Journal of chromatography open最新文献

{"title":"A tutorial on the synthesis and applications of molecularly imprinted polymers in analytical chemistry","authors":"N.P. Kalogiouri ,&nbsp;A. Kabir ,&nbsp;K.G. Furton ,&nbsp;V.F. Samanidou","doi":"10.1016/j.jcoa.2025.100244","DOIUrl":"10.1016/j.jcoa.2025.100244","url":null,"abstract":"<div><div>Molecularly imprinted polymers (MIPs) have emerged as highly selective sorbents with tailor-made recognition sites complementary to target analytes in terms of shape, size, and functional groups. This tutorial provides a comprehensive overview of the principles, synthesis strategies, and analytical applications of MIPs, with a focus on their utility in sample preparation, separation, and detection. Various imprinting techniques—such as bulk, surface, and nanoimprinting—are discussed alongside advances in green synthesis and novel monomer-template systems. Emphasis is placed on the integration of MIPs with chromatographic and spectrometric platforms, particularly in complex matrices relevant to environmental, pharmaceutical, food, and clinical analysis. The tutorial illustrates the practical considerations for designing and optimizing MIP-based methods, including template selection, polymerization conditions, and reusability, reflecting recent and relevant advances of the last ten years This tutorial aims to guide researchers and analysts in leveraging the unique properties of MIPs to enhance selectivity, sensitivity, and sustainability in modern analytical chemistry.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100244"},"PeriodicalIF":3.2,"publicationDate":"2025-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144750367","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":"Quantification of lactate enantiomers in human sweat samples using two-dimensional liquid chromatography","authors":"Kazushi Mori,&nbsp;Makoto Tsunoda","doi":"10.1016/j.jcoa.2025.100243","DOIUrl":"10.1016/j.jcoa.2025.100243","url":null,"abstract":"<div><div>Lactate (LA) is primarily produced by the reduction of pyruvate in the human body and is crucial for energy production via anaerobic glycolysis. Although the D-LA concentration is considerably lower than that of L-LA, a significant increase in D-LA concentration alone has been reported in some diseases. Quantifying LA enantiomers in human biofluids has the potential for disease diagnosis. Sweat has recently been recognized as a novel biological alternative to blood because it can be sampled non-invasively. Therefore, in this study, heart-cutting two-dimensional liquid chromatography (2D-LC) using a highly sensitive fluorescence detection method was developed for the analysis of LA enantiomers in small amounts of human sweat. LA was derivatized with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) and separated from endogenous compounds using an octadecyl silica column in the first dimension. Subsequently, the NBD-PZ-LA peak was fractionated and enantiomerically separated in the second dimension on a chiral column. Sufficient linearities (R² &gt; 0.999) were observed in the ranges of 1–100 and 10–1000 µM for NBD-PZ-D-LA and NBD-PZ-L-LA, respectively. The corresponding limits of quantification were 0.97 and 1.12 µM. The precision values were 1.04 %–12.03 %, and the accuracies were 85.6 %–100.4 %. The developed method was successfully applied to ∼5 µL of human sweat collected from five healthy subjects. The concentrations of D-LA and L-LA in sweat were 30.29 ± 20.18 µM and 23.69 ± 12.15 mM, respectively. The developed 2D-LC system should be clinically applicable to LA enantiomer analysis in human sweat as a non-invasive biomarker.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100243"},"PeriodicalIF":3.2,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144738509","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":"Critical review on the assessments of green metric tools for Olmesartan medoxomil analytical methods: Case studies","authors":"Shaikh Manirul Haque ,&nbsp;Abuzar Kabir","doi":"10.1016/j.jcoa.2025.100242","DOIUrl":"10.1016/j.jcoa.2025.100242","url":null,"abstract":"<div><div>Green chemistry metrics can incorporate features of analytical practices that adhere to the principles of green chemistry. Several metrics are available to evaluate the greenness of the analytical procedure, including the AES, AGREE, and BAGI. The current review article outlines the selected metrics used to determine