Journal of Applied Pharmaceutical Research

Flavonoids as chemopreventive agents: metabolism, apoptosis, and oxidative stress modulation

Faruk Alam — 2026-03-15

Background: Liposomes are widely used as drug delivery systems because of their reduced systemic toxicity. Over the past few decades, numerous drug-loaded liposomes have been approved for clinical use in the treatment of cancer, viral, and fungal infections. Various liposomal formulations have progressed to later phases of clinical trials. Liposomes are spherical vesicles composed of a single or multiple phospholipid bilayers surrounding an aqueous core. Drug-loaded liposomes can exhibit controlled or targeted drug delivery, low immunogenicity, high biocompatibility, biodegradability, prolonged drug half-life, increased efficiency, reduced systemic toxicity, and enhanced pharmacokinetic properties. Methodology: This review article addresses the characteristics and types of liposomes; novel methods for their preparation, such as the Supercritical Anti-solvent Method and the Dual Asymmetric Centrifugation Method; lipid preferences; future directions for liposomes; marketed liposomal formulations; and associated patents. Results and Discussion: It has the potential to protect the drug against degradation. The aforementioned drug delivery system increases in vivo drug distribution toward target sites. PEGylated liposomes can prolong circulation time. It requires expertise in techniques, such as thin-film hydration and reverse-phase evaporation, for preparation. It has been utilized in nanomedicine. This particular delivery system requires characterizations like size, drug loading, drug release, etc. Conclusion: Liposome-embedded delivery systems advance nanotechnology and biopharmaceutics. The role of modern medicine has continued to expand, particularly in the management of chronic diseases.

Advancements in natural and modern treatments for Alzheimer's disease in preclinical and early stages

Ronald Darwin C — 2026-03-15

Background: Alzheimer's disease (AD), the most common cause of dementia, is characterized by memory loss, cognitive decline, and behavioral changes. Current monoclonal antibodies raise concerns about cost and safety, despite modest disease-modifying effects. Methodology: Evidence from preclinical research, clinical trials, and meta-analyses on integrative methods for managing AD is compiled in this narrative review. We assessed natural phytochemicals (from plants like Ginkgo biloba, Bacopa monnieri, Withania somnifera, Curcuma longa, and Salvia officinalis), lifestyle modifications, and new technologies. With an emphasis on preclinical and early-stage AD data through 2025, standardized extracts such as EGb761 and saffron were given priority. Results: Certain plants' phytochemicals have anti-inflammatory, anti-amyloid, antioxidant, and neurotrophic properties that alter important AD pathways. Standardized extracts such as EGb761 and saffron have been shown in clinical trials to provide cognitive benefits comparable to those of prescription medications, with fewer side effects. Pharmacotherapy, natural modulators, stem cell therapy, AI-driven precision medicine, and other integrative strategies have shown promise in slowing disease progression. Discussion: Low bioavailability, formulation variability, and a lack of large-scale trials are some of the issues facing clinical translation. Natural remedies can complement modern treatments, particularly during the early stages of AD. Standardized formulations, sophisticated delivery methods, and thorough verification of synergistic strategies are essential for future success. Conclusion: Modern and natural therapies present promising options for managing AD at an early stage. Integrative care must be advanced through standardized formulations, enhanced delivery techniques, and robust clinical validation of synergistic approaches.

Chitosan-coated solid lipid nanoparticles for nose-to-brain drug delivery: A comprehensive review

Samruddhi Rakshe — 2026-03-15

Background: The blood-brain barrier (BBB) limits the effectiveness of treatment of several central nervous system (CNS) diseases, including Alzheimer's disease, Parkinson's disease, schizophrenia, and bipolar disorder. Because of this limitation, intranasal drug delivery is an effective, non-invasive way to bypass the BBB via the olfactory and trigeminal pathways. Methodology: In this review, we critically evaluate the design, characterization, mechanism of action, and potential therapeutic use of Chitosan-coated Solid Lipid Nanoparticles (SLNs) as a delivery system for drugs from the nasal cavity to the brain. A structured literature search was conducted in PubMed, Scopus, and Web of Science for studies published between 2019 and 2025 using keywords related to chitosan-coated solid lipid nanoparticles and nose-to-brain delivery. Studies reporting pharmacokinetic parameters, such as brain AUC, brain-targeting efficiency (BTE%), and drug-targeting index (DTI), were included. Results and Discussion: Preclinical studies show that some chitosan-coated SLN formulations significantly improve drug pharmacokinetics, increasing the brain area under the curve (AUC) compared with conventional formulations. Reported brain targeting efficiency (BTE%) with drug targeting index (DTI) values confirmed preferential nose-to-brain transport and improved CNS exposure. The improved targeting efficiency is attributed to the mucoadhesive nature of chitosan, enhanced permeability, and prolonged nasal residence time of SLNs. Conclusion: Chitosan-coated Solid Lipid Nanoparticles (SLNs) are an interesting candidate for a nanocarrier system to deliver drugs through the intranasal route into the CNS (Central Nervous System). Future research should emphasize Quality by Design (QbD)-based optimization, advanced surface modifications for targeted delivery, and comprehensive safety and clinical validation to support successful translation into clinical practice.

