Journal of chromatography open最新文献

{"title":"Gas chromatography mass spectrometry based pharmacometabolomic assessment of formulation-dependent metabolic responses of levetiracetam","authors":"Özge Cansın Zeki ,&nbsp;Fatma Ilgaz ,&nbsp;Cemil Can Eylem ,&nbsp;Selin Seda Timur ,&nbsp;Hakan Eroğlu ,&nbsp;Hülya Gökmen-Özel ,&nbsp;Emirhan Nemutlu","doi":"10.1016/j.jcoa.2026.100315","DOIUrl":"10.1016/j.jcoa.2026.100315","url":null,"abstract":"<div><div>This study applied a GC–MS–based pharmacometabolomic approach to investigate formulation-dependent bioavailability and systemic metabolic responses following levetiracetam administration. Plasma samples from rabbits were analyzed after receiving three different formulations: a control formulation in which levetiracetam was mixed with deionized water, a Multithick formulation prepared with an 8% modified maize starch thickener, and a Thicken-Up Clear (TUC) formulation prepared with 3.6% xanthan gum thickener. Although dissolution profiles varied substantially among these formulations, likewise, total levetiracetam exposure, represented by AUC, did not differ significantly among groups (<em>p</em> &lt; 0.05). Untargeted GC–MS analysis was annotated 97 metabolites and multivariate analysis revealed distinct metabolic clustering, with the TUC group showing the most pronounced separation. Correlation analysis identified five metabolites that were positively associated with pharmacokinetic parameters: pyrophosphate, glyceric acid, aconitic acid, methyl-2-butenedioic acid, and itaconic acid. Six amino acid–related metabolites showed negative associations, reflecting enhanced energy metabolism together with reduced excitatory neurotransmission, consistent with the known neuroprotective and antiepileptic actions of levetiracetam. Overall, these results demonstrate that formulation design influences not only dissolution behavior but also systemic metabolic responses. Pharmacometabolomics offers a quantitative framework for integrating chromatographic, pharmacokinetic and biochemical data to support rational formulation development and precision pharmacotherapy.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100315"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147384202","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":"Overcoming fiber degradation: A green headspace solid-phase microextraction method with on-fiber derivatization for urinary amphetamines","authors":"Rukiye Aslan ,&nbsp;İmran Güney Afacan ,&nbsp;Fatma Nil Ertaş ,&nbsp;Hasan Ertaş ,&nbsp;Serap Annette Akgür","doi":"10.1016/j.jcoa.2026.100334","DOIUrl":"10.1016/j.jcoa.2026.100334","url":null,"abstract":"<div><div>Amphetamine-type stimulants (ATS) are the second most abused class of drugs worldwide after cannabis, making their reliable analysis in biological matrices a critical necessity. An eco-friendly headspace solid phase microextraction (HS-SPME) method was developed to sensitive and selective detection of amphetamine (AMP), methamphetamine (MET), 3,4-methylenedioxy amphetamine (MDA), and 3,4-methylenedioxy methamphetamine (MDMA) in urine using gas chromatography-mass spectrometry (GC–MS). The critical effect of derivatization on the fiber was evaluated. The Plackett-Burman design was used to optimize the method by screening the main parameters affecting its performance, and the method was validated. The alignment of the proposed method with the principles of green chemistry was evaluated using the AGREE software.</div><div>Three acylation agents were compared for method performance, and trifluoroacetic anhydride (TFAA) proved the most suitable. Fiber degradation was prevented by using less derivatization reagent, thereby facilitating the complete desorption of analytes from the fiber. 10 min. adsorption time, 80 °C derivatization temperature, and 1 min. of derivatization time gave the optimum results. Sensitive and selective detection of ATS in urine was achieved in the range of 25–2000 ng/mL. Accuracy ranged from 72.8 % to 113.6 %.</div><div>A simple, reliable, and efficient method based on HS-SPME for the simultaneous determination of ATS in urine samples. Deterioration of the polymeric stationary film during the on-fiber derivatization was demonstrated experimentally. The reduced volume of acylation reagent effectively mitigated fiber degradation and yielded well-defined chromatographic peaks for all target ATS, likely by facilitating the desorption of strongly adsorbed analytes.