Journal of chromatography. B, Analytical technologies in the biomedical and life sciences最新文献

{"title":"Recommendation of a milder cleaning-in-place method for Cytiva's second-generation protein L resin MabSelect VL.","authors":"Ju Qu, Yifeng Li, Yan Wan","doi":"10.1016/j.jchromb.2025.124810","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124810","url":null,"abstract":"<p><p>Protein L-based media are a type of affinity resin whose application in antibody purification is only secondary to Protein A resins. As Protein L resins exhibit stronger byproduct separation capabilities than Protein A resins, they are particularly preferred when purifying complex molecules such as bispecific antibodies (bsAbs). Despite the growing use of Protein L resins, comprehensive evaluations of their lifetime and cleaning-in-place (CIP) methods are generally lacking. In the current study, we examined MabSelect VL, Cytiva's second-generation Protein L resin, for its alkaline stability. In specific, the resin's performance was evaluated after 80 cycles of sanitization with 0.1 M sodium hydroxide (NaOH). In addition, the binding behavior of monomer and aggregates to aged resin was studied to understand the change in resin ligands during use. According to the data, 0.1 M NaOH is not well tolerated by MabSelect VL, and its repeated treatment causes performance degradation. Therefore, we recommend a milder sanitization method, 0.05 M NaOH, which can be combined with 1 % benzyl alcohol to improve microbial control.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":"1267 ","pages":"124810"},"PeriodicalIF":0.0,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Retention time prediction of forensic compounds using ensemble machine learning and molecular descriptors.","authors":"Asena Avci Akca, Sefa Akca","doi":"10.1016/j.jchromb.2025.124812","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124812","url":null,"abstract":"<p><p>Retention time (RT) prediction can greatly improve the efficiency of chromatographic workflows in forensic toxicology, especially in high-throughput or non-targeted analytical workflows. In the present study, we compare the performance of four ensemble machine learning models-Random Forest (RF), Extra Trees, XGBoost, and LightGBM-in predicting RTs of 229 structurally diverse forensic compounds. Each compound was represented by a minimal set of RDKit-derived descriptors and an extended feature space that combines Mordred descriptors and Morgan circular fingerprints. All RTs were experimentally measured under standardized reversed-phase liquid chromatographic conditions. Model performance was evaluated using coefficient of determination (R²) and root-mean-square error (RMSE). Results show that models trained on extended descriptors (>2000 molecular features) outperformed those trained on basic descriptors, with XGBoost showing the highest predictive power (R² = 0.718, RMSE = 1.23). Feature importance analysis showed that RTs are not only affected by global molecular properties like hydrophobicity and size but also by topological and electronic features. These results highlight the value of ensemble learning in RT prediction and demonstrate its practical utility in compound screening and chromatographic method development in forensic toxicology.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":"1267 ","pages":"124812"},"PeriodicalIF":0.0,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145234244","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pharmacokinetics and tissue distribution of tripchlorolide in rodents using liquid chromatography tandem mass spectrometry.","authors":"Yanping Deng, Lele Zhou, Zhengyan Gu, Zhou Chen","doi":"10.1016/j.jchromb.2025.124805","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124805","url":null,"abstract":"<p><p>Tripchlorolide is a promising therapeutic compound with potent pharmacological activity and an improved safety profile compared to triptolide. However, its pharmacokinetics and tissue distribution remain poorly characterized. In this study, we developed and validated a rapid and sensitive liquid chromatography-mass spectrometry method for the quantification of tripchlorolide in biological matrices, using triptolide as the internal standard. Quantification was performed in selective ion monitoring mode, following liquid-liquid extraction with ethyl acetate. Chromatographic separation was achieved on a 3.5 μm Agilent ZORBAX Eclipse Plus-C18 column under isocratic elution with a methanol-water mobile phase. Calibration curves were linear over the range of 0.16-200 ng/mL in rat plasma. The method was successfully applied to a pharmacokinetic study in rats and tissue distribution analysis in mice. Tripchlorolide exhibited an absolute bioavailability of 72.97 % after intraperitoneal administration and a half-life of approximately 45 min, with no significant sex-based differences in pharmacokinetic parameters. Tissue distribution following intravenous administration (400 μg/kg) in mice revealed the highest accumulation in the liver, followed by the kidney, spleen, testis, heart, intestine, and brain. These findings provide essential preclinical data for further development of tripchlorolide as a safe and effective therapeutic candidate.