Ethnomedicinal relevance and bioactive potential of manila tamarind: An evidence-based assessment

Sagar Kamble — 2026-03-15

Background: Pithecellobium dulce (Roxb.) Benth., commonly known as Manila tamarind, is a leguminous tree traditionally used in indigenous medicine, nutrition, and agroecological practices. Despite its wide ethnomedicinal usage, its comprehensive bioactive and functional potential remains underexplored in mainstream scientific literature. Methodology: This review systematically compiles and critically analyzes published data from peer-reviewed journals, ethnobotanical records, and experimental studies focusing on the botanical features, phytochemical composition, nutritional attributes, pharmacological activities, and ecological significance of P. dulce. Results and Discussion: The species is rich in phenolics, flavonoids, alkaloids, saponins, and essential nutrients, which collectively contribute to its broad pharmacological spectrum. Reported biological activities include antidiabetic, antihyperlipidemic, anti-inflammatory, antioxidant, antimicrobial, wound-healing, analgesic, and antipyretic effects. Mechanistic evidence suggests modulation of oxidative stress, inflammatory mediators, and metabolic pathways. P. dulce plays a vital environmental role through nitrogen fixation, soil enrichment, carbon sequestration, and biodiversity support. Beyond its medicinal value, the plant shows strong potential for use in food products, animal feed, herbal formulations, and agroforestry systems. However, limitations such as variability in extraction methods, incomplete toxicity profiling, and insufficient clinical validation restrict its translational applications. Conclusion: Pithecellobium dulce represents a promising yet underutilized bioresource with multifaceted therapeutic, nutritional, and ecological benefits. Future research should prioritize standardization, advanced phytochemical characterization, safety evaluation, and clinical studies to support its development into functional foods, phytopharmaceuticals, and sustainable industrial products.

Development of transfersomal buccal patches of galantamine for enhanced bioavailability in Alzheimer’s disease therapy

Pavuluri Chandrasekhar — 2026-03-15

Background: Galantamine, an acetylcholinesterase inhibitor used in Alzheimer’s disease, exhibits low and variable bioavailability due to gastrointestinal metabolism and first-pass hepatic elimination. Buccal drug delivery offers a non-invasive route that bypasses first-pass metabolism and maintains systemic exposure. This work aimed to develop and evaluate transfersomal buccal patches of galantamine to enhance permeation and control release. Methodology: Galantamine-loaded transfersomes were prepared by thin-film hydration using phosphatidylcholine and edge activators, then incorporated into a mucoadhesive polymer matrix (PVA/HPMC/Carbopol/Eudragit L-100) via solvent casting. Formulations were characterized for vesicle size, zeta potential, deformability, entrapment efficiency, physicochemical properties, swelling, mucoadhesion, in vitro release, ex vivo buccal permeation, and stability. Result and Discussion: Transfersomes exhibited a mean size of 126.4 ± 4.8 nm, PDI ≈ 0.21, zeta potential −32.6 ± 1.4 mV, and entrapment efficiency 86.7 ± 2.5%. Optimized patches showed uniform thickness, high folding endurance (>300), drug-content uniformity (≈95–98%), and strong mucoadhesive strength, with PEG-plasticized films demonstrating superior swelling and residence time. Drug release followed diffusion-controlled (Higuchi-type) kinetics with near-zero-order behavior. Ex vivo permeation revealed a 2.5-fold increase in steady-state flux (3.21 ± 0.18 µg/cm²/h) and a higher permeability coefficient compared with the conventional oral formulation. Accelerated and saliva stability studies indicated no significant physicochemical changes. Conclusion: The integration of deformable transfersomes within a mucoadhesive buccal matrix enabled sustained diffusion-controlled delivery, significantly enhancing mucosal permeation. The transfersomal buccal patch represents a promising alternative to oral galantamine therapy, warranting in vivo pharmacokinetic and clinical evaluation.

