Abstract The aim of the proposed study was to build and verify a rapid, sensitive, greener, and stability indicating Ultra-performance liquid chromatography photodiode array detector (UPLC-PDA) method for the estimation of paracetamol (PRL) in marketed formulations. Before developing and validating the UPLC method, a comprehensive evaluation of the quality parameters of the tablet formulations was conducted to ensure their compliance with pharmacopeial standards. This preliminary assessment included general appearance, weight variation, hardness, friability, and disintegration tests. These tests were essential in confirming the overall quality and integrity of the tablets before proceeding with the analytical method development using UPLC and they were found to be within the limits of pharmacopeial standards. The developed UPLC method was validated for several validation parameters and greenness. The greener mobile phase was composed of water and isopropyl alcohol (IPA) (98:2, v/v). The maximum absorbance of PRL was detected at a wavelength of 244.3 nm. The developed UPLC method was found to be linear 5.5–110 ng mL −1 range. And the LOD and LOQ of the PRL were found to be 0.63 ± 0.00 ng mL −1 and 1.91 ± 0.00 ng mL −1 , respectively. In addition, the developed UPLC method for PRL estimation was fast, accurate, precise, robust and highly sensitive. The excellent greenness profile was revealed by the computed values of the analytical Eco-Scale, ChlorTox and AGREE scales for the current technique, which are 93, 0.006 g and 0.82, respectively. We further employed the concept of ‘blueness’ assessment using the recently introduced Blue Applicability Grade Index (BAGI) algorithms. The obtained high BAGI score (77.5) demonstrated excellent applicability of the method. This novel UPLC-PDA approach provides a rapid, environmentally friendly, and highly applicable alternative to conventional techniques for analyzing active constituents in marketed formulations. The degradation of products appears, the current UPLC method was still able to identify PRL, exhibiting its selectivity and stability qualities. Five different marketed formulations named Tbt-1, Tbt-2, Tbt-3, Tbt-4 and Tbt-5 contained 97.35, 96.50, 105.27, 97.65 and 98.46% w/w of PRL, respectively. Moreover, out of five marketed formulations, four were scored, including Tbt-1, Tbt-3, Tbt-4 and Tbt-5 were found to contain 97.93, 98.47, 96.68, and 96.85% w/w of PRL, respectively. These findings exhibited that PRL in marketed formulations can be consistently standardized and analyzed their content by using the proposed UPLC-PDA method.

Abstract The binding interaction between arginine-poly(ester amide) (Arg-PEA), a novel polymeric carrier material, and RNA was investigated using capillary electrophoresis. The peak areas of free RNA molecules were analyzed after the interaction of RNA with various concentrations Arg-PEA in phosphate-buffered saline (pH 7.4) at different temperatures. The binding constant ( K A ) and number of binding sites ( n ) between Arg-PEA and RNA were calculated using the Lineweaver–Burk double reciprocal equation. Furthermore, by calculating the related thermodynamic parameters and performing computer-assisted molecular docking, the types of binding forces were investigated to confirm the mechanism of targeted binding. Results revealed that the unbound RNA molecules were effectively separated and detected by the capillary electrophoresis system. At 37 °C, the K A was 3.956 × 10 6 L·mol −1 and n was 2.48, indicating a powerful interaction between the two materials. The binding of Arg-PEA to RNA was primarily driven by electrostatic forces and supplemented by hydrogen bonding. These findings confirm the RNA-targeting efficacy of Arg-PEA used as a polymeric material for drug carrier construction. This study demonstrates capillary electrophoresis as a novel approach for studying the targeted binding of ligands to receptors, expanding the application scope of this technology.

Abstract Artemether (ART) and lumefantrine (LUM), two key components in antimalarial combination therapy, were assayed simultaneously and tested for dissolution using RP-HPLC. The purpose of this study is to present an eco-friendly RP-HPLC method that is combined with RAPI and MoGSA tools in order to assure an environmentally friendly analysis of both artemether and lumefantrine. In this method, acetonitrile, water, n-propanol, and trifluoroacetic acid were combined in a 50:40:10:1 ratio (v/v), at a flow rate of 1.3 mL min−1, to achieve chromatographic separation. The ART and LUM detections were performed using UV detectors set at 210 and 236 nm, respectively, for the first 6.6 and 21.1 min, respectively. Both analytes were linearly detected over the concentration range of 3–60 μg mL−1, with high recovery rates confirming their accuracy. ICH guidelines were followed to verify method reliability and reproducibility. Under specified dissolution conditions, at least 80% of the active substance of conventional immediate-release tablets was released within 45 min. Over a period of six months, accelerated stability tests confirmed the formulation's stability under stress conditions. Furthermore, MoGSA and RAPI scores classified the method as green and white respectively. In view of its robustness and durability, the method is suitable for routine quality control of ART and LUM in pharmaceutical formulations.

