Journal Of Pharmaceutical And Biomedical Analysis[JOURNAL]
Sun
200 papers
A novel pharmacokinetics-pharmacodynamics strategy for effective components exploration combined with metabolomics and proteomics: Application to Xingbei Zhike granule.
Gao X, Yin M, Liu J … +11 more , Feng J, Jiang Y, He R, Ma P, Fan X, Dang H, Yuan S, Chen X, Cao L, Wang Z, Xiao W
J Pharm Biomed Anal · 2026 Sep · PMID 41990596 · Publisher ↗
Traditional Chinese medicine (TCM) is characterized by its complex and diverse components, difficult to identify the effective components. In this study, a pharmacokinetics (PK)-pharmacodynamics (PD) strategy, integratin...
Traditional Chinese medicine (TCM) is characterized by its complex and diverse components, difficult to identify the effective components. In this study, a pharmacokinetics (PK)-pharmacodynamics (PD) strategy, integrating chemical component profile, PK, metabolomics, and proteomics, was developed to explore the effective components of Xingbei Zhike granule (XBZKG) in treating post-infection cough (PIC). The quality consistency of XBZKG was confirmed by evaluating 78 components across 20 batches. And its good curative effect against PIC was verified through measurements of cough latency, cough frequency, and biochemical assays. Meanwhile, XBZKG was found to ameliorate PIC-induced metabolic disturbances, particularly in amino acid metabolism. Interestingly, the pathological state of PIC itself altered the absorption and metabolism of XBZKG in rats. Among ten components, ephedrine, pseudoephedrine, and amygdalin fitted good dose-effect models with three PD indicators, with the PIC pathological state appearing to enhance their efficacy. Spearman correlation analysis further revealed that 15 components alleviated PIC-induced amino acid imbalances, and some structurally similar components exhibited synergistic effects. Integrative proteomic and metabolomic analysis identified 28 potential pathway of XBZKG in the treatment of PIC, mainly related to inflammation. Finally, in vitro cellular assays confirmed 15 components exert good anti-inflammatory activity. This study provides a reproducible strategy for identifying effective components in TCM and offers new insights into its multi-component, multi-target, and multi-pathway therapeutic mechanisms.
Targeted phytocannabinomics provides new insights into the biosynthetic origin of cis-Δ⁹-THCA in Cannabis sativa L.
Buccolieri L, Paris R, Fulvio F … +3 more , Alberti I, Citti C, Cannazza G
J Pharm Biomed Anal · 2026 Sep · PMID 41990595 · Publisher ↗
Recent studies have reported the occurrence of cis-Δ⁹-tetrahydrocannabinol (cis-Δ⁹-THC) and its carboxylated precursor, cis-Δ⁹-tetrahydrocannabinolic acid (cis-Δ⁹-THCA), minor isomers of the well-known trans counterparts...
Recent studies have reported the occurrence of cis-Δ⁹-tetrahydrocannabinol (cis-Δ⁹-THC) and its carboxylated precursor, cis-Δ⁹-tetrahydrocannabinolic acid (cis-Δ⁹-THCA), minor isomers of the well-known trans counterparts. However, their origin remains unclear and several hypotheses have been proposed. In this work, cis-Δ⁹-THCA and the major phytocannabinoids, including cannabidiolic acid (CBDA), trans-Δ⁹-tetrahydrocannabinolic acid (trans-Δ⁹-THCA), cannabichromenic acid (CBCA), and their biosynthetic precursor cannabigerolic acid (CBGA), were quantified in a large and diverse set of C. sativa accessions using a targeted metabolomics approach based on liquid chromatography coupled to high-resolution Orbitrap mass spectrometry (HPLC-HRMS). Our findings indicate that cis-Δ⁹-THCA does not appear to be chemically derived from other cannabinoids, prompting new considerations regarding its biosynthetic origin. Notably, growth-stage analyses revealed a parallel accumulation pattern between cis-Δ⁹-THCA and CBCA, suggesting the potential involvement of CBCA synthase in its formation. Overall, this study provides new evidence on the distribution, variability, and possible biosynthetic pathways of cis-Δ⁹-THCA, enriching current understanding of cannabinoid diversity in C. sativa.
Simultaneous quantification and aerodynamic particle size distribution analysis of tiotropium bromide and olodaterol in nebulized aerosol using UHPLC-MS/MS.
Dou J, Hong Y, Qian R … +3 more , Chen C, Yuan C, Zhu D
J Pharm Biomed Anal · 2026 Sep · PMID 41990594 · Publisher ↗
Fixed-dose combination (FDC) inhaled aerosols require precise characterization of both active pharmaceutical ingredients (APIs) and aerodynamic properties to ensure therapeutic efficacy and safety. This study established...
