Journal Of Pharmaceutical And Biomedical Analysis[JOURNAL]
Sun
200 papers
Construction of bio-layer interferometry biosensors via cell-free synthesized proteins for fishing bioactive compounds from Chinese herbs.
Wang Y, Wang X, Xia N … +5 more , Chen H, Zhang L, Lu B, Lv D, Cao Y
J Pharm Biomed Anal · 2026 Jun · PMID 41687222 · Publisher ↗
The application of Bio-Layer Interferometry (BLI) is contingent upon the immobilization of highly purified target proteins onto the sensor. The cumbersome and time-consuming nature of traditional protein expression and p...
The application of Bio-Layer Interferometry (BLI) is contingent upon the immobilization of highly purified target proteins onto the sensor. The cumbersome and time-consuming nature of traditional protein expression and purification processes restricts the application of BLI in high-throughput screening of traditional Chinese medicine (TCM). This study aims to develop a rapid and efficient BLI-based platform for screening bioactive components in TCM. An integrated platform combining cell-free protein synthesis (CFPS), BLI, and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) was established for efficient TCM bioactive compound discovery. Functional C-X-C chemokine receptor 4 (CXCR4) was synthesized in vitro using a CFPS system, which were then validated by surface plasmon resonance (SPR) and western blotting. Immobilized CXCR4 on NTA biosensors enabled BLI-based high-throughput screening of TCM extracts, followed by target-specific compound recovery and characterized via mass spectrometry. Three bioactive TCM constituents were successfully fished and identified as coptisine, ligustilide, and senkyunolide A. All of them exhibited negligible cytotoxicity at concentrations ranging from 6.25 to 100 μM). Furthermore, ligustilide and senkyunolide A demonstrated certain affinity for CXCR4 with K of 69.86 μM and 14.7 μM, respectively, and significantly inhibited cell migration. This study is the first identification of ligustilide and senkyunolide A as functional ligands of CXCR4. The established CFPS-BLI-UHPLC-QTOF/MS platform enables efficient discovery of low-toxicity, high-affinity CXCR4-targeting therapeutics from TCM.
Aggregation-induced luminescence probe based lateral flow immunoassay for the simultaneous quantitative detection of IL-6/PCT.
He R, Zhu Y, Zhou W … +7 more , Nawaz MAH, Li Y, Zhu J, Feng H, Nowicka AM, Han W, Yu C
J Pharm Biomed Anal · 2026 Jun · PMID 41678998 · Publisher ↗
The detection of important inflammatory biomarkers possesses substantial advantages in guiding clinical decision-making. In particular, simultaneous detection of interleukin-6 (IL-6) and procalcitonin (PCT) significantly...
The detection of important inflammatory biomarkers possesses substantial advantages in guiding clinical decision-making. In particular, simultaneous detection of interleukin-6 (IL-6) and procalcitonin (PCT) significantly improves the differentiation between bacterial and viral infections, a critical challenge in early-stage diagnostics. To address this need, a reliable lateral flow immunoassay (LFIA) incorporating aggregation-induced luminescent nanoparticles (BTA@PS) was successfully developed. A novel aggregation-induced emission (AIE) probe was designed and encapsulated within polystyrene microspheres, thereby overcoming the limitations of aggregation-caused quenching (ACQ) in conventional fluorescent materials. The resulting BTA@PS nanoparticles, conjugated with specific antibodies against IL-6 and PCT, served as stable and effective immunofluorescent probes for the LFIA platform. Under optimized experimental conditions, the developed BTA@PS-LFIA enabled simultaneous quantification of IL-6 and PCT, demonstrating excellent linearity over the range of 2-8000 pg/mL and 0.04-30 ng/mL, respectively, with coefficient of variation (CV) values of below 5 %. Furthermore, this method demonstrated superior detection capability for PCT and IL-6 in serum, confirming its high potential for rapid clinical diagnostics.
Simultaneous determination of three phosphatidylethanol homologues, 12 drugs and metabolites in whole blood by LC-MS/MS.
Maria MH, Neng NR, Berg T
J Pharm Biomed Anal · 2026 Jun · PMID 41678997 · Publisher ↗
The use of alcohol, legal and illicit substances poses great negative consequences on health and economy worldwide. LC-MS/MS allow simultaneous determination of multiple compounds in biological matrices. The aim of this...
