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Qualitative and quantitative analysis of major components in Abelmoschus manihot flowers treated with different drying methods using UHPLC Q-exactive MS and HPLC-PDA 利用超高效液相色谱 Q-exactive MS 和高效液相色谱-PDA 对采用不同干燥方法处理的马齿苋花中的主要成分进行定性和定量分析。
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-11-02 DOI: 10.1016/j.jpba.2024.116558
Shaoqing Zhu , Wei Wang , Xiang Liu , Chengxue Yi , Li Li , Zhenhua Zhu , Sheng Guo , Jin-ao Duan
The flowers of Abelmoschus manihot (L.) Medic are commonly used in clinical practice in China to cure forms of chronic kidney disease. Despite a long history of traditional use, the flowers obtained by different drying technologies have never been fully chemically characterized, and the ranges of constituents between different drying methods have not been comprehensively reported. To establish a quality control and chemical characterization method, a total of 14 batches of samples corresponding to 14 postharvest treatments were studied. Seven flavonoids were quantified using a HPLC-PDA method. The method was validated in terms of linearity (r > 0.999), precision (intra- and inter-day: 0.7–1.4 %), accuracy (99.90–100.7 %), detection limit (0.34–0.46 µg/mL) and quantification limit (1.15–1.52 µg/mL). The contents of total flavonoids in manihot flowers were as follows in descending order: Infrared Drying (50.96 mg/g) > Microwave Drying (41.84 mg/g) ≈ Hot-air Drying (39.58 mg/g) ≈ Fresh (39.35 mg/g) ≈ Primary Drying (38.95 mg/g). Principal component analysis showed that samples processed with Fresh, Primary Drying, and the investigated three modern drying methods were well classified into three domains, indicating an important difference between drying methods. For the purpose of saving the flavonoids contents, infrared drying under 80–100 °C would be most acceptable. Furthermore, using UHPLC Q-Exactive Orbitrap MS data with targeted and non-targeted approaches, 28 compounds were identified in Abelmoschus manihot samples. Flavonoids were the main group of compounds found in Abelmoschus manihot flowers. The study could provide the scientific evidence for the selection and optimization of appropriate drying method for manihot flowers, and also provide the reference for the formation of generic primary drying processing technology for medicinal flowers containing flavonoids.
在中国的临床实践中,Abelmoschus manihot (L.) Medic 的花通常用于治疗各种慢性肾病。尽管传统使用历史悠久,但通过不同干燥技术获得的花朵从未进行过全面的化学表征,不同干燥方法之间的成分范围也未见全面报道。为了建立一种质量控制和化学特征描述方法,我们对 14 种采后处理方法的 14 批样品进行了研究。采用 HPLC-PDA 方法对七种黄酮类化合物进行了定量分析。该方法的线性(r > 0.999)、精密度(日内和日间:0.7-1.4 %)、准确度(99.90-100.7 %)、检出限(0.34-0.46 µg/mL)和定量限(1.15-1.52 µg/mL)均得到了验证。马尼芦花中总黄酮的含量由高到低依次为红外烘干(50.96 mg/g)>微波烘干(41.84 mg/g)≈热风烘干(39.58 mg/g)≈新鲜(39.35 mg/g)≈一次烘干(38.95 mg/g)。主成分分析表明,用新鲜、初级干燥和所研究的三种现代干燥方法处理的样品被很好地分为三个域,表明不同干燥方法之间存在重要差异。为了保留黄酮类化合物的含量,80-100 °C下的红外干燥是最可接受的。此外,利用超高效液相色谱 Q-Exactive Orbitrap MS 数据,采用靶向和非靶向方法,在马齿苋样品中鉴定出 28 种化合物。黄酮类化合物是在马齿苋花中发现的主要化合物。该研究可为选择和优化适宜的芒果花干燥方法提供科学依据,也可为形成含黄酮类化合物药用花卉的通用初级干燥加工技术提供参考。
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引用次数: 0
Analyte and probe melting temperature guided method development strategy for hybridization LC-MS/MS quantification of siRNAs 分析物和探针熔化温度引导的 siRNA 杂交 LC-MS/MS 定量方法开发策略
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-11-01 DOI: 10.1016/j.jpba.2024.116556
Zifeng Song, Angela Lu, Long Yuan
