Biomedical Chromatography最新文献

Flibanserin is the initial pharmaceutical treatment for hypoactive sexual desire disorder (HSDD). The analysis of urine samples plays a crucial role in the quantitation of flibanserin since a portion of flibanserin is excreted unchanged in the urine. An analytical method was proposed to quantify flibanserin in artificial urine samples (as model matrices). The integration of the spray-assisted fine droplet formation-liquid phase microextraction (SFDF-LPME) method and gas chromatography–mass spectrometry (GC–MS) system was performed for the first time to improve the sensitivity of the GC–MS system for flibanserin. Several parameters, including spraying cycle, extraction solvent type, mixing type and period, and sample volume, were systematically optimized to enhance the signal-to-noise ratio (S/N) of the analyte. After determining the optimal conditions, the analytical performance measurements of the system were figured out. The limit of detection (LOD), the limit of quantification (LOQ), and coefficient of determination (R²) values were 6.91, 23.05 μg kg⁻¹, and 0.9989, respectively. Recovery experiments were performed in artificial urine samples within the specified linear working range of 33.15–505.66 μg kg⁻¹. The SFDF–LPME–GC–MS method was efficiently applied to artificial urine samples by computing the matrix-matching calibration strategy, with percentage recovery values ranging from 90.0% to 105.9%.

Parkinson's disease (PD) is a progressive neurodegenerative disorder marked by the loss of dopaminergic neurons in the substantia nigra and the pathological accumulation of α-synuclein (α-Syn). Rotenone, a mitochondrial complex I inhibitor, reliably reproduces these hallmark features and is therefore widely used to establish experimental PD models. Shiliuwei Kangchanzhijing capsules (SLW-KCZJ capsules), a traditional Chinese medicine (TCM) formulation comprising 16 herbal components, were specifically developed for PD management. This study systematically evaluated the neuroprotective effects and underlying mechanisms of SLW-KCZJ in a rotenone-induced PD mouse model. SLW-KCZJ treatment significantly improved movement distance and balance–coordination, attenuated neuronal loss in the substantia nigra, and markedly suppressed neuroinflammatory responses. At the molecular level, the capsules increased tyrosine hydroxylase expression by approximately 200% and elevated Parkin and protein phosphatase 2A levels by approximately 60% and 80%, respectively. In contrast, they robustly inhibited polo-like kinase 2 expression and reduced α-Syn levels by approximately 60% and 40%. The intervention also increased striatal dopamine (DA) and homovanillic acid levels while lowering neurofilament light chain and oligomeric α-Syn. In summary, this study provides the first evidence that SLW-KCZJ capsules exert neuroprotective effects by modulating the Parkin/PP2A/α-Syn signaling pathway, offering a promising TCM-based therapeutic strategy for PD.

Selpercatinib (SPB) is a selective RET kinase inhibitor used to treat non–small cell lung cancer and thyroid cancer. Separation and identification of impurities formed during the manufacturing or storage of drug substances play a key role in determining their stability under various conditions during the life cycle of the drug. Hence, in the present study, SPB was exposed to different stress conditions like hydrolytic (acid, base and neutral), oxidative, thermal and photolytic stress conditions. A total of five degradation products were formed under the specified conditions. A UHPLC-PDA method employing Agilent Zorbax Extend C18 RRHD (100 × 2.1 mm, 1.8 μm) column with mobile phases of 0.1% formic acid in H₂O (A) and 0.1% formic acid in MeOH: ACN (70:30 v/v) (B) in gradient elution at a flow rate of 0.3 mL/min was developed to separate the potential degradation products. LC-QToF-MS/MS was used for the structural characterisation of the impurities. One of the degradation products (DP-2) was isolated and characterised through 1D and 2D NMR experiments. The developed method was validated as per ICH Q2 (R2) guidelines and provides valuable information about the stability of SPB, which can offer a strong foundation for its formulation development, quality control and regulatory compliance in bulk drug and dosage forms.

