Analytical technologies for body fluids and tissues have advanced substantially over the past decade, particularly in chromatographic, electrophoretic and ion-mobility-based separation strategies designed to address matrix complexity and structural isomerism. Improvements in stationary-phase chemistry, multidimensional liquid chromatography, capillary electrophoresis and mobility-integrated mass spectrometry have enhanced peak capacity, structural discrimination, remains the selection and validation of separation strategies capable of minimizing coelution, ion suppression and quantitative variability in heterogeneous matricesremains the selection and validation of separation strategies capable of minimizing coelution, ion suppression and quantitative variability in heterogeneous matrices. This review examines recent progress in multidimensional chromatography, mobility-resolved workflows, miniaturized separation systems and hybrid imaging-separation platforms, with emphasis on comparative performance, practical implementation constraints and translational reproducibility. Applications across plasma, serum, urine, saliva, cerebrospinal fluid (CSF), tissues and spatially resolved analyses are discussed in the context of analyte chemistry and matrix-dependent method selection. Future progress will depend on standardized multidimensional workflows, validated performance metrics, harmonized reporting frameworks and data-driven optimization strategies that support robust clinical translation. By focusing on separation-centred decision-making and quality assurance, this review provides guidance for analytical scientists seeking reliable molecular characterization of complex human biospecimens.

Cyclic peptides have emerged as particularly potent therapeutic agents due to their distinct structural and functional features. It exhibits remarkable versatility through diverse mechanisms of action, including broad-spectrum antimicrobial activity against multidrug-resistant pathogens, precise immunomodulatory functions that facilitate the transition from inflammatory to proliferative phases, enhanced cellular migration and proliferation through growth factor, angiogenesis promotion via VEGF and integrin pathway activation, and sophisticated extracellular matrix modulation. The combination of cyclic peptides and hydrogel technology is one of possible method for treating chronic wounds, such as diabetic foot ulcers. It has been shown to be highly successful in treating extracellular matrix defects, poor angiogenesis, and chronic inflammation in clinical settings. This thorough review literature include the the current status and future possibilities of cyclic peptide-based wound dressings by emphasizing the scalable manufacturing standardization, complete long-term safety profiles, regulatory pathway optimization, and cost-effective manufacturing processes. Future views include AI-driven peptide design, personalized medicine techniques based on wound-specific biomarkers, smart material integration for temporal control, and combination treatments that target various healing pathways at the same time. The combination of computational design, modern manufacturing, and clinical validation positions cyclic peptide-based wound dressings as transformational medicines with the potential to address significant unmet needs in wound treatment.

Programmed necrosis, a controlled cell death method that bypasses resistance mechanisms that render apoptosis ineffective, is a potential cancer treatment target. Due to their diverse biological activities and low side effects, natural products are being explored as modulators of programmed necrosis pathways. This review highlights the potential of natural compounds to target cancer cells while preserving healthy tissues and their interaction with essential programmed necrosis mechanisms like ferroptosis and necroptosis. Recent developments have identified various types of programmable necrosis, including necroptosis, ferroptosis, pyroptosis, proptosis, mitochondrial permeability transition-driven necrosis, and oncosis. Natural compounds are increasingly being utilized as a primary source of anti-cancer medications, providing new cancer treatments. This review demonstrates the molecular mechanisms behind lipid peroxidation, mixed lineage kinase domain-like protein, and receptor-interacting protein kinases (RIPK1 and RIPK3) inducing cell death. Recent research has identified natural compounds like polyphenols, alkaloids, and terpenoids that can modulate pathways and benefit preclinical cancer models. The review underscores the potential of natural compounds in developing innovative cancer treatments by integrating pharmacology and cellular signaling knowledge. Integrating natural compound studies and programmed necrosis research presents a promising avenue for oncologists to overcome treatment resistance. Natural compounds have shown potential in developing programmed necrosis as a novel cancer treatment approach, enhancing therapeutic effectiveness and minimizing side effects through preclinical research, pharmacology, and molecular biology.

Acquired immune deficiency syndrome(AIDS)is the name used to describe several potentially life-threatening infections and disorders that happen when HIV has severely compromised the immune system.The primary effect of HIV is to decrease host immunity,exposing the host to external pathogens.The development of pharmaceutical drugs that directly cure the infection is crucial because of the current wide-ranging epidemic of HIV.Most therapeutic anti-HIV drugs are nucleosides.However,their high toxicity and potential for drug resistance restrict their use.Many of the most effective clinical drugs used to inhibit HIV,the activation of latent HIV,and AIDS have been obtained from natural sources.This review focuses on potential natural medicinal products for treating and managing HIV and AIDS.Notwithstanding,further clinical research studies are needed to understand the subject and its dynamics.

