Silvia Imre

Bisoprolol and Lisinopril are among the most frequently used substances in pediatric cardiovascular affections. The medication supply to this age population is limited given the lack of appropriate pharmaceutical forms; therefore, pharmaceutical compounding comes to fill in the gap between pediatric therapy needs and availability of adult-approved formulations. The aim of our study was to obtain and quantify two suspensions of Bisoprolol and Lisinopril in small pediatric dosage. The suspensions were obtained using a commercial suspending vehicle and the active substances. We developed the quality control methods according to the USP. Briefly, both methods were developed and validated using a UHPLC Flexar 10 system (Perkin Elmer) equipped with a Phenomenex Luna C8 column (150 × 4.6 mm, 3 μm) under isocratic conditions at a flow rate of 1 mL/min. For bisoprolol, the mobile phase consisted of water (65:35, v/v) with the specified additives, with detection at 273 nm, while lisinopril was analyzed using 10 mM ammonium formate buffer (pH 5) (96:4, v/v), with detection at 210 nm. The calibration ranges were 30–90 μg/mL for bisoprolol and 20–100 μg/mL for lisinopril. We obtained satisfactory results for method validation, and the method was applied for the assay of two types of suspensions. The obtained results ranged between 80 and 86% of the labeled amounts for Bisoprolol, and 106–108% for Lisinopril. The obtained suspensions reflected some issues related to the sample extraction and potential variability in suspension sampling and dosing consistency, which support the need for continuous optimization and quality control of compounded pediatric formulation.

Regional anesthesia techniques such as the ultrasound-guided PECS II (pectoral nerve block) block are increasingly employed to optimize perioperative analgesia while minimizing systemic anesthetic exposure. Ropivacaine is commonly used for its favorable pharmacological profile; however, clinical data on its pharmacokinetics and systemic metabolite behavior following interpectoral administration remain limited. This study aimed to characterize the plasma concentration–time profile of ropivacaine and its main active metabolite, 3-OH-ropivacaine, in patients undergoing interpectoral nerve block, using a validated LC-MS/MS (liquid chromatography coupled with mass spectrometry) method. Venous blood samples were collected from 18 patients at predefined time points (0, 1, 3, 6, and 24 h) following a PECS II block performed with a ropivacaine-lidocaine mixture. Plasma concentrations were quantified via a validated LC-MS/MS protocol in accordance with FDA (Food and Drug Administration) and EMA (European Medicines Agency) guidelines. Pharmacokinetic parameters were derived using non-compartmental analysis. Ropivacaine reached a mean peak plasma concentration (Cmax—maximum concentration) of 167.5 ± 28.3 ng/mL at 1.3 ± 0.2 h (Tmax—maximum time). The metabolite 3-OH-ropivacaine peaked at 124.1 ± 21.4 ng/mL at 2.3 ± 0.3 h. The terminal elimination half-life was 19.4 ± 2.8 h for ropivacaine and 29.2 ± 3.1 h for its metabolite. Plasma levels demonstrated prolonged systemic exposure with predictable pharmacokinetics. The PECS II block using ropivacaine results in sustained systemic levels of both the parent drug and its primary metabolite, supporting its role in prolonged perioperative analgesia. These data provide a pharmacokinetic foundation for personalized regional anesthesia protocols. This strategy facilitates the adaptation of anesthetic protocols to the individual characteristics of each patient, aligning with the principles of personalized medicine, particularly in patients with altered metabolic capacity.

Background: Short-chain fatty acids (SCFAs) and medium-chain fatty acids (MCFAs) are human metabolites which are involved in various biochemical processes and can offer valuable insights and information on various pathological and metabolic issues of patients. Accurate, precise, high-performance bioanalytical methods are important tools in both research and diagnostics of many pathologies, with LC-MS being the most frequently used methodology in modern metabolomics studies. Methods: The current paper describes a complete LC-MS/MS methodology for the accurate quantification of total plasmatic SCFA concentrations in humans using high-resolution QTOF mass spectrometric detection, including sample cleanup, preparation, and derivatization. Results and Conclusions: The method was validated with regard to all relevant parameters (selectivity, sensitivity, accuracy, precision, linearity, recovery, carryover, and reproducibility of sample preparation) according to the current applicable guidelines and tested in an in vivo study to quantify peripheral SCFAs in human patients as biomarkers for gut–brain axis disruption.