Formulation, optimization, and in-vitro diffusion studies of novel niosomal gel of miconazole nitrate

Akash Mummoorthy — 2026-03-15

Background: Miconazole Nitrate exhibits potent antifungal activity; its therapeutic potential is restricted by poor skin permeability and low aqueous solubility. To overcome these limitations, the present study aimed to develop and optimize a niosomal gel delivery system using non-ionic surfactant vesicles to enhance dermal diffusion and therapeutic efficacy. Methodology: The ether injection method was used to create niosomes. The results obtained from applying the Box-Behnken design (BBD) were highly instructive for the PDI, droplet size, and drug release profile. DLS was used to quantify particle size and zeta potential, and SEM was used to characterize particle morphology. The stability and compatibility were examined using DSC and FTIR. Results and Discussion: The optimized niosomal formulation exhibited a particle size of 180.9 ± 1.8 nm, a PDI of 0.1533 ± 0.01, and a zeta potential of –27.28 ± 0.2 mV, indicating excellent colloidal stability. The Box–Behnken model exhibited strong statistical significance (R² = 0.9663 for size, 0.9098 for PDI, and 0.9971 for zeta potential; p < 0.05). The encapsulation efficiency was 91%; the in-vitro diffusion profile followed the Hixson–Crowell model (R² = 0.9959), confirming a controlled-release mechanism. Compared with conventional Miconazole formulations, the optimized niosomal gel demonstrated markedly enhanced diffusion and prolonged release over 8 hours. The Carbopol-based gel exhibited suitable viscosity (3250 cP), spreadability, and pH (5.5 ± 0.1), ensuring dermal compatibility. Conclusion: The niosomal gel displayed characteristics that improved its drug loading, stability, and sustained-release properties. These modifications indicate its potential as a transdermal drug delivery system and improved antifungal activity.

Validated LC-MS/MS method for the quantification of olaparib and abiraterone acetate in human plasma

Bhavik Jani — 2026-03-15

Background: Olaparib and Abiraterone acetate are approved in combination therapy for metastatic castration-resistant prostate cancer (mCRPC). Accurate and simultaneous quantification of these agents in plasma is crucial for pharmacokinetic assessment and drug–drug interaction studies. Method: A rapid, simple, and selective LC-MS/MS method was developed and validated for the simultaneous quantification of Olaparib (25–5000 ng/mL) and Abiraterone acetate (1.25–250 ng/mL) in human plasma. Protein precipitation using chilled acetonitrile was employed for plasma extraction, with Carbamazepine and Abiraterone-D4 as internal standards for Olaparib and Abiraterone acetate, respectively. Validation parameters—linearity, precision, accuracy, recovery, stability, and sensitivity were assessed in accordance with ICH M10, FDA, and EMA bioanalytical guidelines. Results and Discussion: The method exhibited excellent linearity (r² > 0.998) across the specified ranges. LLOQs were 25 ng/mL (Olaparib) and 1.25 ng/mL (Abiraterone acetate), representing improved sensitivity versus earlier assays. Recoveries were 93–99%, with intra- and inter-day precision <10% CV. Stability under bench-top, freeze–thaw, and long-term conditions remained within ±15%. A total run time of 6 min ensured high throughput and reduced solvent consumption. Conclusion: The validated LC-MS/MS method is robust, sensitive, and reproducible, and is well-suited to pharmacokinetic and drug–drug interaction studies in oncology research. This method is suitable for clinical pharmacokinetic and drug–drug interaction studies involving prostate cancer therapies.

Hepatoprotective effects of a polyherbal extract (PHE) containing Averrhoa carambola and Lepidium sativum with trigonelline against CCl4-induced hepatotoxicity in Wistar rats

Suchita Gupta — 2026-03-15

Background: This study was designed to evaluate the hepatoprotective potential of a polyherbal extract (PHE) containing Averrhoa carambola, Lepidium sativum, and trigonelline against carbon tetrachloride (CCl₄)-induced liver injury in Wistar rats. Although individual components possess known antioxidant and hepatoprotective properties, comprehensive pharmacological evaluation of their combined use remains limited. This study aimed to investigate possible synergistic effects on liver function, oxidative stress, and hepatic tissue morphology. Methodology: Seven groups of Wistar rats (n = 6 per group) were administered PHE orally at doses of 400 and 600 mg/kg body weight for 21 consecutive days. On day 22, hepatotoxicity was induced using CCl₄. Hepatoprotective activity was assessed by measuring serum liver biomarkers, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total bilirubin. Oxidative stress parameters, including superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and lipid peroxidation (LPO), were also evaluated. Histopathological examination of liver tissues was performed. Result and Discussion: PHE significantly reduced serum ALT, AST, ALP, and bilirubin levels by 55–65% compared to the toxic control group. Lipid peroxidation decreased by approximately 60%, while antioxidant enzymes SOD and CAT increased by over 70%, demonstrating marked attenuation of oxidative stress. Histopathological analysis revealed substantial preservation of hepatic architecture, comparable to that observed with the standard drug silymarin. Conclusion: The strong antioxidant and hepatoprotective properties of PHE suggest its potential as a holistic alternative to conventional therapies for liver damage. Its ability to restore liver function and structure highlights its effectiveness against chemically induced hepatocyte injury.