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100334"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147797821","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":"Borate complexation-assisted capillary electrophoretic determination of ethylene glycol and propylene glycol using capacitively coupled contactless conductivity detection","authors":"Gabriela A.M.C. Ribeiro ,&nbsp;Layra M. de Faria ,&nbsp;José A. Fracassi da Silva ,&nbsp;Dosil P. de Jesus","doi":"10.1016/j.jcoa.2026.100340","DOIUrl":"10.1016/j.jcoa.2026.100340","url":null,"abstract":"<div><div>A novel, fast, and reliable analytical method for determining ethylene glycol (EG) and propylene glycol (PG) using capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C⁴D) is proposed. The CE separation is based on the anionic complexation of the glycols with borate ions in the background electrolyte (BGE). The borate concentration in the BGE was optimized, and 100 mmol <span>l</span>⁻¹ provided the best compromise between adequate resolution and acceptable Joule heating. Separation of both glycols was achieved in &lt;5 min, with suitable baseline resolution. Good linearity was attained with coefficients of determination higher than 0.99. The limits of detection (LOD) and quantification (LOQ) were 18.8 and 62.7 µg mL⁻¹ for EG and 21.1 and 70.4 µg mL⁻¹ for PG, respectively. The recovery results for the samples analyzed at three concentration levels ranged from 86 % to 113 %, indicating good accuracy. The proposed CE-C⁴D method was successfully applied to determine EG and PG in various samples, including engine coolant, mouthwash, and e-liquid (refill for electronic cigarettes).</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100340"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147797822","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":"Integrated dual-split high capacity sorptive extraction and chemometrics reveal extraction-dependent volatile signatures in Hypericum perforatum","authors":"Valentina Braghin ,&nbsp;Sadia Kanwal ,&nbsp;Davide Barboni ,&nbsp;Brunilda Myftari ,&nbsp;Nicoletta Bianchi ,&nbsp;Paola Tedeschi ,&nbsp;Chiara De Luca ,&nbsp;Alberto Cavazzini ,&nbsp;Simona Felletti ,&nbsp;Martina Catani ,&nbsp;Natasha D. Spadafora","doi":"10.1016/j.jcoa.2026.100339","DOIUrl":"10.1016/j.jcoa.2026.100339","url":null,"abstract":"<div><div><em>Hypericum perforatum</em> (L.), commonly known as St. John’s Worth, is a medicinal plant rich in bioactive compounds including volatile organic ones, which contribute to its antimicrobial, anti-inflammatory, and wound-healing properties. Optimising the extraction and analysis of these compounds is essential for their application in pharmaceutical, cosmetic and nutraceutical industries.</div><div>This study employed high-capacity extraction (HiSorb) probes and Gas Chromatography-Mass Spectrometry (GC–MS) to profile volatile organic compounds (VOCs) from hypericum flowering aerial parts. Four HiSorb phases were evaluated, with the double-phase CWR/PDMS demonstrating superior performance by capturing the broadest range of compounds and yielding the highest total peak area. A dual-split strategy was implemented to prevent detector saturation and ensure retention of low-abundance analytes, enabling comprehensive VOC profiling from a single sample. VOCs were analysed across various extract types, including alcoholic, water infused and oil-based preparations. A total of 108 VOCs were putatively identified, including terpenes, alcohols, and aldehydes, which contribute to the plant’s aromatic and therapeutic properties. Chemometric analyses, including Principal Component Analysis (PCA), hierarchical clustering and Partial least square-discriminant analysis (PLS-DA), identified shared and discriminatory components across extract types. These findings support the selection of optimal extraction strategies tailored to specific industrial applications and highlight the chemical diversity and functional potential of <em>Hypericum perforatum</em>.