Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Potential drug exposure risk assessment of analysts in analytical laboratories through fingernail, hair, and atmospheric environment analyses","authors":"Panpan Chen ,&nbsp;Wu Wen ,&nbsp;Hongxian Yang ,&nbsp;Jianmei Wu ,&nbsp;Xiaoyu Zheng ,&nbsp;Mengxiang Su ,&nbsp;Jie Chen","doi":"10.1016/j.jpba.2025.116779","DOIUrl":"10.1016/j.jpba.2025.116779","url":null,"abstract":"<div><div>Analysts in drug testing laboratories are frequently exposed to drugs as part of their daily work activities. Certain drug components can vaporise and adhere to atmospheric particles. Therefore, the presence of drugs in laboratory settings can affect the health and well-being of the laboratory personnel. This study aimed to highlight possible health consequences of 25 expected drug exposure risks by examining fingernail, hair, and atmospheric environment samples. The results showed that etomidate was detected in the hair samples of eight individuals, with the highest level of 19.73 pg/mg, and no components were detected in the fingernail samples. This indicates that etomidate may adhere to hair but does not significantly affect the human body. An atmospheric particulate sampler was used to collect atmospheric particulate matter and gaseous samples from different laboratory rooms. Methamphetamine, ketamine, etomidate, and other drugs were detected in atmospheric particles and gases. Furthermore, the distribution of these chemicals was evaluated by comparing their concentrations in total suspended particulate matter and gas phase. The development and application of these methods coupled with data analysis, aimed at enhancing the self-protection awareness of laboratory scientists and establish a foundation for future assessments of potential hazards.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116779"},"PeriodicalIF":3.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521017","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":"Exploring efavirenz-DNA interactions: A multidisciplinary approach through electrochemical, toxicological, and in silico investigations","authors":"Manolya Mujgan Yildiz ,&nbsp;Ilker Ates ,&nbsp;Havva Nur Gurbuz ,&nbsp;Mehmet Altay Unal ,&nbsp;Hasan Nazir ,&nbsp;Aytekin Uzunoglu ,&nbsp;Sibel A. Ozkan ,&nbsp;Burcu DOGAN Topal","doi":"10.1016/j.jpba.2025.116763","DOIUrl":"10.1016/j.jpba.2025.116763","url":null,"abstract":"<div><div>Recently, there has been a growing approach that approved drugs have been tested for additional purposes. Efavirenz is a non-nucleoside reverse transcriptase inhibitor used to treat human immunodeficiency virus infection. In addition, it has selective cytotoxic effects against cancer cells. This study constructed an electrochemical dsDNA nanobiosensor to monitor Efavirenz-dsDNA interaction based on the amine-functionalized multi-walled carbon nanotubes. The experimental conditions of the nanobiosensor, such as dropping the volume of nanomaterial suspension, activation of the nanosensor, and dsDNA concentration, were optimized. The peak currents of dsDNA bases were enhanced, and the peak potentials of Efavirenz have shifted to the less positive potential thanks to the modified sensor with amine-functionalized multi-walled carbon nanotubes. The interaction mechanism was also evaluated in incubated solutions. Docking calculations showed that Efavirenz is active in the large cleft regions of DNA that suggest minor groove binding. The effect of efavirenz on the expression profile of particular stress and possible DNA genotoxicity was studied via examining gene polymorphisms in hepatic cells. These findings align with previously released research that shows Efavirenz-treated hepatic cells to have altered mitochondrial function and elevated ROS levels.