Journal of pharmaceutical and biomedical analysis最新文献_第7页

In-depth O-glycosylation characterization and comparison of commercially available etanercept biosimilars by using Electron Activated Dissociation 利用电子激活解离法对市售依那西普生物类似药进行o -糖基化表征和比较

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-24 DOI: 10.1016/j.jpba.2025.117326

Wenhong Fan , Xiang Li , Wentao Wang , Tie Gao , Ji Luo , Zoe Zhang , Bingjie Liu , Chenggang Liang , Hongxu Chen

Glycosylation is one of the important post-translational modifications in protein drugs expressed by eukaryotic cells. It is closely related to the conformation, stability, and solubility of therapeutic proteins and holds vital biological significance. Etanercept contains 26 O-glycosylation sites with high glycosylation heterogeneity, which makes the analysis of O-glycosylation highly challenging. In this paper, using liquid chromatography-mass spectrometry (LC-MS), 11 O-glycosylation on the single chain of etanercept were first identified at the intact protein level, with Core1 and Core2 O-glycans. Second, 11 O-glycosylation sites were identified with the electron activation dissociation (EAD) fragmentation mode at the glycopeptide level, and their glycans were comprehensively characterized. The major O-glycan structures were Core1 and Core2, which were consistent with those observed at the intact protein level. Third, the etanercept innovator (Y1) and six biosimilars or follow-on products (Y2-Y7) from different manufacturers were also compared. Compared with the etanercept innovator, the results showed that etanercept biosimilars or follow-on products from different manufacturers exhibited pronounced differences in O-glycosylation modification due to variation in production processes. Based on a comprehensive assessment of consistency in deconvoluted spectra and the glycosylation ratios, etanercept biosimilar Y2 was the most similar to the innovator Y1. In-depth studies of etanercept’s O-glycosylation can effectively be applied to the etanercept innovator's comparative analysis and biosimilars. Furthermore, the LC-MS method combined with the EAD fragmentation technique can offer detailed and precise data support for the comprehensive quality control of complex glycoprotein drugs and the optimization of production processes.

糖基化是真核细胞表达蛋白药物的重要翻译后修饰之一。它与治疗蛋白的构象、稳定性和溶解度密切相关，具有重要的生物学意义。依那西普含有26个o -糖基化位点，具有很高的糖基化异质性，这使得o -糖基化分析具有很高的挑战性。本文采用液相色谱-质谱（LC-MS）技术，首次在完整蛋白水平上鉴定了依那西普单链上的11个o -糖基化，分别为Core1和Core2 o -聚糖。其次，采用电子激活解离（EAD）裂解模式在糖肽水平上鉴定了11个o -糖基化位点，并对其聚糖进行了全面表征。主要的o -聚糖结构是Core1和Core2，这与在完整蛋白水平上观察到的一致。第三，比较了依那西普创新产品（Y1）和6个不同厂家的生物仿制药或后续产品（Y2-Y7）。结果表明，与依那西普创新产品相比，不同厂家的依那西普生物仿制药或后续产品由于生产工艺的差异，在o -糖基化修饰方面存在显著差异。基于反卷积光谱一致性和糖基化比率的综合评估，依那西普生物仿制药Y2与创新药物Y1最相似。对依那西普o糖基化的深入研究可以有效地应用于依那西普创新者的比较分析和生物仿制药。此外，LC-MS结合EAD碎片化技术可为复杂糖蛋白药物的综合质量控制和生产工艺优化提供详细、精确的数据支持。

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引用次数: 0

Device does matter! Comparison of PEth 16:0/18:1 and PEth 16:0/18:2 blood concentrations determined from three dried blood spot sampling devices 设备很重要！三种干血点取样装置测定的PEth 16:0/18:1和PEth 16:0/18:2血液浓度的比较

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-23 DOI: 10.1016/j.jpba.2025.117325

Jeremai Hose , Paul Stach , Sara Reda , Martin Juebner , Mario Thevis , Hilke Andresen-Streichert