Abstract IntroductionThe standard decoction of Toddalia asiatica (L.) Lam. (FLZX) possesses notable pharmacological properties including anti-inflammatory, analgesic, antimicrobial, and antiviral activities. However, its complex chemical composition necessitates the development of comprehensive quality assessment protocols to ensure consistent therapeutic efficacy.ObjectivesThis study was designed to establish a reliable quality evaluation system for FLZX standard decoction through integrated analytical approaches, thereby facilitating quality control standardization.MethodsFifteen batches of FLZX standard decoctions were subjected to HPLC fingerprint analysis with similarity assessment. Multivariate chemometric analyses including cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were employed to evaluate origin-related variations. Characteristic peaks were identified and three major bioactive constituents were quantitatively determined.ResultsThe established HPLC fingerprints contained eight characteristic peaks, with inter-batch similarities exceeding 0.900. Three bioactive markers (hesperidin, nitidine, and nitidine chloride) were unambiguously identified. Chemometric analysis revealed significant quality variations associated with geographical origin and harvesting period. Quantitative determination yielded concentration ranges of 0.850–3.829 mg g−1 for hesperidin, 3.612–10.251 mg g−1 for nitidine, and 0.087–0.335 mg g−1 for nitidine chloride, with corresponding transfer rates of 7.478–41.458%, 32.927–93.989%, and 3.235–17.392%, respectively. The extraction yields varied from 7.241% to 14.466%.ConclusionThe combination of HPLC fingerprinting with chemometric pattern recognition establishes a robust quality assessment platform for FLZX standard decoction, providing both methodological and practical foundations for quality control in the production of related pharmaceutical preparations.

Abstract A research experiment was conducted at Fruit Research Station, Navsari Agricultural University, Gandevi, Gujarat to study the persistence, dissipation and health risk assessment of spinosad and acephate in sapota fruits. The first spray of insecticides was given at marble size fruit stage followed by 15 days of second spray. About 1.5 kg of sapota fruits were collected from each treatment at 0 (2 h), 1, 3, 5, 10, 20 and 30 days after last spray of insecticides. Analysis of the insecticide residues in sapota fruits was carried out by using the QuEChERS analytical method. On the basis of persistence and dissipation study, the half-life (DT50) of spinosad and acephate in sapota fruits under recommended and double to recommended dose were varied from 1.11 to 1.32 and 2.52–3.73 days, respectively. The waiting period (WP) of spinosad and acephate in sapota fruits under recommended dose were 0.73 and 22.62 days, respectively. Further, Risk Assessment was evaluated and observed no risk for Indian consumer except 1–3 age group of children, due to studied insecticides under recommended doses (RQ < 1) in sapota fruits.

Abstract A rapid, sensitive, and environmentally friendly reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the quantitative determination of osimertinib, a third-generation EGFR tyrosine kinase inhibitor, in pharmaceutical formulations. The method utilizes an isocratic mobile phase of acetonitrile and 0.1 M ammonium acetate (1:1, v/v), with chromatographic separation achieved on a C18 column and detection at 276 nm. Method validation demonstrated excellent linearity over the range of 2.40–20.29 μg mL−1 (R2 = 0.9988), high accuracy (mean recoveries 90–105%), and precision (RSD <1% intra-day, <2% inter-day). Robustness testing confirmed the method's resilience to minor operational changes, and sensitivity was reflected in low limits of detection (0.843 μg mL−1) and quantification (2.55 μg mL−1). Forced degradation studies revealed significant osimertinib instability under acidic, alkaline, and oxidative conditions, but relative stability under thermal stress, confirming the method's stability-indicating capability. Greenness assessment using the AGREE tool yielded a score of 0.82, reflecting strong adherence to green analytical chemistry principles. This validated method is suitable for routine quality control and stability studies of osimertinib, offering both analytical reliability and environmental sustainability.

Abstract Phellopterin, a naturally derived furanocoumarin, is widely distributed in medicinal plants like Angelica dahurica and Toddalia asiatica, as well as in citrus fruits. In this experiment, an UPLC-MS/MS bioanalytical method over the concentration range of 0.725–290 ng mL−1 was established for the quantitative analysis and pharmacokinetic study of phellopterin in rat plasma. Following protein precipitation with acetonitrile, the samples were separated by gradient elution on a UPLC system, using acetonitrile-0.1% formic acid as the mobile phase. Quantification was performed using multiple reaction monitoring (MRM) method in electrospray positive-ion mode (ESI+), with the ion transitions of m/z 301.1 → 233.4 for phellopterin and m/z 209.1 → 194.1 for the internal standard, fraxetin. The accuracy of phellopterin ranged from 87.9 to 112.4%, with the precision between 4.1 and 12.2%. The matrix effect was 92.4–97.4%, and the recovery was over 77.9%. The optimized UPLC-MS/MS bioanalytical method was successfully used to study the pharmacokinetics of phellopterin in rats. The AUC(0−t) values were 74.2 ± 7.9 ng mL−1*h for intravenous and 37.1 ± 5.3 ng mL−1*h for oral administration, while the corresponding half-lives (t1/2z) were 1.1 ± 0.2 h and 3.1 ± 0.7 h, respectively. The bioavailability of phellopterin was determined to be 16.7%.