Small interfering RNA (siRNA) is a novel class of double-stranded oligonucleotide therapeutics rapidly growing in drug research and development. Accurate, sensitive, and reliable quantification of siRNA analytes in biological samples is required to study their pharmacokinetics, toxicokinetics, and biodistribution. Hybridization LC-MS/MS can achieve highly sensitive and specific bioanalysis of single-stranded oligonucleotides, e.g., antisense oligonucleotides (ASOs); however, its application for bioanalysis of siRNA or other double-stranded oligonucleotides is limited. The detailed rationale and principles for assay development are still not well understood. In this work, we systematically evaluated key steps and parameters of hybridization LC-MS/MS assays, including probes (five different types compared), hybridization procedure and temperature, elution temperature, and column temperature using patisiran, an approved siRNA drug, as the test siRNA. Based on the evaluation, a practical and efficient melting temperature (Tm) guided strategy was developed for fast and reliable method development of hybridization LC-MS/MS assays for siRNA bioanalysis. The strategy was successfully applied to siRNA-A, a test siRNA, in mouse plasma over the range of 1.00–1000 ng/mL and the resulting method has been used to support multiple mouse studies. This method-development strategy showed great value as a general approach for other siRNAs or double-stranded oligonucleotides.
小干扰 RNA(siRNA)是一类新型的双链寡核苷酸疗法,在药物研究和开发领域发展迅速。要研究 siRNA 的药代动力学、毒代动力学和生物分布,就必须对生物样本中的 siRNA 分析物进行准确、灵敏和可靠的定量。杂交 LC-MS/MS 可以对单链寡核苷酸(如反义寡核苷酸 (ASO))进行高灵敏度和特异性的生物分析,但在 siRNA 或其他双链寡核苷酸的生物分析中应用有限。人们对检测开发的详细原理和原则仍不甚了解。在这项工作中,我们系统地评估了杂交 LC-MS/MS 检测的关键步骤和参数,包括探针(比较了五种不同类型的探针)、杂交程序和温度、洗脱温度和色谱柱温度。在评估的基础上,开发了一种实用、高效的熔融温度(Tm)指导策略,用于快速、可靠地开发 siRNA 生物分析的杂交 LC-MS/MS 方法。该策略成功地应用于小鼠血浆中1.00-1000纳克/毫升范围内的 siRNA-A(一种测试 siRNA),所得到的方法已用于支持多项小鼠研究。这种方法开发策略作为一种通用方法,对其他 siRNA 或双链寡核苷酸具有重要价值。
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引用次数: 0
Multiparametric LC-MS/MS method for simultaneous determination of eleven antifungal drugs and metabolites in human plasma 同时测定人体血浆中 11 种抗真菌药物及其代谢物的多参数 LC-MS/MS 方法
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-11-01 DOI: 10.1016/j.jpba.2024.116557
Jean-Joseph Bendjilali-Sabiani , Céline Constans , Olivier Mathieu , Yoann Cazaubon
A multiparametric liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous quantification of 11 antifungal drugs and their metabolites in human plasma. This method addresses the critical need for therapeutic drug monitoring in the treatment of invasive fungal infections, which are increasingly prevalent among immunocompromised patients and those in intensive care units. The method quantifies flucytosin, fluconazole, itraconazole, hydroxy-itraconazole, posaconazole, isavuconazole, voriconazole, voriconazole-N-oxide, anidulafungin, caspofungin, and micafungin. Key challenges in method development included optimising mass spectrometer settings, chromatographic conditions, and sample preparation techniques to ensure accurate, sensitive, and specific detection. Validation of this method was conducted in accordance with the guidelines set by the USA Food and drug administration and the European Medicines Agency covering linearity, precision, accuracy, selectivity, matrix effect, and stability. The method exhibited robust performance with intra- and inter-assay precision under 10 % and average accuracy for intra- and inter-assay comparison of −2.35 % and 0.80 %, respectively. Limits of detection (0.002 to 0.110 mg/L) and a quantification range between 0.005 and 200 mg/L make this method suitable for clinical TDM applications. The ability to simultaneously analyse eleven antifungals and their metabolites within a single 5-minute run enhances its utility in clinical settings, particularly for critically ill patients who may experience significant pharmacokinetic variations. The method requires only 100 µL of plasma, demonstrating good analytical performances rendering it a valuable tool for optimising antifungal therapy and improving patient outcomes in ICU management.
