Bearing in mind the centuries-old traditional use of chamomile, but also the increasing demand for its products in modern industry, oriented toward sustainable development, there are increasing efforts for the efficient extraction of high-value compounds of this plant, as well as obtaining its products with added value. With that goal, conventional and contemporary separation techniques were applied in this work. Both hydrodistillation processes (HD), conducted in a traditional manner and coupled with microwave irradiation (MWHD), were used for essential oil isolation. In parallel with those procedures, chamomile lipophilic extracts were obtained by Soxhlet extraction applying organic solvents and using supercritical fluid extraction as a greener approach. The obtained extracts and essential oils were characterized in terms of chemical composition (GC analysis, contents of total phenolics and flavonoids) and biological potential. GC analysis revealed that oxygenated sesquiterpenes and non-terpene compounds were the dominant compounds. α-Bisabolol oxide A (29.71–34.41%) and α-bisabolol oxide B (21.06–25.83%) were the most abundant individual components in samples obtained by distillation while in supercritical and Soxhlet extracts, major compounds were α-bisabolol oxide A and pentacosane. The biological potential of essential oils and extracts was tested by applying a set of analyzes to estimate the inhibition of biologically important enzymes (amylase, glucosidase, acetylcholinesterase, butyrylcholinesterase, tyrosinase) and antioxidant capacity (DPPH, ABTS, CUPRAC, FRAP, chelating and total antioxidant capacity). The results suggested essential oils as better antioxidants, while the extracts were proven to be better inhibitors of the tested enzymes. Principal Component Analysis was conducted using the experimental results of the composition of extracts and EOs of chamomile obtained by different separation techniques, showing clear discrimination between methods applied in correlation with the chemical profile. Molecular docking was applied for the identification of the main active principles present in the essential oil, among which α-bisabolol-oxide B (cp3) showed a higher affinity for tyrosinase. Full article

The enantiomeric separation of antifungal compounds is an arduous task in pharmaceutical and biomedical fields due to the different properties that each diastereoisomer presents. The enantioseparation of a group of fungicides (sulconazole, bifonazole, triadimefon and triadimenol) using supercritical fluid chromatography was achieved in this work. For this goal, four different chiral columns based on polysaccharide derivatives, as well as the effect of different chromatographic parameters such as temperature, type and percentage of organic modifier (methanol, ethanol and isopropanol), were thoroughly investigated. The inversion of the elution order of enantiomers as a result of a change in the stationary phase or organic modifier was also evaluated by employing a circular dichroism detector. The best separation conditions, in terms of the enantioresolution and analysis time, were obtained with the Lux^® Cellulose-2 column using isopropanol as the organic modifier. Full article

The drug 5-fluorouracil (5-FU) is a common cancer chemotherapeutic, presenting toxicity. Mild toxicity is treated with administration of probiotics. The interaction of these probiotics with the drug may have a crucial effect on its therapeutic efficacy. In the present work, a method for the quantification of uracil, 5-FU, and its active metabolite 5-fluorodeoxyuridin monophosphate in cells and culture medium of the probiotic L. lactis is presented. Extraction using H₂O containing 0.05% v/v formic acid (1:5 v/v) was followed by ammonium sulphate protein precipitation and SPE. Analysis was conducted in a Nucleosil column using a gradient of water, formic acid, and acetonitrile. Calibration curves were constructed for 5-FU (5–100 μg/mL), uracil (5–20 μg/mL), and 5-fluorodeoxyuridin monophosphate (5–20 μg/mL) using 5-bromouracil as the internal standard (R² ≥ 0.999). The photodegradation of 5-FU amounted to 36.2% at 96 h. An administration experiment in the dark revealed a decline in 5-FU concentration in the culture media (88.3%) and uptake by the cells, while the uracil and FdUMP levels increased in the cells. The inactive metabolite 5,6 dihydrofluorouracil was detected in the medium. Our results demonstrate that uptake and metabolism of 5-FU in L. lactis cells leads to a decline in the drug levels and in the formation of both the active and the inactive metabolites of the drug. Full article