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100339"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147797823","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":"An alternative easy method for the analysis of volatile compounds from lipid matrices: analysis of terpenes and terpenoids in medical-grade Cannabis sativa L. inflorescences in macerated oils","authors":"Victor Pereira-Francisco ,&nbsp;Muriel Cerny ,&nbsp;Romain Valentin ,&nbsp;Franck Milone-Delacourt ,&nbsp;Marion Alignan","doi":"10.1016/j.jcoa.2026.100316","DOIUrl":"10.1016/j.jcoa.2026.100316","url":null,"abstract":"<div><div>The terpenes and terpenoids present in cannabis are the subject of increasing research, as their interaction with phytocannabinoids appears to enhance their biological activity. In addition, both for health reasons and to meet consumer demand for more natural products, cannabis flowers are increasingly being extracted in vegetable oils, and cannabis extracts are being formulated in vegetable oils. The number of people using cannabis for medical purposes is increasing. In addition to optimizing the combination of active pharmaceutical ingredients (APIs) for the specific indication being treated, it is necessary to know precisely the dose of each important API administered. To achieve this goal, we have developed a new sample preparation method for determining the terpene and terpenoid content of lipid-based plant extracts by GC-FID. The method described has the advantages of being easy to implement, inexpensive, reproducible, and suitable for the quantification of terpenes and terpenoids in oily matrices, an analysis that was generally performed by HS until now. We confirm that the composition of lipid matrices can influence the extraction yields of terpene compounds from medical cannabis. Sesame oil was the most effective lipid matrix (925 μg total terpene compounds/100 mg dry matter) and rapeseed oil was the least effective (762 μg/100 mg dry matter).</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100316"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147384192","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":"Analytical Quality by Design approach in the development of a green reversed-phase ultra-high performance liquid chromatography/high-resolution time-of-flight mass spectrometry method for the simultaneous analysis of synthetic antimicrobial and hypotensive peptides","authors":"Ana Roberta Pereira Johnson dos Anjos ,&nbsp;Márcia Natália da Silveira Pinto ,&nbsp;Dayanne Lopes Porto ,&nbsp;Fabio Santos de Souza ,&nbsp;Rui Oliveira Macedo ,&nbsp;Matheus de Freitas Fernandes Pedrosa ,&nbsp;Cícero Flávio Soares Aragão","doi":"10.1016/j.jcoa.2026.100318","DOIUrl":"10.1016/j.jcoa.2026.100318","url":null,"abstract":"<div><div>The growing therapeutic interest in peptides from <em>Tityus stigmurus</em> scorpion venom and their analogues highlights the need for the development of robust analytical methods for these compounds. However, traditional method development is often inefficient, failing to account for the complex interactions between chromatographic parameters that influence analytical quality. This study aimed to apply a Analytical Quality by Design (AQbD) approach to develop a highly efficient and robust method for the simultaneous separation of four synthetic peptides (Stigmurin, two analogs and TistH). The AQbD framework was used, employing risk assessment to identify critical variables and attributes. Two experimental designs— two-level screening and a three-level optimization—evaluated the impact of column type, mobile phase composition, and flow rate. Statistical analysis generated models and response surface plots to define an optimal operating region. Core-shell particle columns demonstrated superior performance, achieving baseline resolution for all peptides. Statistical models confirmed that mobile phase composition was the most influential factor. The defined design space identified a region for optimal operation: 40–43% (v/v) organic mobile phase, 20–40 °C, and a flow rate of 0.25–0.4 mL/min. Greenness analysis confirmed that it can be considered a green analytical method. The AQbD approach successfully created a reliable ultra-high performance liquid chromatography with time-of-flight mass spectrometry (UHPLC-ToF-MS) green and applicable method for peptide analysis, providing a flexible and high-performance tool for this application. This methodology serves as a valuable blueprint for developing robust analytical methods for other peptide mixtures, ensuring quality and repeatability.