本研究开发了一种多参数液相色谱-串联质谱法,用于同时定量检测人体血浆中的 11 种抗真菌药物及其代谢物。该方法满足了治疗侵袭性真菌感染的治疗药物监测的关键需求,这种感染在免疫力低下的患者和重症监护室患者中越来越普遍。该方法可定量检测氟唑醇、氟康唑、伊曲康唑、羟基伊曲康唑、泊沙康唑、异武康唑、伏立康唑、伏立康唑-N-氧化物、阿尼芬净、卡泊芬净和米卡芬净。方法开发的主要挑战包括优化质谱仪设置、色谱条件和样品制备技术,以确保准确、灵敏和特异的检测。根据美国食品药品管理局和欧洲药品管理局的指导方针,对该方法进行了验证,包括线性、精密度、准确度、选择性、基质效应和稳定性。该方法性能稳定,测定内和测定间精密度均低于 10%,测定内和测定间比较的平均准确度分别为-2.35%和 0.80%。检测限(0.002 至 0.110 mg/L)和定量范围(0.005 至 200 mg/L)使该方法适用于临床 TDM 应用。该方法能在 5 分钟的单次运行中同时分析 11 种抗真菌药物及其代谢物,这增强了它在临床环境中的实用性,尤其适用于可能出现显著药代动力学变化的重症患者。该方法只需要 100 µL 的血浆,具有良好的分析性能,因此是优化抗真菌治疗和改善重症监护室病人治疗效果的重要工具。
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引用次数: 0
Multi-attribute method analysis of therapeutic monoclonal antibodies using an automated sample preparation system 使用自动样品制备系统对治疗性单克隆抗体进行多属性方法分析。
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-30 DOI: 10.1016/j.jpba.2024.116542
Noritaka Hashii , Chihiro Obata , Miho Okada , Shota Nakamura , Keisuke Fukazawa , Shio Watanabe , Akiko Ishii-Watabe
The multi-attribute method (MAM) has attracted increased attention as an alternative strategy for evaluating structural heterogeneity using conventional separation techniques such as ion-exchange chromatography and capillary electrophoresis of therapeutic monoclonal antibodies (mAbs). One of the remaining challenges for the practical use of the MAM is reliable and robust continuous monitoring. In this study, we successfully established an automated sample preparation system as a solution to this issue. Through method optimization, we confirmed that the peptide purification step using a solid-phase extraction column, which is usually performed after the digestion step, was not mandatory and that the addition of methionine as an oxidation inhibitor was able to significantly reduce artificial oxidation. Importantly, the use of our system enabled high-precision analysis for targeted peptide monitoring without relying on the operator’s knowledge and experience with peptide mapping using liquid chromatography/mass spectrometry (LC/MS). Our system could also be useful as a platform approach for targeted peptide monitoring in MAM workflow of traditional IgG-type mAbs. Furthermore, using common samples that were prepared using the automated system, we assessed the compatibility of LC/MS system in the targeted peptide monitoring via a collaborative study with MS vendors. The results showed that there was no significant difference in the mass accuracy, repeatability of the peak retention time and variations of intermediate precision of the measurement values among the four LC/MS systems, suggesting sufficient compatibility among the LC/MS systems. MAM system using our automated sample preparation method, which had high intermediate precision, will be useful for release and stability testing as well as characterization and manufacturing process development.
