Journal of pharmaceutical and biomedical analysis最新文献_第9页

Integrated multi-omics approaches and network pharmacology analysis to explore the diagnosis and regulatory role of active metabolites in allergy 综合多组学方法和网络药理学分析探讨活性代谢物在过敏中的诊断和调节作用

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-08 DOI: 10.1016/j.jpba.2025.117310

Yanhua Gao , Shi Qiao , Jun Li , Ruofan An , Hongpin Hou , Lianmei Wang , Zheng Yuan , Dezhi Kong , Wei Yang

Allergy is a common immune response often triggered by foods, medicine, and pollution, significantly affecting human health. Small-molecule metabolites, as the most direct phenotypic manifestations of biological systems, can directly elucidate gene functions and molecular mechanisms. However, the roles and mechanisms of small-molecule metabolites in allergies remain incompletely understood. In this study, we established an integrated strategy combining metabolomics, network pharmacology, and proteomics to identify active metabolites and clarify their regulatory roles in bovine serum albumin (BSA)-induced allergy. 61 differential metabolites associated with allergy were identified and validated through untargeted and targeted metabolomic analyses of plasma. Combining network pharmacology and proteomics, eight activated differential metabolites affect the complement and coagulation cascades pathway through the proteins F10, F2, and PLG in BSA-induced allergy. This mechanism was further corroborated by proteomic validation of 12 related proteins altered in the complement and coagulation cascades pathway. Through correlation and receiver operating characteristic (ROC) analyses, six diagnostic biomarkers for diagnosing BSA allergy were screened, with area under the ROC curve values exceeding 0.94. Metabolites No. 39 (Glutamylphenylalanine) and 43 (hippuraldehyde sulphate) show excellent sensitivity and accuracy for diagnosing BSA-induced allergy and play a key role in modulating the complement and coagulation cascade. This study found that active metabolites could serve as critical factors and diagnostic biomarkers in allergic responses, offering novel insights into the underlying mechanisms of allergy.

过敏是一种常见的免疫反应，通常由食物、药物和污染引发，严重影响人体健康。小分子代谢物作为生物系统最直接的表型表现，可以直接阐明基因功能和分子机制。然而，小分子代谢物在过敏中的作用和机制仍不完全清楚。在本研究中，我们建立了一种结合代谢组学、网络药理学和蛋白质组学的综合策略，以鉴定活性代谢物并阐明它们在牛血清白蛋白（BSA）诱导的过敏中的调节作用。通过血浆的非靶向和靶向代谢组学分析，鉴定并验证了61种与过敏相关的差异代谢物。结合网络药理学和蛋白质组学，8种活化的差异代谢物通过蛋白F10、F2和PLG影响bsa诱导过敏的补体和凝血级联通路。通过对补体和凝血级联通路中12个相关蛋白的蛋白质组学验证，进一步证实了这一机制。通过相关性和受试者工作特征（receiver operating characteristic， ROC）分析，筛选出6个诊断牛血清白蛋白过敏的诊断性生物标志物，其ROC曲线下面积均大于0.94。代谢物No. 39 （Glutamylphenylalanine）和43 （hippuraldehyde sulfate）在诊断bsa诱导的过敏中表现出极好的敏感性和准确性，并在调节补体和凝血级联中发挥关键作用。本研究发现，活性代谢物可以作为过敏反应的关键因素和诊断生物标志物，为过敏的潜在机制提供了新的见解。

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引用次数: 0

Data fusion and multivariate analysis based on near infrared spectroscopy, electronic noses, and high resolution mass spectrometry: A synergetic approach to boost performance on the authenticity analysis of toxic herbs for Aconitum 基于近红外光谱、电子鼻和高分辨率质谱的数据融合和多变量分析：提高乌头有毒药材真实性分析性能的协同方法

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-08 DOI: 10.1016/j.jpba.2025.117312

Nian Yang , Shengyun Dai , Dongxue Wu , Rui Wu , Xiaoru Hu , Xu Zhao , Jian Zheng