The use of alcohol, legal and illicit substances poses great negative consequences on health and economy worldwide. LC-MS/MS allow simultaneous determination of multiple compounds in biological matrices. The aim of this study was to develop a LC-MS/MS method for the determination of the alcohol biomarker phosphatidylethanol (PEth) - including three homologues (PEth 16:0/18:1, PEth 16:0/18:2, PEth 18:0/18:1) - cocaine and three metabolites, and 8 other drugs in whole blood. Whole blood in K2EDTA tubes was prepared by liquid-liquid extraction using heptane/ethyl acetate/2-propanol (16:64:20, v:v:v). Chromatographic separation was achieved on an Acquity BEH C column (50 × 2.1 mm I.D., 1.7 µm particles). Mobile phase was 0.025 % ammonia, pH 10.7 (Solvent A) and methanol (Solvent B). The method was fully validated with isotope-labelled internal standards for 10 compounds. Inter-assay precision and accuracy were within ± 16 % for all analytes at five to seven tested concentrations. Recovery was within 42-79 % for 14 compounds and 11 % for benzoylecgonine. Matrix effects were within ± 25 % for most analytes. Internal standards compensated for matrix effects for compounds that had their own internal standards. A robust, precise, and accurate LC-MS/MS method for the determinations of three PEth homologues and 12 drugs and metabolites was, developed and validated. The method is valuable, especially for detecting polydrug use and alcohol consumption. To the best of our knowledge, this is the first LC-MS/MS method for the simultaneous determination of three PEth homologues and different drugs and metabolites.
Identification of blood-absorbed components of Qingkui Yuyang decoction and its mechanistic roles in ulcerative colitis based on UPLC-Q-Exactive Orbitrap-MS/MS and network pharmacology.
Chen Y, Liu L, Wei Y … +2 more , Zhang Q, Wang Y
J Pharm Biomed Anal · 2026 Jun · PMID 41678996 · Publisher ↗
Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by recurrent episodes of intestinal inflammation and mucosal injury. Qingkui Yuyang decoction (QKY), a clinically validated traditional Chines...
Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by recurrent episodes of intestinal inflammation and mucosal injury. Qingkui Yuyang decoction (QKY), a clinically validated traditional Chinese medicinal formula, has been widely used in the treatment of UC; however, its pharmacodynamically active constituents and underlying mechanisms of action have not been fully elucidated. In this study, we explored the therapeutic mechanisms of QKY in treating UC by employing a combination of serum pharmacochemistry, network pharmacology, and molecular docking techniques. Initially, using UPLC-Q-Exactive Orbitrap-MS/MS, 28 candidate active compounds in the serum of rats treated with QKY were identified. Subsequently, network pharmacology analysis identified 43 overlapping targets between UC and the active components, and 30 related signaling pathways. Further analysis and molecular docking studies have confirmed that the key active components (Loureirin A, Berberine, Ellagic acid) possess potential for effective therapeutic effects with the core targets (RELA, AKT1). In addition, in vitro experiments demonstrated that QKY significantly downregulated the expression levels of the pro-inflammatory cytokines IL-6 and TNF-α. QKY also markedly reduced the phosphorylation levels of NF-κB p65 and p38 MAPK, as well as the corresponding mRNA expression levels of these signaling molecules. These results suggest that QKY may exert its therapeutic effects on UC by modulating the MAPK and NF-κB signaling pathways, offering a promising strategy for the prevention and treatment of UC.
Determination of Gentamicin C-subtypes in Inner Ear Perilymph Using Liquid Chromatography with Fluorescence Detection.
Dash S, McDevitt MT, Smith DD … +1 more , Steyger PS
J Pharm Biomed Anal · 2026 Jun · PMID 41666750 · Full text
Gentamicin is a broad-spectrum aminoglycoside used frequently to treat gram-positive and gram-negative bacterial infections. In this study, a new, simple, fast, and sensitive isocratic reversed-phase high performance liq...
Gentamicin is a broad-spectrum aminoglycoside used frequently to treat gram-positive and gram-negative bacterial infections. In this study, a new, simple, fast, and sensitive isocratic reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the detection and quantification of fluorescent OPA-ethanethiol derivatized gentamicin in very small biological sample volumes. To our knowledge, there is no report of the use of ethanethiol for the derivatization of gentamicin with OPA, and the simultaneous determination of the four major C-subtypes of gentamicin using OPA-derivatives. Optimum chromatographic conditions were achieved on a C column with a mobile phase consisting of methanol, glacial acetic acid, and an aqueous solution of sodium 1-heptanesulfonate at a flow rate of 1.0 mL/min under ambient conditions. The method was successfully validated according to the acceptance criteria of USP guidelines in terms of selectivity, linearity, accuracy, precision, and sensitivity. The linearity of the method was demonstrated with a concentration range of gentamicin (10-400 ng/mL) prepared in artificial perilymph. The limit of detection was 0.2 ng/mL and the limit of quantification was 10-11 ng/mL for all four major C-subtypes of gentamicin. Finally, due to its high sensitivity, this method was successfully applied to quantify gentamicin concentrations in the small volumes of perilymph present in the inner ear of mice. Thus, this RP-HPLC-fluorescence method for detecting derivatized gentamicin in preclinical models is promising in terms of simplicity and high sensitivity.