Development and validation of a visible spectrophotometric method for the determination of tretinoin in bulk and pharmaceutical formulations

Manikya Sastry Thuttagunta — 2026-03-15

Background: Tretinoin is a widely used dermatological agent, and its accurate quantification is essential for quality control in pharmaceutical formulations. Existing UV-spectrophotometric methods often lack the sensitivity and simplicity required for routine analysis. Methodology: A simple, rapid, and cost-effective visible spectrophotometric method was developed for the determination of tretinoin in bulk and 0.05% cream formulations. The method is based on the formation of a stable ion-pair complex between tretinoin and methylene blue in an alkaline medium (pH 9.8). The resulting complex exhibits maximum absorbance at 650 nm. Method validation was carried out in accordance with ICH Q2(R1) guidelines. Results and Discussion: The method showed linearity in the concentration range of 2–10 µg/mL, with a correlation coefficient (r²) of 0.999. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.0434 µg/mL and 0.1316 µg/mL, respectively. Precision and accuracy results were within acceptable limits. Recovery ranges for 0.05% tretinoin cream formulations are 98.7% to 101.1%, confirming the method’s suitability for real-sample analysis. Compared to existing UV methods, this approach offers enhanced sensitivity and operational simplicity. Conclusion: The proposed spectrophotometric method is a reliable and sensitive alternative for the routine quantification of tretinoin in pharmaceutical formulations. Its simplicity, cost-effectiveness, and compliance with regulatory validation standards make it well-suited for quality control laboratories.

Design and validation of a robust RP-UPLC approach for simultaneous quantification of emtricitabine, tenofovir, and nelfinavir in active pharmaceutical ingredients and finished dosage products

Gayathri N — 2026-03-15

Background: Fixed-dose combinations of antiretroviral agents such as Emtricitabine (EMT), Tenofovir (TEN), and Nelfinavir (NEL) are central to Human Immunodeficiency Virus (HIV) therapy. Reliable quantification of these drugs is essential for pharmaceutical quality assurance and therapeutic drug monitoring (TDM) in clinical research. Methodology: A reverse-phase ultra-performance liquid chromatography (RP-UPLC) method was developed to simultaneously estimate EMT, TEN, and NEL in bulk and combined dosage forms. Separation was performed on a Waters CHS C18 column using potassium dihydrogen phosphate buffer and acetonitrile (65:35 v/v, pH 3.0) at a flow rate of 0.3 mL/min, with UV detection at 260 nm. Method validation was conducted in accordance with ICH guidelines for linearity, accuracy, precision, sensitivity, robustness, and stability. Forced degradation studies were conducted under acidic, alkaline, oxidative, thermal, and photolytic conditions. Results and Discussion: Retention times were 1.004 min (EMT), 1.310 min (TEN), and 1.870 min (NEL), with excellent peak resolution. Recovery studies demonstrated high accuracy, 99.74% (EMT), 99.47% (TEN), and 100.33% (NEL). The method showed strong linearity (r² > 0.999), low LOD/LOQ values, and robustness. Stability-indicating potential was confirmed via forced degradation studies. The technique proved to be rapid, precise, and stability-indicating, making it suitable for routine pharmaceutical quality control. Its sensitivity and robustness suggest potential utility in biomedical applications, including TDM and pharmacokinetic studies in HIV patients. Conclusion: The validated RP-UPLC method enables accurate, precise, and rapid estimation of EMT, TEN, and NEL, supporting its application in both pharmaceutical and clinical research settings.

Extraction and physicochemical–thermal characterization of leaf mucilage from Butea monosperma with potential relevance to biodegradable materials