Aconitum Traditional Chinese medicine, represented by Chuanwu (Aconiti Radix), Caowu (Aconiti Kusnezoffii Radix), Fuzi (Aconiti Lateralis Radix Praeparata), and their processed products ZhiChuanwu (Aconiti Radix Cocta), ZhiCaowu (Aconiti Kusnezoffii Radix Cocta), and ZhiFuzi, constitute a class of Chinese herbs characterized by a "dual nature of toxicity and efficacy." They possess pharmacological effects such as dispelling wind, eliminating dampness, and dispersing cold to relieve pain. Due to their high toxicity, the processed forms—ZhiChuanwu, ZhiCaowu, and ZhiFuzi—are predominantly used in clinical practice. ZhiFuzi is a key herb for warming Yang. ZhiChuanwu specializes in treating cold-bi syndrome (painful obstruction due to cold), and ZhiCaowu excels in pain relief. Therefore, the correct classification of these toxic Aconite materials is crucial for ensuring their quality control and appropriate clinical application. This study proposes a data fusion and multivariate analysis strategy based on near-infrared spectroscopy (NIR), electronic nose (E-nose), and high-resolution mass spectrometry (HRMS) for classifying these toxic Aconite herbs. Traditional chemometric modeling based on a single data source was found incapable of correctly classifying the three types of herbs. However, chemometrics combined with data fusion strategies enhanced the performance of the classification models. Furthermore, to identify the optimal combination of analytical modeling methods, the performance of different classification algorithms under various data training strategies was compared. PCA-Linear Discriminant Analysis (PCA-LDA) and PLS-DA, when combined with the feature-level fused dataset (NIR-E-nose-MS), yielded the best classification results. Overall, the classification strategy established in this study holds significant potential for classifying toxic Aconite medicinal materials.

以“川乌”、“草乌”、“附子”为代表的中药及其制成品“治川乌”、“治草乌”、“治附子”是一类具有“毒效双重性”的中草药。它们具有祛风、祛湿、散寒、止痛等药理作用。由于它们的高毒性，加工后的剂型——知喘乌、知草乌和知附子——主要用于临床。知府子是一种温阳的重要草药。止喘武擅长治疗感冒痹证，止草武擅长缓解疼痛。因此，对这些附子有毒物质进行正确的分类，对于保证其质量控制和临床合理应用至关重要。本研究提出了一种基于近红外光谱（NIR）、电子鼻（E-nose）和高分辨率质谱（HRMS）的数据融合和多变量分析策略，对这些附子有毒药材进行分类。传统的基于单一数据源的化学计量模型不能正确地对三种类型的草药进行分类。然而，化学计量学与数据融合策略的结合提高了分类模型的性能。此外，为了确定分析建模方法的最佳组合，比较了不同分类算法在不同数据训练策略下的性能。pca -线性判别分析（PCA-LDA）和PLS-DA与特征级融合数据集（NIR-E-nose-MS）相结合，分类效果最好。总的来说，本研究建立的分类策略对有毒附子药材的分类具有重要的潜力。

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引用次数: 0

Charge variant analysis of intact monoclonal antibody reference standards using microfluidic capillary zone electrophoresis 用微流控毛细管区带电泳分析完整单克隆抗体标准品的电荷变异

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-07 DOI: 10.1016/j.jpba.2025.117309

Noah Gould , Kendall Johnson , Anne-Lise Marie , Jared R. Auclair , Alexander R. Ivanov