Temperature as the primary risk factor for sevoflurane degradation: Identification and toxicological risk assessment of novel degradants by nuclear magnetic resonance (NMR) spectroscopy.
Jin Y, Li Y, Yang H … +3 more , Yang H, Guo Y, Wu X
J Pharm Biomed Anal · 2026 Jun · PMID 41666749 · Publisher ↗
Sevoflurane is an inhalation anesthetic, which is widely favored by anesthesiologists in modern clinical practice. It is reported that sevoflurane readily degrades under certain conditions. Forced degradation studies are...
Sevoflurane is an inhalation anesthetic, which is widely favored by anesthesiologists in modern clinical practice. It is reported that sevoflurane readily degrades under certain conditions. Forced degradation studies are essential for elucidating its degradation profile under stressed conditions, as recommended by the ICH guidelines. This study systematically assessed the impact of temperature, light, oxidation, and hydrolysis on sevoflurane stability through forced degradation studies. The results indicated that temperature was identified as the primary trigger for degradation, leading to the formation of four degradation products (DPs) (including the pharmacopeial-known Impurity C and three novel sevoflurane degradants H, I, and J) at 60℃ and 95℃. Light, oxidation, and hydrolysis had no significant effect under the tested conditions. The structures of all DPs were confirmed using nuclear magnetic resonance spectroscopy (NMR). Capitalizing on the inherent quantitative proficiency of this technique, we subsequently employed quantitative proton nuclear magnetic resonance spectroscopy (qHNMR) to determine the relative contents of these four DPs, which can further elucidate the degradation profile of sevoflurane. In silico toxicity and metabolic behavior of them were assessed by Derek Nexus and Meteor Nexus software, respectively. DPs H-J exhibited potential skin irritation/corrosion and sensitization effects across species, potentially attributable to alkyl aldehyde functional groups. All of our efforts are expected to provide guidance for the quality control and optimal storage of sevoflurane.
Three-dimensional high-performance liquid chromatographic determination of serine, threonine and allothreonine enantiomers in the d-amino acid oxidase deficient mice and rats.
Oyaide M, Akita T, Ishii C … +5 more , Shimizu Y, Mita M, Konno R, Okamura T, Hamase K
J Pharm Biomed Anal · 2026 Jun · PMID 41655298 · Publisher ↗
The amounts of the serine (Ser), threonine (Thr) and allothreonine (aThr) enantiomers were determined in tissues (cerebrum, cerebellum, pancreas, liver and kidney) and physiological fluids (plasma and urine) of rats and...
The amounts of the serine (Ser), threonine (Thr) and allothreonine (aThr) enantiomers were determined in tissues (cerebrum, cerebellum, pancreas, liver and kidney) and physiological fluids (plasma and urine) of rats and mice with deficiency of d-amino acid oxidase (DAO). DAO is an enzyme metabolizing d-amino acids in mammals and has been implicated in the pathophysiology of several diseases via the alteration of d-amino acids. To determine trace levels of the amino acid enantiomers, a three-dimensional (3D) HPLC system composed of reversed-phase, anion-exchange and chiral separations was designed and utilized. Prior to the 3D-HPLC analysis, the analytes were derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole to enhance the fluorescence detection sensitivity. By using the 3D-HPLC system, the tissues and physiological fluids of F344-Dao rats and B6DAO mice (animals with the DAO deficiency) were analyzed. In both species, d-Ser levels were elevated in the absence of DAO activity except for the cerebrum. The amounts of d-Thr and d-aThr were increased in the cerebellum and kidney with the DAO deficiency while their amounts were almost the same in the other tissues and physiological fluids. These results indicated that the intrinsic d-Ser analogues were metabolized by DAO in mammals and further studies to clarify its physiological significance are expected.
Development and validation of a sensitive and rapid UHPLC-MS/MS method for the simultaneous quantification of CG-0255 and its active metabolite in human plasma and its application to Phase I studies.
Chen H, Li J, Yuan F … +7 more , He G, Wu H, Yan H, Xu H, Liu C, Sheng L, Li X
J Pharm Biomed Anal · 2026 Jun · PMID 41650587 · Publisher ↗
CG-0255, a thiol prodrug of clopidogrel's active metabolite H4 (CG-0236), is a novel thienopyridine P2Y12 receptor antagonist under initial clinical development for the treatment of acute coronary syndromes. Unlike clopi...