The biomarker phosphatidylethanol (PEth) can be determined in whole blood to distinguish between alcohol abstinence and alcohol consumption. Routine analysis is commonly performed using dried blood spots (DBS), as PEth is considered stable for extended time periods in this matrix. However, the coexistence of many DBS devices, coupled with insufficient data regarding their comparability of results, pose a major challenge in forensic and clinical practice. We therefore analysed and compared blood samples using three DBS devices commonly employed for PEth testing (Whatman™ filter paper, Mitra® and Capitainer®B Vanadate). PEth 16:0/18:1 and 16:0/18:2 were extracted from DBS using a water/propan-2-ol mixture and subsequent liquid-liquid-extraction using n-hexane. Analysis was carried out via LC-MS/MS. Ethanol and acetaldehyde concentrations were determined in each blood sample via GC-FID. A total of 100 blood samples were analysed, of which 76 were found to be positive for PEth. These included 24 cases with PEth concentrations < 20 ng/mL, 43 cases with concentrations ≥ 20 ng/mL to < 200 ng/mL and nine cases with concentrations ≥ 200 ng/mL. The Mitra® and Capitainer®B Vanadate systems resulted, on average, in higher PEth concentrations compared to Whatman™ filter paper. The percentage deviations ranged from −8–116 % for PEth 16:0/18:1 and from −14–264 % for PEth 16:0/18:2. Ethanol and acetaldehyde above physiological concentrations (1.42 g/L and 0.727 mg/L, respectively) were observed in one case only. While DBS are a reliable method for PEth analysis, the choice of device can influence results and potentially impact interpretations of alcohol use and abstinence.

生物标志物磷脂酰乙醇（PEth）可以在全血中测定，以区分戒酒和饮酒。常规分析通常使用干血点（DBS）进行，因为在该矩阵中，PEth被认为在较长时间内是稳定的。然而，许多DBS装置的共存，加上关于其结果可比性的数据不足，在法医和临床实践中构成了重大挑战。因此，我们使用三种常用于PEth检测的DBS设备（Whatman™滤纸、Mitra®和Capitainer®B Vanadate）分析和比较血液样本。用水/丙烷-2-醇混合物从DBS中提取PEth 16:0/18:1和16:0/18:2，然后用正己烷进行液-液萃取。采用LC-MS/MS进行分析。通过气相色谱- fid测定每个血液样本中的乙醇和乙醛浓度。总共对100份血液样本进行了分析，其中76份被发现对PEth呈阳性反应。其中24例患者的PEth浓度为<； 20 ng/mL， 43例浓度≥ 20 ng/mL至<； 200 ng/mL， 9例浓度≥ 200 ng/mL。与Whatman™滤纸相比，Mitra®和Capitainer®B钒酸盐系统平均产生更高的PEth浓度。PEth 16:0/18:1的百分比偏差为−8-116 %，PEth 16:0/18:2的百分比偏差为−14-264 %。乙醇和乙醛高于生理浓度（分别为1.42 g/L和0.727 mg/L）仅在一个病例中观察到。虽然DBS是一种可靠的PEth分析方法，但设备的选择会影响结果，并可能影响对酒精使用和戒酒的解释。

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引用次数: 0

Efficient analytical set-up for the monitoring of albumin adduction on Cysteine 34 exposed to mustard agents with optimized digestion and on-line SPE-LC-MS analysis 利用优化的消化和在线SPE-LC-MS分析，建立了高效的分析装置，用于监测暴露于芥菜剂的半胱氨酸34上的白蛋白内聚

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-23 DOI: 10.1016/j.jpba.2025.117324

Lorenzo Avigo , Marine Charpin , Audrey Combès , Charlotte Desoubries , Christine Albaret , Anne Bossée , Emmanuel Joubert , Valérie Pichon