Fixed-dose combination (FDC) inhaled aerosols require precise characterization of both active pharmaceutical ingredients (APIs) and aerodynamic properties to ensure therapeutic efficacy and safety. This study established and validated a sensitive, high-throughput ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of tiotropium bromide (TTP, a long-acting muscarinic antagonist) and olodaterol (ODT, a long-acting β-adrenergic agonist) in nebulized aerosols, coupled with comprehensive aerodynamic particle size distribution (APSD) analysis. The optimized method employed a 50% methanol-water solution as the extraction solvent, with isocratic elution using 0.1% formic acid in water-methanol (50:50, v/v) and multiple reaction monitoring (MRM) in positive ionization mode. The total analysis time was only 3 min, enabling high-throughput analysis. Method validation confirmed excellent linearity for both analytes over the range of 0.5-16 ng/mL (r > 0.998), with a lower limit of quantification (LLOQ) of 0.5 ng/mL. Intra- and inter-batch precision (relative standard deviation, RSD) were < 7.5%, and extraction recoveries ranged from 98.1% to 98.8% (RSD<3%), with good matrix effect tolerance, solution stability (24 h at room temperature/4°C), and dilution integrity. A combination of syringe aspiration and Andersen Cascade Impactor (ACI) sampling was used to characterize aerosol properties at 2 and 8 min of nebulization. Both TTP and ODT exhibited uniform and stable aerosol concentrations (RSD<3.5%), with mass median aerodynamic diameters (MMAD) of 1.87-2.35 μm, geometric standard deviations (GSD) of 2.13-2.44, and fine particle fractions (FPF) exceeding 90%. The proportion of ultrafine particles (<0.41 μm) captured by the micro-orifice collector (MOC) was < 4%, confirming optimal particle size distribution for deep lung deposition. Application of this method to the evaluation of theoretical delivered doses in animal safety assessment tests showed that the doses were 4.7-8.6 times the rat equivalent dose, meeting the design requirements for toxicological studies. This method provides a robust framework for quality control, generic drug consistency evaluation, and preclinical toxicology of TTP/ODT nebulized aerosols, supporting the development of FDC inhaled therapies for chronic obstructive pulmonary disease (COPD).
The drug substance of traditional Japanese Kampo medicines retains exosome-like nanoparticles derived from crude drugs.
Tsuchiya N, Matsumoto T, Hiramoto M … +2 more , Kushida H, Nishi A
J Pharm Biomed Anal · 2026 Sep · PMID 41985257 · Publisher ↗
Recent studies have highlighted the diverse roles of extracellular vesicles, which have been detected in natural products such as ginger and ginseng. This study examined whether traditional Japanese Kampo medicines also...
Recent studies have highlighted the diverse roles of extracellular vesicles, which have been detected in natural products such as ginger and ginseng. This study examined whether traditional Japanese Kampo medicines also contain extracellular vesicles. The drug substance (i.e., spray-dried powder from hot-water extracts of crude drugs) of the Kampo medicine Ninjin'yoeito was suspended in distilled water and fractionated by size exclusion chromatography to isolate nano-sized particles. Nanoparticle tracking analysis and electron microscopy confirmed the particle size distribution and morphology of the product, confirming the presence of exosome-like nanoparticles in the Kampo medicine preparation. Nanoparticle measurements and electron microscopy revealed vesicle-like structures consistent with the known characteristics of exosomes, and proteomic analysis supported their biochemical identity. Lectin array profiling revealed the abundance of high-mannose N-glycans on their surface. These nanoparticles were internalized by cells, and miRNA analysis indicated the presence of multiple miRNA types within the nanoparticles. This is the first report demonstrating the presence and characteristics of exosome-like nanoparticles in Kampo medicines. Although most active components of Kampo medicines have traditionally been considered small molecules, our findings suggest that extracellular vesicles can also serve as potential bioactive components.
PCN-224/β-Cyclodextrin dual-modified magnetic nanocomposite for efficient extraction and determination of nobiletin and tangeretin in Chenpi-based beverages.
Liu R, Su L, Zhang Y … +9 more , Yang Y, Li Y, Wu J, Xie Y, Liu X, Huang Y, Feng T, Wei X, Chen Q
J Pharm Biomed Anal · 2026 Sep · PMID 41985256 · Publisher ↗
Polymethoxylated flavonoids such as nobiletin and tangeretin are the primary bioactive constituents of Chenpi. However, their accurate quantification in complex beverage matrices remains challenging. In this study, a nov...
Polymethoxylated flavonoids such as nobiletin and tangeretin are the primary bioactive constituents of Chenpi. However, their accurate quantification in complex beverage matrices remains challenging. In this study, a novel dual-modified magnetic composite, FeO@PCN-224@β-CD, was synthesized for the efficient extraction of these target analytes. The material integrated the high magnetization of FeO for rapid separation, the porous and π-electron-rich framework of PCN-224 for strong π-π interactions, and a β-cyclodextrin shell for enhanced hydrophobic inclusion and aqueous dispersibility. Coupled with UPLC-UV detection, a magnetic solid-phase extraction method based on this composite was established and optimized through single-factor experiments and response surface methodology. The developed method exhibited excellent linearity (R² ≥ 0.998), low detection limits (0.01-0.02 μg/mL), satisfactory precision and good recoveries (92.59-101.73%). Furthermore, the adsorbent demonstrated outstanding reusability over seven cycles. The high adsorption efficiency was attributed to the synergistic effect of π-π stacking and hydrophobic interactions within the composite. This study provided a robust and environmentally friendly strategy for the selective enrichment and accurate quantification of nobiletin and tangeretin in Chenpi-based beverages, highlighting the potential of PCN-224 and β-CD dual-functionalized magnetic composites for high-efficiency sample pretreatment in complex matrices.