CG-0255, a thiol prodrug of clopidogrel's active metabolite H4 (CG-0236), is a novel thienopyridine P2Y12 receptor antagonist under initial clinical development for the treatment of acute coronary syndromes. Unlike clopidogrel, CG-0255 is converted to the active thiol metabolite H4 (CG-0236) in a single hydrolytic step. Compared with clopidogrel, CG-0255 exhibits more efficient and consistent H4 formation in humans, which can be quantified in plasma following either intravenous or oral administration. In this study, we developed and validated a sensitive, rapid, and robust UHPLC-MS/MS method for the simultaneous quantification of CG-0255 and its derivatized active metabolite (MP-H4, CG-0261) in human plasma. After solid-phase extraction from 94.5 μL of plasma, analytes and isotope-labeled internal standards were separated on an ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using isocratic elution with 0.1 % formic acid in water and acetonitrile (57:43, v/v) at a flow rate of 0.5 mL/min, followed by a 3.5 min column washing and re-equilibration, giving a total analytical run time of 7 min. Baseline separation of CG-0255, CG-0261, and their respective isomers was achieved. Detection was performed using positive electrospray ionization in multiple reaction monitoring mode on a Q-Trap 6500 mass spectrometer. Calibration curves were linear over 0.05-25 ng/mL for both analytes, corresponding to 0.0353-17.65 ng/mL for H4 (CG-0236). Intra- and inter-day precision and accuracy were within ±15 % at all quality-control levels. The validated assay was successfully applied to two phase I clinical studies conducted at our center, characterizing the pharmacokinetics of CG-0255 following single-dose intravenous and multiple-dose oral administration. This UHPLC-MS/MS method provides a reliable platform for the quantitative evaluation of CG-0255 and its active metabolite in human plasma, and is well suited to support further global clinical development.
A streamlined workflow for early phase formulation development of monoclonal antibodies comprising multi-attribute method and ligand binding assay.
Smith R, Guy C, Upton R … +5 more , Clawson S, Fisher H, Nasar MA, Firth D, Watkinson A
J Pharm Biomed Anal · 2026 Jun · PMID 41650586 · Publisher ↗
A streamlined early phase formulation development workflow has been developed for monoclonal antibodies (mAbs) to provide a more efficient process, driving down costs and reducing timelines, without compromising the qual...
A streamlined early phase formulation development workflow has been developed for monoclonal antibodies (mAbs) to provide a more efficient process, driving down costs and reducing timelines, without compromising the quality and hence patient safety. The proposed novel workflow combines liquid chromatography-mass spectrometry multi-attribute method (LC-MS MAM) and sensitive ligand binding using surface plasmon resonance (SPR). By linking the two methodologies it is possible to obtain a comprehensive understanding of a mAb's critical quality attributes (CQAs) and provide a structure/function correlation. As LC-MS MAM cannot address all aspects of degradation, high throughput methods for the analysis of high molecular weight material (HMWM), and conformational and colloidal stability, were also evaluated. The workflow comprises an initial forced degradation study, to verify stability-indication and identify potential degradation routes. Secondly, optimal pH, based on conformational and colloidal stability, is determined. Finally, stabilising excipients are evaluated by design of experiment (DoE). We have verified this workflow using pembrolizumab. In an initial forced degradation study, LC-MS MAM and PD-1 ligand binding could identify the CQAs. Met105 oxidation, located in the CDR3 region, was identified as the major CQA. DoE demonstrated that 25 mM methionine inhibited Met105 oxidation and stabilised PD-1 binding. With this streamlined process, we were able to improve the stability of the protein by formulating in 20 mM histidine, 25 mM methionine, 0.02 % PS80 and 300 mM sucrose, at pH 5.5. The described workflow has the potential to decrease the demand for precious early development material as well as reduce costs and shorten timelines.
Determination of antigen components in inactivated SARS-CoV-2 vaccine using ultra-high-performance liquid chromatography tandem mass spectrometry.
Shi J, Deng F, Li Y … +4 more , Li J, Peng R, Yuan J, Tan L
J Pharm Biomed Anal · 2026 Jun · PMID 41643282 · Publisher ↗
Accurate and reliable quantification of antigen components in vaccines is critical in vaccine quality control and evaluation of immunogenic consistency. However, conventional immunoassays often suffer from limited specif...
