A thin-layer chromatography-densitometry (TLC-densitometry) method was developed and evaluated for the identification of two drugs acting on the central nervous system – trazodone and donepezil. Various chromatographic systems were analysed, selecting optimal separation conditions using 60F254 silica gel plates and a mobile phase consisting of toluene, acetone, ethanol, and ammonia (9:5:1:0.5; v/v/v/v). The resulting conditions enabled effective separation of both compounds, achieving RF values of 0.53 for trazodone and 0.67 for donepezil, respectively. The study assessed the suitability of fourteen visualizing reagents for detection of trazodone and donepezil. Bromophenol blue, rhodamine B, phenol red, metanil yellow, and brilliant green were shown to provide the most effective identification of the tested substances. Bromophenol blue demonstrated particularly favourable properties, allowing both visual differentiation between trazodone and donepezil and maintaining parameters enabling quantitative analysis in the range of 0.1–1.0 µg/spot. Densitometric analysis confirmed good quality of chromatographic separation (Rs = 1.7; α = 1.8), while spectrodensitometric studies demonstrated that the visualizing reagents used significantly affected the number and character of absorption bands. Brilliant green demonstrated the greatest identification potential, generating distinct spectral differences between the tested drugs, including additional bands in the visible light range. The obtained results indicate that TLC-densitometry combined with appropriately selected visualizing reagents is a simple, inexpensive, and effective method for identifying trazodone and donepezil. The developed procedure can be used in routine quality control of medicinal products, pharmaceutical analysis, and screening studies.
A rapid and robust reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and fully validated strictly as a quantitative assay for the high-throughput determination of semaglutide (SMG) in both active pharmaceutical ingredient (API) powder and solutions for injection. Chromatographic separation was achieved on a Phenomenex Aeris™ 3.6 µm Widepore XB-C18 column (150 × 2.1 mm) using a mobile phase consisting of 100 mM ammonium dihydrogen phosphate buffer combined with acetonitrile , with the addition of perchloric acid using an isocratic elution. The advantage of the developed methodology is the compression of the total run time to 6 minutes, which introduces an over three-fold increase in throughput compared to existing methods. The method was successfully validated according to ICH Q2(R2) , showing specificity, linearity (R2 = 0.998 within 70%–130% range), precision (%RSD ≤ 1.73%) and accurate recovery (98.8% – 100.2%) for bulk and final formulation samples.
The escalating antimicrobial resistance crisis demands novel therapeutic agents, with Lucilia sericata (Ls) larval secretion/excretion (S/E) offering potent antibacterial and pro-healing properties for chronic wounds. While high-molecular-weight antimicrobial peptides are well-studied, low-molecular-weight (LMW, <3 kDa) fractions induced by pathogens remain underexplored. This study isolated LMW fractions from S. aureus-induced Ls S/E via ultrafiltration and size-exclusion chromatography, yielding eight fractions (WF1-8). Fractions showed dose- and species-specific bacteriostatic effects, with WF3 strongly inhibiting S. aureus viability (~17% reduction, GLM Wald χ²=46.8, P<0.001) while WF1 and WF4 demonstrated depigmenting P. aeruginosa via potential quorum-sensing disruption. Fractions WF1, WF3, and WF4 potently stimulated HDMEC proliferation (>200% vs. control, dose-dependent r=0.70, P<0.0001, MTS assay) with negligible cytotoxicity, while HUVECs responded modestly to WF1 and WF7, underscoring dermal relevance. LC-MS/MS and MASCOT analysis identified 34 unique peptides (7-30 residues) with high antimicrobial probability, enriched in active fractions. These findings reveal pathogen-inducible LMW synergists for targeted antimicrobials and angiogenesis, advancing maggot-derived therapies; mechanistic validation in vivo is subsequent.
Orodispersible tablets are intended to melt or dissolve in the mouth when taken orally. Montelukast sodium and Loratadine belong to BCS class II i.e. they have low solubility but high permeability characteristics. By merging natural or synthetic polymers, novel co-processed excipients like Ludiflash and Pharmaburst® are formed to enhance both stability and performance of dosage form. The aim of this study was to design and optimize orodispersible tablets (ODTs) of Montelukast sodium/Loratadine while evaluating their pre- and post-compression characteristics along with compatibility. Using direct compression technique, seven different formulations were manufactured by incorporating various excipients in different concentrations, including Ludiflash® and Pharmaburst®. All formulations underwent pre-formulation tests including bulk density, tapped density, angle of repose, etc. followed by post-compression tests like weight variation, thickness; diameter, hardness, wetting time, water dispersion, dissolution and disintegration. Formulation F6 was selected as optimized formulation as all parameters comply with USP standards. The optimized formulation (F6) showed high similarity in dissolution profiles (f1 = 0.27 < 15; f2 ≈ 100), indicating comparable in vitro dissolution performance. A simple, reliable, precise and accurate method of assay was developed by HPLC (Shimadzu, CTO-20AC). All the results found within the specified limits of the BP/USP. By the use of direct compression method, the development of ODTs of montelukast sodium/Loratadine having co-processed excipients, proved to be an effective approach with minimal excipients use. Moreover, the validated HPLC method established a simple, precise and reproducible technique for regular quality control assessments.
Non-alcoholic fatty liver disease (NAFLD) ranks among the most prevalent liver diseases globally. Icariin (ICA) has been shown to improve NAFLD, but the specific molecular mechanisms remain unclear. The study was to probe the underlying mechanisms of ICA in NAFLD. Quantitative real-time PCR (RT-qPCR) was employed to detect gene levels. Enzyme-linked immunosorbent assay (ELISA) was utilized to measure the concentrations of inflammatory factors. The target-binding relationship was validated through a dual-luciferase reporter assay and RNA Immunoprecipitation (RIP) assay. Our findings revealed that ICA dose-dependently decreased the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C), while increasing high-density lipoprotein cholesterol (HDL-C) levels in the NAFLD model. Additionally, it dose-dependently reduced the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-12 (IL-12), and elevated miR-185-3p expression. In free fatty acids (FFA)-treated HepG2 cells, the miR-185-3p inhibitor attenuated the suppressive effect of ICA on inflammatory factors and its promoting effect on miR-185-3p levels. Fractalkine (CX3CL1) levels were diminished in an ICA dose-dependent manner. In FFA-treated HepG2 cells, silencing miR-185-3p elevated the secretion of inflammatory factors, whereas si-CX3CL1 reversed this effect. ICA alleviated liver damage, lipid metabolism disorders, and inflammatory responses via the miR-185-3p/CX3CL1 axis, thereby slowing the progression of NAFLD.
