Analytical and Bioanalytical Chemistry最新文献

{"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":"https://doi.org/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":"Quantitative determination and validation of 96 pesticides in cannabis by LC-MS/MS and GC-MS/MS.","authors":"D A MacKenzie, A M Anyanwu, G McRae, J E Melanson","doi":"10.1007/s00216-025-05918-9","DOIUrl":"https://doi.org/10.1007/s00216-025-05918-9","url":null,"abstract":"<p><p>Canada has established a strict set of testing requirements for pesticides that are unauthorized for use on cannabis, currently holding the most extensive list of analytes in North America to date, listing minimum method performance limits rather than maximum allowable concentration limits. These requirements establish the need for validated analytical methods capable of quantifying pesticides and growth regulators in highly variable cannabis inflorescence prior to distribution and sale. We have developed quantitative LC-MS/MS and GC-MS/MS methods capable of quantifying the 96 pesticides unauthorized in Canada for use on cannabis in dried cannabis flower and hemp. Herein, we report the validation results for linearity, precision, within- and between-sample accuracy, recovery, ion suppression, and limits of quantitation in dried cannabis inflorescence. Accuracy is evaluated in 10 cultivars varying in major cannabinoid content as well as ground whole-plant hemp to demonstrate method performance over a range of sample types. Results of the application of the current method to six cannabis samples seized from illegal storefronts by the Ontario Provincial Police are also reported.</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":"144265003","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":"An LC/MS/MS method for quantifying 25-hydroxyvitamin D₃ in finger-prick plasma sample prepared by DEMECAL^® micro plasma separation device: toward mail-in assessment of vitamin D status.","authors":"Kodai Maeda, Kyoka Sakamoto, Eiko Ito, Shinya Sugimoto, Shingo Tajima, Takahisa Sasahara, Shunji Kawamura, Kazuo Yaegashi, Tatsuya Higashi","doi":"10.1007/s00216-025-05939-4","DOIUrl":"https://doi.org/10.1007/s00216-025-05939-4","url":null,"abstract":"<p><p>A mail-in assessment of vitamin D status will make it possible for the subjects to early detect any vitamin D insufficiency/deficiency at their homes, thus leading to a better management of their health. In this study, a liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) method was developed and validated for quantifying 25-hydroxyvitamin D₃ [25(OH)D₃], which is the established marker of the vitamin D status, in the finger-prick blood plasma sample prepared by the DEMECAL^® device. By using the DEMECAL^® device, the subjects can collect their finger-prick blood at their homes and immediately separate the plasma from the blood by themselves. The DEMECAL^®-based procedure had advantages over the venipuncture- and dried blood spot-based procedures because it is less invasive and a medical professional-free blood collection, and no need to be concerned with the heterogeneity of the analyte distribution in the sample. The developed LC/ESI-MS/MS method enabled the precise and accurate quantification of 25(OH)D₃ and provided the lower limit of quantification of 0.10 ng/mL in the diluted plasma sample due to the derivatization with 4-(4-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione. 25(OH)D₃ was stable in the plasma sample prepared by the DEMECAL^® device at room temperature (23-27 °C) for 7 days (simulated condition in the mail) and at 4 °C for 30 days (simulated condition during storage at a laboratory). The measured 25(OH)D₃ concentrations by the DEMECAL^®-based procedure well agreed with those by the conventional venipuncture-based procedure. These results demonstrated that the derivatization-LC/ESI-MS/MS combined with the DEMECAL^®-based sample preparation is a promising procedure for the mail-in assessment of the vitamin D status.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144265000","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 structure-based surrogate selection for quantitative non-targeted analysis.","authors":"Nathaniel Charest, Shirley Pu, James P McCord, Antony J Williams, Jon R Sobus","doi":"10.1007/s00216-025-05919-8","DOIUrl":"https://doi.org/10.1007/s00216-025-05919-8","url":null,"abstract":"<p><p>Quantitative non-targeted analysis (qNTA) is an important tool for characterizing emerging contaminants in environmental, biological, and product-based samples. While traditional non-targeted analysis (NTA) focuses on chemical identification, qNTA additionally produces chemical concentration estimates. These estimates can inform provisional risk-based decisions and prioritize targets for follow-up analysis. Many common qNTA and \"semi-quantitative\" approaches rely on surrogate chemicals for calibration and model predictions. Despite their importance, surrogates are often chosen based on a combination of intuition and/or availability rather than rational (i.e., structure-based) selection. The lack of rational selection limits the degree to which qNTA can be objectively, mathematically assessed and improved. In this work, we systematically assess the extent to which chemical structure should inform the selection of qNTA surrogates using a dataset from liquid chromatography high-resolution mass spectrometry (LC-HRMS) experiments. First, we calculate a chemical space embedding using available LC-HRMS training data (n=385 chemicals) and 2D molecular descriptors deemed important to electrospray ionization efficiency. Then, using data from EPA's Non-Targeted Analysis Collaborative Trial (ENTACT), we calculate the leverage of measured analytes (n=533 chemicals) within the embedded chemical space. Based on leverage calculations, we implement multiple structure-based surrogate selection strategies and compare those to random selection using qNTA metrics for accuracy, uncertainty, and reliability. Finally, we propose and examine the \"leveraged averaged representative distance\" (LARD) as a means to quantify the coverage of qNTA surrogates within a defined chemical space. Our results show that qNTA models can benefit from rational surrogate selection strategies. They further show that a large enough random surrogate sample can perform as well as a smaller, chemically informed surrogate sample. Researchers are advised to carefully consider these findings when selecting surrogates for future qNTA studies.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144245591","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}