多属性法(MAM)作为使用离子交换色谱法和毛细管电泳法等传统分离技术评估治疗性单克隆抗体(mAbs)结构异质性的替代策略,已引起越来越多的关注。MAM 在实际应用中仍然面临的挑战之一是可靠、稳健的连续监测。在本研究中,我们成功建立了一套自动样品制备系统,作为解决这一问题的方案。通过方法优化,我们证实使用固相萃取柱进行肽纯化步骤(通常在消化步骤后进行)并不是必须的,而且添加蛋氨酸作为氧化抑制剂能够显著减少人工氧化。重要的是,使用我们的系统可以进行高精度分析,进行有针对性的肽监测,而无需依赖操作人员使用液相色谱/质谱(LC/MS)进行肽图绘制的知识和经验。我们的系统还可作为一种平台方法,在传统 IgG 型 mAbs 的 MAM 工作流程中进行靶向肽段监测。此外,我们还利用自动系统制备的普通样品,通过与质谱供应商的合作研究,评估了 LC/MS 系统在靶向肽段监测中的兼容性。结果表明,四种 LC/MS 系统在质量准确度、峰保留时间的重复性和测量值的中间精度变化方面没有明显差异,表明 LC/MS 系统之间具有足够的兼容性。采用我们的自动样品制备方法的 MAM 系统具有较高的中间精度,将有助于释放和稳定性测试以及表征和生产工艺开发。
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引用次数: 0
New metabolites of methyltrienolone by in vitro human microsome and characterized using Liquid Chromatography/High Resolution Mass Spectrometry for doping control purposes 利用液相色谱法/高分辨质谱法鉴定体外人体微粒体中甲基三烯醇酮的新代谢物,用于兴奋剂控制。
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-28 DOI: 10.1016/j.jpba.2024.116552
Xiya Yan , Jinghua Hou , Siyu Yan , Wei Chang , Lisi Zhang , Zhanliang Wang , Sheng Yang
Methyltrienolone (17β-hydroxy-17α-methylestra-4,9,11-trien-3-one) is one of the anabolic androgenic steroids (AAS) banned by the World Anti-Doping Agency (WADA). The biotransformation of methyltrienolone is performed in vitro by human hepatocytes microsomes. Both phase I and phase II experiments are investigated. The incubation samples were extracted and injected to Liquid Chromatography/High Resolution Mass Spectrometry (LC-HRMS), or reacted with methoxylamine (29 Da added) or hydroxylamine (15 Da added), to increase the detection sensitivity. The extracted ion chromatograms of the negative control and the positive samples are compared, and 7 groups of phase I metabolites were found. The metabolic pathways including 17-epimerization, mono-hydroxylation, dihydroxylation, reduced combining mono- or dihydroxylation, as well as 18-nor which are observed and tentatively identified according to the mass spectra by high resolution mass spectrometry. The 18-nor metabolites will potentially be the long-term metabolite in doping control analysis. Its main phase II metabolites, glucuronides of metabolites are all obtained and analyzed.
甲基三烯醇酮(17β-羟基-17α-甲基雌甾-4,9,11-三烯-3-酮)是世界反兴奋剂机构(WADA)禁用的合成代谢雄性类固醇(AAS)之一。甲基三烯醇酮的生物转化是通过人体肝细胞微粒体在体外进行的。对第一阶段和第二阶段实验都进行了研究。孵育样品经提取后注入液相色谱/高分辨质谱仪(LC-HRMS),或与甲氧基胺(添加 29 Da)或羟胺(添加 15 Da)反应,以提高检测灵敏度。比较了阴性对照组和阳性样品的提取离子色谱图,发现了 7 组 I 期代谢物。根据高分辨质谱的质谱图,观察并初步确定了代谢途径,包括 17-环化、单羟基化、二羟基化、还原结合单羟基化或二羟基化,以及 18-去甲代谢物。18-nor 代谢物有可能成为兴奋剂控制分析中的长期代谢物。其主要的第二阶段代谢物、代谢物的葡糖醛酸盐均已获得并进行了分析。
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引用次数: 0
Altered gut metabolites and metabolic reprogramming involved in the pathogenesis of colitis-associated colorectal cancer and the transition of colon "inflammation to cancer" 结肠炎相关性结直肠癌发病机制和结肠 "炎症向癌症 "转变过程中的肠道代谢物变化和代谢重编程过程
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-28 DOI: 10.1016/j.jpba.2024.116553
Dunfang Wang , Lin Zhu , Haifan Liu , Xue Feng , Caijuan Zhang , Bin Liu , Tao Li , Li Liu , Hao Chang , Jingwei Sun , Lei Yang , Weipeng Yang
Colitis-associated colorectal cancer (CAC) is fatal and can develop spontaneously or as a complication of inflammatory bowel diseases. Although co-administration of azoxymethane/dextran sulfate sodium (AOM/DSS) is a classic method for CAC modeling, its limitations need to be addressed. Accordingly, we aimed to optimize the AOM/DSS model to study CAC extensively and further investigate its pathogenic mechanisms relative to microbiota and metabolism. We optimized the CAC model via a single or enhanced injection of AOM combined with different administration modes and varying DSS concentrations. Subsequently, the fecal-microbiota composition was examined using 16S