Timea Frosch

Background: Effective antibiotic therapy in critically ill patients requires precise dosing tailored to individual conditions. However, physiological changes in these patients can complicate drug exposure prediction, leading to treatment failure or toxicity. Therapeutic drug monitoring (TDM) is crucial in optimizing antibiotic therapy, with Raman spectroscopy emerging as a promising method due to its speed and sensitivity. Methods: The utility of resonance Raman spectroscopy in analyzing clinical urine samples was investigated, specifically focusing on piperacillin concentrations. Samples subjected to various preparation techniques, including freezing, centrifugation, and filtration, were analyzed using deep UV resonance Raman spectroscopy. Data analysis involved preprocessing and chemometric modeling to assess concentration changes and the influence of sample matrix. Results: Sample preparation steps induce concentration changes in piperacillin, with freezing having the highest impact. Chemometric modeling reveals that freezing, filtration, and centrifugation, especially when combined, reduce drug concentration. Furthermore, the choice of urine reference for quantification impacts results, with sex-specific urine pools showing better accuracy compared to mixed pools. Conclusions: Resonance Raman spectroscopy effectively quantifies piperacillin concentrations in urine. Freezing, centrifugation, and filtration during sample preparation influence drug concentration. Using sex-specific urine pools as references yields more accurate quantification results. These findings underscore the importance of considering sample processing effects and reference selection in TDM studies, offering insights for optimizing antibiotic dosing in critically ill patients. Further validation on a larger scale is warranted to confirm these observations.

Therapeutic drug monitoring (TDM) at the point of care is of paramount significance in healthcare. It enables timely monitoring of drug concentrations in a patient's body-fluids, especially in the bloodstream, ensuring personalized dosing for optimal therapeutic outcomes while minimizing side effects and toxicity. TDM is crucial for treating sepsis patients, as their altered pharmacokinetics make accurate antibiotic dosing challenging. Additionally, they need quick dose adjustments to match evolving clinical needs. TDM ensures individually optimal antibiotic levels for best efficacy. Recently, we exploited the advantages of deep-UV Raman spectroscopy in drug sensing of antibiotics for TDM. Firstly, deep-UV excitation enhances the Raman scattering, which is particularly useful for compounds with weak Raman signals, including low concentrated drugs. Secondly, it effectively minimizes fluorescence interference, a common challenge in conventional Raman spectroscopy. This makes deep-UV excitation especially suitable for monitoring active ingredients, such as fluoroquinolones and β-lactam antibiotics, in body fluids like urine and plasma at clinically relevant concentrations. Therefore, deep-UV Raman spectroscopy holds significant potential as a tool for personalized antibiotic dosing through rapid TDM at the point of care.

Stress factors caused by inadequate storage can induce the unwanted degradation of active compounds in pharmaceutical formulations. Resonance Raman spectroscopy is presented as an analytical tool for rapid monitoring of small concentration changes of tetracycline and the metabolite 4˗epianhydrotetracycline. These degradation processes were experimentally induced by changes in temperature, humidity, and irradiation with visible light over a time period of up to 23 days. The excitation wavelength λ_exc = 413 nm was proven to provide short acquisition times for the simultaneous Raman spectroscopic detection of the degradation of tetracycline and production of its impurity in small sample volumes. Small concentration changes could be detected (down to 1.4% for tetracycline and 0.3% for 4-epianhydrotetracycline), which shows the potential of resonance Raman spectroscopy for analyzing the decomposition of pharmaceutical products.

Sepsis and septic shock exhibit a rapid course and a high fatality rate. Antibiotic treatment is time-critical and precise knowledge of the antibiotic concentration during the patients’ treatment would allow individual dose adaption. Over- and underdosing will increase the antimicrobial efficacy and reduce toxicity. We demonstrated that fiber enhanced Raman spectroscopy (FERS) can be used to detect very low concentrations of ciprofloxacin in clinically relevant doses, down to 1.5 µM. Fiber enhancement was achieved in bandgap shifted photonic crystal fibers. The high linearity between the Raman signals and the drug concentrations allows a robust calibration for drug quantification. The needed sample volume was very low (0.58 µL) and an acquisition time of 30 s allowed the rapid monitoring of ciprofloxacin levels in a less invasive way than conventional techniques. These results demonstrate that FERS has a high potential for clinical in-situ monitoring of ciprofloxacin levels.