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100318"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147384197","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":"Study on enantioselective recognition of metofoline enantiomers with cyclodextrins using capillary electrophoresis, nuclear magnetic resonance spectroscopy and molecular modeling techniques","authors":"Ketevan Museridze ,&nbsp;Ann Gogolashvili ,&nbsp;Lali Chankvetadze ,&nbsp;Giorgi Jibuti ,&nbsp;Antonio Salgado ,&nbsp;Barbara Sechi ,&nbsp;Paola Peluso ,&nbsp;Bezhan Chankvetadze","doi":"10.1016/j.jcoa.2025.100303","DOIUrl":"10.1016/j.jcoa.2025.100303","url":null,"abstract":"<div><div>Compared to any other analytical technique, the main advantage of using capillary electrophoresis (CE) for studying enantioselective recognition mechanisms in intermolecular interaction is its higher sensitivity to detect weak noncovalent interactions. Furthermore, nuclear magnetic resonance (NMR) and molecular modeling proved to be very effective to complement CE for mechanistic purposes. On the other hand, multiple factors may impact selector-selectand geometries and binding energies affecting the overall enantiorecognition process. In this regard, the validation of computational tools to verify the reliability of proposed models is still a critical issue, and suitable experimental data sets are necessary for this purpose. In this study, we combined CE, NMR spectroscopy and molecular modeling with the aim of investigating the molecular bases of the enantioseparations of metofoline (MF) with β-cyclodextrin (CD), γ-CD, and <em>heptakis</em>(2,3-di-<em>O</em>-acetyl)-β-CD (HDA-β-CD). Based on this strategy and focusing on the reversal of migration order observed by using γ-CD [(<em>R</em>)-(<em>S</em>)] in place of β-CD [(<em>S</em>)-(<em>R</em>)], good agreement between experimental and theoretical data was obtained. The absolute configuration of MF enantiomers was assigned by comparison of experimental and computed chiroptical spectra.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100303"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145924370","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":"Comprehensive characterization of Xuebijing injection and comparative analysis of its systemic exposure in patients with severe pneumonia, sepsis, and acute pancreatitis","authors":"Mingxin Guo ,&nbsp;Shuo Chen ,&nbsp;Xia Wu ,&nbsp;Zhiqiang Hu ,&nbsp;Yanchun Chen ,&nbsp;Baohu Jiang","doi":"10.1016/j.jcoa.2025.100304","DOIUrl":"10.1016/j.jcoa.2025.100304","url":null,"abstract":"<div><div>This study aimed to investigate the chemical constituents of Xuebijing injection and compare differences in its systemic exposure components among patients with severe pneumonia, sepsis, and acute pancreatitis during the steady-state blood concentration phase. Using ultra-high-performance liquid chromatography-mass spectrometry, compounds were identified and analyzed based on retention time, accurate molecular weight, secondary mass spectrometry fragmentation patterns, and comparisons with reference standards and databases. A total of 55 chemical constituents were identified in Xuebijing injection, primarily flavonoids, organooxygen compounds, prenol lipids, and isobenzofurans. In patient plasma, 16 components were detected in severe pneumonia patients, comprising 7 prototypes, 6 Phase I metabolites, and 3 Phase II metabolites; 32 components were found in acute pancreatitis patients, including 12 prototypes, 10 Phase I metabolites, and 10 Phase II metabolites; and 24 components were identified in sepsis patients, consisting of 8 prototypes, 9 Phase I metabolites, and 4 Phase II metabolites. Acute pancreatitis patients exhibited the highest number of systemic exposure components, followed by sepsis and severe pneumonia patients. Hydroxysafflor yellow A, paeoniflorin, and neocnidilide glucuronidation were consistently detected at relatively high concentrations across all groups. This study demonstrates the utility of ultra-high-performance liquid chromatography-mass spectrometry for rapid profiling of the chemical composition of Xuebijing injection and its blood-exposed components, providing a reference for elucidating its pharmacodynamic material basis and mechanism of action.