Monoclonal antibodies (mAbs) are among the most common biotherapeutics, and their clinical applications and global markets have demonstrated increased growth during the last decade, which are expected to grow further due to the shift of therapeutic treatments from general to personalized approaches. As the number and complexity of mAb-based therapeutics grow, there is an urgent need for highly efficient and high-throughput analytical techniques to characterize them and ensure their safety, potency, stability, and efficacy both from a development and regulatory standpoints. A key component in ensuring the methods used for the characterization of antibody-based therapeutics can provide accurate information, with the potential to influence important pipeline decisions, is the availability of diverse commercially available reference standards with well-known and catalogued properties that can serve as benchmarks and validation for the quality assurance of biotherapeutics in development. In this study, we developed and optimized a microfluidic capillary zone electrophoresis electrospray ionization-mass spectrometry (CZE-ESI-MS) method, utilizing the high-mass range MS data acquisition, to characterize three intact mAb reference standards and their proteoforms under near-native conditions using short analysis times (<15 min), low sample amounts (500 picograms), and minimal sample preparation. The developed CZE-MS method identified a number of modifications at the intact mAb level, including relatively high abundance C-terminal lysine clipping (>95 %) and N-linked glycosylation (>99 %), as well as lower abundance modifications, such as deamidation (∼3–7 %) and N-glycan sialylation (<1.5 %). The developed technique is uniquely suited for thorough and reproducible characterization of mAb-based biotherapeutics in industrial settings.

单克隆抗体（mab）是最常见的生物治疗药物之一，其临床应用和全球市场在过去十年中表现出增长，由于治疗方法从一般到个性化的转变，预计将进一步增长。随着以单克隆抗体为基础的治疗方法的数量和复杂性的增加，迫切需要高效和高通量的分析技术来表征它们，并从开发和监管的角度确保它们的安全性、效力、稳定性和有效性。确保用于鉴定基于抗体的治疗方法的方法能够提供准确的信息，并有可能影响重要的管线决定的一个关键组成部分，是提供各种商业上可获得的参考标准，这些标准具有众所周知的和已编目的特性，可作为开发中的生物治疗药物质量保证的基准和验证。在这项研究中，我们开发并优化了一种微流体毛细管区带电泳电喷雾电离质谱（cse - esi -MS）方法，利用高质量范围的MS数据采集，在接近自然条件下，使用短分析时间（<15 min）、低样本量（500 picogram）和最少的样品制备，表征了三种完整的单抗参考标准物及其蛋白形态。开发的cce - ms方法在完整的mAb水平上鉴定了许多修饰，包括相对高丰度的c端赖氨酸剪切（>95 %）和n -链糖基化（>99 %），以及低丰度的修饰，如脱酰胺（~ 3-7 %）和n -糖基化（<1.5 %）。开发的技术是唯一适合于彻底和可重复的表征单克隆抗体为基础的生物疗法在工业环境。

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引用次数: 0

Integrated proteomics and metabolomics reveal NF-κB pathway modulation by Xiangpi Shengji ointment and Qingre Zaoshi compound in anal fistula wound healing 结合蛋白质组学和代谢组学分析，香脾生脂膏和清热燥湿复方对肛瘘创面愈合的NF-κB通路调节作用

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-05 DOI: 10.1016/j.jpba.2025.117306

Mintao Jian , Jiaqing Xiong , Kui Li

This study investigates the molecular mechanisms by which Xiangpi Shengji Ointment (XPSJO), combined with a heat-clearing and dampness-drying compound, accelerates postoperative wound healing in an anal fistula (AF) rat model, employing integrated proteomic and metabolomic approaches. An AF rat model was established and treated topically with the herbal combination. Wound healing progression was monitored postoperatively. Histopathological assessment (hematoxylin and eosin [H&E] staining), enzyme-linked immunosorbent assay [ELISA] quantification of inflammatory markers (e.g., tumor necrosis factor-alpha [TNF-α], interleukin [IL]-6) and vascular endothelial growth factor [VEGF], and proteomic (liquid chromatography-tandem mass spectrometry [LC-MS/MS]) and metabolomic (gas chromatography-mass spectrometry [GC-MS]) analyses identified critical pathways and biomolecules involved. In vitro, lipopolysaccharide (LPS)-stimulated THP-1 macrophages and human umbilical vein endothelial cells (HUVECs) were co-treated with XPSJO and Coptis chinensis-Phellodendron amurense Rupr. Extract (CGE). Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway modulation was examined via reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blot, and functional assays (cell counting kit [CCK]-8, flow cytometry, scratch wound, tube formation). Lentiviral NF-κB overexpression validated mechanistic specificity in vitro and in vivo. Results showed that XPSJO combined with the compound accelerated AF wound closuresuppressed NF-κB signaling pathway activation (evidenced by downregulation of pathway-associated proteins in proteomics and reduced p65 phosphorylation), decreased pro-inflammatory cytokine secretion, and enhanced VEGF expression. Critically, it promoted M2 macrophage polarization (increased Arg-1/CD206, decreased iNOS/CD80) and stimulated HUVEC migration/angiogenesis. Multi-omics analysis further confirmed NF-κB, TNF, and IL-17 pathways as central therapeutic targets. The study concludes that XPSJO combined with a heat-clearing and dampness-drying compound promotes AF healing through pharmacological inhibition of the NF-κB pathway, macrophage reprogramming, and tissue remodeling.