Polyphenols,a diverse group of naturally occurring compounds found in plants,have garnered significant attention for their potential therapeutic properties in treating neurodegenerative diseases(NDs).The Wnt/β-catenin(Wβ C)signaling pathway,a crucial player in neurogenesis,neuronal survival,and synaptic plasticity,is involved in several cellular mechanisms related to NDs.Dysregulation of this pathway is a hallmark in the development of various NDs.This study explores multiple polyphenolic compounds,such as flavonoids,stilbenes,lignans,and phenolic acids,and their potential to protect the nervous system.It provides a comprehensive analysis of their effects on the Wβ C pathway,elucidating their modes of action.The study highlights the dual function of polyphenols in regulating and protecting the nervous system,providing reassurance about the research benefits.This review provides a comprehensive analysis of the results obtained from both in vitro studies and in vivo research,shedding light on how these substances influence the various components of the pathway.The focus is mainly on the molecular mechanisms that allow polyphenols to reduce oxidative stress,inflammation,and apoptotic processes,ultimately improving the function and survival of neurons.This study aims to offer a thorough understanding of the potential of polyphenols in targeting the Wβ C signaling pathway,which could lead to the development of innovative therapeutic options for NDs.

Bimiralisib, a pan-PI3K/mTOR inhibitor, has demonstrated antitumor efficacy in preclinical models. In this study, we present a validated LC-MS/MS method for quantifying bimiralisib from dried blood spots (DBS) in mice. The method was validated in accordance with FDA guidelines. Bimiralisib was extracted from DBS disks using a liquid-liquid extraction technique. Chromatographic separation was achieved on an Atlantis dC18 column (100 × 4.6 mm) using an isocratic mobile phase. The flow rate was set at 0.70 mL/min. Under optimized conditions, the retention times for bimiralisib and the internal standard (IS, Nilotinib) were approximately 1.14 and 1.27 min, respectively, with a total run time of 2.00 min per injection. The monitored MS/MS ion transitions were m/z 412.2 → 141.0 for bimiralisib and 530.4 → 259.0 for the IS. The method employed a broad calibration range (1.00-1434 ng/mL) with a determination coefficient (r2) of 0.996. All validation parameters met the required acceptance criteria, and hematocrit levels had no impact on bimiralisib concentrations in DBS. The validated method was utilized to determine intravenous and oral pharmacokinetic parameters by quantifying bimiralisib in mouse blood, with results correlated to pharmacokinetic data from mice plasma.

We developed and validated a novel analytical methodology for the precise quantification of deucravacitinib, an oral TYK2 inhibitor for treating moderate-to-severe plaque psoriasis in adults. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed in this method for sensitive detection of the compound in rat plasma. Analytical separation was performed utilizing an ACE C18 column (4.6 × 100 mm, 5-μm particle size) with a carefully optimized mobile phase composition of methanol and 2-mM ammonium formate (90:10, v/v), maintained at a consistent flow rate of 0.9 mL/min. Detection was executed in positive ionization mode, targeting multiple reaction monitoring (MRM) transitions of m/z 426.8 → 358.4 for the analyte and m/z 394.1 → 363.2 for the internal standard. The validation of the analytical method encompassed an assessment of selectivity, linearity, accuracy, precision, recovery, and stability. This method demonstrated stability, specificity, and no matrix effect at three concentration levels (1.606, 267.600, 507.780 ng/mL). The method's lower limit of quantification (LLOQ) is 0.556 ng/mL. The calibration curve demonstrates linearity from the LLOQ up to 668.132 ng/mL, exhibiting a high correlation coefficient (r2 = 0.9976). The intraday and interday precisions were less than 6.62% and 5.95%, respectively, with accuracies ranging from 90.68% to 103.80%. The recovery of deucravacitinib ranged from 95.34% to 103.80% and remained stable under different conditions. After successful validation, the method was used for pharmacokinetic profiling of deucravacitinib in rats following oral administration.