Adduction of mustard agents such as sulfur mustard (SM), sesquimustard (Q), and nitrogen mustards (HN-1, HN-2, HN-3) on nucleophilic sites of proteins, particularly Cysteine34 of human serum albumin (HSA), can be monitored in plasma to confirm retrospective exposure. The main purpose of this work is to optimize an on-line solid phase extraction (SPE) of HSA tripeptides containing adducted Cys34, coupled to LC-MS. To improve sensitivity of these biomarkers’ detection and data robustness the impact of sample preparation steps such as protein precipitation and reduction with dithiothreitol (DTT) prior to digestion with proteinase K (Prot. K) were systematically evaluated. Neither improved digestion efficiency, and at low incubation levels of SM, precipitation even reduced signal intensity. Removing these steps decreased variability and consumable use. Washing conditions were optimized to favour the retention of adducted tripeptides on the SPE sorbent while removing more polar digest compounds. Optimal extraction conditions defined using spiked digests involved washing with 7 % (v/v) acetonitrile (ACN) before direct on-line transfer to LC-MS. The optimized method was then applied, as a proof of concept, to plasma incubated with SM and HN-1 at two concentration levels in order to demonstrate its applicability to real biological samples. Limits of quantification (LOQs), determined within this proof-of-concept framework, were established at 10 ng/mL for SM and 40 ng/mL for HN-1 in in vitro adducted plasma.

芥菜制剂，如硫芥菜（SM）、倍半芥菜(Q)和氮芥菜（HN-1、HN-2、HN-3）在蛋白质亲核位点上的内聚，特别是人血清白蛋白（HSA）的半胱氨酸34，可以在血浆中监测，以确认回顾性暴露。本工作的主要目的是优化含有内合Cys34的HSA三肽的在线固相萃取（SPE），并偶联LC-MS。为了提高这些生物标志物检测的敏感性和数据的稳健性，研究了样品制备步骤的影响，如在蛋白酶K （Prot）消化之前用二硫苏糖醇（DTT）沉淀和还原蛋白质。K)进行系统评价。两者都不能提高消化效率，而且在低SM孵育水平下，沉淀甚至降低了信号强度。删除这些步骤减少了可变性和消耗品的使用。优化了洗涤条件，有利于在SPE吸附剂上保留内合的三肽，同时去除更多的极性消化化合物。用加标消化物确定的最佳提取条件为：用7 % （v/v）乙腈（ACN）洗涤，然后直接在线转移到LC-MS。然后将优化后的方法应用于SM和HN-1在两种浓度水平下的血浆中，作为概念验证，以证明其对实际生物样品的适用性。在此概念验证框架内确定的定量限（loq）确定为体外内合血浆中SM为10 ng/mL， HN-1为40 ng/mL。

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引用次数: 0

Multidimensional development of gut-on-a-chip technology: from fabrication processes, models, gut microbiome to gut-organ axis 芯片肠道技术的多维发展：从制造工艺、模型、肠道微生物组到肠道器官轴。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-22 DOI: 10.1016/j.jpba.2025.117322

Danni Chai , Quan Wang , Qian Yong , Chunyan Chen , Yumeng Liao , Ruru Pan , Yujia He , Keshu Sun , Boshi Liu , Rui Liu , Zheng Li

Gut-on-a-chip (GoC) platforms integrate microfluidics and 3D culture to replicate the intestinal microenvironment, offering physiologically relevant alternatives to traditional models. Coupled with multi-organ chips (e.g., gut-brain/gut-liver axes), they unveil microbiome-regulated systemic crosstalk via metabolite signaling—a key yet unresolved mechanism. This review highlights multidimensional advances in organ-on-chip (OoC) technologies for intestinal research, covering fabrication methods (e.g., soft lithography, bioprinting) and their applications in physiological, patient-derived, or indirectly acquired GoC models. We also emphasize breakthroughs in biomimetic intestinal-microbiome symbiosis and spatiotemporal multi-organ integration (e.g., gut-X axis), enabling emulation of complex inter-organ signaling. Yet, critical challenges persist: reproducibility is limited by fabrication variability and cell heterogeneity; standardization lacks universal benchmarks for physiological relevance; and long-term culture stability (e.g. 7–10 days) is constrained by epithelial senescence and microbial imbalance. These gaps highlight needs for standardized protocols, quality control metrics, and strategies to sustain functional homeostasis. By bridging gaps between traditional models and human biology, GoC technologies establish transformative tools for mechanistic studies and therapeutic discovery in gastroenterology and beyond.

