Journal of Chromatography B最新文献

{"title":"Restricted access membrane roll with functionalized silica nanoparticles cross-linked by hydrophilic polymer chains on the surface for the direct extraction of Evodiae Fructus active components from rat plasma","authors":"Hongwu Zhang ,&nbsp;Yuhao Zhong,&nbsp;Lidan Li,&nbsp;Xiaoyu Xin,&nbsp;Jiayi Zhuang,&nbsp;Yue Zhong,&nbsp;Mingfang Zhu","doi":"10.1016/j.jchromb.2025.124589","DOIUrl":"10.1016/j.jchromb.2025.124589","url":null,"abstract":"<div><div>Evodiamine and other indole alkaloids are the main active constituents in Evodiae Fructus (EF), which is widely applied in traditional Chinese medicine. Recently, the toxic side effects of EF caused by the alkaloid evodiamine have gradually attracted the attention of researchers. In this regard, the rapid and accurate detection of these alkaloids in biological body fluids has become a challenging task. Therefore, it has become a priority that the rapid and efficient extraction of evodiamine etc. alkaloids from blood sample. In this research, a novel method was proposed to prepare the restricted access (RA) membrane with the hydrophobic functional silica nanoparticles cross-linked on the surface by hydrophilic poly-hydroxyethyl methacrylate (<em>p</em>-HEMA). The poly-HEMA chains played the role of protein exclusion and the functional nanoparticles played the role of hydrophobic adsorbent. The RA membrane was wound into a roll shape and inserted into a 1 mL medical syringe to assemble a portable SPE device, named as SRAMR-SPE, for the direct and simultaneous extraction of evodiamine (EVO), rutecarpine (RUT) and dehydroevodiamine (DHE) in rat plasma after a simple dilution with PBS. The prepared functional nanoparticles and RA membrane were characterized by Fourier transform infrared (FT-IR) spectrometer, transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM), respectively. HPLC-UV analysis was used to investigate the extraction performance of SRAMR. An associated SRAMR-SPE-HPLC method was constructed and applied to the detection of real rat plasma after the method validation for the verification of the reliability and applicability of the SRAMR-SPE. The experimental results showed that the RA membrane roll (RAMR) had a good ability to exclude plasma proteins and adsorb analytes with multiple reusability. For 3 consecutive cycles of SRAMR-SPE processes, the extraction recoveries of three components at three concentration levels were determined as follows: DHE 93.1–95.1 %, EVO 88.8–91.6 % and RUT 93.6–95.6 %, and the absolute recoveries of the entire SRAMR-SPE-HPLC-UV method were separately as follows: DHE 90.3–92.3 %, EVO 86.2–90.9 %, RUT 91.8–96.4 %. The linear ranges were detected as follows: 0.0600–6.00 μg mL⁻¹ (DHE), 0.105–5.65 μg mL⁻¹ (EVO) and 0.0632–6.32 μg mL⁻¹ (RUT).</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1258 ","pages":"Article 124589"},"PeriodicalIF":2.8,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143850705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advancements in the bioactive alkaloids analysis in plant and biological specimen: From the perspective of activity, sample preparation, and analytical method selection","authors":"Sevilay Erdogan-Kablan ,&nbsp;Seyda Yayla ,&nbsp;M. Mesud Hurkul ,&nbsp;Ahmet Cetinkaya ,&nbsp;Emirhan Nemutlu ,&nbsp;Sibel A. Ozkan","doi":"10.1016/j.jchromb.2025.124592","DOIUrl":"10.1016/j.jchromb.2025.124592","url":null,"abstract":"<div><div>Alkaloids are a diverse group of naturally occurring organic compounds. They are known for their significant pharmacological properties. This review provides an up-to-date analysis of bioactive alkaloids in plant and biological samples, emphasizing their biological activities, extraction techniques, and analytical methods. The study focuses on significant alkaloids such as morphine, codeine, vinblastine, vincristine, berberine, quinine, quinidine, caffeine, nicotine, ephedrine, and atropine, highlighting their chemical structures, therapeutic applications, and mechanisms of action. Recent advances in extraction methods, including conventional and modern green techniques such as supercritical fluid extraction, microwave-assisted extraction, and solid-phase microextraction, are discussed in detail. In addition, the review provides an overview of state-of-the-art analytical techniques used for alkaloid quantification, such as high-performance liquid chromatography, liquid chromatography-mass spectrometry, and novel spectroscopic methods. Emphasis is placed on the challenges associated with alkaloid analysis, including matrix effects, stability, and structural diversity. The results contribute to the growing body of knowledge in alkaloid research, providing insights into their potential therapeutic applications and analytical improvements for more accurate and efficient detection in various biological and plant matrices.