ABSTRACT Gliclazide (antidiabetic) and montelukast (leukotriene antagonist) are frequently co‐prescribed in patients with Type 2 diabetes and coexisting asthma or allergy. However, no analytical approach has been documented for their simultaneous quantification in plasma using a simple, inexpensive RP‐HPLC‐UV assay, nor has their pharmacokinetic interaction been studied in vivo. This study developed and validated a new, rapid, and low‐volume RP‐HPLC for concurrent estimation of gliclazide and montelukast in rat plasma. Chromatographic separation was performed on a C 18 column utilizing isocratic elution with a 20 mM ammonium acetate buffer (pH 5.5) and acetonitrile in a 20:80 (v/v) ratio, at a flow rate of 1.0 mL/min, with a UV detector set at 228 nm. The method was validated in accordance with ICH M10 guidelines, encompassing selectivity, linearity, precision, accuracy, recovery, and matrix effect. The technique was utilized to characterize the pharmacokinetics of gliclazide, with and without montelukast, in male Sprague–Dawley rats. Gliclazide and montelukast eluted at 3.04 and 5.87 min, respectively, with resolution > 2 and tailing factors < 1.6. The method was linear over 5–10 000 ng/mL in plasma, with LOQs of 2.33 ng/mL (gliclazide) and 4.06 ng/mL (montelukast). Recovery exceeded 85%, and no significant matrix effects were observed. All stability conditions met the acceptance criteria. Co‐administration of montelukast significantly increased gliclazide exposure: C max increased by 116% and AUC 0–∞ by 180%, accompanied by prolonged t 1/2 and MRT and reduced elimination rate constant ( K el ), indicating a pharmacokinetic interaction. This study introduces the newly validated RP‐HPLC‐UV method for the concurrent determination of gliclazide and montelukast in plasma, offering the first in vivo evidence of a notable pharmacokinetic interaction between these two medications. This method is efficient, reliable, cost‐effective, and utilizes only 20 µL of plasma, rendering it exceptionally appropriate for routine preclinical drug–drug interaction investigations.

ABSTRACT Voxelotor is the first oxygen‐affinity modulator for hemoglobin. It was demonstrated that voxelotor could potentially modify the disease condition by raising hemoglobin levels and lowering hemolysis markers in sickle cell patients. The voxelotor was exposed to various kinds of degradation conditions, including hydrolysis, oxidation, and photolysis according to the conditions suggested by the International Council for Harmonization (ICH). Two voxelotor degradation impurities were detected during acid hydrolysis and oxidation (H 2 O 2 ). The in silico degradation profile was predicted using Zeneth. The degradation impurities were separated using ammonium formate (pH 6.5) and acetonitrile as the mobile phase with gradient elution and Phenomenex Gemini C18 column (250 × 4.6 mm, 5 µm) as a stationary phase. Liquid chromatography–high resolution mass spectrometry (LC‐HRMS) was used to characterize the separated degradation impurities. The structures of each degradation impurity were ascertained using the accurate masses acquired from LC‐HRMS/MS. A suitable mechanism for the formation of degradation impurities was proposed. The in silico studies for mutagenicity and toxicity were conducted using DEREK and SARAH Nexus.

ABSTRACT Progestins are a representative steroid sex hormone widely used in modern contraception and hormone replacement therapy. The accurate monitoring of its concentration in both biological and environmental matrices is crucial for safeguarding reproductive health, optimizing therapeutic regimens, and assessing environmental pollution. In this study, a magnetic covalent organic polymer was synthesized using a simple one‐step method and employed as an adsorbent for magnetic solid‐phase extraction to efficiently extract progestins from biological and environmental samples. This material was synthesized in just 3 h and provided abundant active sites, which enabled high extraction efficiency and yielded enrichment factors of 37–44 in only 10 min. Furthermore, the material exhibited excellent stability and reusability. The adsorption behaviors of the three target drugs on the material were well‐described by the Freundlich isotherm and pseudo‐second‐order kinetic models. Coupling magnetic solid‐phase extraction with high‐performance liquid chromatography achieved excellent analytical performance, including good linearity ( R 2 > 0.9998), low detection limits (0.15–0.18 ng/mL), and high precision (relative standard deviation ≤5.22%). This method was successfully applied to the rapid determination of progestins in lake water and plasma samples, yielding recoveries of 96.1%–102.6% and 99.0%–103.6%, respectively. In conclusion, the proposed method is highly sensitive, reproducible, and practical, showing great potential for the preconcentration and determination of trace progestins in complex samples.