RNA sequencing, and fecal-colon-metabolome profiles were evaluated via ultra-high performance liquid chromatography-mass spectrometry. Two interval injections of AOM combined with 1.5 % DSS-free drinking resulted in a high tumor formation rate, uniform tumor formation, and low mortality. Based on this model, we innovatively divided the pathogenesis of CAC into three stages, namely inflammation induction, proliferation initiation, and tumorigenesis, and examined the pathological characteristics in each stage. Gut microbial dysbiosis and metabolic alteration drove colorectal tumorigenesis by aggravating inflammation while promoting cell proliferation and carcinogenesis in mice. For the first time, we dynamically demonstrated the process of colon "inflammation to cancer" transformation and provided novel insights to clarify the role of amino acid metabolism in the formation of CAC.
结肠炎相关性结直肠癌(CAC)是一种致命疾病,可自发发病,也可作为炎症性肠病的并发症。尽管联合给药偶氮甲烷/硫酸葡聚糖钠(AOM/DSS)是建立 CAC 模型的经典方法,但其局限性仍有待解决。因此,我们旨在优化 AOM/DSS 模型,以广泛研究 CAC,并进一步研究其与微生物群和新陈代谢有关的致病机制。我们通过单次或加强注射 AOM,结合不同的给药模式和不同浓度的 DSS,对 CAC 模型进行了优化。随后,我们使用16S RNA测序法检测了粪便微生物群的组成,并通过超高效液相色谱-质谱法评估了粪便-结肠代谢组的概况。间隔两次注射 AOM 并饮用不含 1.5 % DSS 的饮料后,肿瘤形成率高、肿瘤形成均匀且死亡率低。在此模型基础上,我们创新性地将CAC的发病机制分为三个阶段,即炎症诱导、增殖启动和肿瘤发生,并研究了各阶段的病理特征。肠道微生物菌群失调和代谢改变通过加重炎症,促进小鼠细胞增殖和癌变,从而推动结直肠肿瘤的发生。我们首次动态展示了结肠 "炎症到癌症 "的转化过程,并为阐明氨基酸代谢在 CAC 形成中的作用提供了新的见解。
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引用次数: 0
Integrating 2D NMR-based metabolomics and in vitro assays to explore the potential viability of cultivated Ophiocordyceps sinensis as an alternative to the wild counterpart 整合基于二维核磁共振的代谢组学和体外试验,探索栽培的冬虫夏草作为野生冬虫夏草替代品的潜在可行性。
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-28 DOI: 10.1016/j.jpba.2024.116551
Xiu Gu , Yanping Li , Yang Li , Xiaohui Duan , Youfan Hu , Jialuo Chen , Huan Du , Jing Bai , Chengyan He , Caihong Bai , Jinlin Guo , Jiahui Yang , Kaifeng Hu
Ophiocordyceps sinensis is widely used to treat various diseases and as a health supplement. The present study comprehensively compared the metabolic differences between wild and cultivated O. sinensis through 2D 1H-13C HSQC-based metabolomics, and assessed their anti-lung cancer activity on A549 cells. To characterize the global metabolic profile, sample preparation was scrutinously optimized, and both polar (1:4 methanol-water) and non-polar (1:4 methanol-chloroform) extracts of O. sinensis were investigated. A total of 47 and 10 metabolites were identified in the polar and non-polar extracts, respectively. Principal Component Analysis (PCA) revealed greater differences between the two types of O. sinensis in the polar extracts than in the non-polar extracts. Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA) together with univariate tests captured 23 and 19 differential spectral features (with 22 and 11 of them assigned) between wild and cultivated O. sinensis in the polar and non-polar extracts, respectively. Meanwhile, the anti-lung cancer activities of both polar and non-polar extracts of wild and cultivated O. sinensis were assessed by MTS assay on A549 cells, and the sterols found in non-polar extracts, such as ergosterol, ergosterol peroxide, and 9,11-dehydroergosterol peroxide, and β-sitosterol, are the active ingredients with potential anti-lung cancer properties. In this study, we introduced a comprehensive strategy integrating 2D NMR-based metabolomics with in vitro assays for comparing the chemical composition and assessing the pharmacological activity of wild and cultivated O. sinensis. Our results provided a scientific basis for the potential viability of cultivated O. sinensis as an alternative to the wild counterpart.
