Analytical and Bioanalytical Chemistry最新文献

{"title":"Understanding the chemical stability of peptidomimetic therapeutics using high-resolution mass spectrometry: a study of terlipressin and its degradation products.","authors":"Ashwini Chawathe, Nitish Sharma","doi":"10.1007/s00216-025-05944-7","DOIUrl":"https://doi.org/10.1007/s00216-025-05944-7","url":null,"abstract":"<p><p>Terlipressin, a synthetic 12-amino acid peptidomimetic of vasopressin, is a critical therapeutic agent for hepatorenal syndrome and oesophageal variceal hemorrhage. The inherent susceptibility of therapeutic peptides to hydrolytic and oxidative degradation necessitates thorough stability profiling. Conformational changes in the peptide, arising from hydrolysis and oxidative degradation, can hinder effective target binding and thereby diminish its capacity to elicit intended downstream effects, leading to reduced efficacy. For synthetic peptides, the most relevant stability testing principles are derived from the parent International Council for Harmonisation (ICH) stability testing guidelines Q1A(R2) and Q5C [1,2]. This study investigated the intrinsic degradation pathways of terlipressin under systematically varied stress conditions, including acidic, basic, neutral, and oxidative (H₂O₂) exposure at room temperature. Terlipressin exhibited sensitivity across all tested conditions, yielding a total of eleven distinct degradation products (DPs). To facilitate the separation of these DPs, a gradient reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed utilizing an XSelect® CSH™ C18 (130 Å, 2.5 µm, 4.6 × 150 mm) column. The analytical assay method was validated according to ICH Q2(R1) guidelines. The intramolecular disulfide linkage between two cysteine residues presented a challenge for DP characterization. To address this, a chemical reduction strategy employing dithiothreitol (DTT) was integrated with ultra-high performance liquid chromatography-high resolution tandem mass spectrometry (UHPLC-HRMS/MS). This approach enabled the successful elucidation of the eleven DPs, revealing modifications such as truncation, deamidation, acetylation, and oxidation. The characterized fragmentation patterns and identified degradation products provide fundamental insights into the stability behavior of disulfide-containing therapeutic peptides, directly contributing to rational formulation design and development.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300866","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a reference material for simultaneous quantitative determination of multiple veterinary drug residues.","authors":"Cheng Ma, Chengping Wu, Yunhua Gao, Lianhua Dong, Jiayi Yang, Hailan Chen","doi":"10.1007/s00216-025-05952-7","DOIUrl":"https://doi.org/10.1007/s00216-025-05952-7","url":null,"abstract":"<p><p>Excessive use of multiple veterinary drugs can lead to the accumulation of residues in food derived from animals, thereby posing potential risks to human health and the environment. Accurate quantification of residues from multiple veterinary drugs is essential for ensuring effective monitoring and regulatory compliance. In this study, a robust high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) method was developed for the simultaneous determination of six veterinary drug residues, including florfenicol (FF), thiamphenicol (TAP), difloxacin (DIF), sulfadimidine (SDM), trimethoprim (TMP), and fenvalerate (FEN). This method was subsequently applied to quantify the multiple veterinary reference material (MVRM). Using the MVRM with assigned values, the varying performance of pre-treatment methods for multiple veterinary drugs in meat samples was evaluated, which may result in differing quantitative outcomes. The application of MVRM is helpful for ensuring food safety and safeguarding human health.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of endogenous SUMOylation sites by click chemistry-based proteomics.","authors":"Ke Ma, Sixian Chen, Jun Zhang, Xinglong Jia, Rufeng Fan, Mingjun Li, Li Dong, Minjia Tan, Wensi Zhao, Dong Xie","doi":"10.1007/s00216-025-05957-2","DOIUrl":"https://doi.org/10.1007/s00216-025-05957-2","url":null,"abstract":"<p><p>SUMOylation, an essential ubiquitin-like modification in eukaryotes, plays vital roles in both physiological and pathological regulation, positioning it as a promising therapeutic target. However, the low abundance of SUMOylation and the high enzymatic activity of sentrin/SUMO-specific proteases (SENPs) complicate the identification of endogenous sites. In this study, we integrated click chemistry, acid cleavage, and SUMOylated peptide enrichment into the workflow and developed a promising methodology for system-wide identification of SUMOylation sites. In total, we identified 962 endogenous SUMOylation sites in HEK293T cells under