Aastha Sharma — 2026-03-15

Background: Growing concerns over the persistence of petroleum-based plastics have renewed interest in plant-derived polysaccharides as sustainable material precursors. Methodology: In this work, mucilage extracted from Butea monosperma leaves was examined using comprehensive physicochemical, rheological, thermal, morphological, and solid-state characterisation to assess its properties. Result and Discussion: The extraction process yielded 10.58±0.42% (n=3) mucilage. Powder flow analysis indicated acceptable handling characteristics, with an angle of repose of 29.17±1.34°, Carr’s index of 17.18±1.02%, and a Hausner ratio of 1.20±0.04. Aqueous dispersions (1.5% w/v) displayed shear-dependent viscosity over the range of 10–150 rpm, consistent with pseudoplastic flow behaviour. Dynamic light scattering measurements showed a mean hydrodynamic diameter of approximately 1085 nm, while the zeta potential value of –11.6 mV suggested moderate electrostatic stability of the dispersion. FTIR spectra confirmed the polysaccharide nature of the mucilage through characteristic hydroxyl, aliphatic, carbonyl, and glycosidic bands. X-ray diffraction revealed a predominantly amorphous structure with limited semi-crystalline domains. Thermal analysis showed minor mass loss below 190°C, followed by a principal degradation stage between 226 and 322 °C with an overall mass loss of ~61.9%, while DSC analysis identified multiple endothermic transitions, including a major event at 275.99°C. Conclusion: Collectively, these findings establish reproducible yield, non-Newtonian rheology, moderate thermal stability, and an amorphous solid-state profile for Butea monosperma leaf mucilage. The observed properties suggest their potential relevance as a plant-based biopolymer and merit further investigation through material fabrication, mechanical and barrier testing, biodegradation assessment, and safety evaluation.

LC-MS profiling and antidiabetic evaluation of ethanolic root extract of Asparagus curillus Buch.-Ham. ex Roxb. in streptozotocin-induced diabetic rats

Anoop Singh Negi — 2026-03-15

Background: Diabetes mellitus is a chronic metabolic disorder characterized by hyperglycemia due to impaired insulin secretion or action and represents a major global health concern, with high prevalence in India. Although several antidiabetic drugs are available, their side effects and limited efficacy highlight the need for safer alternatives, such as traditionally used medicinal plants, such as Asparagus curillus. Methodology: This work used LC-MS to create the phytochemical profile of the ERE of A. curillus and tested for its anti-diabetic potential in STZ-induced diabetic rats. FBG level and biochemical markers, including lipid profile, insulin, HOMA-IR, liver and kidney function, and HbA1c, were measured. Kidney, liver, and pancreatic tissues were examined microscopically. Results & Discussion: LC-MS analysis identified twelve major phytoconstituents, mainly steroidal saponins including Shatavarin I, Shatavarin IV, and sarsasapogenin. The ERE produced dose-dependent changes in the biological parameters. At the dose of 200 mg/kg, ERE reduced FBG by 37% and HbA1c by 22%, while at 400 mg/kg, reductions of 42% in FBG and 25% in HbA1c were observed compared to the diabetic control. Histopathological examination of organs showed corresponding microscopic changes consistent with the biochemical findings. Conclusion: The ERE showed significant antidiabetic activity, improving glycemic indices, insulin sensitivity, lipid profile, and renal parameters, as supported by histopathological findings. LC-MS confirmed the presence of steroidal saponins, suggesting that the extract may serve as a potential natural antidiabetic agent. However, further studies are required to elucidate the mechanism and to validate it in clinical trials.

Safety assessment of multi-drug exposure: acute toxicity study of atorvastatin, ramipril, and BQ-123 combination in rats

Astha Jaiswal — 2026-03-15

Background: Combination cardiovascular therapies such as atorvastatin, ramipril, and endothelin-A receptor antagonists are increasingly used to improve clinical outcomes; however, their concurrent administration may introduce unforeseen pharmacodynamic and pharmacokinetic interactions. To date, no acute toxicity evaluation of this three-drug combination has been reported. Methodology: An acute toxicity study was conducted in female Wistar rats in accordance with OECD Guideline 423. A single intraperitoneal dose of atorvastatin (10 mg/kg), ramipril (1 mg/kg), and BQ-123 (1 mg/kg) was administered to a test group (n = 3), while controls received vehicle only (normal saline, 0.9% NaCl). Animals were monitored for 14 days for clinical signs of toxicity, mortality, body-weight changes, and behavioral changes. Gross necropsy and histopathological examinations of major organs were performed on Day 14. Result and Discussion: No mortality or clinical signs of toxicity were observed throughout the study. Body-weight progression in treated rats was consistent with that of the control group, demonstrating normal physiological growth. Gross pathological examination revealed no organ abnormalities or injection-site reactions. Histopathological analysis confirmed intact tissue architecture in the liver, kidney, heart, lungs, spleen, brain, stomach, and intestines. The absence of clinical, physiological, and histological alterations suggests that the atorvastatin–ramipril–BQ-123 combination does not induce acute systemic or organ toxicity at the tested dose. Findings indicate no synergistic or additive toxicity from co-administration, although further repeated-dose and biochemical assessments are necessary to characterize long-term safety. Conclusion: A single intraperitoneal administration of atorvastatin, ramipril, and BQ-123 was well tolerated in rats, classified as low acute hazard per OECD 423. Additional sub-acute and chronic studies are recommended to support therapeutic safety.