本研究采用蛋白质组学和代谢组学相结合的方法，探讨香皮生脂软膏（XPSJO）联合清热湿燥化合物加速肛瘘（AF）大鼠术后创面愈合的分子机制。建立房颤大鼠模型，采用中药复方局部治疗。术后监测伤口愈合进展。组织病理学评估（苏木精和伊红[H&；E]染色），酶联免疫吸附试验[ELISA]定量炎症标志物（如肿瘤坏死因子-α [TNF-α]，白细胞介素[IL]-6）和血管内皮生长因子[VEGF]，蛋白质组学（液相色谱-串联质谱[LC-MS/MS]）和代谢组学（气相色谱-质谱[GC-MS]）分析确定了关键途径和参与的生物分子。在体外，用XPSJO和黄连-黄柏联合作用于脂多糖（LPS）刺激的THP-1巨噬细胞和人脐静脉内皮细胞（HUVECs）。提取(CGE)。通过逆转录定量聚合酶链反应（RT-qPCR）、Western blot和功能检测（细胞计数试剂盒[CCK]-8、流式细胞术、划伤、成管）检测活化B细胞核因子κB轻链增强子（NF-κB）通路的调节。慢病毒NF-κB过表达在体外和体内验证了机制特异性。结果显示，XPSJO联合复方加速心房纤颤创面闭合，抑制NF-κB信号通路激活（蛋白质组学中通路相关蛋白下调，p65磷酸化降低），减少促炎细胞因子分泌，增强VEGF表达。关键是，它促进M2巨噬细胞极化（增加Arg-1/CD206，降低iNOS/CD80），刺激HUVEC迁移/血管生成。多组学分析进一步证实NF-κB、TNF和IL-17通路是主要的治疗靶点。本研究认为XPSJO联合清热湿燥化合物通过抑制NF-κB通路、巨噬细胞重编程和组织重塑促进AF愈合。

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引用次数: 0

Chemical stability of esomeprazole in biorelevant bicarbonate buffer and an exploratory study for specific degradation products 埃索美拉唑在生物相关碳酸氢盐缓冲液中的化学稳定性及特定降解产物的探索性研究。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-05 DOI: 10.1016/j.jpba.2025.117305

Ayaka Takasusuki , Takahiro Takayama , Koichi Inoue , Kiyohiko Sugano

This study aimed to evaluate the impact of biorelevant bicarbonate buffer solutions (BCB) on drug degradation relative to phosphate buffer solutions (PPB). Esomeprazole (ESM) was selected as a model compound. Stability studies were conducted at 37 °C using 10 mM BCB and 31 mM PPB (both at pH 5.5; buffer capacity (β) = 4.0 mM/pH; ionic strength (I) = 0.14 M). Degradation products were quantified at 4 time points by HPLC. No significant difference was observed in the degradation rate constant of ESM between BCB and PPB. However, LC–MS analysis revealed a distinct tendency that two degradation products with m/z values of 330 and 298 were more abundant in BCB than in PPB. The putative structures of these degradation products matched the previously reported compounds. These findings suggest that buffer species can influence degradation pathways even under equivalent β and I. Consequently, the use of biorelevant buffer systems is essential for accurate assessments of degradation products.