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116763"},"PeriodicalIF":3.1,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527316","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":"Comparative analysis of chemical constituents in the stems and roots of different Ephedra species and their in vitro anti-acute lung injury effects","authors":"Bing Guo ,&nbsp;Hengyang Li ,&nbsp;Qi An ,&nbsp;Lina Yang ,&nbsp;Yongli Liu ,&nbsp;Hao Yuan ,&nbsp;Haiyang Wang ,&nbsp;Yuguang Zheng ,&nbsp;Long Guo ,&nbsp;Dan Zhang","doi":"10.1016/j.jpba.2025.116777","DOIUrl":"10.1016/j.jpba.2025.116777","url":null,"abstract":"<div><div><em>Ephedra</em>, the sole genus in the Ephedraceae family, comprises 75 species distributed across Asia, North America, and South America. Ephedrae herba, a well-established traditional Chinese medicine, has been used in clinical practice for over two millennia. <em>Ephedra</em> is widely used to treat asthma, colds, and coughs. Given the diversity of <em>Ephedra</em> species, studying their chemical composition and biological activity is essential for their comprehensive utilization. The chemical profiles of different <em>Ephedra</em> stem (MHS) and root (MHR) species were analyzed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MSMS). Network pharmacology was applied to investigate the mechanisms by which <em>Ephedra</em> influences acute lung injury (ALI). An <em>in vitro</em> ALI model, induced by LPS, was created to assess the activities of various MHS and MHR species. The anti-inflammatory mechanisms were validated through RT-qPCR and molecular docking studies. The results showed that a total of 27 and 35 constituents were identified in MHS and MHR, respectively, with 12 and 13 compounds screened by chemometric analysis as distinguish chemicals for different species of MHS and MHR Network pharmacology identified 7 core components in MHS and MHR, respectively, that intervene in ALI. Moreover, quercetin-4′-<em>O</em>-glucoside, (+)-catechin, and <em>l</em>-norpseudoephedrine from MHS, along with ephedradine A, epigallocatechin, and (-)-epicatechin from MHR, were identified as key components distinguishing the different parts of <em>Ephedra</em>. A comparison of <em>in vitro</em> anti-inflammatory activities of various MHS and MHR species in an ALI model revealed varying degrees of anti-inflammatory activity. Correlation analyses indicated that apigenin-6,8-<em>C</em>-dihexoside, vitexin-2-<em>O</em>-rhamnoside, quercetin-4′-<em>O</em>-glucoside, and pseudoephedrine from MHS, and ephedradine A, feruloylhistamine, kaempferol-3-<em>O</em>-glucoside-7-<em>O</em>-rhamnoside, and diphenydramine from MHR may be the core compounds contributing to the differences in the activity of various MHS and MHR species. RT-qPCR and molecular docking results indicate that <em>Ephedra</em> may exert anti-inflammatory effects on the MAPK/NF-κB targets through 14 core components. This study serves as a foundational reference for identifying <em>Ephedra</em> species, assessing their quality, and evaluating their pharmacodynamics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116777"},"PeriodicalIF":3.1,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521019","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 high performance liquid chromatography method based on a core-shell octadecyl column to determine Avermectin and its related substances in bulk Avermectin batches","authors":"Shane N. Berger,&nbsp;Abu M. Rustum","doi":"10.1016/j.jpba.2025.116773","DOIUrl":"10.1016/j.jpba.2025.116773","url":null,"abstract":"<div><div>The Avermectins are a family of macrocyclic lactones isolated from the soil bacterium <em>Streptomyces avermitilis</em> that possess broad spectrum activity as anthelmintics and are employed throughout the agricultural and animal health industries. Avermectin is the parent molecule from which several important parasiticides have been derived, but currently the Ph. Eur. and USP do not describe an HPLC monograph for analysis of Avermectin. Here, we detail the development and validation of a reversed-phase HPLC method for determination of Avermectin and its related substances in bulk batches of Avermectin. The separation of all peaks of interest was accomplished by a gradient elution on an ACE UltraCore (core shell) 2.5 Super C18 column (150 mm × 4.6 mm, 2.5 µm particle size) maintained at 45 °C. The mobile phase for the gradient is 5 mM NH₄OAc in water at pH 9.5 (mobile phase-A) and acetonitrile/methanol/dichloromethane (52/40.5/7.5, v/v/v) (mobile phase-B). Avermectin and its related substances were detected and quantified at 245 nm with a quantitation limit of 0.05 % of the analytical concentration. Method validation following the International Conference on Harmonization guidelines was carried out with respect to specificity, linearity, accuracy, intermediate precision, and robustness and all acceptance criteria were met. The new HPLC method will provide reliable analysis of Avermectin and its related substances, while the method development data and forced degradation data herein will provide critical information applicable to the Avermectin family as a whole.