Objective: This study aims to investigate the role of fatty acid ratios, specifically DHA/ARA and EPA/ARA, in predicting severe disability and mortality in stroke patients and compare these ratios with conventional risk factors such as age, sex, hypertension, diabetes, and dyslipidemia. Methods: A prospective study was conducted involving 298 consecutive acute ischemic stroke patients (within 72 h of onset). Fatty acid ratios were measured from plasma, and all patients’ evolution was followed through hospitalization. Binary logistic regression analysis was used to identify predictors of severe disability at discharge (Rankin 4–6) and in-hospital mortality, including fatty acid ratios and conventional risk factors. Results: A higher DHA/ARA ratio was associated with a reduced chance of severe disability (OR = 0.81), while a higher EPA/ARA ratio was associated with an increased chance of severe disability (OR = 1.70). Age was a significant factor, with older age (median 70 years) associated with a lower survivability chance (OR = 0.93) and a higher likelihood of severe disability when surviving. Fatty acid ratios did not significantly affect mortality outcomes. For male patients, EPA/AA ratios showed a powerful association with severe disability (p = 0.045), while no significant effect of fatty acids was observed in females. Conclusions: Fatty acids were significant predictors of severe disability in patients with acute ischemic stroke, independent of conventional risk factors, but without having any effect on in-hospital mortality. Age remained the only significant conventional risk factor predictor of outcome. Integrating fatty acid ratios alongside conventional risk factors may improve predictions of severe post-stroke disability, potentially guiding more personalized interventions for stroke patients.

Cannabidiol (CBD) oils and products have become extremely popular in the last decade. These products are marketed as having different effects and are recommended for many chronic diseases. Various food supplements with CBD are now available on the market, but due to legislative lacks the declared concentration of CBD and other cannabinoids in the products is often significantly different compared with the real concentration. Products that don't meet quality criteria result from a lack of control and standardization. The aim of this study was to develop and validate an LC-MS/MS method for the routine quantification of cannabinoids in herbal drugs and food supplements. An LC-MS/MS method was developed using an UHPLC system coupled with a QTOF mass spectrometer, and the chromatographic separations were performed on a C18 column with isocratic elution, electrospray ionization in negative mode with a run-time of 10 minutes. According to the international guidelines, the method was validated concerning linearity, selectivity, stability, precision, and accuracy. Out of the ten tested products, in three of the supplements, significant differences were noticed in CBD concentration compared to the declared content. Other cannabinoids were also identified in some of the samples. This study raises awareness regarding insufficient controlled food supplements.

Ropivacaine, a widely used regional anesthetic also used for pain management, has been increasingly used in recent years due to its increased efficacy and improved safety compared to similar anesthetics. Biomonitoring of ropivacaine and its metabolites during and after anesthesia is an essential process for ensuring therapeutic efficacy and safe usage for patients. The most useful biomonitoring tool in recent years has been liquid chromatography coupled with mass spectrometry (LC-MS/MS), which offers selectivity, sensitivity, as well as accuracy of measurements. The current manuscript summarizes and discusses the existing liquid chromatographic methods described in the literature, as well as the personal experience with developing bioanalytical and analytical methods for the quantification of ropivacaine in biological samples for clinical applications. It is focused on methodological aspects, recent advancements, challenges, and future perspectives, highlighting the importance of LC-MS/MS techniques in ropivacaine analysis.