本研究旨在评价生物相关碳酸氢盐缓冲液（BCB）相对于磷酸盐缓冲液（PPB）对药物降解的影响。选择埃索美拉唑（ESM）作为模型化合物。稳定性研究在37°C下进行，使用10 mM BCB和31 mM PPB (pH均为5.5，缓冲容量(β) = 4.0 mM/pH；离子强度(I) = 0.14 M)。用高效液相色谱法测定4个时间点的降解产物。BCB和PPB对ESM的降解速率常数无显著差异。但LC-MS分析显示，m/z值为330和298的两种降解产物在BCB中的含量明显高于PPB。这些降解产物的推定结构与先前报道的化合物相匹配。这些发现表明，即使在相同的β和i下，缓冲物种也可以影响降解途径。因此，使用生物相关的缓冲系统对于准确评估降解产物至关重要。

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引用次数: 0

Preparation of Novel magnetic boronic acid-affinity material and investigation of adsorption performance toward ovalbumin 新型磁性硼酸亲和材料的制备及对卵清蛋白吸附性能的研究。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-05 DOI: 10.1016/j.jpba.2025.117304

Xiao Dong , Jianliang Li , Yixuan Ni , Shuna Bao , Kexin Fan , Jingjing Xu , Qiao Deng , Xu Wang

Glycoproteins play a crucial role in various biological processes, including protein folding, information transmission, nerve conduction, and molecular recognition. In this study, a novel magnetic phenylboronic acid-functionalized covalent organic framework core-shell material (Fe₃O₄@BBCOF-BA) was successfully synthesized for the highly selective enrichment of glycoproteins from complex biological samples. Systematic optimization of experimental conditions revealed that under weakly alkaline conditions at pH 8.5, the material specifically recognized glycoproteins through boronate ester bonds. The maximum adsorption capacity for ovalbumin (OVA) reached as high as 469.45 mg/g, representing a two-to threefold improvement over traditional magnetic boronate affinity materials. Adsorption kinetics indicated that equilibrium was reached within 120 min, and rapid separation was achieved within 30 s utilizing the superparamagnetic property. Mild elution was performed using an ethanol-formic acid (90:10, v/v) solution, achieving an OVA recovery rate exceeding 90 %. Selectivity experiments confirmed that the material exhibited significantly higher adsorption of glycoproteins (OVA, HRP) than non-glycoproteins (BSA, Lys). Practical application evaluation demonstrated efficient enrichment of trace glycoproteins from 500-fold diluted egg white and complex protein mixtures. SDS-PAGE analysis confirmed low non-specific adsorption, underscoring its strong potential for application in the pretreatment of complex biological samples.

糖蛋白在蛋白质折叠、信息传递、神经传导和分子识别等多种生物过程中起着至关重要的作用。本研究成功合成了一种新型磁性苯硼酸功能化共价有机骨架核壳材料（Fe3O4@BBCOF-BA），用于高选择性富集复杂生物样品中的糖蛋白。系统优化实验条件发现，在pH 8.5弱碱性条件下，材料通过硼酸酯键特异性识别糖蛋白。对卵清蛋白（OVA）的最大吸附量高达469.45 mg/g，比传统的磁性硼离子亲和材料提高了2 - 3倍。吸附动力学表明，吸附在120 min内达到平衡，利用超顺磁性能在30 s内实现快速分离。使用乙醇-甲酸（90:10,v/v）溶液进行温和洗脱，OVA回收率超过90% %。选择性实验证实，该材料对糖蛋白（OVA， HRP）的吸附明显高于非糖蛋白（BSA, Lys）。实际应用评价表明，从500倍稀释的蛋清和复杂蛋白混合物中有效富集微量糖蛋白。SDS-PAGE分析证实了低非特异性吸附，强调了其在复杂生物样品预处理方面的强大应用潜力。