Accurate and reliable quantification of antigen components in vaccines is critical in vaccine quality control and evaluation of immunogenic consistency. However, conventional immunoassays often suffer from limited specificity, trace-level antigen concentrations, and indirect quantification. In this study, we demonstrated an ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the determination of effective antigen components in inactivated SARS-CoV-2 vaccines. Specifically, the vaccine samples were denatured and digested with trypsin to generate tryptic peptides. Then, the signature peptides derived from the nucleocapsid protein and their stable isotope-labeled internal standards were selectively captured and separated using anti-peptide antibody-conjugated magnetic beads. The signature peptides were characterized by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, which confirmed their amino acid sequence and multi-charged ionization states. Quantitative analysis was then performed using UHPLC-MS/MS in positive electrospray ionization mode with multiple reaction monitoring. Chromatographic separation of the signature peptides was achieved on an ACQUITY Premier Peptide BEH C₁₈ column using 0.1 % formic acid in water and 0.1 % formic acid in acetonitrile as the mobile phases. The method was validated and exhibited excellent linearity for the signature peptides over the concentration range of 1-60 μg/L, with correlation coefficients higher than 0.999. The recovery ranged from 75.1 % to 86.3 %, with intra-day precision (RSD) of 0.8-1.0 % and inter-day precision of 1.3-3.6 %. Finally, the method was successfully applied to determine the effective antigen components in inactivated SARS-CoV-2 vaccine samples. The concentrations of the signature peptide ADETQALPQR ranged from 4.95 to 12.95 µg/L across the three vaccine batches analyzed, corresponding to 4.38-11.47 nmol/L of nucleocapsid protein. The results indicated that the method exhibited great promise for the determination of active antigenic proteins in inactivated SARS-CoV-2 vaccine samples and provided an alternative analytical platform for vaccine quality control.
Fecal metabolic biomarkers associated with insomnia severity: A study on 5-hydroxyindoleacetic acid, octopamine, oleoylethanolamide, and elaidic acid.
Wu Y, Li X, Feng H … +3 more , Chen Y, Wu W, Wang S
J Pharm Biomed Anal · 2026 Jun · PMID 41638038 · Publisher ↗
Insomnia is increasingly recognized as a disorder with complex metabolic underpinnings. We investigated the changes in fecal metabolite levels of 5-hydroxyindoleacetic acid (5-HIAA), octopamine (OA), oleoylethanolamide (...
Insomnia is increasingly recognized as a disorder with complex metabolic underpinnings. We investigated the changes in fecal metabolite levels of 5-hydroxyindoleacetic acid (5-HIAA), octopamine (OA), oleoylethanolamide (OEA), and elaidic acid (EA) in patients with sleep disorders, as well as their correlations with insomnia severity. Sixty participants were divided into two groups, with thirty patients with sleep disorders hospitalized in the Department of Neurology, Zhongda Hospital, Southeast University (October 2024-March 2025) and 30 healthy controls recruited during the same period. Fecal samples were collected from all participants, and metabolite levels were analyzed via untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS). The Chinese version of the Insomnia Severity Index (C-ISI) was employed to evaluate insomnia severity, and the correlations between insomnia severity and these four metabolites were subjected to statistical analysis. Both univariate and multivariate analyses revealed significant metabolic differences between groups. The experimental group showed significantly lower levels of 5-HIAA (FC = 0.947, P = 0.020) and OA (FC = 0.953, P < 0.001), but higher OEA (FC = 1.101, P < 0.001) and EA (FC = 1.026, P < 0.001). C-ISI scores correlated negatively with 5-HIAA (r = -0.380, P = 0.003) and OA (r = -0.448, P < 0.001), and positively with OEA (r = 0.500, P < 0.001) and EA (r = 0.408, P = 0.001). These fecal metabolites associate with insomnia severity and may serve as potential biomarkers for understanding its pathophysiology and developing interventions.
Detection and localization of single-nucleotide mutations in synthetic oligonucleotides by ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry.
Gab-Allah MA, Hwang H, Kim M … +7 more , Tran NT, Kim BJ, Kim M, Han JH, Kim Y, Choi BY, Kim J
J Pharm Biomed Anal · 2026 Jun · PMID 41638037 · Publisher ↗
Accurate detection of point mutations in mitochondrial DNA (mtDNA) is crucial for diagnosing various mitochondrial disorders. In this study, we developed an ultra-high-performance liquid chromatography coupled with high-...
Accurate detection of point mutations in mitochondrial DNA (mtDNA) is crucial for diagnosing various mitochondrial disorders. In this study, we developed an ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) method for the direct, label-free identification and localization of single-nucleotide mutations using synthetic 20- and 49-mer oligonucleotides as model fragments representing the pathogenic mtDNA point mutation (mt.3243 A>G). Three mobile phase systems, including ammonium bicarbonate (ABC), triethylamine/hexafluoroisopropanol (TEA/HFIP), and tributylamine/HFIP (TBA/HFIP), were systematically evaluated to assess their effects on oligonucleotide retention behavior and duplex stability under denaturing and non-denaturing conditions. The ABC buffer provided optimal performance for maintaining partial duplex integrity, while TEA/HFIP offered superior ionization efficiency for single-stranded analysis. Deconvoluted mass spectra revealed accurate monoisotopic mass differences between wild-type and mutant oligonucleotides, including ∼ + 16 Da for the sense strand (A>G), ∼ -15 Da for the antisense strand (T > C), and ∼ + 1 Da for the duplex, enabling confident mutation discrimination at the intact molecular level. High-resolution MS achieved excellent mass accuracy within ±3 ppm, and high-energy collision dissociation (HCD) MS/MS enabled sequence-specific fragmentation that localized the mutation site with high confidence when compared with theoretical fragments. Overall, this study establishes a reliable analytical framework for mutation detection in oligonucleotide models and highlights the potential of UHPLC-HRMS/MS as a complementary tool for targeted mtDNA fragment analysis.