Pirfenidone is an antifibrotic and anti-inflammatory drug used for the management of idiopathic pulmonary fibrosis. The current oral delivery of PD has multiple drawbacks, including first-pass metabolism and gastrointestinal discomfort. Efforts have been made to create nanostructured lipid carriers (NLCs) using solid lipids, liquid lipids, and surfactants through an emulsification process followed by ultrasonication to achieve sustained drug release. A central composite design (CCD) utilizing response surface methods (RSMs) was employed to develop and optimize the formulation. The assessed characteristics included particle size distribution, surface topography, drug entrapment efficiency, in vitro drug release, and kinetic profiles in animal models. Cytotoxicity experiments were performed on HepG2 and Caco-2 cell lines and compared with that of PD-NLCs. The optimized formulation yielded a particle size of 159.8 ± 3.46 nm and an encapsulation efficiency of 81.4 ± 7.1% after 10 freeze-thaw cycles of homogenized lipid carriers. In vitro tests assessing various tested flow rates revealed that over 95% of the released drug was retrieved. In vitro studies showed that the PD-loaded nanostructured lipid carrier (NLC) was more cytotoxic to HepG2 and Caco-2 cells than a pure aqueous solution of the drug. Using 25% w/w sorbitol as a cryoprotectant, the findings showed no variation in the properties of NLC before and after freeze-drying. PD-NLCs carriers were shown to have better bioavailability, longer retention time in the lung, and a 15.94-targeting factor related to the PD aqueous solution. Hence, the outcomes confirmed the potential of the PD-NLCs formulation to improve the efficacy of the drug in inhalation therapy.

In treating type 2 diabetes, avoiding glucose reabsorption (glucotoxicity) and managing hyperglycemia are also important. A metabolic condition known as diabetes (type-2) is characterized by high blood sugar levels in comparison to normal Bilosomes (BLs) containing Dapagliflozin (Dapa) were formulated, optimized, and tested for oral therapeutic efficacy in the current investigation. Used the Box Behnken design to optimize the Dapa-BLs, formulated via a thin-film hydration technique. Bile salts (X1) concentration, edge activator (X2) in mg, and non-ionic surfactant (X3) were the independent variables. The Entrapment Efficiency (Y1), Particle size (PS), polydispersity index (PDI), and zeta potential (ZP), were selected as dependent variables. To get the optimal formula, use Design-Expert® software for numerical optimization. The optimal bilosomal formula was selected by boosting %EE, ZP (absolute value), and in vitro drug release while also considering decreasing PS and PDI. Ex vivo skin permeation, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and scanning electron microscopy (SEM) were evaluate the optimized formulation. The in vivo pharmacodynamics activities of the optimized formula were examined on rats and compared to that of the oral Dapa solution. The optimized Dapa-BLs were shown a particle size of 155.36±2.48 nm and an entrapment efficiency of 86.37±2.6%. The SEM image showed a spherical particle with sharp boundaries. The drug release study revealed a significant enhancement in Dapa release (75.31 ± 2.68%) from Dapa -BLs as compared to drug solution (30.46 ± 3.64%). The results of the exvivo permeation and pharmacokinetic studies revealed a 4.49 times higher flux and 3.41 folds higher AUC0-t than drug solution. The antidiabetic activity results showed significant (P < 0.05) enhancement in therapeutic efficacy than drug solution. The results also showed marked improvement in biochemical parameters. Our findings suggested, the prepared Dapa loaded bilosomes was found to be an efficient delivery in the therapeutic efficacy in diabetes.

One of the major global causes of early mortality and illness is diabetes. Diabetes mellitus, a global metabolic condition, affects almost every age group in the world's population. The primary cause of diabetes is insufficient pancreatic function, which occurs when the organ fails to produce enough insulin or fails to adequately use the insulin it produces. The aim of this study is to identify a lead drug as a potent alpha-amylase inhibitor to fight T2DM. We determine the pharmacological alpha amylase target (PDB ID: 3BAX) through extensive analyses and reviews of the available literature. We used a pharmacophore query to search the Natural Products Atlas library for a potent inhibitor. The PyRx version of AutoDock Vina is utilized for the docking process. We use the Desmond software to identify the stability of complexes in physiological environments. According to our analysis, the primary substances NPA016689 and NPA011565 are potent inhibitors against alpha-amylase (3BAX). After determining the lead, we identified the NPA016689 and NPA011565 compounds as the most reactive, with binding affinities of-9.3 kcal/mol and-9.2 kcal/mol, respectively. TYR_A:62, TYR: A_151, LYS: A_200, HIS: A_201, ILE: A_235, and HIS: A_305 residues critically interacted with the NPA016689 ligand. Similarly with ligand NPA011565, the binding residues were TRP: A_59, THR: A_163, LEU_A:165, ARG: A_195, ASP_A:197_A:198, GLU: A_233, and HIS: A_299. This study concluded that NPA016689 and NPA011565 have the potential to modulate the activity of alpha-amylase enzyme, making them potential lead molecules for the design or development of additional anti- alpha-amylase compounds.