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1258 ","pages":"Article 124592"},"PeriodicalIF":2.8,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143826341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantification of isavuconazole from dried blood spots: Applicability in therapeutic drug monitoring","authors":"Jens Martens-Lobenhoffer ,&nbsp;Stefan Angermair ,&nbsp;Stefanie M. Bode-Böger","doi":"10.1016/j.jchromb.2025.124590","DOIUrl":"10.1016/j.jchromb.2025.124590","url":null,"abstract":"<div><div>Dried blood spots (DBS) is a convenient method of blood sampling for biomedical quantification of various drugs. Compared to conventional venipuncture and subsequent plasma measurement, DBS provides advantages in less invasive sampling and easy and safe shipping of samples. The main drawback is the difficult calculation of precise plasma concentrations from the DBS measurements. In this study, a method for the quantification of the antimycotic drug isavuconazole from DBS was developed and its applicability in therapeutic drug monitoring (TDM) was evaluated by comparing the DBS quantification results with the corresponding plasma values.</div><div>DBS were produced by spotting 15 μL of EDTA-blood onto DBS cards. The whole DBS spots were extracted in methanol:water 10:1, and the extracts were analyzed by an established HPLC method using fluorescence detection. Isavuconazole proved to be stable in DBS over 3 weeks at room temperature or refrigerated at 6 °C. Intra-day precision and accuracies of the quantification from DBS were better than 5 %, while the inter-day results were better than 12 %.</div><div>Fourteen plasma samples from intensive care patients showing isavuconazole concentrations of &lt;0.1 μg/mL to 3.52 μg/mL (median 1.42 μg/mL) were compared to the results obtained from corresponding DBS samples. All concentration values were covered by the calibration range (0.1–20 μg/mL) of the analytical method. Differences between plasma and DBS results were less than 0.2 μg/mL, with an underestimation of less than 6 % in the DBS values. As these differences were of no therapeutic relevance, DBS could be considered a viable matrix for TDM of isavuconazole in intensive care patients.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1258 ","pages":"Article 124590"},"PeriodicalIF":2.8,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143820453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"QbD and green metric based HPLC method development and validation of bromocriptine mesylate in bulk product and biological samples","authors":"Sumant Saini ,&nbsp;Ripandeep Kaur ,&nbsp;Ranjot Kaur ,&nbsp;Monica Gulati ,&nbsp;Bhupinder Singh ,&nbsp;Afzal Hussain ,&nbsp;Abdul Ahad ,&nbsp;Mohhammad Ramzan ,&nbsp;Mudassar Shahid ,&nbsp;Arusha Ayub","doi":"10.1016/j.jchromb.2025.124566","DOIUrl":"10.1016/j.jchromb.2025.124566","url":null,"abstract":"<div><div>Bromocriptine mesylate (BM) is a well-established antiparkinsonian drug, yet its analytical methodologies remain limited in terms of efficiency, cost-effectiveness, and reproducibility from commercial product and biological samples. We aimed at systematic analytical quality by design-assisted method development for routine quantitative assessment. Initially, various plausible variables (peak area, retention time, tailing factor, and plates) were screened using Ishikawa fishbone method. Following risk estimation studies, factor screening exercise was conducted employing a Taguchi design (TgD) for selecting various significant variables. Furthermore, a central composite-design (CCD) was conducted to delineate operable conditions and design space verification using Monte Carlo simulations. In the optimization, flow rate (X₁ as A) and the injection volume (X₂ as B) were two significant factors against four responses (Y₁: peak area, Y₂: retention time, Y₃: Tailing factor and Y₄: Theoretical plate count). The method was validated by estimating validation parameters as per ICH guidelines under environmentally sustainable conditions (using AGREE tools). Finally, optimized mobile phase composition (Acetonitrile (ACN): Trifluoroacetic acid (TFA) 70:30) and the flow rate (0.9 mL.min⁻¹) were set as the optimal chromatographic conditions. The method was found to be highly robust, precise, and sensitive, with a wide linearity range of 0.2–100 μg/mL. Conclusively, AQbD paradigms coupled with simulations enabled in selecting environmentally sustainable and influential method variables, and arriving at the “best possible” chromatographic conditions for effective quantification of BM.