Development and validation of an UPLC-MS/MS method for simultaneous determination of meropenem and its open-ring metabolite in human serum and cerebrospinal fluid with application to clinical samples.
Chen X, Xu J, Wang C … +7 more , Yang L, Fan J, Li T, Zhang Q, Yu Y, Tang L, Huang S
J Pharm Biomed Anal · 2026 Jun · PMID 41638036 · Publisher ↗
Central nervous system (CNS) infections require adequate drug exposure at the site of action, yet antibiotic meropenem (MER) shows limited cerebrospinal fluid (CSF) penetration and easily undergoes non-enzymatic degradat...
Central nervous system (CNS) infections require adequate drug exposure at the site of action, yet antibiotic meropenem (MER) shows limited cerebrospinal fluid (CSF) penetration and easily undergoes non-enzymatic degradation to an inactive open-ring metabolite (ORM). In this study, we developed a simple, sensitive, and fast liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of MER and ORM in human serum and CSF. Chromatographic separation was accomplished on an Agela Venusil MP C18 column, with MER-d6 and ORM-d6 as internal standards. Methanol was found to promote methanolysis, yielding a characteristic product (m/z 416.2). Therefore, acetonitrile was selected as both the organic phase and the protein-precipitation solvent. Method validation was conducted according to the ICH M10 guideline. Follow validation, the method was successfully applied to 57 serum and 16 CSF samples. ORM concentrations in human CSF were reported for the first time. This method provides a valuable tool to support MER monitoring in patients with CNS infections.
Cost-effective routine pharmaceutical testing using radial flow stream splitting HPLC columns: Quantitative analysis and performance metrics in the analysis of over-the-counter drugs.
McDermott M, Sargeant Z, Karlsen CE … +3 more , Li F, Shalliker RA, Cravino JA
J Pharm Biomed Anal · 2026 Jun · PMID 41638035 · Publisher ↗
The demand for rapid and reliable analytical methods in the pharmaceutical industry continues to grow, with Ultra-/High-Performance Liquid Chromatography (U/HPLC) remaining the gold standard for impurity profiling, quant...
The demand for rapid and reliable analytical methods in the pharmaceutical industry continues to grow, with Ultra-/High-Performance Liquid Chromatography (U/HPLC) remaining the gold standard for impurity profiling, quantification of active pharmaceutical ingredients, and degradation product analysis. However, traditional HPLC methods are often constrained by pressure limitations at higher flow rates, which can hinder analytical throughput. While recent advancements in column technology have improved performance, they typically exacerbate pressure-related challenges. In this study, we evaluate a novel column technology designed to address these limitations by enabling high-resolution separations at reduced pressures and increased flow rates. Our findings demonstrate that the column, when operated in Radial Flow Stream Splitting (RFS) mode, maintains quantitative accuracy and repeatability while achieving up to a 120 % improvement in separation efficiency and a 30 % reduction in backpressure compared to conventional operation. By way of assaying over-the-counter medication, we have found no difference in the quantitative reliability of the assay when in RFS vs stock mode, despite reducing the analysis time by up to 40 %.
Sequential digestions enable identification and quantification of rapid aspartic acid isomerization in the CDR of a monoclonal antibody light chain.
Jonveaux J, Faudon M, Heymes P … +4 more , Lucchini V, Estrada MFZ, Jahn M, Zarei M
J Pharm Biomed Anal · 2026 Jun · PMID 41638034 · Publisher ↗
Isomerization of aspartic acid (Asp) to isoaspartic acid (isoAsp) within the complementarity-determining regions (CDRs) of monoclonal antibodies (mAbs) can lead to conformational changes that decrease antigen-binding aff...