肠道芯片（GoC）平台集成了微流体和3D培养来复制肠道微环境，为传统模型提供了生理学相关的替代方案。结合多器官芯片（如肠-脑/肠-肝轴），它们揭示了微生物组通过代谢物信号调节的系统性串音——一个关键的尚未解决的机制。本文综述了用于肠道研究的器官芯片（OoC）技术的多方面进展，包括制造方法（如软光刻、生物打印）及其在生理、患者衍生或间接获得的GoC模型中的应用。我们还强调在仿生肠道-微生物共生和时空多器官整合（如肠道- x轴）方面的突破，使复杂的器官间信号模拟成为可能。然而，关键的挑战仍然存在：可重复性受到制造可变性和细胞异质性的限制；标准化缺乏生理相关性的普遍基准；长期培养稳定性（如7-10天）受到上皮衰老和微生物失衡的限制。这些差距突出了对标准化方案、质量控制指标和维持功能稳态策略的需求。通过弥合传统模型和人类生物学之间的差距，GoC技术为胃肠病学和其他领域的机制研究和治疗发现建立了变革性工具。

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引用次数: 0

Integrated metabolomics and transcriptomics reveal biomarkers for detecting fentanyl analog abuse through machine learning 综合代谢组学和转录组学揭示了通过机器学习检测芬太尼类似物滥用的生物标志物。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-22 DOI: 10.1016/j.jpba.2025.117323

Tao Wang , Xiaoxue Zhou , Tingli Qu , Zixiang Kai , Rongyu Yang , Hongliang Su , Bo Hao , Rui Gao , Haiyan Cui , Meng Hu , Zhe Chen , Zhiwen Wei , Keming Yun

Fentanyl analogs’ (FA) rampant proliferation causes widespread abuse, fatalities, and severe social issues globally. Their high toxicity, rapid metabolism, and poor detectability make them a major anti-drug challenge. This study aimed to establish an accurate method for identifying their abuse. Firstly, a rat model of FA (fentanyl, remifentanil, and sufentanil) abuse was constructed using the conditioned place preference (CPP) paradigm. Subsequently, dual-platform (Liquid and Gas Chromatography - High Resolution Mass Spectrometry) based metabolomic and transcriptomic data were used for weighted gene co-expression network analysis (WGCNA) and trait-module correlation analysis, key genes and metabolites associated with overall FA abuse were identified. Thirdly, we established a FA abuse identification model (100 % accuracy, AUC=1) based on 31 metabolites and 12 hub genes. Finally, we established an accurate classification model of different FA (77.27 % accuracy; AUCs: 1, 1, 0.981, 0.979 for control/fentanyl/remifentanil/sufentanil groups) using ten genes and five metabolites. The identified biomarkers and computational models offer valuable tools for clinical diagnosis and drug regulation, which are critical for addressing the ongoing opioid crisis.

芬太尼类似物（FA）的猖獗扩散导致了广泛的滥用、死亡和严重的全球社会问题。它们的高毒性、快速代谢和不易检测性使它们成为一个主要的抗药物挑战。本研究旨在建立一种准确的方法来识别它们的滥用。首先，采用条件位置偏好（CPP）范式构建芬太尼、瑞芬太尼和舒芬太尼滥用大鼠模型。随后，基于双平台（液相和气相色谱-高分辨率质谱）的代谢组学和转录组学数据被用于加权基因共表达网络分析（WGCNA）和性状-模块相关性分析，确定了与FA滥用相关的关键基因和代谢物。第三，我们基于31种代谢物和12个枢纽基因建立了FA滥用鉴定模型（准确率为100 %，AUC=1）。最后，我们利用10个基因和5种代谢物建立了不同FA的准确分类模型（准确率77.27 %；对照/芬太尼/瑞芬太尼/舒芬太尼组的auc分别为1,1,0.981,0.979）。确定的生物标志物和计算模型为临床诊断和药物监管提供了有价值的工具，这对于解决持续的阿片类药物危机至关重要。

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引用次数: 0

Systemic exposure characteristics and pharmacokinetic studies of intranasally delivered qinhao nasal drops by UHPLC-Q/TOF-MS and UHPLC-MS/MS 用UHPLC-Q/TOF-MS和UHPLC-MS/MS研究秦芩滴鼻给药的暴露特征和药代动力学

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-19 DOI: 10.1016/j.jpba.2025.117320

Maojie Zhou , Mireyi Bahatijiang , Wenkang Liu , Junyi Wang , Yan Mao , Zhengyi Gu , Changhong Wang