Diabetes mellitus is a global health burden, necessitating effective combination therapies. The fixed-dose formulation of sitagliptin phosphate (SGP, 100 mg) and lobeglitazone sulfate (LGS, 0.5 mg) offers synergistic benefits by enhancing insulin sensitivity and glucose-dependent insulin secretion. However, simultaneous quantification remains challenging due to their dosage disparity and the narrow therapeutic index of LGS. This study reports a validated, stability-indicating RP-HPLC method for concurrent estimation of SGP and LGS in a synthetic mixture. Separation was achieved on a Pursuit 5 Diphenyl column (150 × 4.6 mm, 5 μm) using a gradient of 0.1% formic acid and acetonitrile at 1.0 mL/min, with PDA detection at 254 nm. The method complied with ICH Q2(R2) guidelines, showing specificity, linearity, accuracy, precision, and robustness. Forced degradation studies revealed marked instability of SGP under acidic and alkaline conditions, whereas LGS remained stable. LC-MS/MS analysis identified a major degradation product at m/z 234, and probable fragmentation pathways were proposed. In silico ADMET and toxicity profiling predicted high gastrointestinal absorption for all degradation products, with PI-1 and PI-2 flagged for carcinogenicity and all except PI-3 showing nephrotoxicity. This integrated analytical–computational approach provides a reliable tool for quality control and safety assessment of this antidiabetic formulation.

Enasidenib (ESD), an oral medication, was used for the treatment of patients with refractory or relapsed Acute Myeloid Leukemia. No prior UPLC-MS/MS method with combined green metrics and in silico metabolism analysis for ESD was reported, so the research sought to establish an ultrafast UPLC-MS/MS method for estimating ESD in the HLMs matrix. The MS/MS analysis was applied applying positive ESI ionization mode, and chromatography using an Eclipse Plus C8 column (reversed phase) within one minute. The UPLC-MS/MS method exhibited a good degree of greenness as approved by the MoGAPI score (68.0) and the AGREE score (0.66). The linearity range was form 1 ng/mL (LOQ) to 4000 ng/mL. The precision and accuracy for intraday and interday measurements varied from −3.11% to 10.67% and −7.22% to 12.33%, respectively. The in vitro t_1/2 and the intrinsic clearance (Cl_int) were determined to be 82.52 min (long) and 9.83 mL/min/kg (low), respectively that is considered a low clearance drug. Slight structural modifications to the 2-methyl propan-2-ol moiety and pyridine ring in drug design could upsurge the ESD metabolic stability. The combined in vitro/in silico method delivers a resource-efficient strategy for early-stage metabolic screening and progressing innovative drug research focused on improving metabolic stability.

Zuclopenthixol was reported to treat schizophrenia, psychoses, and acute bipolar mania. 2-Chlorothioxanthon is reported to be zuclopenthixol official Impurity C (IMP C), and it is considered to be zuclopenthixol synthetic precursor. IMP C was reported to have toxicity on mRNA and protein expression. Environmentally validated HPTLC and HUPLC were developed for the simultaneous analysis of zuclopenthixol and IMP C in the analytical standard form and in pharmaceutical form, according to ICH guidelines. Regarding the HPTLC method, zuclopenthixol and its toxic impurity were separated using a mobile phase consisting of dichloromethane:ethyl acetate:triethylamine (6:4.5:0.4, v/v), and the scanning wavelength was 257 nm. For the HUPLC method, the simultaneous separation was completed within 2 min using Waters, CORTECS C₈ (4.6 mm × 150, 2.7 μm) as a stationary phase. The applied mobile phase was an isocratic mixture of methanol:water (80:20, v/v), pH was 3.25 ± 0.02 using orthophosphoric acid, pumping at 3 mL/min, and the detection wavelength was 257 nm. To evaluate the greenness value of the presented methods, the Eco-Scale Assessment method, Analytical Greenness Metric, and Green Analytical Procedure Index were applied. The practicality of the analytical methods was evaluated using the Blue Applicability Grade Index. Additionally, the analytical whiteness was determined via the RGB algorithm.