Isomerization of aspartic acid (Asp) to isoaspartic acid (isoAsp) within the complementarity-determining regions (CDRs) of monoclonal antibodies (mAbs) can lead to conformational changes that decrease antigen-binding affinity. Although isomerization can significantly alter the chromatographic and electrophoretic profiles, precise localization of this modification requires a mass spectrometry-based approach, such as peptide mapping. In this work, we present a case study that investigates various analytical strategies to identify the root cause of significant changes observed in the chromatographic and electrophoretic profiles of an mAb during formulation development. LC-MS analysis of reduced mAb using high-resolution mass spectrometry, peptide mapping using trypsin digestion, and fraction collection of the newly identified peak followed by trypsin digestion suggested that isomerization occurs within the CDR of the mAb. However, due to the presence of three Asp residues within a single tryptic peptide, this modification could not be precisely localized. To overcome this limitation, we developed a sequential enzymatic digestion strategy, utilizing trypsin followed by Asp-N digestion, which enabled accurate localization and quantification of the isomerization site. The resulting data indicated that the main isoAsp signal originated from isomerization at the DS motif that increased substantially over time in the liquid formulation, while no significant change was observed in the lyophilized formulation. The level of isomerization determined through this sequential digestion method correlated well with the LC-UV quantitation data of the reduced mAb.
Corrigendum to 'A liquid biopsy-RNAseq method for monitoring the expression of genes involved in drug disposition: proof-of-concept application to cholestatic liver disease' [J. Pharm. Biomed. Anal. (2025) 269: 117244].
Dahal A, Sierra T, Hayes CM … +4 more , Assis DN, Rostami-Hodjegan A, Ghonem NS, Achour B
J Pharm Biomed Anal · 2026 Jun · PMID 41633090 · Publisher ↗
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Comprehensive pharmacokinetic and tissue distribution study of α,ω-dipropionic acid polyethylene glycol (PA-PEG₁₂-PA) in rats using a validated UPLC-MS/MS method.
Liu M, Yu Y, Jing Y … +6 more , Guo Y, Wang C, Xue H, Liu H, Yin L, Shi M
J Pharm Biomed Anal · 2026 May · PMID 41619531 · Publisher ↗
While polyethylene glycol (PEG) is an extensively utilized pharmaceutical polymer with established biocompatibility and regulatory acceptance, the in vivo pharmacokinetics of its bifunctional derivatives, such as α,ω-dip...
While polyethylene glycol (PEG) is an extensively utilized pharmaceutical polymer with established biocompatibility and regulatory acceptance, the in vivo pharmacokinetics of its bifunctional derivatives, such as α,ω-dipropionic acid polyethylene glycol (PA-PEG-PA), remain largely unexplored. This study presents the development and validation of a highly sensitive and selective UPLC-MS/MS method for the accurate quantification of PA-PEG-PA in various biological matrices, utilizing a straightforward protein precipitation protocol. After intravenous administration in rats (10 mg/kg), PA-PEG-PA demonstrated rapid clearance (t = 3.99 ± 1.06 h). Tissue distribution analysis revealed a pronounced affinity for renal accumulation, with kidney concentrations (16473 ± 881 ng/g at 2 h) substantially surpassing those in the liver and lungs, followed by rapid depletion within 24 h. Excretion studies indicated renal clearance as the dominant pathway, with 55.93 % of the administered dose recovered unchanged in urine over 72 h, while fecal excretion was minimal (1.76 %). This work provides the first comprehensive pharmacokinetic and biodistribution profile of PA-PEG-PA, underscoring its renal-driven clearance and tissue disposition. The findings offer crucial insights for the rational design of PEGylated drug delivery systems, and the robust UPLC-MS/MS assay established herein serves as a valuable tool for characterizing polymeric excipients in biological environments.
Chemical profiling and multi-dimensional pharmacokinetic analysis of shengmaiyin oral liquid for cardiac dysfunction.
Zhao L, Wu S, Yu X … +5 more , Huang Z, Liu L, Cheng X, Yuan Z, Li Y
J Pharm Biomed Anal · 2026 May · PMID 41616564 · Publisher ↗
Shengmaiyin oral liquid (SMY), formulated with schisandra chinensis, red ginseng, and Ophiopogon japonicus, is widely used for treating cardiac dysfunction (CD). However, its functional chemical basis and pharmacological...
Shengmaiyin oral liquid (SMY), formulated with schisandra chinensis, red ginseng, and Ophiopogon japonicus, is widely used for treating cardiac dysfunction (CD). However, its functional chemical basis and pharmacological profiles remain insufficiently explored. This study employed advanced analytical strategies to characterize the bioactive components of SMY and investigate their in silico pharmacodynamics and in vivo pharmacokinetics, providing mechanistic insights into their roles in CD treatment. Using UPLC-Q-TOF-MS/MS, 132 compounds were identified in SMY, with 31 detected in the plasma of SMY-treated mice. Among these, 21 chemicals (12 lignans and 9 saponins) were identified as key bioactives against CD. Network pharmacology and molecular docking revealed their multi-target interactions and varied binding affinities with CD-related proteins. Pharmacokinetic (PK) analysis showed that 5 compounds had high plasma exposure and rapid elimination in CD model mice. All 21 chemicals exhibited significant tissue distribution following prolonged SMY administration. The globally pharmacokinetic seeking (GPS) box analysis revealed two distinct fast and slow PK patterns among the chemicals. Notably, a highly tissue-specific and time-dependent alteration of lignan and saponin clusters was observed in the hearts of CD mice within 8 h post-administration. This study highlights the GPS box as an innovative platform for multi-dimensional PK analysis. These findings advance the integration of traditional herb medicine with modern analytical methodologies, offering new perspectives for developing precision medicine approaches in ethnopharmacology.