heat shock conditions, which showed good complementarity with previous studies. Our approach uncovered 105 potentially new sites, including SSRP1-K248/K319/K612, DHX9-K806, and ILF3-K241, which showed high conservation and were located in functionally important domains. The overlap between SUMOylation sites and the known ubiquitination or acetylation sites suggested the potential PTM crosstalks. KEGG analysis further suggested SUMOylated proteins were associated with carbon metabolism and biosynthesis of amino acids pathways. Collectively, this study provides a valuable tool for systematically identifying SUMOylation sites, advancing further biological understanding of their dynamic regulatory networks and pathophysiological mechanisms.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Direct detection of CRISPR-Cas9 ribonucleoprotein gene doping using RNA immunoprecipitation and quantitative PCR.","authors":"Kentaro Akiyama, Atsushi Momobayashi, Masato Okano","doi":"10.1007/s00216-025-05959-0","DOIUrl":"10.1007/s00216-025-05959-0","url":null,"abstract":"<p><p>Gene doping, using technologies such as CRISPR-Cas9, poses a considerable threat to the integrity of sports. In 2018, the World Anti-Doping Agency implemented a ban on genome editing, which highlighted the need for sensitive and specific detection methods. Detection techniques that are currently available have shown effectiveness in specific contexts, but are limited by low sensitivity and short detection windows. To overcome these limitations, this study presents a new detection method for CRISPR-Cas9 ribonucleoprotein (RNP) complexes, termed RNA immunoprecipitation followed by quantitative PCR (RIP-qPCR). The primary focus of this research was the in vitro development of a detection method targeting genes critical for doping, including myostatin (MSTN), α-actinin 3 (ACTN3), erythropoietin receptor (EPOR), and erythropoietin (EPO), with in vivo proof-of-concept demonstrated using MSTN. The RIP-qPCR method demonstrated sensitive performance, with a limit of quantification of 0.1 ng/mL in plasma. This method successfully detected single guide RNA targeting MSTN, ACTN3, EPOR, and EPO, along with two types of Cas9 proteins in RNP complexes in vitro. Additionally, the detection capabilities of RIP-qPCR were maintained for up to 30 days when plasma samples were stored at 4 °C. In vivo experiments were performed where RNPs were administered via intramuscular and intravenous injections to target the murine Mstn gene. CRISPR-Cas9 RNPs remained detectable for up to 24 h following intramuscular injection and 12 h after intravenous injection. This study underscores the potential of RIP-qPCR as a powerful tool for anti-doping analysis, with future efforts on expanding the target gene panel to enhance the detection of gene editing in sports doping.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Examining the effects of analytical replication on data quality in a non-targeted analysis experiment.","authors":"Troy M Ferland, Heather D Whitehead, Timothy J Buckley, Alex Chao, Jeffrey M Minucci, E Tyler Carr, Greg Janesch, Safia Rizwan, Nathaniel Charest, Antony J Williams, James P McCord, Jon R Sobus","doi":"10.1007/s00216-025-05940-x","DOIUrl":"https://doi.org/10.1007/s00216-025-05940-x","url":null,"abstract":"<p><p>Non-targeted analysis (NTA) methods are integral to environmental monitoring given their ability to expand measurable chemical space beyond that of traditional targeted methods. Such vast quantities of NTA data are generated that exhaustive manual review is generally unfeasible. Computational tools facilitate automated data processing, but cannot always distinguish real signals (i.e., originating from a chemical in a sample) from artifacts. Replicate analysis is recommended to aid data review, but as NTA studies become larger, the cost of analytical replication becomes untenable. A need therefore exists for examination of information penalties associated with reduced replication. To investigate this issue, using an existing NTA dataset, we performed over 70,000 simulations of variable replication designs and calculated false discovery rates (FDRs) and false negative rates (FNRs) for NTA features and occurrences. We used regression models to explore associations between replication percentage and FDR/FNR, and to test whether rates were affected by NTA feature attributes. Inverse relationships were generally observed between replication percentage and FDR/FNR, such that lower replication yielded higher information penalties. Significant increases in FDR/FNR were observed for suspected per- and polyfluoroalkyl substances (PFAS) compared to non-PFAS, highlighting the potential for differences in information penalties across feature groups. Specific quantitative information penalties are expected to be unique for each NTA study based on sample type and workflow. The methods presented here can support future pilot-scale investigations that will inform the required level of replication in full-scale studies.