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引用次数: 0

Electrochemical multi-drug sensors: Current advances, challenges, and future perspectives 电化学多药物传感器：当前进展、挑战和未来展望

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-04 DOI: 10.1016/j.jpba.2025.117303

Vildan Sanko , Mustafa Ali Güngör , Filiz Kuralay

The rapid growth of the pharmaceutical industry has highlighted the importance of controlling both the quality control process and appropriate dosing in drug purchasing to a critical level. In addition to this, pharmaceutical pollutants can harm the environment, especially water resources, creating a need for low-cost, sensitive, selective, and portable technologies to monitor personal health and environmental applications. Because monitoring more than one drug simultaneously provides convenience in terms of diagnosis and treatment, it increases the drug monitoring capacity and features. Therefore, various electrochemical methods and micro/nanomaterials integration have been employed to achieve well-separated peaks for drug molecules and enhance sensitivity. However, detecting more than one drug using an electrochemical method remains a challenging research topic. This review discusses electrochemical multi-drug (bio)sensing techniques, focusing on future trends. For this purpose, drugs were categorized as anticancer, anti-inflammatory, antidepressant, antibacterial, antiviral, and antifungal according to the area they act on. Moreover, integration of herbal drugs, which are highlighted by their rising therapeutic importance and intricate phytochemical compositions, has found place in electrochemical sensing. Current electrochemical (bio)sensors that detect multiple drugs for each group were discussed. In addition, mixed-type multi-drugs containing different drug groups were also discussed in a separate section.

制药工业的快速发展突出了在药品采购中控制质量控制过程和适当剂量的重要性。除此之外，药物污染物可能危害环境，特别是水资源，因此需要低成本、敏感、选择性和便携式技术来监测个人健康和环境应用。由于同时监测多种药物在诊断和治疗方面提供了便利，增加了药物监测的能力和特点。因此，利用各种电化学方法和微纳米材料集成来实现药物分子的良好分离峰，提高灵敏度。然而，使用电化学方法检测多种药物仍然是一个具有挑战性的研究课题。本文综述了电化学多药物（生物）传感技术，并对其发展趋势进行了展望。为此，根据药物作用的区域，将药物分为抗癌、抗炎、抗抑郁、抗菌、抗病毒和抗真菌。此外，草药的整合，由于其日益增长的治疗重要性和复杂的植物化学成分，已经在电化学传感中找到了一席之地。讨论了目前电化学（生物）传感器检测每组多种药物的方法。此外，包含不同药物组的混合型多药也在另一节进行了讨论。

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引用次数: 0

A universal LC-HRMS workflow integrating targeted and untargeted strategies for rapid and comprehensive metabolite profiling of oligonucleotide-based therapeutics 整合靶向和非靶向策略的通用LC-HRMS工作流程，用于快速和全面的基于寡核苷酸的治疗的代谢物分析。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-04 DOI: 10.1016/j.jpba.2025.117298

Yuqing Zhao , Yue Yang , Mingshe Zhu , Chongzhuang Tang

Metabolite profiling and identification of oligonucleotide-based therapeutics (OBTs) is important in drug discovery and development. Conventional liquid chromatography-high resolution mass spectrometry (LC-HRMS) methods (using ProMass, BioPharma Finder) are effective for targeted analysis of predictable nuclease-derived metabolites but often miss unexpected non-nuclease-mediated ones. This study aimed to develop a universal analytical workflow for comprehensive metabolite profiling of multiple classes OBTs by integrating BioPharma Finder for targeted identification with a Background Subtraction Filter (BSF) for untargeted detection of unexpected biotransformation products. Inclisiran, a GalNAc-conjugated siRNA, was used as a model compound. Samples incubated with rat, monkey, and human liver S9 fractions were analyzed by the LC-HRMS workflow. As a result, a total of 25 (22 predicted +3 unpredicted) inclisiran metabolites were identified across the three species. BioPharma Finder enabled rapid and sensitive identification of 22 predicted metabolites, supported by automated fragment ion assignment. The BSF data processing revealed 22 metabolites not present in control samples, including SS-1GalNAc, SS-2GalNAc, and SS-3GalNAc, which were missed by BioPharma Finder and likely formed via β-N-acetylglucosaminidase-mediated biotransformation. The structures of BSF-detected metabolites were confirmed by comparison with BioPharma Finder-identified metabolites or characterized through manual MS/MS interpretation. Product ion spectra of these metabolites showed weak or absent diagnostic ion (m/z 94.936), a characteristic fragment of phosphorothioate-containing oligonucleotides, suggesting that the product ion filtering of this ion exhibits limited utility in discovering inclisiran metabolites. Overall, the integrated LC-HRMS workflow shows strong potential as a universal platform for biotransformation studies of OBTs.