Qinhao nasal drops (QHND) is a formulation developed based on the integration theory of ethnomedical theory and traditional Chinese medicine, which plays a significant role in the treatment of acute, chronic, and allergic rhinitis. However, its primary active constituents and in vivo pharmacokinetic profile remain unclear. This study aimed to characterize the chemical composition of QHND using ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry, as well as to identify the systemic circulating components following intranasal administration of QHND. The results showed that 78 chemical constituents were identified in QHND, including flavonoids, terpenoids, alkaloids, and others. Notably, 19 prototype compounds and their 12 phase I/II metabolites were detected in rat plasma. Further, a sensitive, robust, and accurate ultra-high-performance liquid chromatography-tandem mass spectrometry method was established for the quantitative determination of rupestonic acid (RA), casticin (CT), baicalin (BC), wogonin (WG), ephedrine (EP), and pseudoephedrine (PE) in rat plasma. The method underwent comprehensive validation (including selectivity, linearity, precision, accuracy, matrix effect, extraction recovery, stability, and dilution integrity), confirming its suitability for the intended applications. With this validated method, the plasma concentration-time profiles of eight target components were monitored following both intranasal and intragastric administration of QHND. The results revealed that intranasal delivery significantly reduced the onset time of pharmacological effects, facilitated more efficient and rapid systemic absorption, and subsequently enhanced the bioavailability of key active constituents (RA, EP, and PE) in rats.

秦好滴鼻剂（QHND）是一种基于民族医学理论与中医相结合的制剂，对急性、慢性和变应性鼻炎具有重要的治疗作用。然而，其主要活性成分和体内药代动力学特征尚不清楚。本研究旨在利用超高效液相色谱-四极杆飞行时间质谱法表征芪黄芩苷的化学成分，并鉴定经鼻给药芪黄芩苷后的体循环成分。结果表明，黄芪黄酮类、萜类、生物碱等化学成分共鉴定出78种。值得注意的是，在大鼠血浆中检测到19种原型化合物及其12种I/II期代谢物。建立了灵敏、稳健、准确的超高效液相色谱-串联质谱法定量测定大鼠血浆中芦皮酸（RA）、蓖麻素（CT）、黄芩苷（BC）、枸杞素（WG）、麻黄碱（EP）和伪麻黄碱（PE）的方法。该方法经过了全面的验证（包括选择性、线性度、精密度、准确度、基质效应、萃取回收率、稳定性和稀释完整性），确认其适用于预期的应用。通过这种验证的方法，在鼻内和胃内给药QHND后，监测了8种目标成分的血浆浓度-时间谱。结果表明，鼻内给药可显著缩短药理作用的起效时间，促进更有效和快速的全身吸收，并随后提高大鼠体内关键活性成分（RA， EP和PE）的生物利用度。

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引用次数: 0

Revolutionizing ELISA development: The transformative impact of automation on monoclonal antibody potency assays 革命性ELISA发展：自动化对单克隆抗体效价测定的变革性影响

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-19 DOI: 10.1016/j.jpba.2025.117319

Xiaoran Xin , Douglas Pung , Amy Hsu , Monisha Dey , Qian Wu , Bo Feng , Dengyun Sun , Kevin Gurney , Anka Ehrhardt

Enzyme-linked immunosorbent assays (ELISA) are a fundamental technique for determining the potency of monoclonal antibodies (mAbs), yet the traditional ELISA development process is often time-consuming and labor-intensive. In this study, we explored how automation and miniaturization in a 384-well format can revolutionize ELISA development, significantly enhancing throughput and accuracy. By employing automated liquid dispensing technology, we streamlined the optimization of assay parameters, leading to faster and more reliable potency measurements. Our findings not only demonstrate the feasibility of this innovative approach but also highlight its potential to transform the landscape of analytical assays in biopharmaceutical development.