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144293161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-residue environmental method for seed treatment insecticides, neonicotinoid degradation products, and fungicides using liquid chromatography tandem mass spectrometry with a new ionization source.","authors":"Jascika A A Maclean, Shannon Bartelt-Hunt, Joyce Cristale, Sathaporn Onanong, Daniel D Snow","doi":"10.1007/s00216-025-05927-8","DOIUrl":"https://doi.org/10.1007/s00216-025-05927-8","url":null,"abstract":"<p><p>Few methods provide simultaneous determination of multiple pesticides and degradation products in environmental samples using liquid chromatography tandem mass spectrometry (LC-MS/MS). As LC-MS/MS method performance is significantly influenced by the type and design of the ion source, we compared three ion sources: electrospray ionization (ESI) source, atmospheric pressure chemical ionization (APCI) source and UniSpray™ ionization source. A gain in sensitivity was observed with UniSpray™ and ESI as compared to APCI source on the same instrument. Matrix effects in the three interfaces were evaluated in reagent water and wastewater extracts. UniSpray™ showed the lowest matrix effect among the three sources, with APCI exhibiting more pronounced signal enhancement. A solid-phase extraction method using the UniSpray™ source provides method detection limits (MDLs) ranging from 0.00189 to 0.0209 µg/L in extracts from water samples. Recoveries in water ranged up to 94.35%, with above 60% of the pesticides having an average recovery exceeding 70%.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144281931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction mechanism for Raman spectroscopy in emulsions.","authors":"Erik Spoor, Matthias Rädle, Jens-Uwe Repke","doi":"10.1007/s00216-025-05925-w","DOIUrl":"https://doi.org/10.1007/s00216-025-05925-w","url":null,"abstract":"<p><p>When the concentration of a fluid mixture is measured with Raman spectroscopy in emulsions instead of pure liquids, the signal strength is influenced by the light scattering of the droplets which gives wrong results. This work investigates this influence in the example of a water-toluene-acetone emulsion. For this purpose, the Raman spectroscopy is supported by a scattered light probe, which is intended to quantify the light losses when the dispersed toluene phase increases. The scattered light probe is aligned with the focal point of the Raman probe and detects the light from the 785 nm laser scattered by droplets. The aim is to determine the effects of emulsions on Raman spectroscopy dependent on the concentration of the disperse phase and to determine the acetone concentration of the mixture. The Raman signal decreases with increasing turbidity due to the disperse phase and the concentration of acetone can than no longer be determined from the signal. However, the increase in droplets increases the scattering of the excitation light, whereby a reduction in signal strength is detected. These measurements can be correlated to create a correction function. This makes it possible to correct the measured data of the acetone up to an RMSEP of 1.5 wt%.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144265002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of current use pesticides in flowers, pollen provision, and wild bees: HPLC-ESI-MS/MS method development and field implementation.","authors":"Carolina Honert, Katharina Wifling, María José Lazo Hernández, Carsten A Brühl","doi":"10.1007/s00216-025-05935-8","DOIUrl":"https://doi.org/10.1007/s00216-025-05935-8","url":null,"abstract":"<p><p>Synthetic pesticide use is a major driver of pollinator declines in agricultural landscapes. To understand the impact of pesticides, it is essential to quantify residues in food resources and in insects themselves. We developed simple, fast, and cost-effective multiresidue methods for the simultaneous quantification of up to 83 current use pesticides (CUPs) in flowers (0.5 g sample weight) and 71 CUPs in pollen provision (0.1 g sample weight) via liquid chromatography-tandem mass spectrometry. Additionally, methods were developed for individual wild bees (Osmia bicornis), enabling the analysis of 65 CUPs in 0.02 g samples (females) and 45 CUPs in 0.01 g samples (males). The extractions used acidified acetonitrile (2.5% formic acid), with phase separation assisted by ammonium formate and clean-up via freeze-out. The validation showed limits of quantification between 0.00025 mg/kg and 0.05 mg/kg for flowers, 0.0002 mg/kg to 0.052 mg/kg for pollen provision, 0.0002 mg/kg to 0.08 mg/kg for female bees, and 0.00008 mg/kg to 0.1 mg/kg for male bees. The methods were applied to flowers, pollen provision, and post-pupal bees from agricultural sites. In total, 47 CUPs were detected in flowers, 35 in pollen provision, and 4 in post-pupal bees, with herbicides being most prevalent. This study highlights the exposure of pollinators to CUP mixtures, including emerging bees that have not yet been active in the environment. Our methods provide practical tools for monitoring CUP residues in small environmental samples, supporting the assessment of exposure in plant-insect matrices.