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116773"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521018","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":"Fast quality assessment and origin identification of Gentianae Macrophyllae Radix using fourier transform infrared photoacoustic spectroscopy coupled with chemometrics","authors":"Xiang Liu ,&nbsp;Jing Huang ,&nbsp;Wenlong Li ,&nbsp;Rongqin Chen ,&nbsp;Liuye Cao ,&nbsp;Tiantian Pan ,&nbsp;Fei Liu","doi":"10.1016/j.jpba.2025.116774","DOIUrl":"10.1016/j.jpba.2025.116774","url":null,"abstract":"<div><div>This study explores the application of Fourier Transform Infrared Photoacoustic Spectroscopy (FTIR-PAS) combined with chemometric analysis for rapid quality assessment and origin species verification of <em>Gentianae Macrophyllae Radix</em> (Qinjiao). Qinjiao, a traditional Chinese medicinal herb, faces significant challenges in quality control due to frequent adulteration and substitution. In this study, 195 Qinjiao samples were analyzed using FTIR-PAS, and chemometric techniques were employed for both classification and regression analysis. The results demonstrate that FTIR-PAS, integrated with chemometric methods, effectively differentiates Qinjiao samples by species. The combination of Detrending (DT) preprocessing and a K-Nearest Neighbors (KNN) classification model achieved an accuracy of 97.4 % in species identification. For quantitative analysis, Savitzky-Golay (SG) smoothing was the optimal preprocessing method for gentiopicroside, while DT was best for loganic acid, and no preprocessing was necessary for roburic acid. Additionally, the Competitive Adaptive Reweighted Sampling (CARS) algorithm, combined with models such as XGBoost and Random Forest (RF), significantly improved the prediction accuracy of key active components, with the highest <span><math><msubsup><mrow><mi>R</mi></mrow><mrow><mi>P</mi></mrow><mrow><mn>2</mn></mrow></msubsup></math></span> reaching 0.84. This research underscores the potential of FTIR-PAS as a rapid, non-destructive approach for the quality assessment and authentication of traditional Chinese medicinal materials.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116774"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527314","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":"Integrating 16S rDNA sequencing analysis and targeted metabolomics to explore the mechanism of Xiexin Tang in treating atherosclerosis mice induced by high-fat diet","authors":"Junling Li ,&nbsp;Qianru Gao ,&nbsp;Hongtao Liu ,&nbsp;Songlin Liu ,&nbsp;Yanchun Wang ,&nbsp;Xiongjie Sun ,&nbsp;Junping Zheng ,&nbsp;Huabing Yang ,&nbsp;Baifei Hu","doi":"10.1016/j.jpba.2025.116760","DOIUrl":"10.1016/j.jpba.2025.116760","url":null,"abstract":"<div><div>Xiexin Tang (XXT) is a classic Chinese medicine formula that can be used to treat Atherosclerosis (AS). This study aimed to investigate the mechanism by which XXT regulated AS lipid levels. Firstly, the mixture components of XXT were analyzed by High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Then, the AS model based on Apolipoprotein E knockout (ApoE^-/-) mice was established. Cytokines related to lipid metabolism and bile acid metabolism were detected by Quantitative Real-time PCR (qRT-PCR). 16S rDNA gene sequencing was performed to analyze differential bacterial populations, and the mechanism of XXT regulation of bile acids affecting lipid metabolism was further explored by targeted metabolomics. Further, antibiotic-treated mice were used to investigate the role of gut microbiota in the anti-AS effect of XXT. The results showed that XXT attenuated the lipid levels and reversed the abnormal elevation of cytokines, such as hepatic lipid metabolism and inflammatory reaction in AS mice. XXT also repaired the gut barrier damage and reversed gut microbiota disorders in AS mice. Furthermore, the metabolic levels of bile acids were reshaped by XXT. Whereas, in the absence of gut microbiota, XXT failed to attenuate lipid levels and inhibit the expression of cytokines related to inflammation and bile acid metabolism in AS mice and failed to play a role in ultimately treating AS. In conclusion, XXT could effectively inhibit the inflammatory reaction and lipid accumulation in AS mice, and this effect was closely related to its remodeling of gut microbiota to regulate bile acid metabolism.