Antiproliferative effects of pachymic acid on thyroid cancer cells and characterization of its metabolites.
Wang F, Li J, Hu B … +2 more , Tang M, Chen J
J Pharm Biomed Anal · 2026 Sep · PMID 41985255 · Publisher ↗
Thyroid cancer (THCA) is the most prevalent endocrine malignancy, underscoring the need for novel therapies. Pachymic acid (PA), a key bioactive triterpenoid from P. cocos, possesses documented anticancer activity, yet i...
Thyroid cancer (THCA) is the most prevalent endocrine malignancy, underscoring the need for novel therapies. Pachymic acid (PA), a key bioactive triterpenoid from P. cocos, possesses documented anticancer activity, yet its role in thyroid cancer is poorly understood. In this study, we evaluated PA's effects on THCA in vitro and in vivo. Our results revealed that PA potently suppressed THCA cell proliferation, migration and tumor growth. Metabolite profiling identified seven metabolites and their formation pathways. Network pharmacology predicted 147 therapeutic targets, primarily enriched in PI3K-AKT, MAPK, and EGFR signaling, and PA was shown to exhibit the strong and stable binding affinity with AKT, EGFR, HSP90AA1, which was functionally validated by Western blot showing inhibition of AKT (Ser473) and EGFR (Tyr1068) phosphorylation. Crucially, hydrolytic metabolites M4-M7 exhibited superior binding affinity to AKT and EGFR. In summary, PA inhibits thyroid cancer cell proliferation likely through blocking AKT and EGFR signaling, and its hydrolytic metabolites M4-M7 may mediate enhance antitumor activity via more potent target engagement.
A rapid 2-minute LC-MS/MS method with simple protein precipitation for simultaneous determination of tryptophan and its two metabolites kynurenine and kynurenic acid in human serum.
Yan H, Lu H, Huang S … +5 more , Li Y, Xia H, Huang W, Shang D, Zhang M
J Pharm Biomed Anal · 2026 Sep · PMID 41985254 · Publisher ↗
Dysregulation of the tryptophan (TRP) metabolic pathway is closely linked to the pathophysiology of neuropsychiatric disorders, such as depression. This study aimed to develop and validate a sensitive, rapid, and robust...
Dysregulation of the tryptophan (TRP) metabolic pathway is closely linked to the pathophysiology of neuropsychiatric disorders, such as depression. This study aimed to develop and validate a sensitive, rapid, and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of TRP and its metabolites, kynurenine (KYN) and kynurenic acid (KYNA), in human serum. Analytes were extracted from 100 µL of serum via simple protein precipitation with acetonitrile. Chromatographic separation was achieved on an Agilent ZORBAX HILIC Plus column (4.6 mm×100 mm, 3.5 µm) using isocratic elution with a mobile phase of methanol: acetonitrile containing 5 mM ammonium formate. Quantification was performed using an electrospray ionization source in positive ion multiple reaction monitoring (MRM) mode, with a total run time of only 2.0 min.The linear ranges were 1-50 µg/mL for TRP, 0.1-5 µg/mL for KYN, and 1-50 ng/mL for KYNA, covering clinically relevant levels. A weighting factor of 1/x² provided the best fit for calibration curves (R² > 0.99). Extraction recoveries ranged from 88.23% to 99.39%, and mean internal standard-normalized matrix effects were 81%-100%. Accuracy and precision values met bioanalytical acceptance criteria. Stability assessments confirmed that samples were stable at -20°C and -80°C for 31 days and through three freeze-thaw cycles. This validated method was successfully applied to analyze serum samples from 103 adolescent patients with first-episode depression.
Heterologous internal calibration for multiplexed internal quantification in targeted LC-MS/MS bioanalysis.
Strassel O, Feinberg M, Visconti G … +2 more , Boccard J, Rudaz S
J Pharm Biomed Anal · 2026 Sep · PMID 41980398 · Publisher ↗
Absolute quantification is crucial in bioanalysis for both clinical and non-clinical applications but remains challenging, especially for endogenous metabolites. In external calibration, the absence of a true blank matri...
