Biomedical Chromatography最新文献

The present study aims to evaluate the in vivo drug interactions between sotorasib and ondansetron to provide critical data for accidental or self-medication cases to ensure safe and effective concurrent use. A rapid UHPLC-MS/MS technique was developed on a Zorbax Eclipse XDB-C18 column (3.0 × 100 mm, 3.5 μm) using ammonium bicarbonate and methanol. The MRM analysis identified distinct transitions for sotorasib, ondansetron, and the internal standard (IS) at m/z 561.2 → 134.0, 317.0; m/z 294.0 → 184.0, 170.0; and m/z 529.2 → 326.1, 160.9, respectively. Target analytes were extracted from rat plasma by organic solvent precipitation. The UHPLC-MS/MS method revealed linearity as R² > 0.99, sensitivity at 0.4 ng/mL concentration, accuracy as 99.15%–99.87%, and precision as 1.62%–6.45% for LQC, MQC, and HQC. In silico predictive assessments revealed moderate and serious risk levels. Preclinical pharmacokinetic study revealed that ondansetron exhibited diminished C_max (0.69-fold), AUC (0.45-fold), and half-life (0.75-fold), alongside increased clearance (2.04-fold) when administered concurrently. Study findings are likely due to CYP3A4 induction by sotorasib, overriding its P-gp inhibition effects. To alleviate these effects, physicians must consider dose adjustments or stringent alternative antiemetic strategies.

Cerebral ischemia–reperfusion injury (CIRI) is classified under the category of stroke. Sanhua Decoction (SHD) is a classic prescription for the treatment of stroke, but its pharmacodynamic material basis and mechanism of action have not been fully elucidated. This study aimed to elucidate the bioactive components and therapeutic mechanisms of SHD against CIRI by integrating serum pharmacochemistry, network pharmacology, and metabolomics. Using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry, a 22 absorbed SHD prototype compounds were identified from rat serum. Through the application of network pharmacology and molecular docking technology, it was concluded that the key active ingredients, such as marmesin and aloe-emodin, mainly play a therapeutic role through key targets such as steroid receptor coactivator (SRC) and protein kinase B1 (AKT1). Molecular docking showed that these targets had strong affinity with key compounds. The results of pharmacodynamics showed that SHD could improve the neurological function score of CIRI rats, reduce the infarct area, and improve the pathological changes of brain tissue. Metabolomics analysis showed that SHD may play a therapeutic role by regulating steroid hormone synthesis and ubiquinone-related pathways. This study provides a methodological framework for exploring the role of Chinese herbal compounds in the treatment of CIRI.

Honeybees are frequently exposed to a wide range of environmental contaminants during foraging, leading to the inadvertent transfer of contaminants into honey. To assess pesticide contamination, 30 honey samples were collected and analyzed between 2022 and 2023 for pesticide residues. The QuEChERS method, followed by GC–MS/MS analysis, was employed to quantify pesticide levels. In the honey samples, a total of 15 pesticides were identified, with organophosphate compounds being the most prevalent. Insecticides, including omethoate and chlorpyrifos-methyl, are extensively applied in intensive apple orchards. Analysis of honey samples collected from apiaries located in these regions revealed significantly elevated concentrations of these pesticides. This study underscores the widespread presence of pesticide residues in honey collected from migratory apiaries in the Kashmir region. Although human health risk assessments indicate no immediate concerns in most cases, precautionary measures should be considered, particularly due to the potential risks associated with honey consumption by infants and the cumulative effects of these chemicals over time.

An adapted QuEChERS extraction protocol coupled with LC–MS/MS analysis was established to enable concurrent quantification of 11 different pesticides in tomato specimens obtained from Dhaka, Bangladesh. Acetonitrile (ACN) was used for pesticide extraction, and among different sorbent combinations tested, the 50 mg primary secondary amine, 10 mg C₁₈, and 3 mg graphitized carbon black with minimal matrix effects was selected for d-SPE cleanup. The pesticides exhibited good linearity over a wide linear range of 0.1–200 μg/L, with R² ≥ 0.9925 in solvent and matrix-matched calibration. The method exhibited high sensitivity with limit of detection = 0.10 to 1.10 μg/kg and limit of quantitation = 0.33 to 3.33 μg/kg. The newly developed method showed excellent results in recovery studies (80.0%–91.6%) at five (n = 5) different fortification levels (10, 20, 50, 100, and 200 μg/kg) with relative standard deviations ≤9.0%. Azoxystrobin and difenoconazole were the most frequently detected pesticides, with residue levels ranging from 0.41 to 97.33 μg/kg and 1.58 to 177.2 μg/kg, respectively. The hazard quotient and acute health index values remained below 1 for all pesticides, indicating no significant health risk. This study confirms that tomato samples from Dhaka were largely compliant with MRLs, supporting safer local production but indicating the need for regular monitoring.