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116760"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510527","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":"Illuminating green fluorescent protein: Characterizing tri-peptide fluorescent chromophore, probing reactivity of cysteines, and unveiling site-directed modifications through mass spectrometry","authors":"Jianmin Zhang,&nbsp;Bing Wang","doi":"10.1016/j.jpba.2025.116771","DOIUrl":"10.1016/j.jpba.2025.116771","url":null,"abstract":"<div><div>Bioconjugation technologies enable covalent attachment of diagnostic or therapeutic effectuators onto biological targets, allowing for the precise delivery of desired drugs to the intended targets with enhanced potency, selectivity, specificity, and prolonged duration of action. As the number of bioconjugation techniques has grown enormously, identification and in-depth characterization of in-process products play a critical role in the development of covalent drug conjugates. This is especially significant in light of the increased complexity of novel biotherapeutics derived from biological matrices. This paper describes liquid chromatography-mass spectrometry (LC-MS/MS)-based studies that have contributed to the development of site-specific genetic incorporation of non-natural amino acids (nnAAs) into proteins. A holistic approach was implemented to characterize a wild type green fluorescent protein (wtGFP) and an enhanced green fluorescent protein (eGFP). By using the wtGFP as a pilot and model system, the reactivity of cysteine residues was investigated under different sample processing conditions, followed by a stability evaluation using intact mass measurement. The subsequent complementary proteolytic peptide mappings were performed to achieve full sequence coverage of the proteins, identification of predominant modifications, and granular details of the fluorescent chromophore. The developed method was successfully applied to isolate the eGFP incorporated with nnAA from cells. This enables the verification of the specific site of nnAA incorporation, and the characterization of complex variants using <em>de novo</em> sequencing techniques. MS studies demonstrated that <em>p</em>-azido-phenylalanine <em>(p</em>AzF) was specifically incorporated into the desired site of eGFP with high efficiency and fidelity.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116771"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527318","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":"The active metabolite hydroxyitraconazole has substantially higher in vivo free fraction and free concentrations compared to itraconazole","authors":"Motoshi Iwao ,&nbsp;Ryota Tanaka ,&nbsp;Yosuke Suzuki ,&nbsp;Ryosuke Tatsuta ,&nbsp;Takehiro Hashimoto ,&nbsp;Kazufumi Hiramatsu ,&nbsp;Hiroki Itoh","doi":"10.1016/j.jpba.2025.116775","DOIUrl":"10.1016/j.jpba.2025.116775","url":null,"abstract":"<div><div>Itraconazole (ITCZ) is a triazole antifungal agent that is metabolized to many products. Hydroxyitraconazole (OH-ITCZ) is the major metabolite with antifungal activity comparable to that of ITCZ. Protein-free drug concentration has been reported to be a better biomarker for pharmacodynamics compared with total drug concentration. We developed an assay for quantification of free ITCZ and free OH-ITCZ concentrations using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) with equilibrium dialysis. The assay fulfilled the requirements of the US Food and Drug Administration guideline for assay validation, with lower limit of quantification of 0.025 and 0.25 ng/mL for free ITCZ and OH-ITCZ, respectively, than previous studies. Using this validated assay, we measured plasma free ITCZ and OH-ITCZ concentrations in 18 samples of 11 adult patients who received oral ITCZ between July 2016 and November 2022 at Oita University Hospital, and compared the <em>in vivo</em> percent free fraction and free concentration between the two compounds. Average plasma free concentrations and percent free fraction in 18 samples were, respectively, 0.188 ± 0.123 ng/mL and 0.024 ± 0.016 % for ITCZ, and 1.449 ± 1.017 ng/mL and 0.251 ± 0.109 % for OH-ITCZ, indicating that OH-ITCZ was 8.52-fold higher in percent free fraction and 10.42-fold higher in free concentration compared to ITCZ. Given that OH-ITCZ and ITCZ have similar <em>in vitro</em> antifungal activity, OH-ITCZ may contribute more to <em>in vivo</em> antifungal efficacy than ITCZ, suggesting that monitoring OH-ITCZ would be more beneficial.