Absolute quantification is crucial in bioanalysis for both clinical and non-clinical applications but remains challenging, especially for endogenous metabolites. In external calibration, the absence of a true blank matrix complicates calibration curve preparation. Multi-targeted internal calibration (IC) using stable isotope-labeled standards (SILs) as one-point calibrants offers an attractive alternative, yet its use is limited by the availability and cost of homologous SILs. To address this, heterologous SILs can serve as surrogate calibrants for multiple analytes, relying on response factors (RFs) established between each analyte and selected SILs in the matrix of interest. This study assessed the suitability of such an alternative isotopic-dilution strategy, referred to as heterologous internal calibration (H-IC), within an LC-MS/MS method developed for chronic kidney disease (CKD) research. A panel of 18 metabolites and their corresponding SILs was initially analyzed to generate reference values, and the performance of various analyte-SIL combinations was compared with that of the original homologous internal calibration procedure. Validation indicated satisfactory accuracy and precision for most analytes, with trueness generally within 70-130% and precision below 10%. Only a few low-abundance metabolites showed minor deviations. Finally, reducing the calibration panel to five heterologous SILs for quantifying all 18 analytes in 30 patient samples produced results comparable to those obtained using individual homologous SILs. These findings demonstrate that H-IC can substantially reduce SIL requirements and analytical costs while maintaining acceptable quantitative performance for metabolite measurement in human plasma.
GC-MS method development and validation for the determination of Short Chain Fatty Acids in human feces.
Vasiliki G, Konstantina F, Olga B … +3 more , Georgios T, Helen G, Chistina V
J Pharm Biomed Anal · 2026 Sep · PMID 41967141 · Publisher ↗
Short Chain Fatty Acids (SCFAs), the end products of microbial fermentation of dietary fibers, appear to be key mediators of the beneficial effects elicited by the gut microbiome and have been shown to exert multiple eff...
Short Chain Fatty Acids (SCFAs), the end products of microbial fermentation of dietary fibers, appear to be key mediators of the beneficial effects elicited by the gut microbiome and have been shown to exert multiple effects on metabolism. In this study, we developed and validated a sensitive, accurate, and reproducible GC-MS method for the simultaneous quantification of SCFAs (Acetic acid (C), propionic acid (C), butyric acid (C), isobutyric acid and isovaleric acid) in human feces. Sample preparation was simplified while maintaining robustness, following systematic evaluation of homogenization, extraction solvents, and acidification conditions. The optimized method demonstrated high analytical performance, with limits of detection ranging from 0.01 to 0.52 μmol/g and good precision and accuracy in accordance with FDA and EMA bioanalytical guidelines Stability studies revealed that SCFAs remain stable in acidified fecal samples for up to 10 days without cold-chain requirements, while -80 °C storage was optimal for long-term preservation and 4 °C suitable for short-term handling. The applicability of the method was confirmed through analysis of samples collected from healthy volunteers. Overall, the developed approach provides a practical, high-throughput, and scalable tool for SCFA analysis, supporting applications in clinical research, metabolomics, and large-scale microbiome studies.
Methyl orange-enhanced DNA hybridization: A gene sensor approach for hepatitis delta detection.
Moraes DD, Luz LFG, Soares MMCN … +4 more , Dall'Acqua D, Oliveira RJ, Madurro JM, Madurro AGB
J Pharm Biomed Anal · 2026 Sep · PMID 41967140 · Publisher ↗
Hepatitis delta is an infection triggered by the hepatitis delta virus (HDV), a defective virus that requires the hepatitis B virus (HBV) to replicate. Coinfection with both viruses can result in more severe complication...
Hepatitis delta is an infection triggered by the hepatitis delta virus (HDV), a defective virus that requires the hepatitis B virus (HBV) to replicate. Coinfection with both viruses can result in more severe complications, including progressive chronic liver damage. In this study, we present the development of an electrochemical device for molecular diagnosis of HDV in the blood serum of infected patients, employing screen-printed carbon electrodes functionalized with ethylenediamine and sensitized through the use of a specific DNA probe. The interaction with specific genomic RNA was evaluated using Differential Pulse Voltammetry, Electrochemical Spectroscopy Impedance, Scanning Electron Microscopy, Atomic Force Microscopy and molecular docking. The genosensor efficiently detected genomic RNA within a linear range from 9 copies/mL to 3 × 10 copies/mL, with a detection limit of 9 copies/mL. EIS revealed that the sensor's resistance to charge transfer increased exponentially with increasing concentration of the target genomic RNA. This platform offers several advantages for application in real samples, including high sensitivity, selectivity, and low sample volume requirements, making it particularly suitable for the diagnosis of hepatitis delta virus in blood serum samples. Furthermore, the simple and low-cost modification via direct drop-casting highlights the potential of this platform for integration into portable point-of-care devices.
Sorbent-coated metal discs: A hybrid active/passive exhaled breath sampling alternative for early lung cancer screening.
Lundberg R, Lundeberg T, Andrasko J … +2 more , Lagesson-Andrasko L, Dahlén J
J Pharm Biomed Anal · 2026 Sep · PMID 41967139 · Publisher ↗
In this new breath sampling method, a metal disc with carbopack X sorbent was exposed to prolonged exhalation. Sampled volatile organic compounds (VOCs) were transferred to a solid phase micro-extraction (SPME) fiber for...
