No abstract

In the context of this work we evaluated a multisensory, noninvasive prototype platform for shake flask cultivations by monitoring three basic parameters (pH, pO2 and biomass). The focus lies on the evaluation of the biomass sensor based on backward light scattering. The application spectrum was expanded to four new organisms in addition to E. coli K12 and S. cerevisiae [1]. It could be shown that the sensor is appropriate for a wide range of standard microorganisms, e.g., L. zeae, K. pastoris, A. niger and CHO-K1. The biomass sensor signal could successfully be correlated and calibrated with well-known measurement methods like OD600, cell dry weight (CDW) and cell concentration. Logarithmic and Bleasdale-Nelder derived functions were adequate for data fitting. Measurements at low cell concentrations proved to be critical in terms of a high signal to noise ratio, but the integration of a custom made light shade in the shake flask improved these measurements significantly. This sensor based measurement method has a high potential to initiate a new generation of online bioprocess monitoring. Metabolic studies will particularly benefit from the multisensory data acquisition. The sensor is already used in labscale experiments for shake flask cultivations.

A novel online sensor system for noninvasive and continuous monitoring of cell growth in shake flasks is described. The measurement principle is based on turbidity measurement by detecting 180°‐scattered light and correlation to OD by nonlinear calibration models. The sensor system was integrated into a commercial shaking tablar to read out turbidity from below the shake flasks bottom. The system was evaluated with two model microorganisms, Escherichia coli K12 as prokaryotic and Saccharomyces cerevisiae as eukaryotic model. The sensor allowed an accurate monitoring of turbidity and correlation with OD600 ≤ 30. The determination of online OD showed relative errors of about 7.5% for E. coli K12 and 12% for S. cerevisiae. This matches the errors of the laborious offline OD and thus facilitates to overcome the drawbacks of the classical method as risk of contamination and decreasing volumes through sampling. One major challenge was to ensure a defined, nonvarying measurement zone as the rotating suspension in the shake flask forms a liquid sickle which circulates round the flasks inner bottom wall. The resulting alteration of liquid height above the sensor could be compensated by integration of an acceleration sensor into the tablar to synchronize the sensor triggering.

20Online pH control during microbial shake flask cultivation has not been established due to the 21 lack of a practical combination of an online sensor system and an appropriate control unit. The 22 objective of this investigation was to develop a minimum scale dosage apparatus, namely shake 23 hich can control the pH during a complete cultivation and serves as 24 technical example for the application of small liquid dispensing lab devices. A well evaluated 25 optical, chemosensor based, noninvasive, multisensory platform prototype for online DO 26 (dissolved oxygen)-, pH-and biomass measurement served as sensor. The SFC was designed as 27 cap-integrated, semi-autarkical control unit. Minimum scale working parts like the commercial 28 mp6 piezoelectric micropumps and miniature solenoid valves were combined with a selective 29 laser sintering (SLS) printed backbone. In general it is intended to extend its application range 30 on the control of enzymatic assays, polymerization processes, cell disruption methods or the 31 precise dispense of special chemicals like inducers or inhibitors. It could be proved that pH 32 control within a range of 0.1 pH units could be maintained at different cultivation conditions. A 33 proportional-integral-derivative-(PID) controller and an adaptive proportional controller were 34 successfully applied to calculate the balancing solution volume. SLS based 3D printing using 35 Page 2 of 31 A c c e p t e d M a n u s c r i p t 2 polyamide combined with state-of-the-art micro pumps proved to be perfectly adaptable for 1 minimum size, autoclavable lab devices.2

Several studies indicate that the terpene trilactones (TTL) of EGb 761® are responsible for most of its pharmacological action in the brain . Therefore, we investigated the ability of the TTL to cross the blood brain barrier in rats after a single oral administration (600 mg/kg) of EGb 761® and compared it with the plasma levels. In addition, we checked the pharmacokinetic characteristics of an application of EGb 761® against a similar amount of pure substances. For this purpose, we developed a sensitive HPLC-(APCI)-MS method for the determination of the Ginkgo biloba TTL (ginkgolide A [GA], B [GB], C [GC] and bilobalide [Bb]) in plasma as well as in brain tissue. The following animal study shows that the oral application of 600 mg/kg EGb 761® results in significant GA, GB, and Bb concentrations in plasma as well as in the CNS of the rodents, while the GC concentration was below the detection limit of the analytical method in both matrices. GA, GB, and Bb brain concentrations showed a rapid increase up to 55 ng/g, 40 ng/g, and 98 ng/g with no difference of the characteristic after extract or pure substance application. Regarding the plasma levels, significant higher C(max) and AUC values were detected after application of the extract EGb 761®. These results allow for the first time a discussion of pharmacological effects with the knowledge of the pharmacokinetic behavior of the TTL in target tissues.