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116775"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509853","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 of pH-adjusted NMR methodology for quantitation of caffeoylquinic acid derivatives and evaluation of their antimalignant pleural mesothelioma potential","authors":"Hau Thi Hong Bui ,&nbsp;Yoon-Jin Lee ,&nbsp;Trung Huy Ngo ,&nbsp;Punam Thapa ,&nbsp;Yun-Seo Kil ,&nbsp;Chang Yeol Lee ,&nbsp;Hyukjae Choi ,&nbsp;Kyu Joon Lee ,&nbsp;Joo-Won Nam","doi":"10.1016/j.jpba.2025.116776","DOIUrl":"10.1016/j.jpba.2025.116776","url":null,"abstract":"<div><div>Quantitative analysis of caffeoylquinic acids (CQAs) is challenging due to their structural similarities and susceptibility to isomerization and degradation. To overcome these limitations, a pH-adjusted quantitative ¹H NMR (qHNMR) method is proposed for evaluating different CQAs in the extracts using a MeOD/HEPES-<em>d</em>_<em>18</em> buffer. This method was applied to measure the concentrations of chlorogenic acid (<strong>1</strong>) and 3,5-di-CQA (<strong>4</strong>) in <em>Cuscuta japonica</em> extract, and of chlorogenic acid (<strong>1</strong>) in green coffee bean extracts. A comparison of quantum mechanics (QM)-based and non-quantum mechanics-based approaches in qHNMR has shown that the application of QM-qHNMR improves the precision and sustainability of quantitative NMR. In addition, <em>C. japonica</em> extract and its isolated CQAs (<strong>1</strong>−<strong>7</strong>) were investigated for anticancer effects against NCI-H2452 cells, showing that methyl di-CQA esters (<strong>5</strong> and <strong>7</strong>) inhibit proliferation and induce apoptosis of NCI-H2452 cells, indicating their potential as therapeutic agents for treating malignant pleural mesothelioma.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116776"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527315","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":"Machine learning enabled protein secondary structure characterization using drop-coating deposition Raman spectroscopy","authors":"Jeremy Peters ,&nbsp;Chunguang Jin ,&nbsp;Anna Luczak ,&nbsp;Brendon Lyons ,&nbsp;Ravi Kalyanaraman","doi":"10.1016/j.jpba.2025.116762","DOIUrl":"10.1016/j.jpba.2025.116762","url":null,"abstract":"<div><div>Protein structure characterization is critical for therapeutic protein drug development and production. Drop-coating deposition Raman (DCDR) spectroscopy offers rapid and cost-effective acquisition of vibrational spectral data characteristic of protein secondary structures. Amide I region (1600 −1700 cm⁻¹) and amide II region (1500–1600 cm⁻¹) of DCRD Raman spectra measured for model proteins of varying molecular size and structural distribution were first analyzed by peak fitting for their proportions of six secondary structure motifs: α-helices, 3₁₀-helices, β-sheets, turns (β-turns and γ-turns), bends, and random coil. The high spectral resolution and superior signal-to-noise of DCDR spectra made it possible to estimate all six structural motifs at accuracy comparable to X-ray crystallographic measurement. The ease of DCDR measurement was further explored by introducing machine learning algorithm to spectroscopic data analysis. Partial Least Squares (PLS) regression modeling was used as a machine learning tool to predict the protein secondary structural composition from the amide I band of model proteins. Once developed on a training sample set, the PLS model was tested by applying to a sample set that was not used previously for model development. Low prediction errors were achieved at 1.36 %, 0.78 %, 0.42 % 0.41 %, 0.81 %, and 0.52 %, respectively for the six structural component, α-Helix, β-Sheet, 3₁₀-helices, random, turns, and bends. The PLS model was further tested on an independent sample set that contains three IgG proteins. The proportion ofα-Helix, β-Sheet, 3₁₀-Helix were estimated with an error of 3.1 %, 2.3 % and 2.8 %, respectively.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116762"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}