ABSTRACT Dual drug delivery systems have transformed breast cancer therapy by enhancing efficacy and reducing side effects. While Doxorubicin (DOX) is highly effective against breast cancer, its clinical utility is limited by substantial adverse effects. Co‐delivery with Baicalein (BCL) in a liposomal formulation offers a promising strategy to mitigate these effects while improving therapeutic outcomes. However, a reliable analytical method is essential for accurate quantification of both drugs in complex formulations. A robust and validated high‐performance liquid chromatography (HPLC) method was developed to simultaneously facilitate DOX and BCL quantification. The method exhibited high sensitivity, with a limit of quantification below 0.3 µg/mL and excellent linearity over a range of 0.0195 to 10 µg/mL. Liposomal formulation co‐loaded with DOX and BCL was prepared using the thin‐film hydration method to validate the method's applicability. The developed HPLC method successfully estimated the entrapment efficiency of both drugs. In addition, in‐vitro drug release studies confirmed the stability of liposomes in acidic conditions and demonstrated sustained drug release in the intestine. This validated method, compliant with International Council for Harmonization guidelines, provides a reliable tool for analyzing and optimizing dual drug delivery systems, paving the way for advanced therapies in breast cancer treatment.

ABSTRACT Vonoprazan is an innovative potassium competitive acid blocker and has emerged as an effective alternative to proton pump inhibitors for the treatment of acid‐related disorders. High‐performance liquid chromatography was performed using a Symmetry C18 column (150 mm × 4.6 mm ID, 3.0 µm) with a mobile phase flow rate of 1.0 mL/min. Mobile phase A consisted of ammonium formate, pH adjusted to 3.0 and mobile phase B was ethanol. The analysis employed a gradient elution method. This method was used for the quantification of degradant impurities, including impurity A and impurity C, originating from vonoprazan. The accuracy results demonstrated that the results for impurities A and C ranged from 100.6% to 102.6%. Additionally, the sample solution was confirmed to remain stable under all evaluated conditions for up to 24 h. Using analytical quality by design principles and design expert software, method robustness was assessed in terms of mobile phase flow rate, column temperature, and mobile phase buffer pH variability. Moreover, the method was assessed for environmentally sustainable behavior using the analytical eco‐scale evaluations, analytical greenness, and green analytical procedure index.

ABSTRACT This study aimed to develop and validate a straightforward HPLC method for simultaneously estimating mesalamine and quercetin in both pure forms and solid lipid nanoparticle formulations. Following International Council for Harmonisation Q2(R2) guidelines, the analytical method was validated. Furthermore, this optimized method was applied to measure the percentage entrapment efficiency and in vitro drug release in central composite design‐based optimized solid lipid nanoparticle formulations. The chromatogram showed the peak of mesalamine and quercetin at retention times of 2.28 and 8.79 min, respectively. For mesalamine, limits of detection and quantification were found to be 1.64 and 4.97 µg/mL, respectively. Whereas for quercetin, limits of detection were 2.10 µg/mL and limit of quantification was 6.36 µg/mL, respectively. The entrapment efficiency of mesalamine and quercetin in solid lipid nanoparticles was 85.27 ± 1.74% and 78.62 ± 2.38%, respectively. Furthermore, the in vitro drug release study indicated 98.83 ± 2.94% release of mesalamine and 96.56 ± 1.92% release of quercetin from solid lipid nanoparticles over 24 h. The results demonstrated that the HPLC method developed was accurate, precise, and robust, maintaining its reliability despite minor changes in flow rate and wavelength. Consequently, this method was suitable for estimating mesalamine and quercetin in various formulations.