Off-label practice in pediatrics requires relentless engagement from all the health professionals involved. Community pharmacists are the last ones in the prescribing–dispensing chain; therefore, they have the key responsibility of verifying the correctness of a treatment. A cross-sectional study was conducted for assessing the awareness and views of Romanian community pharmacists, regarding off-label drugs in the pediatric population, through a 28-item questionnaire comprising five sections of different topics (general knowledge, frequency of prescribing and dispensing off-label medication, views, and attitudes). The sample size was 236 questionnaires with a response rate of 41.11%. A statistical analysis of the obtained data was performed with GraphPad Prism v.9. The results indicate that 55.1% of the community pharmacists have a good general knowledge and awareness regarding the off-label practice, although the legal frame is unclear. The responses highlight a high frequency of prescribing and request of medication for respiratory conditions (45.3%) and antibiotics (23.5%), with a concerning gap regarding the adverse events related to the off-label treatments (56.7%). A very small percentage of pharmacists (7.1%) contact a fellow healthcare professional when encountering an off-label prescription. In conclusion, in addition to the pharmacist’s conduct towards the best interest of the patient, there is a clear need to improve the doctor–pharmacist collaboration in order to make an off-label treatment successful.

Introduction Benign prostatic hyperplasia (BPH) is a prevalent condition among aging men that affects their life quality due to urinary symptoms. Current pharmacologic treatments, often lead to sexual dysfunction, so dietary supplements (DS) containing plant -based compounds such as beta-sitosterol (SIT) are preferred. DS are highly accessible and widely used, but poorly regulated, so often patients are victims of fraud. The use of DS to treat BPH symptoms is questionable, and this may be due not to the efficacity of the active compound but to the quality of commonly available DS. Aim This study aimed to assess the concentration of SIT in DS available on the market and evaluate whether the concentration of the active compound at the recommended dosage is sufficient to elicit beneficial effects in BPH. Method An HPLC-UV method based on direct saponification and acid hydrolysis was developed for the quantification of free and conjugated SIT in DS. The concentration of SIT in various DS was determined and compared with the one declared on the label. Results The chromatographic analysis confirmed the presence of SIT in all the DS but also showed a considerable variability of SIT content among DS, with only one product meeting the necessary concentration to bring potential benefits in BPH. Conclusion The study highlights inconsistencies in SIT content among DS and the importance of DS containing a standardized extract of SIT. Quality control measures are imperative to ensure that consumers receive effective and safe SIT -based DS to manage BPH symptoms. Further research is needed to establish standardized dosages and to evaluate their long-term efficacy and safety.

Background: LC-MS is an ever-increasingly used methodology for clinical applications. Due to the superior selectivity and sensitivity, in certain situations, it can offer an advantage or be the only option for diagnostics and biomonitoring applications. Methods: A high selectivity sensitive LC-MS/MS method was developed for direct quantification of free plasma polyunsaturated fatty acids as well as conjugated membrane polyunsaturated fatty acids, using isocratic reverse phase elution. A quick and simple sample purification method was used in order to ensure high-throughput analysis of biological samples. The method was validated with regard to selectivity, sensitivity, linearity, accuracy, precision, carryover, and recovery, as well as other relevant parameters. Results and Conclusions: The method was developed and validated with respect to all relevant parameters and was successfully used in a number of clinical diagnostics and biomonitoring applications. The simple sample purification process allowed for an easy learning curve for analysts and other users, while ensuring a low chance of systematic or random errors and thus reliable results usable in a clinical setting.

Carvedilol, a highly protein-bound beta-blocker, is used in therapy as a racemic mixture of its two enantiomers that exhibit different pharmacological activity. The aim of this study was to evaluate the stereoselective nature of its binding to the two major plasma proteins: albumin and alpha-1-acid glycoprotein. The determination of the plasma protein-binding degree for carvedilol and its enantiomers was achieved using ultrafiltration for the separation of the free fraction, followed by LC-MS/MS quantification, using two different developed and validated methods in terms of stationary phase: achiral C18 type and chiral ovomucoid type. Furthermore, molecular docking methods were applied in order to investigate and to better understand the mechanism of protein-binding for S-(-)- and R-(+)-carvedilol. A difference in the binding behavior of the two enantiomers to the plasma proteins was observed when taken individually, with R-(+)-carvedilol having a higher affinity for albumin and S-(-)-carvedilol for alpha-1-acid glycoprotein. However, in the case of the racemic mixture, the binding of the S enantiomer to alpha-1-acid glycoprotein seemed to be influenced by the presence of its antipode, although no such influence was observed in the case of albumin. The results raise the question of a binding competition between the two enantiomers for alpha-1-acid glycoprotein.