代谢物分析和鉴定基于寡核苷酸的疗法（OBTs）在药物发现和开发中具有重要意义。传统的液相色谱-高分辨率质谱（LC-HRMS）方法（使用ProMass， BioPharma Finder）对于可预测的核酸酶衍生代谢物的靶向分析是有效的，但经常错过意想不到的非核酸酶介导的代谢物。本研究旨在开发一种通用的分析工作流程，用于多种obt的综合代谢物分析，方法是将用于靶向鉴定的BioPharma Finder与用于非预期生物转化产物非靶向检测的背景减除过滤器（BSF）集成在一起。Inclisiran是一种GalNAc-conjugated siRNA，被用作模型化合物。用大鼠、猴和人肝脏S9部分孵育的样品采用LC-HRMS工作流程进行分析。结果，在三个物种中共鉴定出25个（22个预测的+3个未预测的）inclisiran代谢物。BioPharma Finder能够快速、灵敏地识别22种预测的代谢物，并支持自动片段离子分配。BSF数据处理显示22种代谢物在对照样品中不存在，包括SS-1GalNAc、SS-2GalNAc和SS-3GalNAc，这些代谢物可能是通过β- n -乙酰氨基葡萄糖酶介导的生物转化形成的。bsf检测到的代谢物的结构通过与BioPharma finder鉴定的代谢物进行比较来确认，或者通过手动MS/MS解释来表征。这些代谢物的产物离子谱显示弱或缺失诊断离子（m/z为94.936），这是含硫代磷酸酯寡核苷酸的特征片段，表明该离子的产物离子过滤在发现含硫代磷酸酯代谢物方面的应用有限。总的来说，LC-HRMS集成工作流显示出强大的潜力，作为obt生物转化研究的通用平台。

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引用次数: 0

Simultaneous identification of 50 illegal adulterants in dietary supplements using high-performance liquid chromatography–single quadrupole mass spectrometry 高效液相色谱-单四极杆质谱法同时鉴定膳食补充剂中50种非法掺假物质。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-03 DOI: 10.1016/j.jpba.2025.117300

Takayuki Sakai , Misa Tanaka , Yuki Azuma, Yusuke Sakamoto, Takaomi Tagami, Akiko Asada, Takahiro Doi

Health hazards associated with the ingestion of dietary supplements containing illegal adulterants have been reported. High-performance liquid chromatographysingle quadrupole mass spectrometry (LC-MS) is useful for identification of illegal adulterants in dietary supplements because of its moderate sensitivity and wide accessibility. Previously, we reported a method for the simultaneous identification of 18 illegal adulterants in dietary supplements using LC-MS. Considering numerous analogs of pharmaceutical ingredients recently identified, developing a more convenient screening method to identify a broader range of such compounds is essential. In this study, we introduced another LC-MS-based method to simultaneously identify 50 illegal adulterants in dietary supplements, including newly identified compounds such as descarbonsildenafil and N-phenylpropoxyphenylcarbodenafil. The proposed method achieved clear separation of the targeted peaks in their extracted ion chromatograms. All coefficients of determination (r²) exceeded 0.9794, with most compounds showing good linearity above 0.99. Recovery tests confirmed the specificity of detection for each compound, with recovery rates ranging from 74.9 % to 127.7 % and relative standard deviations below 21.3 %. Validation results demonstrated the method’s suitability for identifying all 50 targeted compounds and quantifying 32 compounds with acceptable accuracy and precision. The method also successfully detected illegal adulterants in positive samples. Thus, the proposed method may be useful for identifying—and partially quantifying—illegal adulterants present in dietary supplements.