The discovery of pharmacodynamic material basis and mechanism of Shuganning injection in treating damp-heat jaundice syndrome: Combining metabolomics and serum pharmacochemistry.
He Y, Lin X, Zhong W … +5 more , Liu X, Yang X, Yao Y, Li M, Zhang J
J Pharm Biomed Anal · 2026 May · PMID 41616563 · Publisher ↗
Shuganning injection (SGNI) is derived from Yinchenhao decoction and has the functions of promoting diuresis, reducing jaundice and protecting the liver. It is primarily utilized in the treatment of liver diseases, parti...
Shuganning injection (SGNI) is derived from Yinchenhao decoction and has the functions of promoting diuresis, reducing jaundice and protecting the liver. It is primarily utilized in the treatment of liver diseases, particularly damp-heat jaundice. However, the pharmacodynamic material basis and specific mechanisms remain unclear. Therefore, this study aims to elucidate the pharmacodynamic material basis and mechanism of SGNI in the treatment of damp-heat jaundice syndrome (DHJS). High-throughput metabolomics was employed to identify biomarkers associated with DHJS. The migratory prototype components and metabolites present in the blood of SGNI were characterized through serum pharmacochemistry. Correlation analysis was conducted to associate the biomarkers of DHJS with the migrating components in the blood of SGNI, clarifying the pharmacodynamic basis of SGNI in treating DHJS. Furthermore, metabolic pathway enrichment analysis was performed to explore the endogenous core metabolites and key metabolic pathways, providing insights into the mechanism of SGNI in treating DHJS from the metabolic perspective. Finally, a total of 25 metabolites were identified as biomarkers for DHJS, along with 24 prototype migratory components and 38 migratory metabolites of SGNI. It was established that the pharmacodynamic substance basis of SGNI in the treatment of DHJS comprises 15 components. These components primarily exert their effects through key metabolic pathways, including pentose and glucuronate interconversions, arachidonic acid metabolism, and primary bile acid biosynthesis. In conclusion, SGNI demonstrates a significant therapeutic effect on DHJS. It has preliminarily elucidated the pharmacodynamic substance basis and mechanism of action at the metabolic level, offering a scientific foundation for the clinical application and further development of SGNI.
The mechanism of acetyl-L-carnitine on colorectal cancer and its metabolomic study.
Ning J, Cai C, Fan X … +2 more , Liao C, Shen M
J Pharm Biomed Anal · 2026 May · PMID 41610723 · Publisher ↗
Colorectal cancer (CRC) is a prevalent malignancy worldwide, with increasing incidence and mortality rates. Acetyl-L-carnitine (ALC), a natural form of L-carnitine, possesses anti-inflammatory, free radical-scavenging, a...
Colorectal cancer (CRC) is a prevalent malignancy worldwide, with increasing incidence and mortality rates. Acetyl-L-carnitine (ALC), a natural form of L-carnitine, possesses anti-inflammatory, free radical-scavenging, and mitochondrial membrane-stabilizing properties. However, its precise mechanism of action in CRC remains unclear. To articulate the mechanism of action of ALC in CRC, we investigated its biological activity in HCT116 cells and a nude mouse xenograft model. The effects of ALC on HCT116 cells using CCK-8, colony formation, migration, invasion, and cell cycle assays. And detected oxidative stress-related indexes including reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD). The inhibitory effect of ALC on colorectal tumor growth was further validated in a nude mouse xenograft model, and untargeted metabolomic analysis was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry. The results showed that ALC can inhibit the proliferation and migration of HCT116 cells, induce cell cycle arrest and apoptosis, and affect the changes of oxidative stress indexes, and exacerbating cellular oxidative stress damage. In vivo experiments demonstrated that ALC effectively suppressed colorectal tumor growth in a dose-dependent manner. Plasma metabolomic analysis identified 10 differential metabolites between the ALC group and the control group, including meisoindigo, indole-3-acetaldehyde, etc., and 4 tumor differential metabolites, including L-proline, 3,4,5-Trimethoxytoluene, etc. Pathway enrichment analysis revealed that ALC altered 6 metabolic pathways in plasma, including tryptophan metabolism and glycine, serine, and threonine metabolism, and 5 pathways in tumor tissue, including arginine and proline metabolism and the sphingolipid signaling pathway. These findings provide novel theoretical evidence for the antitumor mechanisms of ALC in CRC.
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