Formulation and characterisation of polyherbal syrup against aldose reductase enzyme

Dipak Babaso Bhingardeve — 2026-03-15

Background: Traditional medicinal plants have been suggested to help regulate blood glucose levels and inhibit glycation. This investigation aims to formulate and evaluate the potential of herbal formulations containing plant extracts that may inhibit Advanced glycation end product (AGE) formation and provide therapeutic advantages in the management of diabetes. Methodology: The herbal syrup containing polyherbal extracts of onion, garlic, and cinnamon was developed at varying concentrations with a viscosity modifier, preservative, flavoring agent, and other excipients. The developed syrup formulations were physicochemically characterised, including antioxidant potential, an ALR (aldose reductase) inhibition assay, and a stability study. Results and Discussion: The formulated herbal syrup exhibited a dark brown colour with a slightly Pungent taste. The pH of the syrup was measured at 6.48, indicating a mildly acidic to neutral nature. The viscosity of the prepared herbal syrup formulations (F1–F4) ranged from 3.71 to 3.80 cP. All formulations exhibited concentration-dependent antioxidant activity. F3 demonstrated the highest antioxidant activity and aldose reductase inhibition with 59.58% and 66.44% inhibition at 1000 μg/mL, respectively. Overall, the findings suggest that the herbal syrup formulation exhibited excellent physical and chemical stability over 3 months. Conclusion: F3 formulation may be a valuable candidate for the prevention and management of oxidative stress-related disorders, including diabetic complications, owing to its notable aldose reductase enzyme activity and potent antioxidant potential.

Integrated LC–MS/MS bioanalysis for the simultaneous quantification of metformin HCl, pioglitazone HCl, and teneligliptin HBr hydrate in human plasma

Sejal H. Pandya — 2026-03-15

Background: Combination therapy is widely prescribed in Type II diabetes mellitus to maintain effective glycemic control. The rising use of multidrug regimens demands selective and reliable bioanalytical methods capable of simultaneously quantifying multiple antidiabetic agents in human plasma for pharmacokinetic and bioequivalence studies. Methodology: A rapid, sensitive, and cost-effective LC–MS/MS method was developed and validated in accordance with ICH M10, USFDA, and EMA guidelines for the simultaneous estimation of metformin hydrochloride, teneligliptin hydrobromide hydrate, and pioglitazone hydrochloride. The assay enabled triple-drug quantification within a single 7-minute chromatographic run, showing an estimated 12–40% reduction in analysis time versus previously reported 8–15-minute single- or dual-analyte methods. Separation was achieved on a Cosmosil CN column (150 × 4.6 mm, 5 μm) using 10 mM ammonium acetate and acetonitrile (40:60 %v/v). Plasma samples were prepared by protein precipitation followed by liquid–liquid extraction, and detection was performed in positive electrospray ionization multiple-reaction-monitoring mode. Results and Discussion: Strong linearity was obtained for all analytes (r² > 0.995). LLOQs were 10.0 ng/mL for metformin, 1.25 ng/mL for teneligliptin, and 5.0 ng/mL for pioglitazone. Metformin-D6 served as the internal standard for metformin, while saxagliptin was used as the internal standard for teneligliptin and pioglitazone to ensure appropriate normalization across chemical classes. Precision remained below 10% CV, recovery was consistent, and stability stayed within ±15% under tested conditions. Reduced runtime and unified multi-analyte detection improved analytical throughput and minimized solvent consumption without compromising regulatory compliance. Conclusion: The validated LC–MS/MS method provides a reliable, resource-efficient platform for concurrent quantification of combined antidiabetic drugs in pharmacokinetic, bioequivalence, and clinical studies.

Robust and ICH-validated UV-Vis spectrophotometric method for the determination of benfotiamine in bulk substance, formulated products, and plasma using Bratton-Marshall chemistry

