust gas during aerobic yeast fermentation was performed by two photometric gas analyzers, and dry yeast biomass was monitored using a fiber optic backscatter set-up. The optical sensors could be easily fitted to the bioreactor and exhibited high robustness during measuring. The ethanol content of the fermentation broth was monitored on-line by measuring the ethanol concentration in the fermentation exhaust and applying a conversion factor. The vapor/liquid equilibrium and the associated conversion factor strongly depend on the process parameter temperature but not on aeration and stirring rate. Dry yeast biomass was determined in-line by a backscattering signal applying a linear calibration. An on-line balance with a recovery rate of 95-97% for carbon was achieved with the use of three optical sensors (two infrared gas analyzers and one fiber optic backscatter set-up).

MIR-ATR sensor for process monitoring

A mid-infrared attenuated total reflectance (MIR-ATR) sensor has been developed for chemical reaction monitoring. The optical setup of the compact and low-priced sensor consists of an IR emitter as light source, a zinc selenide (ZnSe) ATR prism as boundary to the process, and four thermopile detectors, each equipped with an optical bandpass filter. The practical applicability was tested during esterification of ethanol and formic acid to ethyl formate and water as a model reaction with subsequent distillation. For reference analysis, a Fourier transform mid-infrared (FT-MIR) spectrometer with diamond ATR module was applied. On-line measurements using the MIR-ATR sensor and the FT-MIR spectrometer were performed in a bypass loop. The sensor was calibrated by multiple linear regression in order to link the measured absorbance in the four optical channels to the analyte concentrations. The analytical potential of the MIR-ATR sensor was demonstrated by simultaneous real-time monitoring of all four chemical substances involved in the esterification and distillation process. The temporal courses of the sensor signals are in accordance with the concentration values achieved by the commercial FT-MIR spectrometer. The standard error of prediction for ethanol, formic acid, ethyl formate, and water were 0.38 mol L − 1, 0.48 mol L − 1, 0.38 mol L − 1, and 1.12 mol L − 1, respectively. A procedure based on MIR spectra is presented to simulate the response characteristics of the sensor if the transmission ranges of the filters are varied. Using this tool analyte specific bandpass filters for a particular chemical reaction can be identified. By exchanging the optical filters, the sensor can be adapted to a wide range of processes in the chemical, pharmaceutical, and beverage industries.

Nicht-invasive Prozesssonde zur Inline-Ramananalyse durch optische Schaugläser

Zusammenfassung Trotz der bekannten Vorteile der Raman-Spektroskopie, wie bspw. eine höhere chemische Selektivität gegenüber Messmethoden im nahen Infrarot (NIR) oder die im Vergleich zum mittleren Infrarotbereich (MIR) niedrigen Matrixeinflüsse des Wassermoleküls, ist diese optische Messtechnik in der Online-Prozessanalysentechnik nicht weit verbreitet. Ein wesentliches Problem besteht in einem oftmals kostenintensiven Nachrüsten einer Messstelle durch den Einbau sogenannter Immersionssonden in eine produktführende Rohrleitung oder einen Behälter. Eine praktikable Alternative stellt das hier entwickelte neuartige Sondensystem dar, welches eine Strahlführung über Linsen mit relativ großen Durchmessern beinhaltet, da dieses an vorhandene Schauglasarmaturen angekoppelt werden kann. Mit diesem robusten Sondenaufbau sind Brennweiten weit über 25 mm möglich, welche Echtzeit-Messungen von außerhalb der produktführenden Leitungen durch optische Schaugläser gestatten. Die dadurch entstehenden Messoptionen werden exemplarisch am Nachweis von Ethanol durch Schaugläser unterschiedlicher Dicken sowie bei einer quantitativen Echtzeit-Verfolgung eines Propylencarbonat-Wasser-Gemisches durch eine Schauglasarmatur (Nenndruck PN 16, Nennweite DN 50) im Technikumsmaßstab untersucht. Die vorgestellte Raman-Sonde hat durch einfache Adaption an bereits vorhandene Armaturen industrieller Anlagen das Potential einer preiswerten und kontaktlosen Inline-Messlösung mit hoher Standzeit in der Prozessanalysentechnik (PAT).

Non-contact Raman spectroscopy for in-line monitoring of glucose and ethanol during yeast fermentations

The monitoring of microbiological processes using Raman spectroscopy has gained in importance over the past few years. Commercial Raman spectroscopic equipment consists of a laser, spectrometer, and fiberoptic immersion probe in direct contact with the fermentation medium. To avoid possible sterilization problems and biofilm formation on the probe tip, a large-aperture Raman probe was developed. The design of the probe enables non-contact in-line measurements through glass vessels or inspection glasses of bioreactors and chemical reactors. The practical applicability of the probe was tested during yeast fermentations by monitoring the consumption of substrate glucose and the formation of ethanol as the product. Multiple linear regression models were applied to evaluate the Raman spectra. Reference values were determined by high-performance liquid chromatography. The relative errors of prediction for glucose and ethanol were 5 and 3%, respectively. The presented Raman probe allows simple adaption to a wide range of processes in the chemical, pharmaceutical, and biotechnological industries.