ABSTRACT Liu Shenqu (Massa Medicata Fermentata [MMF]) is a natural fermented product of flour (and/or wheat bran) and five medicinal herbs, Prunus armeniaca , Vigna umbellata , Artemisia annua , Polygonum hydropiper , and Xanthium strumarium . Unprocessed, stir‐fried, and charred MMF has been used therapeutically in China for over a thousand years to treat conditions such as dyspepsia, abdominal distension, and diarrhea. Variations in quantities, fermentation conditions, and processing procedures across different production regions can influence the quality of MMF products, ultimately affecting their efficacy. Therefore, it is essential to establish robust qualitative and quantitative methods for the quality control of MMF. In this study, betaine, a bioactive compound, was identified via TLC, isolated using preparative TLC and analyzed using NMR spectroscopy in MMF. Ultrasonic extraction of betaine from MMF samples was optimized using orthogonal experimental design. Following aluminum oxide (OH − type) solid‐phase extraction, a simple, rapid, and accurate HPLC method was developed for the quantification of betaine, utilizing a HILIC column. Analysis of 58 commercially available MMF batches revealed that betaine concentrations in 49 batches (84.5%) ranged from 0.75 to 4.0 mg/g, whereas only 9 samples contained concentrations below 0.5 mg/g. Notably, betaine levels were significantly lower in charred products compared with unprocessed ones ( p < 0.001), suggesting that betaine may serve as a potential biomarker for the quality control of MMF.

ABSTRACT Coreopsis tinctoria Nutt. (CT) has the effects of clearing heat and detoxifying, which can be used to treat red‐eye swelling. At present, there are studies on CT treatment of diabetic retinopathy (DR). Diabetes can affect the function of the eyes, which is closely related to the liver. The connection between the liver and the eyes is valued as the liver–eye axis. There are no reports on the mechanism of CT action on the liver in DR. This study analyzed the differences in the liver in DR rats before and after administration using network pharmacology, LC‐MS‐based non‐targeted metabolomics, and energy substances metabolism. Then, a joint analysis of hepatomics, plasma metabolomics, energy metabolism, and network pharmacology results was conducted to discover the overall biological axis pattern. Non‐targeted metabolomics revealed the pentose phosphate pathway (PPP) and riboflavin pathway are the main enriched metabolic pathways, closely related to energy metabolism. Energy substance metabolism revealed CT regulates liver energy metabolism, and the tricarboxylic acid cycle is the main enriched metabolic pathway. Integrated analysis revealed the PPP is the upstream key pathway. The research reveals the mechanism of CT treatment for DR based on the liver and provides a reference for studying the liver–eye axis.

ABSTRACT Sunitinib belongs to the tyrosine kinase inhibitors class of medical drugs prescribed to treat various types of cancers, including renal cell carcinoma, gastrointestinal stromal tumors, and pancreatic neuroendocrine tumors by blocking the action of enzymes that promote cancer cell growth. Forced degradation studies are crucial for assessing drug stability and will be helpful in drug discovery and development. In this study, sunitinib underwent different stress testing according to ICH guidelines to identify and analyze its degradation products (DPs). The stress study findings proved that sunitinib is highly sensitive to acid (2DPs), base (1 DP), and oxidation (1 DP) degradation but remains stable in other stress conditions with significantly less degradation. The separation of sunitinib and its impurities and four DPs was achieved using a Waters XBridge C18 column (250 mm × 4.6 mm, 5 µm) with 0.7 mL/min gradient flow of ammonium formate buffer (pH 4.8) and acetonitrile with a detection wavelength of 258 nm. These conditions produce well‐retained and resolved peaks corresponding to sunitinib at 8.4 min, whereas 4.8, 3.5, and 1.9 min are noticed for impurities 1, 2, and 3, respectively. These study findings provide a comprehensive analysis of sunitinib, its impurities, and DPs, offering crucial insights into its stability and toxicity that can enhance drug development and safety profiling. The study also detailed the degradation pathway of sunitinib, its impurities, and the fragmentation patterns of DPs, which had not been previously reported in any other studies. All these four DPs belong to Class 4 (moderate) toxicity with distinct pharmacokinetic profiles. Hence, it can be concluded that these study findings provide a comprehensive analysis of sunitinib, its impurities, and DPs, offering crucial insights into its stability and toxicity that can enhance drug development and safety profiling.