case studies of Olmesartan, employing various analytical techniques, including UV spectrophotometry, HPLC, HPTLC, UPLC, LC-MS, and UPLC-MS, based on specific criteria. These criteria also include the advantages, disadvantages, and limitations of each technique. The analytical eco–scale, AGREE and BAGI values were in the range 79–86, 72–84, 66–68, 77, 75–89, 67–75; 0.6–0.71, 0.55–0.65, 0.58–0.65, 0.62, 0.65–0.66, 0.68–0.69; 77.5–82.5, 72.5–82.5; 75–77.5, 80, 75 – 80 and 80 respectively for UV spectrophotometry, HPLC, HPTLC, UPLC, LC-MS and UPLC-MS. The results are well-organised according to the criteria and have produced an excellent greener methodology. The new researchers could understand and use all the assessment tools for different purposes. Additionally, they can utilise it to develop new procedures, which could demonstrate a greener methodology for quantifying Olmesartan in various matrices.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100242"},"PeriodicalIF":0.0,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144686445","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":"Ionic liquids in liquid chromatography","authors":"Sini Santasalo ,&nbsp;Susanne K. Wiedmer","doi":"10.1016/j.jcoa.2025.100239","DOIUrl":"10.1016/j.jcoa.2025.100239","url":null,"abstract":"<div><div>The use of ionic liquids (ILs) in liquid chromatography (LC) is reviewed. ILs have several different types of functionalities, which means that they are prone to multiple simultaneous interactions with stationary phases in LC. ILs have been investigated by LC as analytes and they have been used as mobile phase additives and column stationary phase modifiers in LC. All these applications include a diverse range of interactions with different LC stationary phases. Common reasons for using ILs as mobile phase additives have been to reduce unwanted interactions between analytes and free silanols on the column stationary phase, to act as ion pair reagents to separate polar compounds with reversed phase LC, and to act as UV absorption reagents to detect compounds that do not absorb UV light. The review covers the most important works between 2003–2025.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100239"},"PeriodicalIF":0.0,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144655275","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 thermodynamic study of relative extractability of ethanol to blood simulating solvent in extractables and leachables analysis of medical devices by Abraham’s Solvation Parameter Model","authors":"Jianwei Li","doi":"10.1016/j.jcoa.2025.100238","DOIUrl":"10.1016/j.jcoa.2025.100238","url":null,"abstract":"<div><div>A thermodynamic study of over-extraction of blood contacting medical devices by ethanol to ethanol/water binary cosolvent mixture (50/50, v/v), as a blood simulating solvent, in chemical analysis of medical devices is evaluated by Abraham’s solvation parameter model using five representative materials (low density polyethylene or LDPE, silicone, polyurethane or PU, polyoxymethylene or POM, and polyacrylate or PA). The Abraham’s model is initially used to calculate the material-solvent partition system coefficients by the corresponding partition system constants and representative extractables compounds between five materials and ethanol/water (and methanol/water) cosolvent mixtures at different volume fractions. The partition system constants are indirectly derived by a “thermodynamic circle conversion” method, based on material-water partition systems and solvent-water water partition systems. The material-solvent (mixture) partition coefficient, <span><math><msub><mi>P</mi><mrow><mi>M</mi><mo>/</mo><mi>S</mi><mi>o</mi><mi>l</mi><mi>v</mi><mi>e</mi><mi>n</mi><mi>t</mi></mrow></msub></math></span>=<span><math><mrow><msub><mi>C</mi><mi>M</mi></msub><mo>/</mo><msub><mi>C</mi><mrow><mi>S</mi><mi>o</mi><mi>l</mi><mi>v</mi><mi>e</mi><mi>n</mi><mi>t</mi></mrow></msub></mrow></math></span>, defined as the concentration in the material phase divided by the concentration in the solvent phase, is used as an indicator of the solvent extraction strength. <span><math><mrow><mi>log</mi><mo>(</mo><msub><mi>P</mi><mrow><mi>M</mi><mo>/</mo><mi>S</mi><mi>o</mi><mi>l</mi><mi>v</mi><mi>e</mi><mi>n</mi><mi>t</mi></mrow></msub><mo>)</mo></mrow></math></span> values are computed for all material-solvent pairs using the representative extractables compounds, mostly from Wayne