The particle shape, size and distribution of active pharmaceutical ingredients (API) are relevant quality indicators of pharmaceutical tablets due to their high impact on the manufacturing process. Furthermore, the bioavailability of the APIs from the dosage form depends largely on these characteristics. Routinely, particle size and shape are only analyzed in the powder form, without regard to the effect of the formulation procedure on the particle characteristics. The monitoring of these parameters improves the understanding of the process; therefore, higher quality and better control over the biopharmaceutical profile can be ensured. A new fiber-array-based Raman hyperspectral imaging technique is presented for direct simultaneous in-situ monitoring of three different active pharmaceutical ingredients- acetylsalicylic acid, acetaminophen and caffeine- in analgesic tablets. This novel method enables a chemically selective, noninvasive assessment of the distribution of the active ingredients down to 1 µm spatial resolution. The occurrence of spherical and needle-like particles, as well as agglomerations and the respective particle size ranges, were rapidly determined for two commercially available analgesic tablet types. Subtle differences were observed in comparison between these two tablets. Higher amounts of acetaminophen were visible, more needle-shaped and bigger acetylsalicylic acid particles, and a higher incidence of bigger agglomerations were found in one of the analgesic tablets.

The fight against counterfeit pharmaceuticals is a global issue of utmost importance, as failed medication results in millions of deaths every year. Particularly affected are antimalarial tablets. A very important issue is the identification of substandard tablets that do not contain the nominal amounts of the active pharmaceutical ingredient (API), and the differentiation between genuine products and products without any active ingredient or with a false active ingredient. This work presents a novel approach based on fiber-array based Raman hyperspectral imaging to qualify and quantify the antimalarial APIs lumefantrine and artemether directly and non-invasively in a tablet in a time-efficient way. The investigations were carried out with the antimalarial tablet Riamet^® and self-made model tablets, which were used as examples of counterfeits and substandard. Partial least-squares regression modeling and density functional theory calculations were carried out for quantification of lumefantrine and artemether and for spectral band assignment. The most prominent differentiating vibrational signatures of the APIs were presented.

This work reports the development and application of two liquid-core microstructured polymer optical fibers (LC-mPOF) with different microstructure sizes. They are used in a Fiber Enhanced Raman Spectroscopy (FERS) sensing platform, with the aim of detecting glucose in aqueous solutions in the clinically relevant range for sodium-glucose co-transporter 2 (SGLT2) inhibitor therapy. The sensing platform is tested for low concentration glucose solutions using each LC mPOF. Results confirm that a significant enhancement of the Raman signal is achieved in comparison to conventional Raman spectroscopy. Additional measurements are carried out to obtain the valid measurement range, the resolution, and the limit of detection, showing that the LC-mPOF with 66-μm-diameter central hollow core has the highest potential for future clinical applications. Finally, preliminary tests successfully demonstrate glucose identification in urine.

Fiber-enhanced Raman spectroscopy was developed for the chemically selective and sensitive quantification of the important antibiotic cefuroxime in human urine. A novel optical sensor fiber was drawn and precisely prepared. In this fiber structure, light is strongly confined in the selectively filled liquid core, and the Raman scattered signal is collected with unprecedented efficiency over an extended interaction length. The filling, emptying, and robustness are highly improved due to the large core size (>30 μm). Broadband step-index guidance allows the free choice of the most suitable excitation wavelength in complex body fluids. The limit of detection of cefuroxime in human urine was improved by 2 orders of magnitude (to μM level). The quantification of cefuroxime was achieved in urine after oral administration. This method has great potential for the point-of-care monitoring of antibiotics concentrations and is an important step forward to enable clinicians to rapidly adjust doses.

Innovations in Raman spectroscopic techniques provide a potential solution to current problems in pharmaceutical drug monitoring. This review aims to summarize the recent advances in the field. The developments of novel plasmonic nanoparticles continuously push the limits of Raman spectroscopic detection. In surface‐enhanced Raman spectroscopy (SERS), these particles are used for the strong local enhancement of Raman signals from pharmaceutical drugs. SERS is increasingly applied for forensic trace detection and for therapeutic drug monitoring. In combination with spatially offset Raman spectroscopy, further application fields could be addressed, e.g. in situ pharmaceutical quality testing through the packaging. Raman optical activity, which enables the thorough analysis of specific chiral properties of drugs, can also be combined with SERS for signal enhancement. Besides SERS, micro‐ and nano‐structured optical hollow fibers enable a versatile approach for Raman signal enhancement of pharmaceuticals. Within the fiber, the volume of interaction between drug molecules and laser light is increased compared with conventional methods. Advances in fiber‐enhanced Raman spectroscopy point at the high potential for continuous online drug monitoring in clinical therapeutic diagnosis. Furthermore, fiber‐array based non‐invasive Raman spectroscopic chemical imaging of tablets might find application in the detection of substandard and counterfeit drugs. The discussed techniques are promising and might soon find widespread application for the detection and monitoring of drugs in various fields.

Fiber enhanced resonance Raman spectroscopy (FERS) is introduced for chemically selective and ultrasensitive analysis of the biomolecules hematin, hemoglobin, biliverdin, and bilirubin, as well as intact red blood cells.