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":"1267 ","pages":"124805"},"PeriodicalIF":0.0,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145246285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Navigating atypical equilibrium dynamics of protein aggregates for biopharmaceutical release and stability monitoring.","authors":"Yiting Zhang, Ruojia Li, Hangtian Song, Andrew McClain, Richard Ludwig, Li Tao, Jeff Beckman, Ming Zeng","doi":"10.1016/j.jchromb.2025.124804","DOIUrl":"https://doi.org/10.1016/j.jchromb.2025.124804","url":null,"abstract":"<p><p>High molecular weight species (HMW) is a critical quality attribute in biopharmaceuticals requiring comprehensive characterization and accurate quantification throughout drug development and manufacturing to ensure product efficacy and patient safety. In this study, the primary goal was to develop a robust size exclusion chromatography (SEC) method to quantify HMW varying in relative proportion to its parent molecule, an Fc-fusion protein therapeutic (FC1) for quality control release and stability testing. However, a unique \"partially reversible\" HMW kinetics behavior was observed, with dissociation towards equilibrium occurring slowly, up to several days, with rates dependent on time, temperature, concentration, and formulation components. This kinetic behavior posed challenges to process optimization and quality control testing, as the \"true\" quantitative value of HMW was highly variable based on these conditions, varying by up to an order of magnitude in relative proportion. To address this, mathematical models were built to describe HMW equilibrium kinetics which supported the development of a robust SEC test method that accurately quantifies this attribute under relevant conditions. The developed method was validated and implemented in quality control laboratory for drug release and stability testing. This work also provides new insights into the need to control protein aggregation dynamics as it relates to process optimization and drug product release requirements.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":"1267 ","pages":"124804"},"PeriodicalIF":0.0,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biological monitoring and analytical toxicology in occupational and environmental medicine.","authors":"Michael Bader,&nbsp;Thomas Göen","doi":"10.1016/j.jchromb.2010.08.025","DOIUrl":"https://doi.org/10.1016/j.jchromb.2010.08.025","url":null,"abstract":"","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"2465-6"},"PeriodicalIF":3.0,"publicationDate":"2010-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2010.08.025","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40061683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunogold-silver staining-on-a-chip biosensor based on cross-flow chromatography.","authors":"Il-Hoon Cho,&nbsp;Sung-Min Seo,&nbsp;Eui-Hwan Paek,&nbsp;Se-Hwan Paek","doi":"10.1016/j.jchromb.2009.07.016","DOIUrl":"https://doi.org/10.1016/j.jchromb.2009.07.016","url":null,"abstract":"<p><p>Immunogold-silver staining (IGSS) was adopted in cross-flow chromatographic analysis in which immunological reactions and silver intensification were sequentially conducted in the vertical and horizontal directions, respectively. Factors controlling the performance, except the silver substrate solution, were optimized to increase the signal-to-background ratio in measurements of cardiac troponin I as a model analyte. In generating the signal, the size of colloidal gold catalyst was critical; the smallest size (5-nm diameter) in the selected range yielded the highest colorimetric signal. To maintain the low background, two processes, blocking the remaining surfaces of membrane after antibody immobilization and washing the residual tracer after immunological reaction, were necessary. Self-nucleation of silver ions also caused a background signal and was controlled to some degree by decreasing the hydrodynamic force that arose when the substrate solution was supplied in the horizontal direction. Finally, a new chip (IGSS-on-a-chip; IOC) that allowed for convenient, efficient IGSS was produced by injection molding of plastic. This method enhanced the detection capability by 51-fold compared to the conventional rapid test kit using 30nm-sized colloidal gold as the tracer. The IOC biosensor results also showed that silver intensification yield via cross flow after immunological reaction was 19% higher than that by traditional incubation.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"271-7"},"PeriodicalIF":3.0,"publicationDate":"2010-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2009.07.016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40008592","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Total on-line analysis of a target protein from plasma by immunoextraction, digestion and liquid chromatography-mass spectrometry.","authors":"A Cingöz,&nbsp;F Hugon-Chapuis,&nbsp;V Pichon","doi":"10.1016/j.jchromb.2009.07.032","DOIUrl":"https://doi.org/10.1016/j.jchromb.2009.07.032","url":null,"abstract":"<p><p>A total on-line analysis of a target protein from a plasma sample was made using a selective immunoextraction step coupled on-line to an immobilized enzymatic reactor (IMER) for the protein digestion followed by LC-MS/MS analysis. For the development of this device, cytochrome c was chosen as model protein due to its well-known sequence. An immunosorbent (IS) based on the covalent immobilization of anti-cytochrome c antibodies on a solid support was made and an immunoextraction procedure was carefully developed to assess a selective extraction of the target protein from plasma. For the first time, IS was easily coupled on-line with a laboratory-made IMER based on pepsin. The whole on-line device (IS-IMER-LC-MS/MS) allowed the quantification of cytochrome c from 8.5pmol to 1.7nmol in buffer medium. Finally, this device was applied to the analysis of only 85pmol of cytochrome c from plasma with a RSD value lower than 10% (n=3).