State University experimental descriptor database (WSUEDD). The predictive <span><math><mrow><msub><mi>log</mi><mn>10</mn></msub><mrow><mo>(</mo><msub><mi>P</mi><mrow><mi>M</mi><mo>/</mo><mi>S</mi><mi>o</mi><mi>l</mi><mi>v</mi><mi>e</mi><mi>n</mi><mi>t</mi></mrow></msub><mo>)</mo></mrow></mrow></math></span> values between LDPE and silicone materials and ethanol/water (and methanol/water) cosolvent mixtures are compared with the available experimental values to assess the model’s predictive accuracy. Afterward, the predictive <span><math><mrow><msub><mi>log</mi><mn>10</mn></msub><mrow><mo>(</mo><msub><mi>P</mi><mrow><mi>M</mi><mo>/</mo><mi>S</mi><mi>o</mi><mi>l</mi><mi>v</mi><mi>e</mi><mi>n</mi><mi>t</mi></mrow></msub><mo>)</mo></mrow></mrow></math></span> values of ethanol to ethanol/water binary cosolvent mixture (50/50, v/v) are used to calculate relative extractability (<em>RE</em>), as an indicator of over-extraction. Several conclusions can be drawn from this study. First, the predicted partition coefficients are confirmed by available experimental values (LDPE and silicone). Second, the over-extraction of medical devices by ethanol to the blood simulating solvent is less important ","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100238"},"PeriodicalIF":0.0,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144655274","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":"Chemical analysis using miniaturized and Portable 3D printed systems: Where are we now?","authors":"Miryam Perrucci ,&nbsp;Imran Ali ,&nbsp;Fotouh R. Mansour ,&nbsp;Halil I. Ulusoy ,&nbsp;Songül Ulusoy ,&nbsp;Abuzar Kabir ,&nbsp;Ornella Abollino ,&nbsp;Agnese Giacomino ,&nbsp;Paolo Inaudi ,&nbsp;Marcello Locatelli ,&nbsp;Erika Maria Ricci","doi":"10.1016/j.jcoa.2025.100241","DOIUrl":"10.1016/j.jcoa.2025.100241","url":null,"abstract":"<div><div>With the growing emphasis on green chemistry (GC) and green analytical chemistry (GAC), significant efforts have been directed toward minimizing sample size and simplifying sample preparation procedures. Concurrently, ongoing research has enabled the miniaturization of instrumental configurations used for quantitative analysis, aligning with the principles of sustainability.</div><div>One major challenge lies in the inherent complexity of liquid chromatography systems, which comprise multiple interconnected components. Each component has been subject to miniaturization, with careful attention to preserving analytical sensitivity, selectivity, and accuracy when applied to real samples. Interest in miniaturized systems is further fueled by the theoretical principle that reducing flow rates enhances the surface-to-volume ratio, thereby improving detection sensitivity.</div><div>Building on this theoretical foundation—particularly the goals of portability, efficient sampling, and <em>on site</em> analysis—innovative approaches such as microfluidics and 3D printing have been employed to develop compact fluidic systems. These technologies not only broaden the scope of liquid chromatography but also open new frontiers, such as drug testing on engineered tissues, and the development of new devices suitable for space missions.</div><div>This review aims to present a comprehensive overview of the challenges encountered and the advancements achieved in the miniaturization of instrumental setups and portable systems, encompassing both pre- and post-analytical stages. Furthermore, a discussion on 3D technologies available for LC-chip and miniaturized systems was also presented.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100241"},"PeriodicalIF":0.0,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144633153","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":"Advances in miniaturized liquid sample preparation techniques for environmental analysis: A special look towards green solvents and automation","authors":"Rafael Oliveira Martins,&nbsp;Camila Will,&nbsp;Maria Flávia Assunção Magalhães,&nbsp;Fernando Mauro Lanças","doi":"10.1016/j.jcoa.2025.100240","DOIUrl":"10.1016/j.jcoa.2025.100240","url":null,"abstract":"<div><div>The continuous advancements in agricultural and industrial practices have led to the widespread introduction of various chemicals across key production sectors. While these substances offer significant benefits, their improper disposal has led to their accumulation in environmental compartments, raising global concerns about their potential