The traditional Chinese medicine Astragali Radix (AR) shows therapeutic effects on nephrotic syndrome (NS), with AR n-butanol (AR-B) particularly ameliorating podocyte injury. Nevertheless, the molecular mechanisms are unclear. This study investigates how serum containing AR-B (AR-B-S) mitigates podocyte injury. Using rats as serum donors, we established an adriamycin (ADR)-induced injury model. The findings indicated that the optimal preparation protocol for AR-B-S was as follows: using serum collected 3.5 h after administration, incubating it at 56°C for 30 min, and employing a final concentration of 2.5%. Subsequently, compositional analysis using UPLC-MS technology and integrated network pharmacology predicted potential pathways, with more than 30% of AR-B-S targets for amelioration of podocyte injury related to the cell cycle. Furthermore, ADR was shown to facilitate the transition of MPC5 cells from a quiescent to a differentiated state. Additionally, ADR was demonstrated to have the ability to cause mitotic arrest in podocytes, which in turn leads to apoptosis. Conversely, intervention with AR-B-S was found to reverse ADR-induced mitotic arrest through the PI3K/Akt pathway, reducing podocyte apoptosis. This study showed AR-B-S reduces podocyte damage by regulating the cell cycle, providing a methodological reference for herbal serum preparation and suggesting a new therapeutic target for kidney disease.

Metal–organic frameworks (MOFs) offer a revolutionary platform for addressing the inherent toxicity–efficacy paradox of bioactive alkaloids in traditional Chinese medicine (TCM). This review summarizes recent advances in MOF-based strategies for detoxification and potentiation of alkaloid drugs by integrating TCM holism principles with modern material science. We emphasize a methodology that combines network pharmacology for in silico prediction of multi-target mechanisms with chromatographic techniques (e.g., HPLC, LC–MS) for experimental validation of key alkaloids. Using Coptis chinensis against influenza A virus (H1N1) as an example, network pharmacology identifies modulation of TNF-α and IL-6 pathways as core mechanisms, while chromatographic analysis confirms and quantifies the predicted bioactive compounds (e.g., berberine and coptisine). This validated multi-target profile provides a rational basis for designing MOF-based systems capable of targeted co-delivery and controlled release of alkaloids, thereby achieving synergistic toxicity reduction and efficacy enhancement. It presents a robust, methodology-rich framework that bridges computational prediction, analytical validation, and precision delivery for modernizing TCM alkaloid research.

This study aimed to establish an UPLC-MS/MS method for determining memantine (MM) in rat plasma and to investigate its pharmacokinetic variations at different altitudes. MM was separated on a Hypersil Gold C18 column using a gradient elution method and detected by selected reaction monitoring (SRM). Sixteen fasted SD rats were randomized into low- and high-altitude groups (n = 8 each) and orally administered MM at 5 mg/kg. Serial blood samples were collected at 0-, 0.08-, 0.25-, 0.5-, 0.75-, 1-, 2-, 4-, 8-, 12-, 24-, 36-, and 48-h postdose for pharmacokinetic analysis. Compared with the low-altitude group, the high-altitude group exhibited a 72.15% increase in the area under the plasma concentration-time curve (AUC(0-∞)), a 39.97% reduction in apparent volume of distribution (Vz/F), the apparent clearance (CLz/F) decreased by 41.99%, the peak concentration (Cmax) increased by 131.52%, while the half-life (t_1/2) and time to peak concentration (Tmax) showed no significant changes. In conclusion, high-altitude hypoxia significantly affects MM metabolism in rats, potentially posing a risk of drug accumulation in plasma. The research findings provide a reference for the rational use of MM in high-altitude regions.