ABSTRACT The approach provides a reliable framework for standardization and quality control of Daribu‐5. Qualitative profiling was conducted on high‐performance liquid chromatography coupled with quadrupole–orbitrap high‐resolution mass spectrometry (HPLC‐Q‐Exactive‐MS). Quantitative analysis was performed on HPLC–MS/MS. Eight marker constituents, namely, geniposide, liquiritin, rutin, quercetin, isorhamnetin, glycyrrhizic acid, costunolide, and dehydrocostus lactone, were quantified across 15 batches of Daribu‐5 by multiple reaction monitoring (MRM) in negative ion mode. The resulting data were analyzed using the Wei Sheng Xin platform and SIMCA 14.1. Quality‐discriminating markers were identified as variables with variable importance in projection (VIP) >1.0. The entropy‐weighted TOPSIS method was then applied to rank the overall quality of the batches. Seventy‐nine components were identified in Daribu‐5. The eight marker compounds exhibited excellent linearity within their respective ranges ( r ≥ 0.9990) cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares‐discriminant analysis (OPLS‐DA) classified the 15 batches into two distinct groups: S3, S8–S9, and S13 formed one cluster, whereas S1–S2, S4–S7, S10–S12, and S14–S15 formed another. Four differential markers, glycyrrhizic acid, isorhamnetin, quercetin, and rutin, were identified as the main contributors to quality variation (VIP > 1.0). Entropy‐weighted TOPSIS analysis revealed Euclidean closeness values ranging from 0.182 to 0.663, indicating significant batch‐to‐batch variability. Among the samples, S2, S4, S11, and S15 ranked highest in quality. The established method is rapid, reliable, and effective for both qualitative and quantitative evaluation of Daribu‐5. Chemical pattern recognition enables the identification of key quality markers, and when combined with entropy‐weighted TOPSIS, provides a comprehensive and objective framework for batch quality assessment and quality control of Daribu‐5.

ABSTRACT Pesticides are used to control, prevent, or eliminate pests that threaten crops, public health, and stored products. Malathion, an organophosphate pesticide, remains a priority contaminant across water, food, and soil, leading to environmental contamination and bioaccumulation in ecosystems. Their widespread and often indiscriminate use has raised significant concerns. This comprehensive review presents recent advances in the detection of Malathion using spectrometric, chromatographic, and biosensor techniques. Spectrometric methods deliver rapid screening with minimal sample preparation. Chromatographic workflows provide confirmatory quantification and separation, whereas immunoassays are required for pesticide residue quantification or screening. Electroanalytical sensors and advanced biosensors can be categorized into different types, such as non‐enzymatic, colorimetric, fluorescence‐based, including metallic organic framework (MOF‐based), aptamer‐based, and molecular imprinted polymer (MIP‐based), offering low‐cost, real‐time monitoring of samples. Future advancements in Malathion detection by integrating nanomaterials, microfluidics, and aptamer or antibody‐based recognition elements in biosensors, enabling faster, more selective, and ultra‐sensitive detection. The convergence of spectrometric, chromatographic, and biosensor technologies promises comprehensive tools for environmental monitoring, food safety, and health diagnostics in the upcoming years.

ABSTRACT Oxalis corniculata has a long application history in traditional medicines and exhibits diverse pharmacological activities. This work established an effective and efficient ultra‐high performance liquid chromatography‐electrospray ionization‐quadrupole‐time of flight/high‐resolution mass spectrometry (UHPLC‐ESI‐Q‐TOF/HRMS), with precise and rapid separation yield. The method was applied to the qualitative and quantitative analysis of loliolide in the herb and biological samples taken from rats after a single oral administration of the herb decoction, for the first time. The bioanalytical validation of this UHPLC‐ESI‐Q‐TOF/HRMS was completed successfully. The correlation coefficients of calibration curves were more than 0.997 in the concentration range from 10 to 1000 ng/mL. The average matrix factor was 0.953. The intra‐day and inter‐day accuracy varied from 85.0% to 110.4%, with relative error from − 15.03% to 10.39% and relative standard deviation (RSD) from 0.38% to 14.41%. The recovery ranged from 83.8% to 113.9% with RSD from 0.20% to 6.99%. The lower limit of quantification was 10 ng/mL. The content of loliolide in the dry herb was detected to be 50.2 mg/kg. UHPLC‐ESI‐Q‐TOF/HRMS analysis indicated that loliolide could be absorbed into the rat systemic circulation with a C max at 1.5 h. Loliolide could be distributed into major organs and reached its C max at 5 min after oral administration, except in the spleen at 15 min. The in vivo distribution of loliolide in rats exhibited concentration fluctuation after reaching C max . The maximum brain uptake index of 98.8 ng/g in wet weight provided a solid material basis for its neuropharmacological effects. Furthermore, a series of complex metabolic schemes in rats were deduced from their molecular performance in HRMS and HRMS/MS, including dehydration, deoxygenation, demethylation, dehydrogenation, hydrogenation, glucuronidation, and hydrolysis.