冬虫夏草被广泛用于治疗各种疾病和作为保健品。本研究通过基于二维 1H-13C HSQC 的代谢组学,全面比较了野生和栽培的冬虫夏草代谢差异,并评估了它们对 A549 细胞的抗肺癌活性。为了描述总体代谢特征,对样品制备进行了严格的优化,并研究了野生中华皂苷的极性(1:4 甲醇-水)和非极性(1:4 甲醇-氯仿)提取物。在极性和非极性提取物中分别鉴定出 47 种和 10 种代谢物。主成分分析(PCA)显示,极性萃取物与非极性萃取物之间的差异更大。正交偏最小二乘法判别分析(OPLS-DA)和单变量检验分别捕获了极性和非极性提取物中野生和栽培中药材的 23 个和 19 个差异光谱特征(其中 22 个和 11 个为指定特征)。同时,利用 MTS 法对 A549 细胞进行了抗肺癌活性评估,结果表明非极性提取物中的麦角甾醇、过氧化麦角甾醇、9,11-脱氢过氧化麦角甾醇、β-谷甾醇等甾醇类化合物是具有潜在抗肺癌活性的有效成分。在这项研究中,我们引入了一种综合策略,将基于二维核磁共振的代谢组学与体外检测相结合,用于比较野生和栽培的中华皂苷的化学成分并评估其药理活性。我们的研究结果为栽培的中华皂苷替代野生中华皂苷的潜在可行性提供了科学依据。
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引用次数: 0
Nanomaterial-based magnetic solid-phase extraction in pharmaceutical and biomedical analysis 基于纳米材料的磁性固相萃取在制药和生物医学分析中的应用
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-28 DOI: 10.1016/j.jpba.2024.116543
Jingxin Hou , Cong Hu , Hanyin Li , Hongmei Liu , Yangjiayi Xiang , Gou Wu , Yan Li
Magnetic solid-phase extraction (MSPE) holds significant scientific and technological interest as a novel sample preparation method for complex samples due to its easy operation, swift separation, high adsorption efficiency, and environmental friendliness. As the core of MSPE, magnetic sorbents have captured tremendous attention in recent years. Various promising nanomaterials, such as metal-organic frameworks and covalent organic frameworks, have been synthesized and utilized as sorbents in pharmaceutical and biomedical analysis. This review intends to (1) summarize recent progress of magnetic sorbents applied in this area and discuss their advantages, disadvantages, possible interaction mechanisms with the target substances; (2) explore their innovative applications in the analysis of pharmaceuticals, proteins, peptides, nucleic acids, nucleosides, metabolites, and other disease biomarkers from 2021 to 2024; (3) present the integration of MSPE with emerging analytical technologies; and (4) discuss the current challenges and future perspectives. It is expected to provide references and insights for the development of novel magnetic sorbents and their applications in bioanalysis.