In this new breath sampling method, a metal disc with carbopack X sorbent was exposed to prolonged exhalation. Sampled volatile organic compounds (VOCs) were transferred to a solid phase micro-extraction (SPME) fiber for gas chromatography-mass spectrometry (GC-MS) analysis. The performance of these discs was compared to breath biomarker extraction by SPME from Tedlar® bags. Ten commonly reported breath biomarkers were selected as targets (acetone, isoprene, toluene, hexanal, benzaldehyde, decane, octanal, nonanal, dodecane and decanal). The new method proved significantly better at extracting the latter eight of the ten target compounds. The two methods were also evaluated by exposing the sorbent disc and an SPME fiber to a mixture of seven VOCs. The sorbent discs extracted the standard compounds more efficiently, yielding 2-9 times larger GC-MS signals for the four last-eluting compounds than direct SPME. In conclusion, the new sorbent disc method can, with minimal effort from patients, extract VOCs from breath more efficiently than the current state-of-the-art.
A holistic strategy for the preparation and quality control of sugar-free Fufang Ciwujia granules: Comprehensive quality evaluation through fingerprint and QAMS.
Zhan Z, Chen Z, Lim X … +5 more , Ding Z, Li Z, Fan M, Zhang T, Li L
J Pharm Biomed Anal · 2026 Sep · PMID 41962349 · Publisher ↗
Fufang Ciwujia granule (FCG), a herbal extract formulation effective for treating insomnia and dream-disturbed sleep, is limited for sugar-restricted populations due to its high sucrose content. Furthermore, its quality...
Fufang Ciwujia granule (FCG), a herbal extract formulation effective for treating insomnia and dream-disturbed sleep, is limited for sugar-restricted populations due to its high sucrose content. Furthermore, its quality control (QC), reliant on a single marker compound, is inadequate for representing the multi-component nature of this traditional Chinese medicine formulation. This study developed a sucrose-free FCG formulation suitable for populations requiring glycemic control, and established a comprehensive QC strategy to ensure its quality equivalence to the original product. The Analytic Hierarchy Process-Entropy Weight (AHP-Entropy) combined with Box-Behnken response surface methodology (RSM) optimized the granulation process, identifying an optimal composite filler system (soluble starch, dextrin, maltodextrin) and 10% ethanol as the wetting agent. Pilot-scale batches demonstrated excellent formability (>96%). A high-performance liquid chromatography (HPLC) fingerprinting method and quantitative analysis of multi-components by a single marker (QAMS) were established. Comparative analysis revealed high fingerprint similarity (>0.98), minimal variation in the content of eight marker components (RSD<4%), and equivalent dissolution profiles (f>50) between the sucrose-free and sucrose-based granules, which provides a holistic framework, from formulation to comprehensive quality assessment, for developing sucrose-free TCM granules, demonstrating successful preservation of critical quality attributes despite significant excipient reformulation.This study covers the entire workflow from formulation development and quality evaluation to consistency verification. It provides a complete technical framework for the clinical translation of sugar-free FCG and the development of other sugar-free TCM granule formulations.
Biosensor based on hybrid composite and Concanavalin a lectin for microorganisms differentiation.
Souza AMN, Silva-Junior AG, Costa MP … +3 more , Lima GMS, Oliveira MDL, Andrade CAS
J Pharm Biomed Anal · 2026 Sep · PMID 41955779 · Publisher ↗
For the treatment of candidemia, especially in critical care settings, rapid identification of Candida species remains essential. Hybrid compounds have emerged as a powerful strategy to address challenges in microbiologi...
For the treatment of candidemia, especially in critical care settings, rapid identification of Candida species remains essential. Hybrid compounds have emerged as a powerful strategy to address challenges in microbiological detection and control. By integrating organic and inorganic components within a single architecture, these materials exhibit synergistic physicochemical and biological properties that are useful for sensor platform strategies. Lectins can act as bioreceptors due to their affinity for carbohydrates, and the addition of nanomaterials increases the platform's surface area and sensitivity. This work developed an electrochemical biosensor using a self-assembling monolayer of 4-mercaptobenzoic acid (MBA) and a hybrid nanocomposite composed of chitosan-coated magnetite nanoparticles capped with gold (FeO@Chit@Au) and the lectin Concanavalin A (ConA) as the biorecognition element. The hybrid architecture was designed to enhance the microorganism-sensor interface by increasing surface area and improving lectin immobilization. Topographic analysis confirmed the effective assembly of the sensing layer on gold electrodes. Selective responses to Candida albicans and Candida tropicalis were observed by electrochemical analysis, whereas bacterial species such as Staphylococcus aureus, Klebsiella pneumoniae, and Bacillus subtilis exhibited reduced signals. The stronger response for Candida species is consistent with the high affinity of ConA for mannan residues present in fungal cell walls. The biosensor demonstrated a limit of detection of 0.71 CFU.mL⁻ and a limit of quantification of 2.39 CFU.mL⁻ for C. albicans within a working range of 10-10 CFU.mL⁻. These findings demonstrate how lectin-based electrochemical platforms can identify and distinguish clinically significant microorganisms.