K. Bhavya Sri — 2026-03-15

Background: Benfotiamine is widely used in the treatment of diabetic neuropathy and metabolic disorders. For the determination of pharmaceutical formulations and biological matrices, Spectrophotometry is widely employed in pharmaceutical analysis due to its simplicity and cost-effectiveness. The Bratton–Marshall reagent is used for Chromogenic reactions involving aromatic amines, producing colored complexes suitable for spectrophotometric quantification. The study aimed to develop and validate a simple, rapid, accurate, and precise UV–Visible spectrophotometric method for quantitative estimation of benfotiamine in bulk drug and human plasma using the Bratton–Marshall reagent. Methods: Benfotiamine was dissolved in 0.1 M hydrochloric acid and reacted with the BM reagent. In this reaction, sodium nitrite in an acidic medium diazotizes the primary aromatic amino group of benfotiamine, producing a pink Chromogenic exhibiting maximum absorbance at 550 nm. The Chromogenic method was further extended to a bioanalytical application for plasma samples. Protein precipitation was carried out using Acetonitrile to remove plasma proteins, followed by vortex mixing and centrifugation at 1000 rpm for 15 minutes. The resulting supernatant was analyzed spectrophotometrically. Results and Discussion: The developed method demonstrated linearity in the concentration range of 3–21 µg/ml with a correlation coefficient (R²) of 0.998. Accuracy ranged between 98.44% and 99.64%. Precision studies indicated %RSD values below 2% and remained within acceptable limits. The recovery of benfotiamine from human plasma was 98.91%, and the matrix effect showed acceptable variability. Conclusion: The proposed method is rapid, precise, and reliable for the quantification of benfotiamine in pharmaceutical and plasma samples.

Formulation and in vitro evaluation of crisaborole-loaded solid lipid nanoparticles - topical system for atopic dermatitis

Nisha V Kalayil — 2026-03-15

Background: Designing and testing a new way to deliver drugs through the skin using Solid Lipid Nanoparticles (SLNs) for sustained as well as enhanced topical delivery of Crisaborole. Methodology: Crisaborole was incorporated into SLNs using the Double Emulsification Solvent Evaporation Method. The SLNs were enhanced using a Box-Behnken design. The best-performing formulation was evaluated after optimization for particle size, etc, along with entrapment efficiency and in vitro release. Results: The SLNs exhibited a remarkable entrapment efficacy of 69.8 ± 1.1% to 80.6 ± 0.3%, and their particle size varied from 222.1 ± 8.5 nm to 440 ± 7.5 nm. In the in vitro release study of Crisaborole-SLNs, 92.7 ± 1.17 % was observed at 12 hours. The developed SLN formulation displays no significant change after three months of stability study under accelerated conditions. Conclusion: These results suggest that crisaborole-loaded solid lipid nanoparticles (SLNs) represent a promising vehicle for the topical delivery of crisaborole, with potential for enhanced dermal residence due to controlled release and nanoscale size within the skin, while minimizing systemic exposure.

LC-MS/MS based bio analysis of desidustat: from development to application

Shaik Ayesha Ameen — 2026-03-15

Background: Desidustat's potential as a medicinal treatment for anaemia has generated considerable interest. For pharmacokinetic analysis, therapeutic monitoring, and drug development research, precise quantification of Desidustat in biological matrices is crucial. Hence, more advanced methods are necessary for detection at minute concentrations (ng/mL level) in plasma. Methodology: A reverse-phase C18 column with an isocratic elution technique utilising acetonitrile and an aqueous buffer as mobile phase to accomplish chromatographic separation. To produce characteristic ion transitions for quantification, after electrospray ionization (ESI), tandem mass spectroscopy in positive ion mode was chosen for detection. The technique's linearity, sensitivity, precision, and accuracy were systematically improved and verified, and the validated method was successfully applied to pharmacokinetic studies. Results and Discussion: The developed method’s validation results confirmed good selectivity, sensitivity (LLOQ 6 ng/ml), excellent linearity, and acceptable precision and accuracy, with a remarkably low lower limit of quantification (LLOQ). Recovery and matrix effect studies showed minimal ion suppression and reproducible extraction efficiency. The stability of desidustat is demonstrated in rat plasma. Intra-day and inter-day precision and accuracy values were well within the acceptable limits prescribed by regulatory guidelines, confirming the method's reliability and reproducibility. The optimized LC–MS/MS conditions ensured high sensitivity and selectivity, making the method suitable for bioanalytical applications. Conclusion: A novel, highly sensitive, and reliable LC–MS/MS bioanalytical method for estimation of Desidustat in rat plasma was successfully developed and validated. The method provides a robust analytical tool to support pharmacokinetic studies, therapeutic monitoring, and future research related to Desidustat.