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100304"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145924371","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":"Analytical approaches for the identification and quantitation of nitazenes: a review","authors":"Roberto Mandrioli ,&nbsp;Michele Protti ,&nbsp;Roberta Di Lecce ,&nbsp;Laura Mercolini","doi":"10.1016/j.jcoa.2025.100305","DOIUrl":"10.1016/j.jcoa.2025.100305","url":null,"abstract":"<div><div>Nitazenes, a class of high-potency novel synthetic opioids containing a benzimidazole moiety, are currently being reported across seized materials and drug-checking submissions, human biological matrices (postmortem and clinical/forensic) and community-level samples including wastewater. The analytical literature converges on a layered, matrix-oriented toolkit. Rapid presumptive strategies (vibrational fingerprints and ambient-ionization techniques) support triage in solids and field-adjacent contexts, while chromatographic separations coupled to mass spectrometry (HPLC-MS/MS/HRMS and less frequently GC–MS/MS/HRMS) deliver selectivity and structure-aware confirmation for closely related analogues. In human biofluids, both targeted triple-quadrupole panels and HRMS suspect screening workflows are present for identification; validated quantitative methods consistently address calibration model and range, sensitivity (LOD/LLOQ), precision and accuracy, selectivity, carry-over and matrix effects, often with explicit attention to isomer resolution. Cross-cutting issues recurring in the field include reference standard availability, isomeric/isobaric interferences, immunoassay cross-reactivity and the promise of miniaturised sampling formats. Organised by matrix and analytical objective, this review provides enough information to enable efficient extraction of details at the method level, while ensuring a comprehensive coverage of the most important topics.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100305"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145924372","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":"A size exclusion chromatography- and ultrafiltration-based two-step process for purifying phage for therapeutic applications","authors":"Rahul Patel ,&nbsp;Dhruvrajsinh Rathod ,&nbsp;Kyle Jackson ,&nbsp;Mrunal Ingawale ,&nbsp;Yating Xu ,&nbsp;Zeinab Hosseinidoust ,&nbsp;Raja Ghosh","doi":"10.1016/j.jcoa.2025.100288","DOIUrl":"10.1016/j.jcoa.2025.100288","url":null,"abstract":"<div><div>The rise in the number of antibiotic-resistant bacteria has created the need for alternative therapeutic approaches, including bacteriophage therapy. Bacteriophages are viruses that infect and lyse bacteria and therefore offer a highly targeted and adaptable solution to combat multidrug-resistant pathogens. However, the scalable purification of bacteriophages, while achieving high yield of product and efficient removal of impurities such as endotoxins, remains a significant barrier to their therapeutic adoption. This study presents a two-step purification process combining size-exclusion chromatography (SEC) and ultrafiltration as an efficient and scalable method for bacteriophage purification. T7 was used as the model bacteriophage. Preparative SEC using a cuboid device served as the first step for removing endotoxin and other impurities from bacterial culture filtrate. The SEC-purified phage sample was then processed using a carrier phase ultrafiltration and backflow recovery technique to further purify and enrich the bacteriophage. Phage titer, endotoxin level, and protein concentration were quantified at each stage to evaluate purification performance. The endotoxin level was substantially reduced, decreasing from 1.44 EU/mL in the initial culture filtrate to 0.49 EU/mL in the final purified phage sample, the FDA's acceptable limit for products that directly or indirectly contact the cardiovascular and lymphatic systems being 0.50 EU/mL. The phage titer was enhanced from 5.33×10⁹ PFU/mL (in the cell culture filtrate) to 9.33×10⁹ PFU/mL in the final product. The overall protein removal achieved was 95.8 % while the overall endotoxin removal was 92.5 %. These findings suggest that the proposed two-step purification process could be developed further and adopted for scalable production of high-purity bacteriophage preparations suitable for therapeutic applications.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"9 ","pages":"Article 100288"},"PeriodicalIF":3.2,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145618380","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}