磁性固相萃取(MSPE)作为一种新型的复杂样品制备方法,以其操作简便、分离迅速、吸附效率高和环境友好等优点,在科学和技术领域备受关注。作为 MSPE 的核心,磁性吸附剂近年来备受关注。各种有前景的纳米材料,如金属有机框架和共价有机框架,已被合成并用作药物和生物医学分析中的吸附剂。本综述旨在:(1) 总结应用于该领域的磁性吸附剂的最新进展,讨论其优缺点以及与目标物质可能的相互作用机制;(2) 探讨 2021 年至 2024 年其在药物、蛋白质、肽、核酸、核苷、代谢物和其他疾病生物标记物分析中的创新应用;(3) 介绍 MSPE 与新兴分析技术的整合;以及 (4) 讨论当前挑战和未来展望。希望能为新型磁性吸附剂的开发及其在生物分析中的应用提供参考和启示。
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引用次数: 0
Metabolomics of Withania somnifera L. extracts by an integrated LC-MS and NMR approach and evaluation of their tyrosinase inhibitory activity 采用 LC-MS 和 NMR 综合方法研究睡茄提取物的代谢组学并评估其酪氨酸酶抑制活性
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-24 DOI: 10.1016/j.jpba.2024.116520
Luciana Maria Polcaro , Antonietta Cerulli , Milena Masullo , Sonia Piacente
Withania somnifera L. (Solanaceae), for over 3000 years, has been considered an essential herb in Ayurvedic medicine. The roots of W. somnifera contain metabolites mainly belonging to steroidal lactones called withanolides, which possess various pharmacological activities such as neuroprotective, cardioprotective, anti-diabetic, antioxidant and anti-inflammatory. Since the demand on the market for W. somnifera extracts is increasing, with the aim to find an ecological and environmentally friendly strategy of extraction, the roots were submitted to different extraction techniques (macerations, ultrasound-assisted extraction and solid-liquid dynamic extraction) using EtOH:H2O 50:50, 75:25, 100:0. W. somnifera extracts were investigated by an integrated LC-ESI/QExactive/MS/MS and NMR approach to obtain comprehensive metabolite profiles. Principal Component Analysis of LC-MS and NMR data revealed how the extraction method and the solvent can affect the chemical profile of the extracts. Extracts obtained by maceration exhibited the highest amount of withanolides and withanosides, while the SLDE-Naviglio EtOH extract showed the highest amount of metabolites as benzoic acid, tropane alkaloids and sarcosine, reported for their CNS activity. Moreover, based on the use of this plant in the treatment of neurological disorders, the tyrosinase inhibitory activity of all the extracts was herein tested by spectrophotometric assay, showing IC50 values in a range of 32.86–85.36 µg/ml.
3000 多年来,睡茄(茄科)一直被视为阿育吠陀医学中的一种重要草药。睡茄的根部含有主要属于甾类内酯的代谢物,称为睡茄内酯,具有多种药理活性,如神经保护、心脏保护、抗糖尿病、抗氧化和抗炎。由于市场对索曼叶草提取物的需求不断增加,为了找到一种生态和环境友好的提取策略,研究人员采用不同的提取技术(浸渍、超声辅助提取和固液动态萃取)对索曼叶草根进行了提取,提取物的EtOH:H2O比例分别为50:50、75:25和100:0。采用 LC-ESI/QExactive/MS/MS 和 NMR 综合方法对索姆尼佛拉提取物进行了研究,以获得全面的代谢物概况。LC-MS 和 NMR 数据的主成分分析揭示了提取方法和溶剂如何影响提取物的化学特征。浸渍法提取物中的山奈酚苷和山奈苷含量最高,而 SLDE-Naviglio EtOH 提取物中的苯甲酸、托烷生物碱和肌氨酸等代谢物含量最高,据报道它们具有中枢神经系统活性。此外,基于该植物在治疗神经系统疾病方面的用途,我们通过分光光度法测试了所有提取物的酪氨酸酶抑制活性,结果显示 IC50 值在 32.86-85.36 µg/ml 之间。
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引用次数: 0
Integration of four-dimensional proteomics and network pharmacology to reveal molecular mechanisms of multi-components multi-targets effects of Sini decoction on myocardial infarction 四维蛋白质组学与网络药理学相结合,揭示西尼煎剂对心肌梗死多成分多靶点作用的分子机制
IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-23 DOI: 10.1016/j.jpba.2024.116526