Neurodegenerative diseases (NDs) such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis pose significant challenges due to their complex pathophysiology and lack of effective treatments. Green tea, rich in the epigallocatechin gallate (EGCG) polyphenolic component, has demonstrated potential as a neuroprotective agent with numerous medicinal applications. EGCG effectively reduces tau and Aβ aggregation in ND models, promotes autophagy, and targets key signaling pathways like Nrf2-ARE, NF-κB, and MAPK. This review explores the molecular processes that underlie EGCG's neuroprotective properties, including its ability to regulate mitochondrial dysfunction, oxidative stress, neuroinflammation, and protein misfolding. Clinical research indicates that EGCG may enhance cognitive and motor abilities, potentially inhibiting disease progression despite absorption and dose optimization limitations. The substance has been proven to slow the amyloidogenic process, prevent protein aggregation, decrease amyloid cytotoxicity, inhibit fibrillogenesis, and restructure fibrils for synergistic therapeutic effects. The review highlights the potential of EGCG as a natural, multi-targeted strategy for NDs but emphasizes the need for further clinical trials to enhance its therapeutic efficacy.

ABSTRACTPaxalisib is a dual PI3K/mTOR inhibitor, being used in advanced cancer treatment. In this research, we report a validated LC–MS/MS method for quantifying paxalisib from mouse dried blood spot (DBS). We validated the method in‐line with the FDA guidelines. Liquid–liquid extraction technique was used to extract paxalisib from the DBS discs. We used a Chromolith RP‐18 end cap (100 × 4.6 mm) column and isocratic mobile phase for the chromatographic separation of paxalisib and the internal standard (IS, dasatinib). The flow was 0.80 mL/min. In the optimized chromatographic conditions, the retention of paxalisib and the IS was ~2.13 and 2.06 min, respectively. Each injection total run time was 2.50 min. The MS/MS ion transitions monitored were m/z 383.2 → 309.1 and 488.1 → 410.1 for paxalisib and the IS, respectively. We have used a broad calibration range (1.24–3762 ng/mL) with a determination coefficient (r2) of 0.995. All the validation parameters assessed met the acceptance criteria, and hematocrit had no effect on DBS Paxalisib concentrations. We have used the validated method to derive the intravenous and oral pharmacokinetic parameters by quantifying paxalisib in mouse blood and correlated with mice pharmacokinetic data.

Neurological disorders (NDs) encompass a range of debilitating conditions that affect the nervous system, including prevalent illnesses such as Alzheimer's disease, Parkinson's disease, and ischemic stroke. Despite significant ongoing studies, effective therapeutic strategies to halt or slow down the progression of these illnesses are still lacking. Stilbenes, a class of natural polyphenols, have shown potential as candidates for therapeutic strategies due to their capacity to protect the nervous system. Preclinical studies have provided strong evidence that stilbenes can regulate many cellular pathways implicated in neurodegeneration, with resveratrol being a well-studied compound that has shown the ability to reduce oxidative damage, promote neurogenesis, and enhance mitochondrial function - crucial for maintaining brain health. In preclinical animal models, initial research has also shown promise in additional substances such as piceatannol and pterostilbene. Furthermore, clinical studies have explored the therapeutic benefits of stilbenes in NDs. Despite promising results in preclinical research, the use of stilbenes in clinical trials is currently limited, with most studies focusing on resveratrol. Although several clinical studies have demonstrated the beneficial impact of resveratrol supplementation on brain health and degenerative consequences, other investigations have yielded ambiguous findings, underscoring the urgent need for more comprehensive and precisely planned clinical research. This study delves into the potential benefits of stilbenes as neuroprotective agents for NDs. It emphasizes the need for more clinical research to enhance our understanding of their therapeutic effectiveness in specific patient groups.

Zolmitriptan is the primary drug for the treatment of Migraine. However, the bioavailability of the drug is low and requires repetitive administration leading to side effects. Zolmitriptan's bioavailability can be improved by incorporating it into liposomes as a topical intranasal gel. The formulation was developed using a Central composite design employing a response surface approach. The new formulations were tested for particle size, shape, drug entrapment efficiency, and in vitro drug release. Permeation experiments and histopathology in rats were also conducted to determine the formulation's safety. The vesicle size was found to be in the range of 103.82±7.16 to 694.38±1.02 nm, zeta potential --19.28 to -32.8 mV, Entrapment Efficiency from 55.49±1.37 to 99.12±0.36 %, and cumulative drug release from 59.71±6.94 to 99.38±0.13 % respectively. In-vitro drug release of G1 and G3 gel formulations showed a non-Fickian released pattern during the studies. A comparison of the permeation coefficient of G1 (0.539 μg/cm2) and G3 (5.3 μg/cm2) showed a slight variation in the drug release rate after 24 hrs. For the liposomal gel and its solution, we found a significant difference in drug penetration of p0.05 after 12 hours compared to the control gel. There were substantial differences in bioavailability and pharmacokinetics between the optimal Liposomal Gel Formulation and other formulations, including the drug solution, liposomal suspension, and optimized formulation F12. The liposomal gel is non-irritating and safe for topical administration by histopathological investigations. Therefore, the study demonstrated that Zolmitriptan Liposomal gel has better efficacy, good tolerability, and enhanced bioavailability, making it an optimal treatment for acute Migraine.