</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"213-21"},"PeriodicalIF":3.0,"publicationDate":"2010-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2009.07.032","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40021564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stable-isotope dilution LC-ESI-MS/MS techniques for the quantification of total homocysteine in human plasma.","authors":"Michela Tomaiuolo,&nbsp;Gennaro Vecchione,&nbsp;Maurizio Margaglione,&nbsp;Daniela Pisanelli,&nbsp;Elvira Grandone","doi":"10.1016/j.jchromb.2009.07.024","DOIUrl":"https://doi.org/10.1016/j.jchromb.2009.07.024","url":null,"abstract":"<p><p>Homocysteine is an endogenous sulphydryl aminoacid irreversibly catabolized by transsulfuration to cysteine or remethylated to methionine. Increased plasma levels of homocysteine are an independent risk factor for atherosclerosis and cardiovascular disease. Accurate and reliable quantification of this amino acid in plasma samples is essential in clinical practice to explore the presence of a hyperhomocysteinemia, for instance after an ischemic event, or to control a possible adjunctive risk factor in patients at higher risk. In this review, LC-ESI-MS/MS methods are discussed and compared with other analytical methods for plasma homocysteine. LC-ESI-MS/MS is a technique combining the physicochemical separation of liquid chromatography with the analysis of mass spectrometry. It is based on stable-isotope dilution and possesses inherent accuracy and precision. Quantitative analysis is achieved by using commercially available homocystine-d(8) as an internal standard. Taking advantage of the high sensitivity and specificity, approaches involving LC-ESI-MS/MS require less laborious sample preparation, no derivatization and produce reliable results.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"3292-9"},"PeriodicalIF":3.0,"publicationDate":"2009-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2009.07.024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40008590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Capillary electrophoresis in the evaluation of aminothiols in body fluids.","authors":"Filippo Carlucci,&nbsp;Antonella Tabucchi","doi":"10.1016/j.jchromb.2009.07.030","DOIUrl":"https://doi.org/10.1016/j.jchromb.2009.07.030","url":null,"abstract":"<p><p>Thiols play a fundamental role in cell biology, biochemistry and pharmacology. Altered thiol levels in body fluids are linked to specific pathological conditions. Glutathione is the most abundant intracellular low-molecular-mass thiol, playing an essential role in protecting cells from toxic species; other relevant thiol-containing compounds are homocysteine (Hcy), cysteine (Cys), cysteinylglycine (CysGly). Plasma aminothiols can be bound to proteins but they also occur free in the disulfide (symmetrical and mixed) and in the reduced forms. The simultaneous determination of these aminothiols, their precursor and metabolites is a useful tool in studying oxidative stress, metabolic and redox regulation. Many capillary electrophoresis methods have been proposed for this purpose, the aim of the present review is to support researchers in the choice of suitable methods for the determination of thiols in body fluids evaluating the different approaches and technologies proposed from the literature.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"3347-57"},"PeriodicalIF":3.0,"publicationDate":"2009-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2009.07.030","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40008591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HPLC analysis of human erythrocytic glutathione forms using OPA and N-acetyl-cysteine ethyl ester: evidence for nitrite-induced GSH oxidation to GSSG.","authors":"Jan Thomas Michaelsen,&nbsp;Sabine Dehnert,&nbsp;Daniela Giustarini,&nbsp;Bibiana Beckmann,&nbsp;Dimitrios Tsikas","doi":"10.1016/j.jchromb.2009.06.043","DOIUrl":"https://doi.org/10.1016/j.jchromb.2009.06.043","url":null,"abstract":"<p><p>Glutathione exists in biological samples in the reduced form (GSH), as its disulfide (GSSG) and as a mixed disulfide (GSSR) with thiols (RSH). GSH is the most abundant low-molecular-mass thiol and plays important roles as a cofactor and as a main constituent of the intracellular redox status. Due to its own sulfhydryl (SH) group, GSH reacts readily with o-phthaldialdehyde (OPA) to form a highly stable and fluorescent isoindole derivative (GSH-OPA), which allows for sensitive and specific quantitative determination of GSH in biological systems by HPLC with fluorescence (FL) detection. In the present article we report on the utility of the novel, strongly disulfide bond-reducing thiol N-acetyl-cysteine ethyl ester (NACET) for the specific quantitative analysis of GSH and GSSG in the cytosol of red blood cells (RBC) as GSH-OPA derivative with FL (excitation/emission 338/458nm) or UV absorbance (338nm) detection. Unlike in aqueous solution, the derivatization of GSH in RBC cytosol yielded two closely related derivatives in the absence of NACET and only the GSH-OPA derivative in the presence of NACET. The HPLC method was optimized and validated for human RBC and applied to measure GSH and GSSG in RBC of healthy subjects. Basal GSH and GSSG concentrations were determined to be 2340+/-350microM and 11.4+/-3.2microM, respectively, in RBC of 12 healthy young volunteers (aged 23-38 years). The method was also applied to study the effects of nitrite on the glutathione status in intact and lysed human RBC. Nitrite at mM-concentrations caused instantaneous and considerable GSSG formation in lysed but much less pronounced in intact RBC. GSH externally added to lysed RBC inhibited nitrite-induced methemoglobin formation. Our findings suggest that nitric oxide/nitrite-related consumption rate of GSH, and presumably that of NADH and NADPH, could be of the order of 600micromol/day in RBC of healthy subjects.</p>","PeriodicalId":520661,"journal":{"name":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","volume":" ","pages":"3405-17"},"PeriodicalIF":3.0,"publicationDate":"2009-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jchromb.2009.06.043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40021565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}