In order to meet the analytical requirements of the European Medicines Agency (EMA), a new HPLC-FLD method was successfully developed using dansyl chloride for the derivatization and determination of the genotoxic impurities N-Nitrosodimethylamine (NDMA) and N-Nitrosodiethylamine (NDEA) in Lisinopril API and its final product. Samples’ pretreatment includes liquid–liquid microextraction, denitrosation, and derivatization steps. To optimize the process, the parameters contributing to high sensitivity and yielding reliable results were thoroughly studied and optimized using one-factor-at-a-time and experimental design approaches. The analytes were pre-column derivatized with Dansyl-Cl and analyzed by HPLC-fluorescence (λ_em/λ_em = 340/530) using a C₁₈ column and a mixture of phosphate buffer (pH = 2.8; 20 mM)/acetonitrile 55:45 v/v as the mobile phase. The six-level concentration calibration was shown to be linear, with R equal to 0.9995 for both analytes. The limit of detection (LOD) was satisfactory and equal to 4.7 and 0.04 ng/mL for NDMA and NDEA, respectively. Precision was less than 13.4% in all cases, and the average recoveries were equal to 109.2 and 98.1% for NDMA and NDEA, respectively. The proposed procedure is relatively easy, rapid, and suitable for the determination of the two nitrosamines in routine analysis tests. Full article

Solid organ transplantation is a life-saving intervention for patients suffering from end-stage organ failure. Although improvements in surgical techniques, standards of care, and immunosuppression have been observed over the last few decades, transplant centers have to face the problem of an insufficient number of organs for transplantation concerning the growing demand. An opportunity to increase the pool of organs intended for transplantation is the more frequent use of organs from extended criteria and the development of analytical methods allowing for a better assessment of the quality of organs to minimize the risk of post-transplant organ injury and rejection. Therefore, solid-phase microextraction (SPME) has been proposed in various studies as an effective tool for determining compounds of significance during graft function assessment or for the chemical profiling of grafts undergoing various preservation protocols. This review summarizes how SPME addresses the analytical challenges associated with different matrices utilized in the peri-transplant period and discusses its potential as a diagnostic tool in future work. Full article

In recent years, the scientific community has turned its attention to the further study and application of green chemistry as well as to sustainable development in reducing the consumption of raw materials, solvents, and energy. The application of green chemistry aims to ensure the protection of the environment and to also, consequently, improve the quality of human life. It offers several benefits, both socially and economically. In the last few decades, new alternative non-conventional green extraction methodologies have been developed for the purposes of the extraction of active ingredient compounds from various raw products. The main objective of this literature review is to present the current knowledge and future perspectives regarding the green extraction of tea species in respect of the isolation of safe active biomolecules, which can be used as commercially available products—both as dietary supplements and pharmaceutical formulations. More specifically, in this literature review, the intention is to investigate several different extraction techniques, such as ultrasonic-assisted extraction, ultrasonic-assisted extraction with DESs, the microwave assisted-extraction method, and the reflux method. These are presented in respect of their role in the isolation of bioactive molecules regarding different tea species. Furthermore, following the literature review conducted in this study, the commonly used green extraction methods were found to be the ultrasound-assisted method and the microwave-assisted method. In addition to these, the use of a green solvent, in regard to its role in the maximum extraction yield of active ingredients in various species of tea, was emphasized. Catechins, alkaloids (such as caffeine), gallic acid, and flavonoids were the main extracted bioactive molecules that were isolated from the several tea species. From this literature review, it can be demonstrated that green tea has been widely studied at a rate of 52% in respect of the included research studies, followed by black tea at 26%, as well as white tea and oolong tea at 11% each. Regarding the determination of the bioactive molecules, the most utilized analytical method was found in the combination of high-performance liquid chromatography (HPLC) with a photodiode array detector (PDA) and mass spectrophotometry (MS) at a usage rate of about 80%. This method was followed by the utilization of UPLC and GC at 12% and 8%, respectively. In the future, it will be necessary to study the combination of green extraction techniques with other industry strategies, such as an encapsulation at the micro and nano scale, for the purposes of preparing stable final products with antioxidant properties where, finally, they can be safely consumed by humans. Full article

The analysis of the secoiridoid type of phenolic compounds present in virgin olive oil has become a challenging area of research since the first evidence of their presence in the polar fraction of the oil. Separation techniques, mainly liquid chromatographic ones, prevailed over the years of application toward elucidation of their structure, content determination and collection of evidence on cultivar, origin, processing and storage conditions dependence. One of the latest challenges in their analysis was related to the need to address the requirement set by EC Regulation 432/2012 for the implementation of the health claim on ‘olive oil polyphenols’. The present work considers in a chronological order the original articles, viewpoints, review articles and other published efforts that appeared in the literature after the issuing of the relevant EFSA scientific opinion in 2011. The EFSA health claim created a lot of expectations among producers of virgin olive oil and boosted research for the development of a ‘fit for the purpose’ analytical protocol. Emphasis is given to the dedicated separation protocols that have been developed in the last 10 years and to the progress in their validation in comparison to the features of the method that were recently adopted by the International Olive Council. Full article