Xin Ding , Min Xu , Ya Zhang , Cuiping Long , Xuemei Su , Yang Zhang , Yan Qiao , Xingxing Zhang , Qian Zhou , Guangguo Tan , Jing Ma
Sini Decoction (SND) has been proven to be an effective formula to alleviate cardiac injury of myocardial infarction (MI). However, the potential mechanism of SND remains unclear. In this study, the MI rat model was established by ligating the left anterior descending coronary artery. A total of 17 SND-distributed components in heart were identified by using ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOFMS). The combination of four-dimensional (4D) proteomics and network pharmacology was employed to find the potential targets for therapeutic intervention, and molecular docking and cellular thermal shift assay (CETSA) were used to reveal the interactions between the potential targets and the potential active components distributed in heart of SND. 33 SND-effected proteins were identified by 4D proteomics, which was involved in carbon metabolism, fatty acid metabolism, valine, leucine and isoleucine degradation, tricarboxylic acid (TCA) cycle and PPAR signaling pathway. 17 potential SND-targeted direct proteins were screened by comparing SND-effected proteins generated from 4D proteomics with the MI-related proteins obtained from disease database. The potential relationships between 17 components and 17 potential SND-targeted direct proteins were established by molecular docking analysis, in which songorine, benzoylhypaconine, hypaconine, formononetin, and liquiritigenin could be bound to the surrounding amino acid residues in the binding pocket of Mtor, Parp1, Acadm, Crat, and Aldh2. Then, CETSA analysis further confirmed that songorine and benzoylhypaconine could increase the heat stability of Mtor and Parp1 in cardiac tissue lysate, respectively, which suggested that there existed direct interactions between songorine and Mtor, and benzoylhypaconine and Parp1. In summary, this work concluded that SND produced cardioprotective effects mainly through preserving energy metabolism, also demonstrated that the combination of 4D proteomics and network pharmacology was a promising tool for uncovering the molecular mechanisms of multi-components multi-targets effects of TCM.
西尼煎剂(SND)已被证明是缓解心肌梗死(MI)心脏损伤的有效配方。然而,SND 的潜在机制仍不清楚。本研究通过结扎左前降支冠状动脉建立了心肌梗死大鼠模型。利用超高效液相色谱-四极杆飞行时间质谱(UHPLC-Q-TOFMS)鉴定了心脏中的17种SND分布成分。结合四维(4D)蛋白质组学和网络药理学,找到了潜在的治疗干预靶点,并利用分子对接和细胞热转移分析(CETSA)揭示了潜在靶点与分布在SND心脏中的潜在活性成分之间的相互作用。通过4D蛋白质组学鉴定了33个受SND影响的蛋白质,它们参与了碳代谢、脂肪酸代谢、缬氨酸、亮氨酸和异亮氨酸降解、三羧酸(TCA)循环和PPAR信号通路。通过将 4D 蛋白组学中生成的 SND 影响蛋白与疾病数据库中获得的 MI 相关蛋白进行比较,筛选出了 17 个潜在的 SND 靶向直接蛋白。通过分子对接分析,确定了17种成分与17种潜在的SND靶向直接蛋白之间的潜在关系,其中松果菊碱、苯甲酰基乌头原碱、次乌头原碱、formononetin和liquiritigenin可与Mtor、Parp1、Acadm、Crat和Aldh2结合口袋中的周围氨基酸残基结合。然后,CETSA分析进一步证实,松果菊碱和苯甲酰紫堇碱可分别增加Mtor和Parp1在心脏组织裂解液中的热稳定性,这表明松果菊碱与Mtor、苯甲酰紫堇碱与Parp1之间存在直接的相互作用。综上所述,本研究认为SND主要通过保护能量代谢产生心脏保护作用,同时也证明了四维蛋白质组学与网络药理学的结合是揭示中药多成分多靶点效应分子机制的一种很有前途的工具。
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Journal of pharmaceutical and biomedical analysis
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