Development and validation of an online extraction HPLC-HRMS method for the quantification of carbapenems in FecalSwab™ samples.
Holub M, Mizrahi A, Genty T … +3 more , Pilmis B, Le Monnier A, Leveque N
J Pharm Biomed Anal · 2026 Sep · PMID 41955778 · Publisher ↗
Quantifying carbapenems in fecal samples is analytically challenging due to their instability and the complexity of intestinal matrices. We developed and validated a novel HPLC-HRMS method enabling direct quantification...
Quantifying carbapenems in fecal samples is analytically challenging due to their instability and the complexity of intestinal matrices. We developed and validated a novel HPLC-HRMS method enabling direct quantification of imipenem and meropenem in human fecal material collected with the FecalSwab™ system. Sample processing consisted of minimal handling, involving clarification and direct online extraction using TurboFlow™ prior to chromatographic separation on a reversed-phase column. Detection was performed using Orbitrap high-resolution mass spectrometry in full-scan mode. Because only a small fraction of parenterally administered carbapenems reaches the intestinal lumen, but even low concentrations can exert a strong ecological pressure on gut microbiota and promote selection of resistant Enterobacterales, sensitive measurement of residual fecal levels is essential to understand antibiotic-driven dysbiosis and resistance emergence. The method was validated according to ICH M10 guidelines. Linearity was achieved over 1.5-750 ng/L for imipenem and 2.9-1500 ng/L for meropenem (R² > 0.995). Trueness and precision were within ±15% at all QC levels (±20% at LLOQ). Matrix effects were controlled using isotopically labeled internal standards, yielding normalized matrix factors with CV < 12%. Stability studies confirmed rapid degradation of imipenem and moderate stability of meropenem in fecal medium, prompting the identification and semi-quantification of their hydrolysis products. The method was successfully applied to 40 clinical samples, detecting carbapenems or their degradation products in 17/20 imipenem-exposed and 18/20 meropenem-exposed patients. This validated workflow provides a robust and clinically applicable approach for assessing intestinal exposure to carbapenems and supports ecological investigations of antibiotic-microbiota interactions.
Discovery of anti-renal fibrosis quality markers from Piperis longi fructus: An integrated strategy of hollow fibre cell trapping method coupled with liquid chromatography-mass spectrometry and in vitro efficacy verification.
Zhang XL, Li Q, Wang RQ … +4 more , Xing RR, Chen X, Liu JF, Hu S
J Pharm Biomed Anal · 2026 Sep · PMID 41950810 · Publisher ↗
Modern pharmacological research has shown that the extract of traditional Chinese medicinal herb of Piperis longi fructus has anti-renal fibrosis activity, which made it potentially useful as a therapeutic medicine for r...
Modern pharmacological research has shown that the extract of traditional Chinese medicinal herb of Piperis longi fructus has anti-renal fibrosis activity, which made it potentially useful as a therapeutic medicine for renal fibrosis. To enhance the quality control of Piperis longi fructus, a hollow fiber cell trapping (HFCT) was established to investigate its anti-renal fibrosis quality markers (Q-markers). Human kidney 2 cells or transforming growth factor-β (TGF-β)-induced renal fibrosis model cells were loaded into the lumen of the hollow fiber with scanning electron microscopy confirming the cells adherence and proliferation on the inner wall of the fiber. Then the fiber was immersed in the extract of Piperis longi fructus to screen the active components. Cell counting kit-8 assay and flow cytometry were used to detect the effects of Piper longum extract on cell proliferation and HFCT procedure on apoptosis, respectively. The critical variables were investigated and optimized to ensure cell viability and reliable active screening. Trapped components were separated and identified using liquid chromatography-mass spectrometry, and cell trapping factor with an asterisk (CTF*) was proposed to assess the binding strength between the model cell and the active ingredients. Western blot analysis was used to study the active effects of high-CTF* components (piperlonguminine, piperlongumine and piperine) on renal fibrosis-related proteins including collagen type I (COL-I) and α-smooth muscle actin (α-SMA). Finally, piperlonguminine and piperlongumine were determined as anti-renal fibrosis Q-markers of Piperis longi fructus. This study demonstrated that HFCT is a simple and cost-effective technique for rapid screening of quality markers in traditional Chinese medicines.
Uncovering the active substances of Modified Fangjihuangqi Decoction in reducing cardiomyocytes' mitochondrial oxidative damage.
Chen H, Luo Y, Lin S … +3 more , Zhao X, Zhao X, Yang Z
J Pharm Biomed Anal · 2026 Sep · PMID 41946241 · Publisher ↗
Modified Fangjihuangqi Decoction (MFJHQD) is a formula with clear clinical efficacy for relieving heart failure (HF). However, its potential mechanisms and active substances on HF are rarely reported. In this study, ultr...