ABSTRACT The pharmaceutical combination of Racecadotril (RAC), Ofloxacin (OFL), and Ornidazole (ORN) is a commonly prescribed treatment for diarrhea and dysentery in pediatric patients. A rapid and highly sensitive reverse phase–high‐performance liquid chromatography–diode array detector (RP‐HPLC–DAD) method has been established for the concurrent analysis of all three drugs in a combined oral suspension. Drugs were separated by chromatography using a Sapphire octadecylsilane (ODS) C18 (150 × 4.6 mm, 5 µm) column, operated at 30°C. The elution solvent consisted of a mixture of acetonitrile and 0.1% triethylamine solution (70:30 v/v), with the pH adjusted to 3.1 using 10% orthophosphoric acid. The flow rate was maintained at 1 mL/min, and all three drugs were monitored at 230 nm. The method showed linearity within the ranges: RAC (3–9 µg/mL), OFL (10–30 µg/mL), and ORN (25–75 µg/mL). Retention times were 2.03 min for OFL, 2.99 min for ORN, and 6.99 min for RAC. The drug content for RAC, OFL, and ORN ranged from 100.92% to 101.39%, 98.55% to 100.76%, and 99.33% to 101.22%, respectively, with recoveries of 96.33%–102.16%, 97.50%–101.32%, and 98.25%–100.76%, and %RSD values consistently below 2 in within‐day, between‐day, and repeatability studies, demonstrating method precision. The projected chromatography technique has been authenticated in line with International Council on Harmonization Q2 (R1) guidelines. This approach offers a short runtime of 10 min, providing an economically efficient chromatographic technique for the concomitant assessment of RAC, OFL, and ORN in a combined oral suspension preparation.

ABSTRACT Menatetrenone, a form of vitamin K (MK‐4) is used for various pharmaceutical and agrochemical applications. Monitoring and control of process impurities is crucial in the manufacturing of pharmaceutical‐grade Menatetranone. In this article, we report a gas chromatography (GC)‐based analytical method for the detection and quantification of trace levels of five process‐related impurities in MK‐4 originating from the starting materials and reagents. A GC‐flame ionization detector (GC‐FID)‐based method is presented for simultaneous quantitative estimation of five structurally diverse compounds, some of which are conventionally analyzed by liquid chromatography. After a series of method development trials, an optimized method with a mid‐polar column dura bond‐624 (6% cyanopropyl/phenyl and 94% polydimethylsiloxane) with initial column oven temperature 150°C, detector temperature 280°C was finalized for the separation of these impurities. The developed GC‐FID‐based method was validated for specificity, the limit of detection (LOD), the limit of quantification (LOQ), linearity, range, precision, accuracy, and robustness as relevant to the International Council for Harmonization guidelines. The method was found to be highly sensitive for all the constituents with an LOD ranging from 0.05 to 0.43 pg and a limit of quantification of 0.17–1.41 pg, as estimated by the signal‐to‐noise ratios. This is the first report of an analytical method for the simultaneous quantitative determination of five process impurities of MK‐4.

ABSTRACT A herbo‐dental formulation, Dant‐Kanti natural toothpaste consists of 13 potent medicinal herbs. The present study aims to comprehensively identify the signature phytometabolites present in Dant‐Kanti natural toothpaste using high‐performance thin layer chromatography (HPTLC), ultra‐high‐performance liquid chromatography (UHPLC), and gas chromatography–mass spectrometry (GC–MS) methods. Among the identified phytometabolites, gallic acid, piperine, and anethole, were quantified by UHPLC, whereas camphor, menthol, thymol, and eugenol were quantified using GC–MS. An inductively coupled plasma‐mass spectrometry was used to estimate the mineral contents in Dant‐Kanti natural toothpaste. To the best of our knowledge, these studies are the first‐ever to identify all the signature components in an herbal dental formulation. These developed and validated methodologies were then used across 25 distinct batches of Dant‐Kanti natural toothpaste, in oder to ascertain the consistency and homogeneity. Both the UHPLC, and GC–MS methods used in this study were validated as per ICH Q2 (R1) guidelines, and were found to be suitable, linear ( r > 0.99), and precise. In the cell‐biological study, Dant‐Kanti natural toothpaste showed a concentration‐dependent reduction in proinflammatory cytokine release, in human macrophages induced with Porphyromonas gingivalis lipopolysaccharide. This robust study methodology, validation, and results provide valuable insights into the complex composition of Dant‐Kanti natural toothpaste, and contribute to the understanding of its potential oral health benefits.