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1258 ","pages":"Article 124566"},"PeriodicalIF":2.8,"publicationDate":"2025-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143855037","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A simple and efficient approach to overcome the purification bottleneck of polar unstable prodrugs & New Chemical Entities (NCEs)","authors":"V. Venkataramasubramanian,&nbsp;K. Kalyan Motamarri,&nbsp;Balamurali Rajagopal,&nbsp;Varsha Agarwal,&nbsp;Umesh Waman Mali,&nbsp;Raja K. Rit,&nbsp;P. Satish,&nbsp;Subhendu K. Mohanty,&nbsp;Santoshkumar N. Patil","doi":"10.1016/j.jchromb.2025.124588","DOIUrl":"10.1016/j.jchromb.2025.124588","url":null,"abstract":"<div><div>High-Performance Liquid Chromatography (HPLC), a sophisticated form of column chromatography, remains a workhorse technique that is commonly used in the pharmaceutical industry and academia for the purification of organic substances from complicated mixtures. The HPLC technique uses a high-pressure pump through a chromatographic column packed with small particles with a mobile phase buffer system which could be acidic, basic, or neutral and is chosen based on the compatibility requirements of the compound to be purified. However, purification of prodrugs that have highly sensitive cleavable linkers remains challenging via HPLC conditions. The sensitivity of the linkers is due to their instability to either basic or acidic conditions. Even though a neutral buffer method is developed for such compounds, the majority of compounds often degrade due to their longer contact time with the aqueous mobile phase during prep. HPLC and lyophilization. Towards this, we propose a simple method of purification of such compounds which is efficient and cost-effective. This method utilizes hydrophobic, reverse-phase, silica-bound C18 cartridges on a bench-top setup. We compared the effectiveness of purification of some sensitive compounds using these cartridges with the conventional prep-HPLC purification. Our method reduces cost, minimizes solvent consumption, and reduces purification time drastically. The compounds were obtained with a good recovery and purity using cartridges as shown in <span><span>Fig. 1</span></span>. This method was suitable for purifying compounds that had stability issues during prep-HPLC purification.</div><div>In addition, we hope this method being versatile and simple would promote routine purification and catalyze innovation in colleges and institutes around the globe that cannot afford preparative HPLC for routine purification. Even in industry, our learning is that this method when used in tandem with preparative HPLC as it reduces TAT (turnaround time), is cost-effective, and prolongs prep HPLC column life. Also, we demonstrated that this cartridge method can be used in tandem with prep. HPLC in industry. This reduces net purification machine time, reduces cost, and prolongs HPLC column life.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124588"},"PeriodicalIF":2.8,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143807423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid analysis in small-volume urine: Optimizing LC-MS/MS for free glucocorticoids","authors":"Ikumi Endo ,&nbsp;Mikiko Tokiya ,&nbsp;Kazuhiro Kawamoto ,&nbsp;Yasuyuki Maeda ,&nbsp;Sudarma Bogahawaththa ,&nbsp;Masayoshi Ichiba ,&nbsp;Akiko Matsumoto","doi":"10.1016/j.jchromb.2025.124579","DOIUrl":"10.1016/j.jchromb.2025.124579","url":null,"abstract":"<div><div>Glucocorticoids, regulators of energy metabolism and immune responses, are known biomarkers of stress response and disease. Downscaling the required sample volumes facilitates large-scale studies and accelerates the generation of valid results. To meet this expectation, we modified our LC-MS/MS method using 100 μL of urine to that using 20 μL, following the same steps as in the original protocol with the addition of cortisone-d8 as an internal standard. Calibration was performed as in the original protocol but using 1/5 diluted standards, resulting in a good linearity of 2–200 ng/mL in the log-log plot, indicating a lower limit of quantitation (LLOQ) of &lt;2 ng/mL from the residue. The coefficients of variation of the matrix factors of the 15 urine samples and water were within 15 % for 10 and 300 ng/mL cortisol and cortisone. The matrix factor of the downscaled method was larger than that of the original method, especially for urine extracts with high specific gravity (up to 2.1-fold), indicating the advantage of the downscaled method; although the theoretical LLOQ is 5-fold with the 1/5 downscaling, the effect on the practical LLOQ may be reduced by the suppressed matrix effect. The current method showed good recovery, accuracy, reproducibility, and agreement with the original method (regression coefficient close to 1.0). After sample freezing, cortisol and cortisone were detected at higher and lower levels, respectively, within 25 %. The downscaled method of urine-free cortisol and cortisone may contribute to the advancement of related research.