Modified Fangjihuangqi Decoction (MFJHQD) is a formula with clear clinical efficacy for relieving heart failure (HF). However, its potential mechanisms and active substances on HF are rarely reported. In this study, ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry was used for identifying chemical composition of MFJHQD and compounds migrating to blood. Overall, 239 compounds were identified in MFJHQD, while 79 prototype compounds and 41 metabolites were recognized in serum. Network pharmacology analysis showed that MFJHQD may modulate oxidative stress to treat HF. Subsequently, an oxidative stress model was established using tert-butyl hydroperoxide (tBHP) stimulation, and a phenotype-anchored hierarchical strategy (the formula, the medicinal herbs, and the representative compounds) was employed to identify active substances against mitochondrial oxidative damage. Experimental studies demonstrated that MFJHQD could alleviate tBHP-induced mitochondrial oxidative injury in cardiomyocytes. Among medicinal herbs of MFJHQD, Stephaniae Tetrandrae Radix, Glycyrrhizae Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma, and Descurainiae Semen exhibited great activity in resisting mitochondrial oxidative damage. Tetrandrine, fangchinoline, calycosin 7-O-glucoside, isoliquiritigenin, caffeic acid, tanshinone IIA, cryptotanshinone, and quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside exhibited significant activity against mitochondrial oxidative injury. This work provides a transferable framework for investigating complex herbal formulas, and reveals the active substances in MFJHQD that alleviate mitochondrial oxidative damage, laying a foundation for its further development.
Development of a facile and novel strategy for N-glycan analysis: Stable isotope labeling via Endo‑CC N180H‑catalyzed transglycosylation for quantitative glycomics.
Li Y, Wang S, Shi H … +5 more , Cong S, Zheng M, Li XL, Todoroki K, Min JZ
J Pharm Biomed Anal · 2026 Sep · PMID 41946240 · Publisher ↗
The Endoglycosidase CC N180H mutant (Endo-CC N180H) is extensively utilized in the synthesis of homogeneous glycopeptides and glycoconjugates due to its high transglycosylation activity. However, its potential in analyti...
The Endoglycosidase CC N180H mutant (Endo-CC N180H) is extensively utilized in the synthesis of homogeneous glycopeptides and glycoconjugates due to its high transglycosylation activity. However, its potential in analytical glycomics remains underexplored. In this study, various structural acceptors were synthesized using N-acetylglucosamine (GlcNAc) as a substrate, and their transglycosylation reactions catalyzed by Endo-CC N180H were investigated. Modifications at the C6 position of GlcNAc completely abolished transglycosylation activity, while alterations at the C1 position were well tolerated. Among these acceptors, d0-Bn-β-GlcNAc exhibited the highest transglycosylation efficiency, with optimal reaction conditions identified as 150 mU/mL Endo-CC N180H at 45 °C for 36 h. Leveraging these conditions, d5-Bn-β-GlcNAc was further synthesized, enabling the development of a stable isotope-labeled chemoenzymatic strategy for N-glycan relative quantification. Using sialylated glycopeptides as a model, the method achieved excellent linearity over a 1:10-10:1 molar ratio range (R = 0.9991), high precision (CV = 0.22-6.24%), and a limit of detection of 50 fmol. Moreover, the strategy was successfully applied to the profiling and relative quantification of high-mannose N-glycans in ribonuclease B. Collectively, this work expands the analytical utility of Endo-CC N180H and establishes a robust and sensitive platform for quantitative glycomics for relative quantification of selected N-glycan species in glycomics research.
Characterization of forced degradation products of tyrosine kinase inhibitor apatinib mesylate by LC-MS/MS and NMR with in silico toxicity assessment.
Shaikh N, Khemchandani R, Shevatkar J … +1 more , Samanthula G
J Pharm Biomed Anal · 2026 Sep · PMID 41946239 · Publisher ↗
Apatinib (APA) a potent tyrosine kinase inhibitor used to treat gastric carcinoma. The study presents the systematic forced degradation behaviour of APA and characterization of the formed degradation products (DPs) by LC...
Apatinib (APA) a potent tyrosine kinase inhibitor used to treat gastric carcinoma. The study presents the systematic forced degradation behaviour of APA and characterization of the formed degradation products (DPs) by LC-MS/MS and NMR. The stability of APA was evaluated under hydrolytic (acid, base, and neutral), oxidative, photolytic (UV and Visible), and solid thermal conditions. The resulting DPs were separated chromatographically by Waters XBridge C18 (250 × 4.6 mm, 5μm) column utilizing the mobile phase of 0.1% trifluoroacetic acid as solvent A and acetonitrile as solvent B. The potential DPs were characterized by interpreting the fragmentation pathway of APA based on LC-MS/MS results. The enriched DP, namely A-6, was isolated by HPLC and further characterized by 1D and 2D NMR data. A mechanistic pathway for the formation of DPs of APA was proposed. In line with ICH Q2 (R2), the developed HPLC assay method of APA was validated and established as linear, precise, specific, accurate, and robust in nature. In silico toxicity profile for APA and its DPs was evaluated by ADMET Predictor®, revealing the positive alerts across multiple toxicity endpoints. The present research provides the framework for drug development based on the stability behaviour of APA, and the developed HPLC method was utilized for routine stability and quality control analysis.