In recent years, quantum dots (QDs) have emerged as a promising nanomaterial with the potential to revolutionize diagnostic applications due to their unique optical and electrical capabilities. In this setting, there has been a lot of interest in incorporating QDs into composite structures to create cutting-edge diagnostic tools and methods. This summary summarizes the present status of diagnostic composites based on Quantum Dots. It describes the fundamentals of QDs and how their fluorescence can be tuned and their properties vary depending on their size, making them promising candidates for sensitive and multiplexed detection. The synthesis, functionalization, and applications of QDs-based composites across multiple diagnostic platforms are all covered in this article. Their importance in improving the efficiency of in vitro and in vivo diagnostic techniques, such as biosensing, immunoassays, cellular imaging, and targeted drug delivery, is highlighted in particular. The difficulties and factors related to QDs-based composites' biocompatibility, toxicity, and clinical translation are also discussed in the abstract. Overall, this chapter highlights the revolutionary potential of Quantum Dots based composites in diagnostics, allowing for more precise, efficient, and individual monitoring of disease.

The term “antivirals” has been associated with COVID-19 during the period when the usage of this specific class of medications has increased. Nirmatrelvir, a protease inhibitor, is used to treat mild to moderate COVID-19 symptoms by stopping SARS-COV-2 reproduction. With deucravacitinib serving as the internal standard, liquid chromatography-tandem mass spectroscopy in human plasma was used to provide a quick, simple, innovative, trustworthy, and sensitive approach. Liquid-liquid extraction was used to separate nirmatrelvir and the internal standard. The extracted sample was then run through a chromatographic system with ACE-C18 column (4.6 × 100 mm, 5 μm); and mobile phase with methanol and 2 mM ammonium formate in a ratio of 80:20, and a flow rate of 1.00 mL/min. The system operates for three minutes in multiple reaction monitoring mode at the ABSCIEX API 4000 mass spectrometer using electron spray ionization. Nirmatrelvir ion transitions are 500.10 to 110.10, while deucravacitinib are 426.30 to 358.20. The validation was conducted using a concentration range of 5.00 to 4000 ng/mL, and the results showed that the selectivity, accuracy, precision, linearity, and selectivity were all within the acceptability limits.

After lung cancer, breast cancer (BC) is the second most frequent malignancy in women globally. Surgery followed by chemotherapy is the conventional treatment plan for BC. However, both are unsuccessful in treating BC because of the harmful effects that these treatments have on healthy tissues and organs. Many polymeric nanoparticles (PNPs) have been discovered and created recently to selectively aim cancer cells without harming normal cells. As an outcome, drug delivery systems (DDS) mediated by NPs have developed as a possible method to treat BC. PNPs have several special qualities that make them ideal for cancer treatment due to their tunable surface functions and choosiness to target tumor cells and minimize side effects

The term “anticancer drugs” evokes memories of a time when the development of medications in that category was still necessary. Capmatinib, for example, is a kinase inhibitor that targets the C-Met receptor tyrosine kinase in the treatment of non-small cell lung cancer, which involves tissue repair and organ regeneration.1 Thus, utilizing liquid chromatography-tandem mass spectroscopy (LC-MS/MS) in human plasma in accordance with United States Food and Drug Administration (USFDA) bioanalytical technique validation criteria, a quick, simple, specific, dependable, and sensitive approach was created using deucravacitinib as an internal standard. Capmatinib and the internal standard were separated using liquid-liquid extraction. The extracted sample run through a chromatographic system with an ACE-C18 column (4.6 × 100 mm, 5 μm); the mobile phase consisted of methanol and 2 mM ammonium formate in an 80:20 ratio at a flow rate of 1.00 mL/min. The system operates for three minutes in multiple reaction monitoring mode at the ABSCIEX API 4000 mass spectrometer using electron spray ionization. For capmatinib, the ion transitions are 413.10 to 382.10 and 426.30 to 358.20 for deucravacitinib, with a concentration range of 5.00 to 4000 ng/mL, the accuracy, precision, linearity, selectivity, and selectivity were validated and found to be within the acceptability limits.