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124579"},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143817235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the mechanism of action of Qianlie Jindan tablets in rats with chronic prostatitis using non-targeted metabolomics","authors":"Tengfei Chen ,&nbsp;Qian Zhang ,&nbsp;Zhichao Jia ,&nbsp;Zhuozhuo Shi ,&nbsp;Junguo Ma ,&nbsp;Zhaowang Gao ,&nbsp;Chongfu Zhong","doi":"10.1016/j.jchromb.2025.124577","DOIUrl":"10.1016/j.jchromb.2025.124577","url":null,"abstract":"<div><h3>Objective</h3><div>To explore the mechanism of action of Qianlie Jindan tablets by analyzing the metabolomic changes in prostate tissues obtained from rats with chronic prostatitis/chronic pelvic pain (CP/CPPS).</div></div><div><h3>Materials and methods</h3><div>Male SD rats were randomly divided into three groups: blank control (BC), model control (MC), and treatment (QLJD) groups, with 10 rats in each group. The model was induced using Complete Freund's adjuvant (CFA) and prostate protein purification solution, and the corresponding drug intervention was given. At the end of the experiment, pathological changes in the prostate tissues were observed using hematoxylin and eosin (HE) staining. Differential metabolites were determined by ultra-high-performance liquid chromatography and tandem electrostatic field orbital trap mass spectrometry (UHPLC-Q Exactive HFX), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation was performed. The results were then verified by quantitative real-time polymerase chain reaction (PCR).</div></div><div><h3>Results</h3><div>QLJD reversed the histopathological damage induced by CP/CPPS. Metabolomics analysis showed that UDP-GlcNAc was the key differential metabolite, and can activate the PI3K/AKT/mTOR pathway. The PCR analysis revealed that the mRNA expression levels of PI3K, AKT and mTOR in the MC group were significantly lower than those in the BC group (<em>P</em> &lt; 0.05). These parameters were increased in the QLJD group compared to the MC group (P &lt; 0.05), validating our metabolomics results.</div></div><div><h3>Conclusion</h3><div>QLJD exerts a therapeutic effect on CP/CPPS by activating the PI3K/AKT/mTOR pathway through the regulation of UDP-GlcNAc levels.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124577"},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791268","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of a stability-indicating reversed phase high-performance liquid chromatography method for quantifying rivaroxaban (XARELTO)","authors":"Aktham Mestareehi","doi":"10.1016/j.jchromb.2025.124573","DOIUrl":"10.1016/j.jchromb.2025.124573","url":null,"abstract":"<div><div>Rivaroxaban is an anticoagulant medication that targets a key stage in the blood clotting process, preventing the formation and growth of clots. It is commonly used to prevent thrombosis or inhibit the enlargement of existing clots. Rivaroxaban functions as a Factor Xa inhibitor and is indicated for: reducing the risk of stroke and systemic embolism in patients with non-valvular atrial fibrillation, treating deep vein thrombosis (DVT) and pulmonary embolism (PE), as well as reducing the risk of recurrent DVT and PE, and prophylaxis of DVT, which may lead to PE in patients undergoing knee or hip replacement surgery.</div><div>A robust, precise, and selective reversed-phase high-performance liquid chromatography (HPLC) method was developed and validated for analyzing Rivaroxaban in raw materials. Isocratic elution at a flow rate of 1 mL/min was performed using a Thermo ODS Hypersil C18 column (4.6 × 250 mm, 5 μm) at ambient temperature. The mobile phase consisted of monobasic potassium phosphate at pH 2.9 and acetonitrile in a 70:30 (<em>v</em>/v) ratio, with UV detection at 249 nm. Linearity was established in the concentration range of 50–1000 ppm (r² = 0.999), and the retention time for Rivaroxaban was approximately 12 min. The percentage relative standard deviation (RSD) for precision and accuracy was consistently below 2.0 %. Rivaroxaban was subjected to forced degradation under various conditions, including acid and base hydrolysis, hydrogen peroxide oxidation, heat, and UV light exposure. The developed method was validated for specificity, robustness, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ), following International Conference on Harmonisation (ICH) guidelines. The LOD for impurities and degradants was found to be 0.3 ppm, with an LOQ of 1 ppm.