Computational insights into taxifolin's therapeutic mechanisms for Alzheimer’s disease

Ashif Anjukandan — 2026-03-15

Background: Alzheimer's disease (AD) poses a significant challenge for research. Taxifolin is a natural flavonoid and has potential in protecting against Alzheimer’s by inhibiting oxidative stress. This study aims to elucidate the multi-target effects of Taxifolin on Alzheimer’s disease by computational techniques. Materials and Methods: The target genes were mapped into a PPI network using the STRING database. Hub genes were identified using Cytoscape software. Gene Ontology and KEGG pathway analyses were performed to identify AD-related pathways. Finally, docking analysis was performed using CDOCKER and Autodock, and GROMACS molecular dynamics simulations were performed to study the ligand's behaviour. Results and Discussion: The results revealed promising drug-like properties, as assessed by Lipinski’s rule and ADMET predictions. A total of 47 human genes showed significant similarity (≥ 0.70), with 10,234 targets linked to Alzheimer’s disease. Of these, 673 genes were highly associated with the disease (GDA > 0.1). A Venn diagram identified 16 overlapping genes, including BACE1, DPP4, PIK3CA, MTOR, ESR2, and APP. Network analysis revealed interactions among Taxifolin, MTOR, SERPINE1, ESR2, PIK3CA, and NOS3. Taxifolin docking against Alzheimer’s targets identified 5T4B as the best hit, with the lowest binding energy in CDOCKER (−43.57 kcal/mol) and AutoDock Vina (−8.8 kcal/mol). A 100-ns MD simulation confirmed a stable 5T4B–taxifolin complex, showing structural stability (RMSD/RMSF), compactness (Rg), solvent exposure (SASA), and persistent hydrogen bonds. MM-PBSA analysis supported strong binding, primarily driven by van der Waals and electrostatic interactions. Conclusion: Hence, the study offers insights into the multi-target mechanisms of Taxifolin against AD.

Phytochemical characterization and antibacterial potential of Spirulina platensis

Arpitha MP — 2026-03-15

Background: Algae are among the most diverse types of organisms on the planet. They have many advantages for humans. The most significant one is blue-green algae, which contain bioactive compounds, such as those found in Spirulina platensis. Methodology: The present study examines the antibacterial properties and phytochemical composition of Spirulina platensis, isolated from different freshwater bodies in the Chitradurga district, Karnataka, India. Results & Discussions: The anti-bacterial activity of the selected bacterial strains was investigated, and some appropriate results were found, which are discussed in the following sections. The methanolic extraction has revealed the maximum activity and the highest inhibition zones against E. coli and P. aeruginosa, followed by the acetone and the hexane extracts. The GC-MS profile examined has shown a range of bioactive compounds, viz., hexadecenoic acid, methyl esters, n-hexadecenoic acid, and glycerine. The occurrence of these compounds represents glycerine, ester compounds & lipid-derived metabolites, and the aromatic metabolites produced have the potential to exhibit antibacterial activity by S. platensis. It has the highest potential in the pharmaceuticals, nutraceuticals, and biotechnologies. Conclusions: The presence of these substances suggests that glycerine, esters, and aromatic compounds act in concert to produce potentially broad antibacterial activity by S. platensis. It can be inferred that the methanolic extract has shown high bioactivity.

QbD assisted green and white analytical UPLC method quantification of pharmacopeia impurities of Pravastatin in bulk drug and pharmaceutical formulations

N. Usha Rani — 2026-03-15

Background: A QbD-assisted green and white UPLC method was optimized for the simultaneous quantification of pravastatin and its EP impurities B, C, and D. Methods: A Design of Experiments (DoE) approach was used to evaluate the influence of critical method parameters (CMPs) on key chromatographic responses. Results and discussion: The statistical modeling using quadratic response surface methodology yields excellent regression coefficients (R² = 0.9399 for Pravastatin–impurity C and R² = 0.9758 for impurities C-D) and significant F-values (p < 0.0001), confirming robust model performance. The optimized chromatographic conditions were ethanol and 0.1% formic acid (65:35, v/v) at 0.3 mL/min as the mobile phase, with detection at 239 nm and a Waters ACQUITY BEH C18 column (100 × 2.1 mm, 1.7 µm). These conditions provide sharp, symmetrical peaks at retention times of 0.31, 0.89, 1.16, and 1.43 min for impurity B, pravastatin, impurity C, and D, respectively. Method was validated in accordance with ICH Q2(R2) guidelines, and results demonstrate excellent linearity (r² > 0.999), precision (%RSD < 1%), recovery (98.17–101.09%), and sensitivity (LOD = 0.015 µg/mL; LOQ = 0.05 µg/mL). The AGREE metric yields the greenness score of 0.81, and the GAPI pictogram confirms the minimum environmental impact (E-factor = 7.0 × 10⁻²). Furthermore, RGB 12 whiteness assessment provides a whiteness brilliance (MB) score of 80.6%, indicating an optimal balance among analytical performance, eco-friendliness, and operational efficiency. Conclusion: The proposed method was robust, rapid, environmentally sustainable, and practically efficient for routine impurity profiling and quality control of pravastatin formulations.