impacts on ecosystems and human health. In quantitative and qualitative analysis, liquid miniaturized techniques have been widely applied to these matrices to ensure the detection and quantification of these compounds in environmental samples. Many of their advantages are rooted in key principles of green analytical chemistry, such as reduced sample and solvent consumption, as well as their versatility in enabling diverse extraction designs. However, the use of conventional organic solvents and labor-intensive extraction procedures still limits the full potential of these techniques in environmental applications. Hence, the present review provides an overview of current applications of miniaturized liquid-phase extraction protocols in environmental analysis, highlighting innovative approaches. Moreover, we discuss the use of green solvents and the potential for automation in these methods. Through this review, we aim to encourage researchers to adopt miniaturized liquid techniques for environmental monitoring, thereby advancing the implementation of more sustainable and efficient analytical strategies.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100240"},"PeriodicalIF":0.0,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144596592","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":"Metabolite patterns of the carbonic anhydrase inhibitors brinzolamide and dorzolamide: potential markers for the route of application","authors":"Y Jin ,&nbsp;A Thomas ,&nbsp;S Gochard ,&nbsp;P Delahaut ,&nbsp;M Thevis","doi":"10.1016/j.jcoa.2025.100236","DOIUrl":"10.1016/j.jcoa.2025.100236","url":null,"abstract":"<div><div>Brinzolamide (BA) and dorzolamide (DA) are carbonic anhydrase inhibitors (CAIs), and are commonly used therapeutics for glaucoma treatment via topical application. According to the regulations of the World Anti-Doping Agency (WADA), both drugs are classified under \"Diuretics and Masking Agents\", prohibiting their use in- and out-of-competition, except for ophthalmic application. Despite ophthalmic use being permitted, systemic absorption occurs, enabling BA and DA detection in blood and urine samples after topical administration. Thus, distinguishing between legitimate (e.g. topical) and prohibited (e.g. oral) drug application routes is critical for sports drug testing result management.</div><div>This study investigated the elimination and metabolic profiles of BA and DA following topical and systemic administration in male piglets. Three piglets received BA or DA ophthalmic suspensions, while another three received an oral dose. Urine and blood samples were collected over one week, and all samples were analysed using liquid chromatography-high-resolution tandem mass spectrometry (LC<img>HRMS MS). In vitro experiments yielded five phase I metabolites for DA and BA. After method validation, the approach was confirmed to detect DA and BA, with Limits of Detection (LODs) of 55 pg/mL and 75 pg/mL in urine, 110 pg/mL and 180 pg/mL in red blood cells, and 380 pg/mL and 910 pg/mL in plasma. BA and DA metabolites were primarily found in the red blood cell fraction, with only trace amounts detectable in plasma. N-desethylation was observed as the main metabolic reaction for both drugs, and metabolite-to-parent drug ratios were determined in all collected post-administration samples alongside drug concentration levels. The combined consideration of analyte ratios and drug concentrations appears to be indicative of time and dose of drug use (under the chosen routes of administration), which might assist in sports drug testing result management.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100236"},"PeriodicalIF":0.0,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144563681","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":"Cutaneous volatile and semi-volatile organic compounds as markers of malaria-infection by wearable samplers and two-dimensional gas chromatography—time-of-flight-mass spectrometry","authors":"Daniel T. Pretorius ,&nbsp;Egmont R. Rohwer ,&nbsp;Yvette Naudé","doi":"10.1016/j.jcoa.2025.100233","DOIUrl":"10.1016/j.jcoa.2025.100233","url":null,"abstract":"<div><div>Malaria has been found to alter normal cutaneous volatile organic compound (VOC) profiles, suggesting their potential application as markers of <em>Plasmodium</em> infection. The cutaneous VOCs and semi-VOCs (SVOCs) of malaria-negative and -positive individuals, who visited two local