A robust, precise, accurate, and stability-indicating reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the quantitative estimation of levothyroxine sodium in soft gelatin capsule dosage forms across a wide strength range (13 to 200 μg). The objective of this study was to establish a single, specific method capable of assessing both assay with high sensitivity, particularly at a low-test concentration of 1 ppm, while ensuring specificity toward all known impurities listed in the levothyroxine drug substance monograph. Chromatographic separation was achieved using an InertSustain AQ C18 column (250 × 4.6 mm, 5 μm) maintained at 60°C. The mobile phase system comprised 90% phosphate buffer and 10% acetonitrile (Mobile Phase A) and 90% acetonitrile with 10% water (Mobile Phase B), delivered at a flow rate of 1.2 mL/min. Detection was carried out at 225 nm with a total runtime of 25 min. The retention time of levothyroxine was approximately 7.1 min. The method was validated across a linearity range of 0.5 to 2.0 ppm (50% to 200% of the test concentration), yielding a correlation coefficient (r²) > 0.999. The method demonstrated excellent precision, with %RSD values of 0.2% for standard solutions and 0.8% for six sample preparations. Limits of detection (LOD) and quantification (LOQ) were determined to be 0.05 and 0.1 ppm, respectively. The method was found to be specific to all 14 known impurities, robust, accurate, and stability-indicating. This validated method is suitable for routine quality control analysis and regulatory submissions, meeting industrial standards for accuracy, precision, and specificity across all marketed strengths of levothyroxine sodium capsules.

Depression, affecting approximately 300 million people globally, is characterized by low mood, cognitive dysfunction, and neuroendocrine issues. Rhodiola rosea (R. rosea) has shown significant antidepressant activity, but its mechanisms are unclear. This study identified 62 chemical constituents in R. rosea extract, with 21 bioavailable compounds detected by UHPLC-Orbitrap Fusion TMS. A total of 3830 depression-related targets were collected from GeneCards, ChEMBL, DisGeNET, and OMIM, and 290 predicted targets for the orally bioavailable components were obtained from TCMSP, SwissTargetPrediction, and PharmMapper. Venny analysis found 157 core targets and was used to construct a PPI network. KEGG analysis showed these targets enriched in endocrine resistance, lipid and atherosclerosis, and AGE-RAGE signaling pathways. Molecular docking revealed salidroside, davidioside, 3,5-digalloylepicatechin, and kenposide A bound strongly to STAT3, HSP90AA1, ESR1, PRKACA, and EGFR. Molecular dynamics simulations confirmed complex stability. In vitro, R. rosea dose-dependently improved viability of CORT-induced HT-22 cells and reversed CORT-mediated downregulation of TPH2, Ht1b, Ht2a, and 5-HT. Notably, this regulatory effect was abolished by the ESR1 inhibitor fulvestrant, confirming R. rosea protects the serotonergic system via ESR1 in the endocrine resistance pathway. This study demonstrates R. rosea's antidepressant effect through multicomponent, multitarget, multipathway synergy, supporting its clinical use.

Triclabendazole (TCLZ) is a halogenated benzimidazole anthelmintic widely used in humans and animals to treat fascioliasis which acts by disrupting microtubule-dependent processes in the species of Fasciola. This study aims to characterize TCLZ's degradation profile under different degradation conditions and assess the toxicological potential of its degradation products (DPs). TCLZ was subjected to forced degradation as required by the ICH Q1A(R2) and Q1B guidelines, including oxidation (H₂O₂), photolysis (UV and visible light), thermal, and hydrolysis (acidic, basic, and neutral) conditions. Reverse-phase high-performance liquid chromatography (RP-HPLC) was used to resolve DPs with Agilent Eclipse C18 column (4.6 × 250 mm, 5 μm) and the mobile phase consisting of 0.1% acetic acid (Solvent A) and methanol (Solvent B). TCLZ is susceptible to oxidative and photolytic (UV) degradation, leading to four DPs. The observed DPs were structurally characterized by liquid chromatography-high-resolution mass spectrometry (LC-HRMS), and the corresponding fragmentation pathways were proposed. In silico tools like Zeneth, DEREK Nexus, and ADMET Predictor were used in the study. Zeneth was used to predict the probable degradation pathway of TCLZ. Three experimentally observed DPs matched with the Zeneth predictions. The toxicity of TCLZ and its DPs was predicted using in silico tools like DEREK Nexus and ADMET Predictor.

Nitrosamine impurities, which are categorized as mutagenic and probable human carcinogens, have been widely detected in various drug substances and products. This study investigates the formation of the N-nitrosopiperazine (NNPZ) genotoxic impurity during the manufacturing of cetirizine dihydrochloride (CTZ), originating from piperazine, a key starting material. A sensitive and selective LC–MS method was developed for the accurate quantification of NNPZ in the CTZ drug substance. The allowable intake limit for NNPZ is 26.5ng/day and maximum daily dose 880mg/day as per US FDA. This analytical method was developed and validated in accordance with ICH Q2R2 guidelines. It demonstrated a low detection limit (LOD) of 0.03 ng/mL and a quantitation limit (LOQ) of 0.1 ng/mL. The linearity study yielded a strong regression coefficient of 0.9974. With a high recovery rate of 96.56% to 102.35% for spiked NNPZ, the method's accuracy was well-established. The overall results confirm that this methodology is reliable, precise, and reproducible, and it can therefore be used for the determination of NNPZ-genotoxic impurity in CTZ drug substance samples via LC–MS.