Abstract In accordance with the International Coordinating Committee's recommendations, an acceptable UPLC-MS method is being developed for analyzing Relugolix stability under various stress scenarios. Stress conditions included exposure to 0.1 and 1 N acid, alkali, hydrolysis, peroxide, light, reduction, and heat. The mobile phase for this analysis was prepared by blending acetonitrile and a 0.1% orthophosphate solution in a 50:50 (v/v) ratio. A C18 Column with dimensions of 100 × 2.1 mm and a particle size of 1.7 µm was employed as the stationary phase. The wavelength at which detection was performed was 247 nm, and the flow rate was maintained stable at 0.5 mL min−1. Strong linearity was shown by this method across a concentration range of 1.25 μg mL−1 to 7.5 μg mL−1. The method's reliability was shown by ensuring all of the validation parameters fell within the acceptable range.Furthermore, forced degradation studies were conducted using standard drugs. Different stress conditions led to the formation of degradation products, which were successfully separated from Relugolix using the C18 column in combination with an auto-purification mass spectrometer. Among the stress conditions, alkaline conditions showed a lower degradation rate. Interestingly, seven distinct types of degradation products were identified from the degradation studies under acid, alkali, hydrolysis, hydrogen peroxide, light, reduction, and thermal conditions. Remarkably, these seven degradation products have not been reported in previous literature, making this study the first to document their existence.

Abstract A simple, rapid, and green high-performance liquid chromatography-mass spectrometry (HPLC-MS) method was developed for determination of tauroursodeoxycholic acid (TUDCA), taurocholic acid (TCA), and taurochenodeoxycholic acid (TCDCA) in bio-transformed Jindanfen (BTJDF), which is obtained from chicken bile through a bioconversion process. The sample was prepared using water. The HPLC separation was operated on a poroshell 120 EC-C18 column with a 2.0 min gradient elution procedure. Detection was performed on a single quadrupole mass spectrometer in negative mode with selected ion monitoring mode (SIM). This developed HPLC-MS method presented good linearity (r > 0.997) and sensitivity (limit of quantification, 30.0–80.0 pg) for three analytes. The relative standard deviations (RSDs) for precision, repeatability, and stability were all below 3.00%. The matrix effects and average recoveries of three analytes were 91.2–97.9% (RSDs < 1.50%) and 95.4–103% (RSDs < 3.00%), respectively. The average contents of TUDCA, TCA, and TCDCA in ten batches of samples were 33.8, 13.2, and 20.5%, respectively. Finally, the greenness of the developed method was validated by Analytical Eco-Scale and Complex-GAPI. The developed method was proved to be an eco-friendly, effective, and reliable approach for assaying the three cholic acids in BTJDF, which is help to improve the quality evaluation level of the BTJDF industry.

Abstract Apples are among the most common fruits, produced on the territory of North Macedonia. Before they reach the market, it is essential to be tested for pesticide residues, which are used for protection against pests for this culture, and in order to ensure the customer safety. For this reason, a novel and simple method for simultaneous determination of captan, folpet, difenoconazole and chlorpyrifos in apple samples has been developed and validated. Acetone is used for extraction of the pesticide residues, followed by liquid-liquid (LLE) and solid–phase extraction (SPE). Separation and quantification of analytes is achieved on reversed-phase high performance liquid chromatography (RP-HPLC) with UV diode array detector (UV-DAD). The best results are obtained using analytical column LiChrospher 60 RP-select B (250 mm × 4 mm, 5 µm), with isocratic elution and acetonitrile/0.1 % acetic acid in water (70:30, V/V) as a mobile phase. The flow rate is 1 mL min−1, and UV detection is performed at 220 and 230 nm. The linearity of the method is tested in the range of 1.50–3.60 mg kg−1 for captan and folpet, and 0.35 – 0.60 mg kg−1 for difenoconazole and chlorpyrifos. The obtained values for recovery and RSD ranged from 94.94 to 114.63 %, and 0.09–9.25 %, respectively. The validated method is successfully applied to apple samples for the determination of the investigated pesticide residues.