Network pharmacology and experimental validation reveal the multi-target mechanism of Jianpi XiaoxianFormula (BYTQF) against Allergic Rhinitis in mice.
Xiao J, Xie ZF, Zhou JR … +4 more , Huang S, Ma PL, Ban JF, Zheng JK
J Pharm Biomed Anal · 2026 Sep · PMID 41946238 · Publisher ↗
To explore the potential active components and mechanisms of Jianpi XiaoxianFormula (BYTQF) in treating allergic rhinitis (AR) using network pharmacology, molecular docking, and experimental validation. Network pharmacol...
To explore the potential active components and mechanisms of Jianpi XiaoxianFormula (BYTQF) in treating allergic rhinitis (AR) using network pharmacology, molecular docking, and experimental validation. Network pharmacology was employed to screen the active components of BYTQF with therapeutic effects on AR. Molecular docking was used to investigate the mechanism of BYTQF in treating AR. Liquid chromatography-mass spectrometry (LC-MS) was applied to identify the chemical components of BYTQF, and a high-performance liquid chromatography (HPLC) method was established for quality control using neochlorogenic acid, chlorogenic acid, and 1,3-dicaffeoylquinic acid as indicators. An AR mouse model was constructed to verify the efficacy of BYTQF through nasal symptom observation and pathological staining. Network pharmacology identified 71 active components in BYTQF, corresponding to 560 targets, while 2193 AR-related targets were retrieved. A total of 186 overlapping targets were obtained. Through PPI network topological analysis, 10 core targets and 10 key components were identified. Molecular docking showed strong binding activity between key components and core targets such as PTGS2 and MMP2. KEGG pathway enrichment analysis revealed significant enrichment of TNF signaling pathway, AGE-RAGE signaling pathway, etc. LC-MS identified 40 chemical components. HPLC analysis of 10 batches of BYTQF showed the contents of neochlorogenic acid, chlorogenic acid, and 1,3-dicaffeoylquinic acid as 0.3147 ± 0.0057 mg/ml, 0.6263 ± 0.0091 mg/ml, and 0.1709 ± 0.0032 mg/ml, respectively. In vivo experiments demonstrated that BYTQF effectively reduced nasal mucosal epithelial thickness and alleviated inflammatory cell infiltration compared to the model group. BYTQF exerts therapeutic effects on AR by inhibiting inflammatory mediators through the interaction of key components (e.g., chlorogenic acid) with core targets (e.g., MMP2) and regulating pathways such as AGE-RAGE, providing a basis for further research and developmentc.
Metabolism and efficacy-oriented screening of Q-markers for platycodon grandiflorum: A comprehensive evaluation framework of inflammation-specific metabolites and pharmacologically active components in chronic pharyngitis treatment.
Wang X, Chen Y, Wang W … +7 more , Luo L, Bai Z, Li K, Zhang J, Hou Y, Liu D, Bai G
J Pharm Biomed Anal · 2026 Sep · PMID 41946237 · Publisher ↗
Platycodonis Radix (Jiegeng, JG), the dried root of Platycodon grandiflorum (Jacq.) A. DC., is a traditional Chinese medicine frequently utilized for respiratory ailments. Due to the complexity of its saponin constituent...
Platycodonis Radix (Jiegeng, JG), the dried root of Platycodon grandiflorum (Jacq.) A. DC., is a traditional Chinese medicine frequently utilized for respiratory ailments. Due to the complexity of its saponin constituents, the problems related to quality control indicators are yet to be resolved. This research formulates an integrated approach to screen quality markers (Q-markers) for JG in the treatment of chronic pharyngitis (CP). Initially, the anti-inflammatory efficacy of the JG ethanol extract (JGEE) was verified in a CP rat model through histopathology, inflammatory factor assays, and non-targeted metabolomics. Subsequently, metabolic profiling based on UPLC-Q-TOF-MS identified 41 parent compounds (including 36 platycodins) and 57 metabolites in biological specimens (plasma, urine, feces), uncovering remarkable metabolic disparities between the normal and CP states. A quantitative HPLC-ELSD method was developed to evaluate eight major platycodins, meeting the measurability criterion for Q-markers. In vitro screening in RAW264.7 cells identified seven platycodins with significant anti-inflammatory activity. Pharmacokinetic investigations of these active compounds further emphasized that deapio - platycodin E, platycodin E, platycodin D3, and platycodin D demonstrated pathology - dependent pharmacokinetic behavior, corroborating their association with efficacy (effectiveness) and systemic exposure (transmissibility). In summary, this study constructs a comprehensive "efficacy-metabolism-pharmacokinetics" framework and identifies Deapio - platycodin E, Platycodin E, Platycodin D3, and Platycodin D as potential Q - markers for JG in CP treatment, based on specificity, effectiveness, transmissibility, and measurability.
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