据报道，与摄入含有非法掺假物的膳食补充剂有关的健康危害。高效液相色谱-单四极杆质谱法（LC-MS）具有灵敏度中等、可及性广的优点，可用于膳食补充剂中非法掺杂物的鉴别。此前，我们报道了一种使用LC-MS同时鉴定膳食补充剂中18种非法掺杂物的方法。考虑到最近发现的许多药物成分类似物，开发一种更方便的筛选方法来识别更广泛的此类化合物是必不可少的。在这项研究中，我们引入了另一种基于lc - ms的方法来同时鉴定膳食补充剂中的50种非法掺杂物，包括新鉴定的化合物，如去碳化西地那非和n -苯基丙氧苯基卡地那非。所提出的方法在其提取的离子色谱中实现了目标峰的清晰分离。所有测定系数（r2）均大于0.9794，大部分化合物在0.99以上线性良好。回收率为74.9 % ~ 127.7 %，相对标准偏差小于21.3 %。验证结果表明，该方法可用于鉴定所有50种目标化合物，并对32种化合物进行定量，准确度和精密度可接受。该方法还成功地检测出阳性样品中的非法掺杂物。因此，提出的方法可能有助于识别和部分量化存在于膳食补充剂中的非法掺假。

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引用次数: 0

Development of a reliable and convenient lateral flow immunoassay for dihydromyricetin detection in Ampelopsis grossedentata samples 建立一种可靠、便捷的横向流动免疫分析法检测长叶葡萄样品中的二氢杨梅素。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-03 DOI: 10.1016/j.jpba.2025.117299

Xueyan Cui , Zhen Cao , Hailong Yu , Hui Li , Ying Ying , Ting Luo , Jing Wang

Dihydromyricetin, a flavonoid abundant in Ampelopsis grossedentata distributed across southern China, exhibits diverse pharmacological activities. In this study, a precise, sensitive, rapid, and reliable lateral flow immunoassay (LFIA) was developed for dihydromyricetin quantification in Ampelopsis grossedentata. A dihydromyricetin hapten was synthesized by introducing a carboxylic acid spacer via a four-carbon chain at the 3-position. A hybridoma cell line (2H3) secreting a highly specific monoclonal antibody (mAb) against dihydromyricetin was established. Dihydromyricetin demonstrated excellent stability in most organic solvents and acidic conditions. The LFIA exhibited a sensitivity (IC₅₀) of 33.61 ng/mL and a linear range of 4.54–281.83 ng/mL. The recovery rates of dihydromyricetin were in the range of 70.83–96.17 %. These findings indicate that LFIA, leveraging the high specificity of the dihydromyricetin mAb, offers a simple, rapid, and accurate method for dihydromyricetin analysis in Ampelopsis grossedentata.

二氢杨梅素是分布在中国南方的藤属葡萄中富含的一种类黄酮，具有多种药理活性。本研究建立了一种精确、灵敏、快速、可靠的横向流动免疫分析法（LFIA），用于测定长齿葡萄中二氢杨梅素的含量。通过在3位的四碳链上引入羧酸间隔物，合成了二氢杨梅素半抗原。建立了一株分泌抗二氢杨梅素高特异性单克隆抗体（mAb）的杂交瘤细胞株（2H3）。二氢杨梅素在大多数有机溶剂和酸性条件下表现出良好的稳定性。LFIA的灵敏度（IC₅₀）为33.61 ng/mL，线性范围为4.54-281.83 ng/mL。二氢杨梅素的回收率为70.83 ~ 96.17 %。这些结果表明，LFIA利用双氢杨梅素单抗的高特异性，提供了一种简单、快速、准确的双氢杨梅素分析方法。

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