ABSTRACT Promethazine hydrochloride is a phenothiazine derivative and is used for the treatment of antihistamine, antipsychotic, antiemetic, analgesic, anticholinergic, and sedative. During the routine analysis of promethazine hydrochloride, an unknown broad peak appeared using the related substances method indicated in the US Pharmacopeia. The separation method was optimized, and an unknown peak at a level of 0.11% was detected. Further study showed that the impurity could be detected in both USP reference standard and tablet samples, with the levels of 0.01% and 0.11%, respectively. In order to identify and characterize the impurity, high performance liquid chromatography‐tandem mass spectrometry technology was conducted to propose the chemical structure. 1D and 2D NMR were used to confirm the chemical structure of the unknown impurity as 1‐(10 H ‐[3,10′‐biphenothiazin]‐10‐yl)‐ N , N ‐dimethylpropan‐2‐amine. The formation of the impurity during the synthesis route was illustrated. The current study provided an optimized method for the quality control of promethazine hydrochloride drug substance and dosage forms.

ABSTRACT Radix scutellariae and Licorice are representative herbs in traditional Chinese medicine, widely employed for the management of inflammatory disorders owing to their well‐recognized anti‐infective and anti‐inflammatory properties. In clinical practice, they are often prescribed in combination; however, their precise mechanisms of action and metabolic interactions remain insufficiently elucidated. In the present study, an ultra‐high‐performance liquid chromatography‐quadrupole time‐of‐flight‐tandem mass spectrometry approach was utilized to comprehensively profile the in vivo metabolites of Radix scutellariae , Licorice , and their co‐administration in rats. Biological matrices, including bile, urine, and feces, were examined to identify biotransformation pathways and to compare metabolic signatures between single‐herb and combined administration. A total of 87 metabolites were characterized, encompassing a wide array of transformations such as oxidation, hydrogenation, sulfation, and methylation. Notably, the herb pair facilitated distinct metabolic conversions and reshaped the overall metabolite spectrum relative to individual herbs. These results provide mechanistic insights into the metabolic basis of this classical herb pair and suggest potential directions for further pharmacological research and clinical application.

ABSTRACT The phytochemical constituents of a Thai traditional anti‐hemorrhoidal formulation (TTAHF) remain insufficiently characterized, despite their importance for quality control. Therefore, this study aimed to investigate the liquid chromatography–mass spectrometry (LC‐MS) profiling and quantification of key compounds in TTAHF. LC‐MS analysis revealed that the most abundant compounds in the negative ionization mode were acteoside, emodin, quercitrin, kaempferol‐3‐O‐α‐L‐rhamnopyranoside, and chrysophanol. In contrast, the predominant compounds in the positive ionization mode were acteoside, chrysophanol‐8‐O‐glucoside, chrysophanol, acacetin, and stigmasterol. Three key compounds—acteoside, emodin, and chrysophanol—were selected as marker compounds for TTAHF. A high‐performance liquid chromatography (HPLC) method was validated for the quantification of these markers. The method demonstrated linearity, specificity, precision, and accuracy. Quantitative analysis of the TTAHF extract using the validated HPLC method showed that it contained 0.51% acteoside, 0.11% emodin, and 0.57% chrysophanol. In conclusion, this study provides LC‐MS profiling data and successfully establishes a validated HPLC method for the quantification of three marker compounds in TTAHF, which can be applied for the quality control of this traditional formulation.

ABSTRACT Prismatomeris tetrandra is a commonly used medicinal material for preventing and treating liver diseases. We use internal transcribed spacer 2 ( ITS2 ) barcode technology to provide molecular identification methods and sequences for P. tetrandra , optimize its high‐performance liquid chromatography (HPLC) fingerprint and multi‐component content determination method, and then evaluate the quality of different origins. In terms of ITS2 barcode identification, morphological and molecular identifications were performed on samples from 13 different origins, and a total of 28 ITS2 sequences were obtained, confirming that all the samples tested were authentic P. tetrandra . Since its sequences were not included in the GenBank database, we created 28 sequences and assigned a unique ID number to each sequence. A phylogenetic neighbor‐joining evolutionary tree was constructed, and its haplotypes were analyzed. The phylogenetic tree shows that all the P. tetrandra can be clearly clustered and identified, and are clearly distinguished from the other four medicinal materials of the same genus. In addition, a quality evaluation method for the simultaneous determination of three chemical components by HPLC was optimized and established. This method employed methanol as the extraction solvent, ultrasonic extraction for 40 minutes, a detection wavelength of 277 nm, and gradient elution using acetonitrile‐water as the mobile phase. The measured contents of scopolamine, 1‐rubiacin, and rubiacin were 0.009–0.359, 2.638–94.901, and 0.061–2.649 mg/g, respectively. It is speculated that the significant differences in content may be due to differences in growth environment and year. This study provides new insights for the accurate identification and quality evaluation of P. tetrandra .