Mettler Toledo

The development and existence of bioburden in pharmaceutical water systems is often misunderstood. Microorganisms are always present and have a keen ability to adapt to their environment. This is especially true in a Water-for-Injection (WFI) system, where microbial attachment and biofilm growth will occur regardless of flow rate, material of construction, turbulent flow and low nutrient conditions. While industry makes every effort to control and eliminate bioburden, traditional sanitization methods are not one-hundred-percent effective at accomplishing this objective. Additionally, because of the limitations and time to result delay of conventional plate counting, we may be at a disadvantage for assessing bioburden, causing us to use water at risk. This poster explores real-life examples of biofilm in pharmaceutical water systems, risk mitigation strategies, and how real-time microbial detection could be used as a tool for improved risk management and process control.

Bacterial endospores represent a significant challenge to the pharmaceutical industry due to their presence in the environment, resistance to many commonly used inactivation procedures, and difficulty in culturing using traditional plating methods. This may result in the inadvertent release of contaminated products that may present health concerns for the patient. As a result, the use of a bio-fluorescent particle counter (BFPC) may prove advantageous for the detection of both water-borne and air-borne spores as their detection is not dependent on traditional culturing methods. In this study, we investigate the ability of an online water bioburden analyzer (OWBA), a specific class of BFPC, to detect Bacillus subtilis spores in pharmaceutical-grade water and present the results as auto-fluorescence units (AFU's) per B. subtilis spore. The spores were a commercial grade spore preparation and were previously quantified per manufacture recommendations. The results show that the OWBA can detect B. subtilis spores with an accuracy of 1.25 AFU per spore. The limit of detection was determined to be 1 spore/mL with a linearity greater than 0.9025 up to a concentration of 100 spores/mL. This data shows that OWBA's are a rapid and effective tool for the detection of bacterial endospores in pharmaceutical waters.

This study investigates the mechanisms underlying the elastocaloric (eC) effect in thermoplastic polyurethaneurea elastomers, focusing on the contributions of amorphous chain orientation and strain-induced crystallization (SIC) of the soft segments. Two types of TPU systems-strain induced crystallizable (PTMO-based polyurethaneurea (TPUU)) and non-crystallizable (PEO-based polyurea (TPU))-were synthesized with different amounts of HMDI hard segments (20 or 30 wt%) and subjected to mechanical loading under in situ wide-angle X-ray scattering (WAXS) and infrared thermography. The amorphous chain orientation is found to evolve much faster with strain in the TPUUs and SIC is found to initiate in these materials above a critical value of the Hermann's orientation factor of 0.7-0.75. SIC in these systems is found to enhance the eC effect via latent heat, but amorphous orientation is also found to significantly contribute through thermoelastic entropy changes, as attested by high heat sources of 15 MW m-3 for crystallizable TPUU before SIC (at 100 % of deformation) as compared to 5 MW m-3 for non-crystallizable TPUs. Under continuous cycles, TPUU samples demonstrate stronger eC responses and greater reversibility than TPU, resulting in higher coefficients of performance (Best COP = 13 for the TPUU20), particularly after cyclic pre-conditioning. This interpretation of molecular orientation and crystallization in relation with eC properties opens new design strategies for polymer-based solid-state cooling, where microstructural tailoring-beyond relying solely on crystallization-can optimize elastocaloric performance in practical applications.

Thermochemical energy storage has a very high potential to significantly decrease greenhouse gases using seasonal heat storage for heating applications. The conversion behavior of the thermochemical energy-carrier system CuSO4.5H2O was investigated relevant to continuously operated 3-phase suspension reactor conditions. The temperature in the reactor was measured continuously, and samples were taken periodically and analyzed with XRD, which revealed conversions of up to 95 % from CuSO4.5H2O to CuSO4.H2O. Investigating single particles in oil under the microscope while heating them up showed that the dehydration reaction is strongly dependent on the heating rate. Reaction temperatures from CuSO4.5H2O to CuSO4.3H2O started already at 87 degrees C and from CuSO4.3H2O to CuSO4 center dot H2O at 118 degrees C. In-situ XRD shows the mechanism of the reaction from CuSO4.5H2O to CuSO4 center dot H2O, where the intermediate phase CuSO4.3H2O is formed, starting at 30 degrees C. Furthermore, in-line IR spectroscopy, which allows instantaneous live tracking of the conversion, not only confirms the results obtained from in-situ XRD but also proves useful as a tool for process monitoring and control. The CuSO4 particles have been analyzed with SEM, showing the change of structure and shape of the used CuSO4 particles in contrast to the original CuSO4 particles, where the used CuSO4 particles break into smaller particles but form small agglomerates of an average of 3.5 mm in diameter. The specific surface area measured by BET shows that the specific surface area of the used CuSO4 particles was reduced by a factor of 3.6, but no loss in performance was observed.

Automation of metabolite control in fermenters is fundamental to develop vaccine manufacturing processes more quickly and robustly. We created an end-to-end process analytical technology and quality by design-focused process by replacing manual control of metabolites during the development of fed-batch bioprocesses with a system that is highly adaptable and automation-enabled. Mid-infrared spectroscopy with an attenuated total reflectance probe in-line, and simple linear regression using the Beer-Lambert Law, were developed to quantitate key metabolites (glucose and glutamate) from spectral data that measured complex media during fermentation. This data was digitally connected to a process information management system, to enable continuous control of feed pumps with proportional-integral-derivative controllers that maintained nutrient levels throughout fed-batch stirred-tank fermenter processes. Continuous metabolite data from mid-infrared spectra of cultures in stirred-tank reactors enabled feedback loops and control of the feed pumps in pharmaceutical development laboratories. This improved process control of nutrient levels by 20-fold and the drug substance yield by an order of magnitude. Furthermore, the method is adaptable to other systems and enables soft sensing, such as the consumption rate of metabolites. The ability to develop quantitative metabolite templates quickly and simply for changing bioprocesses was instrumental for project acceleration and heightened process control and automation. ONE-SENTENCE SUMMARY:Intelligent digital control systems using continuous in-line metabolite data enabled end-to-end automation of fed-batch processes in stirred-tank reactors.