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144265001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Green sample preparation for anthocyanin extraction from purple corn: analytical evaluation of pressurized liquid and ultrasound-assisted extraction using sustainable solvents.","authors":"Jorge A Custodio-Mendoza, Alexandra Rangel Silva, Patryk Pokorski, Havva Aktaş, Marcin A Kurek","doi":"10.1007/s00216-025-05951-8","DOIUrl":"https://doi.org/10.1007/s00216-025-05951-8","url":null,"abstract":"<p><p>This study presents an analytical comparison of two extraction techniques-pressurized liquid extraction (PLE) and ultrasound-assisted extraction (UAE)-for the recovery of anthocyanins from purple corn using sustainable solvents (o-phosphoric acid, ethanol, and water). Twelve anthocyanins were identified using high-performance liquid chromatography with ultraviolet detection and tandem mass spectrometry, confirming method specificity and selectivity. Both extraction methods were optimized using multivariate experimental designs and validated following U.S. Food and Drug Administration guidelines for analytical methods in food matrices. The protocols demonstrated excellent linearity (coefficient of determination ≥ 0.9992), low detection limits (0.30-1.70 mg/kg), and high precision and accuracy (relative standard deviation ≤ 5.4%, recoveries between 97.1 and 101.9%) at 50, 100, and 150 mg/kg. To assess their environmental and operational performance, both methods were evaluated using two quantitative tools: AGREEprep, which measures greenness in sample preparation workflows, and the Blue Applicability Grade Index (BAGI), which evaluates practical applicability. PLE achieved a higher sample throughput and lower detection limits, while UAE minimized waste and required less energy. AGREEprep scores (0.73 for PLE, 0.76 for UAE) and BAGI scores (77.5 for PLE, 72.5 for UAE) confirmed both techniques as sustainable and viable for routine analysis. These results demonstrate that both PLE and UAE provide validated, efficient, and environmentally friendly alternatives for anthocyanin extraction from plant-based matrices. The study contributes to the advancement of green sample preparation and highlights the use of structured assessment tools in optimizing analytical workflows for bioactive compound determination in food analysis.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144273887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of process analytical technology for real-time monitoring of synthetic co-culture bioprocesses.","authors":"Nicole A Dambruin, Jack T Pronk, Marieke E Klijn","doi":"10.1007/s00216-025-05949-2","DOIUrl":"https://doi.org/10.1007/s00216-025-05949-2","url":null,"abstract":"<p><p>Synthetic microbial co-cultures can enhance bioprocess performance by division-of-labor strategies that, through spatial segregation of product-pathway modules, circumvent or mitigate negative impacts of the expression of an entire product pathway in a single microorganism. Relative abundance of the microbial partners is a key parameter for the performance of such co-cultures. Population control strategies based on genetic engineering have been explored, but the required interventions may impose an additional metabolic burden and thereby negatively affect co-culture performance. Regulation of co-culture composition by controlled substrate feeding strategies or temperature control requires real-time population monitoring. Process analytical technology (PAT) is an approach for real-time monitoring and control of processes, enabling continuous observation of co-cultivation that may serve as a foundation for population control strategies. In this review, we discuss PAT methods for monitoring synthetic co-cultures, either through direct biomass measurements or by tracking soluble or volatile metabolites. We discuss advantages, limitations, and applications of established as well as emerging technologies and conclude that leveraging PAT for precise, real-time population control has the potential to enhance stability, efficiency, and industrial scalability of synthetic co-cultures.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144245590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}