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124573"},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A rapid and sensitive LC–MS/MS method for the simultaneous determination of chlorfenapyr and its major metabolite tralopyril in human plasma: From method development, validation to clinical application","authors":"Shiyuan Tang ,&nbsp;Shunjie Zhang ,&nbsp;Hongling Du ,&nbsp;Xia Yang ,&nbsp;Qunmei Yao ,&nbsp;Aihua Peng ,&nbsp;Minghai Tang ,&nbsp;Yu Cao","doi":"10.1016/j.jchromb.2025.124580","DOIUrl":"10.1016/j.jchromb.2025.124580","url":null,"abstract":"<div><div>Chlorfenapyr (CFN), widely used as an insecticide, is a pro-insecticide that acts after being metabolized to the active metabolite tralopyril (TLP). In clinical practice, CFN poisoning is characterized by atypical symptoms, delayed toxicity and high mortality. The therapeutic importance of assessing TLP levels in the management of CFN poisoning has been demonstrated in recent clinical and toxicokinetic studies. Unfortunately, the simultaneous quantification of CFN and TLP poses technical challenges and is still lacking in clinical practice. In this study, a novel LC-MS/MS method was developed for rapid and accurate simultaneous analysis of CFN and TLP in human plasma within 3 min. Samples were extracted by acetonitrile protein precipitation. Stable CFN anionic adducts [M + HCOO]⁻ were selected to provide satisfactory MS signals. To improve the signal and peak shape, the mobile phase was preferred to consisting of 2 mM ammonium formate containing 0.01 % ammonia and acetonitrile. The optimized multiple reaction monitoring transitions of <em>m</em>/<em>z</em> 451.0 → 346.9, 349.9 → 78.9 and 307.1 → 161.1 were selected in the negative ionization mode to detect CFN, TLP and IS (warfarin), respectively. The method demonstrated satisfactory linearity with low LLOQ (5 ng/mL for CFN and 1 ng/mL for TLP, respectively), and the precision and accuracy were acceptable. Both compounds showed good stability under relevant conditions. Furthermore, the method was successfully applied to 72 clinical plasma samples from 20 patients with CFN poisoning. The results showed that the concentration of TLP in plasma was significantly higher than that of CFN (<em>P</em> &lt; 0.0001), and all five deaths demonstrated high TLP concentrations (&gt;1000 ng/mL). These data demonstrate the potential relationship between toxins concentration and toxicity in CFN poisoning patients.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124580"},"PeriodicalIF":2.8,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143777477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Size exclusion chromatography: An efficient tool for hsEV isolation from insect Haemolymph","authors":"Jaspreet Kaur ,&nbsp;Janvi Aggarwal ,&nbsp;Mukesh Behera ,&nbsp;Tannu Rachna Dahiya ,&nbsp;Dhiraj Kumar ,&nbsp;Narender K. Dhania","doi":"10.1016/j.jchromb.2025.124570","DOIUrl":"10.1016/j.jchromb.2025.124570","url":null,"abstract":"<div><div>Extracellular vesicles (EV) serve an important role in biological system as they can change the phenotype and function of the cell. The discovery of EVs has led to the development of novel vaccines, associated with immunosuppressive and immunostimulatory effects during disease progression. To address the opportunities in EVs, here we used insect as model system. The insect also has medicinal properties as it produces antimicrobial peptides which are found circulating fluid called as hemolymph. Hemolymph consists of defensins, gloverins, proline-rich peptides, attacins, cecropins, moricins, anionic antimicrobial peptides, and several other components that create robust defense mechanisms against a broad spectrum of pathogens. Here, the insect hemolymph was used for EV isolation which encapsulates such kinds of proteins and acts as cargo transporters. As the biogenesis was unknown, we have defined it as hemolymph-derived small extracellular vesicles (hsEV). The present paper deals with the appropriate methodology for the isolation of hsEV. The hemolymph was processed for isolation via size exclusion chromatography. Molecular and physical characterization was performed using western blotting utilizing anti-CD63, anti-Flotillin-2, TEM, FESEM, confocal microscopy, and DLS. The isolated hsEV was used to assess the antibacterial property by measuring bacterial growth.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124570"},"PeriodicalIF":2.8,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143783397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}