clinics in the Vhembe district of Limpopo Province, South Africa, were extracted into wearable silicone rubber (polydimethyl siloxane [PDMS]) sampling bands adhered to the surface of the epidermis. After sampling of epidermal VOCs from participants the samplers were analysed by thermal desorption-comprehensive two-dimensional gas chromatography-time-of-flight-mass spectrometry (TD-GC × GC-TOFMS). Individual cutaneous VOCs and SVOCs profiles were constructed from these complex chromatographic profiles in order to identify potential signatures of <em>Plasmodium</em> infection. Fatty acid compounds associated with rancid malodour, and previously reported as mosquito attractants, were found at an overall greater abundance in chemical profiles of malaria-positive cases. A targeted analysis was performed for compounds previously reported to be associated with <em>Plasmodium</em> infection, viz., heptanal, (E)-2-octenal, 2-octanone, octanal, nonanal and (E)-2-decenal. The linearity (R²) range was 0.93–0.99 for a matrix matched (simulated cutaneous sampling) calibration range of 2.5–60 ng. Limits of detection (LOD) ranged from 0.4 pg (2-octanone) to 6.3 pg ((E)-2-octenal), whilst limits of quantification (LOQ) ranged from 1.4 pg to 21.1 pg. The mean percentage recoveries (<em>n</em> = 2) ranged from 77.8 % ((E)-2-decenal) to 118.9 % (2-octanone). The percentage relative standard deviations ( %RSDs; <em>n</em> = 2) ranged from &lt; 1 % for 2-octanone, octanal and nonanal to 27.1 % for (E)-2-octenal. We found that this particular suite of compounds, previously reported as indicators of malaria, was in fact non-specific for <em>Plasmodium</em> infection when compared to control subjects with comorbidities. A previously unreported (in a malaria-infection context) compound, (E)-2-octen-1-ol, correlated with malaria-positive participants, but was also observed for two malaria-negative participants, which could indicate latent malaria. In chronic cases, <em>Plasmodium vivax</em> can occur in reservoirs outside of the bloodstream, and thus blood-based diagnostic tests can miss latent infection. A key advantage of the epidermal sampler over blood tests is that the former collects whole-body organic compounds, and is therefore not limited to blood-borne markers of infection. As such it appears to be feasible for future investigations.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100233"},"PeriodicalIF":0.0,"publicationDate":"2025-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144306606","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":"Optimization of immobilized metal affinity chromatography for a recombinant protein expressed in CHO cells","authors":"Xiaoyong Jiang ,&nbsp;Rongqiu Qin ,&nbsp;Tengfei Ma,&nbsp;Kexin Xu,&nbsp;Fengjuan Lv","doi":"10.1016/j.jcoa.2025.100230","DOIUrl":"10.1016/j.jcoa.2025.100230","url":null,"abstract":"<div><div>Immobilized metal affinity chromatography (IMAC) is widely used in research laboratories to purify His-tagged proteins expressed in Escherichia coli (<em>E. coli</em>) by enabling direct capture and moderate removal of process-related impurities, such as host cell proteins (HCPs) and DNA. However, its application for purifying recombinant proteins secreted by mammalian cells is limited due to incompatibility issues between cell culture media and IMAC resins. In this study, we purified a CHO-expressed and secreted recombinant protein with His-tag using a Ni Sepharose excel resin, resistant to EDTA and reducing agents, and optimized loading, washing, and elution conditions to maximize protein recovery and HCP clearance. The resin demonstrated a maximum load capacity of 10 mg/mL. Results show that incorporating Triton X-100, PS80 &amp; TnBP, or 2-propanol in washing buffers significantly improves HCP removal while maintaining low salt concentrations in the elution buffer enhances both yield and product quality. Resin lifetime studies conducted under optimal conditions showed acceptable yields, stable product quality attributes (PQAs), and effective cleaning over 54 purification cycles. This protocol provides a robust IMAC purification strategy, potentially broadening its industrial applications.</div></div>","PeriodicalId":93576,"journal":{"name":"Journal of chromatography open","volume":"8 ","pages":"Article 100230"},"PeriodicalIF":0.0,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144272210","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}