酶联免疫吸附试验（ELISA）是测定单克隆抗体（mab）效价的基本技术，但传统的ELISA开发过程往往耗时费力。在这项研究中，我们探索了384孔格式的自动化和小型化如何彻底改变ELISA的发展，显著提高通量和准确性。通过采用自动化液体分配技术，我们简化了分析参数的优化，从而实现更快，更可靠的效价测量。我们的发现不仅证明了这种创新方法的可行性，而且还强调了它在改变生物制药开发中分析分析的前景方面的潜力。

引用次数: 0

Development and validation of a rapid LC-MS/MS method for methylphenidate, atomoxetine, and their metabolites with application to pediatric pharmacokinetics 盐酸哌甲酯、托莫西汀及其代谢物的LC-MS/MS快速测定方法的建立与验证及其在儿童药代动力学中的应用

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-19 DOI: 10.1016/j.jpba.2025.117321

Duygu Eryavuz Onmaz , Merve Kuz , Fatih Hilmi Cetin , Halit Necmi Ucar , Serhat Turkoglu , Rabia Coban , Ali Unlu

Therapeutic drug monitoring of attention-deficit/hyperactivity disorder (ADHD) pharmacotherapy is challenged by substantial interindividual variability and the lack of rapid, multiplex analytical platforms. We developed and validated a fast, integrated 5-minute LC–MS/MS assay for the simultaneous quantification of stimulant (methylphenidate, ritalinic acid) and non-stimulant (atomoxetine, 4-hydroxy-atomoxetine) ADHD medications in human serum. The method employs a single-step protein precipitation and requires only 200 µL of serum. Validation demonstrated excellent linearity, precision, accuracy, minimal matrix effects, and robust analyte stability in accordance with FDA and CLSI guidelines. Application to pediatric ADHD patients revealed formulation- and dose-dependent concentration differences for methylphenidate and atomoxetine, along with strong parent–metabolite correlations. No significant associations were observed with age, sex, or body mass index. This rapid and reliable LC–MS/MS platform supports clinically actionable therapeutic drug monitoring and provides a practical framework for personalized ADHD treatment.

注意缺陷/多动障碍（ADHD）药物治疗的药物监测受到大量个体间差异和缺乏快速、多元分析平台的挑战。我们开发并验证了一种快速、集成的5分钟LC-MS/MS分析方法，用于同时定量人血清中兴奋剂（哌甲酯、利他酸）和非兴奋剂（阿托西汀、4-羟基阿托西汀）多动症药物。该方法采用单步蛋白沉淀法，仅需200 µL血清。验证证明了良好的线性，精密度，准确度，最小的基质效应，和稳健的分析物稳定性符合FDA和CLSI指南。应用于小儿多动症患者发现哌甲酯和阿托西汀的配方和剂量依赖性浓度差异，以及强烈的父母代谢物相关性。未观察到与年龄、性别或体重指数有显著关联。这种快速可靠的LC-MS/MS平台支持临床可操作的治疗药物监测，并为个性化ADHD治疗提供实用框架。

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引用次数: 0

Recycling of active pharmaceutical ingredients Metformin hydrochloride and Losartan potassium from expired medications and crystal structure of a new Losartan polymorphic form. 从过期药物中回收有效药物成分盐酸二甲双胍和氯沙坦钾及一种新的氯沙坦多晶型的晶体结构。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-15 Epub Date: 2025-08-05 DOI: 10.1016/j.jpba.2025.117090

Laura Galindo-Leon, Juan D Galofre-Benitez, Julián Corredor-Gamba, Deissy N Jaramillo, Johan D Lozano, Mario A Macías, Elizabeth Jiménez-Díaz

The disposal of expired pharmaceutical tablets, particularly those from household sources, is a growing environmental concern due to the accumulation of pharmaceutical contaminants in water ecosystems. Despite FDA recommendations to discard expired medications, studies have shown that many of these pills retain up to 90 % of their original potency long after their expiration date. This opens up the possibility for recycling and reusing the Active Pharmaceutical Ingredients (APIs) contained in these expired pills. This study presents a simple yet effective extraction protocol for recovering APIs from expired tablets of Metformin hydrochloride and Losartan potassium. The method achieved recovery yields of 88 % for Losartan and 37 % for Metformin. Structural characterization via NMR, HPLC, IR, HRMS, and both powder, and single-crystal X-ray diffraction confirmed the integrity and high purity of the recovered APIs compared to chemical standards. Biological assays carried out in L929 cell line revealed that the extracted APIs retained comparable activity to their chemical-standard counterparts, suggesting their potential for reuse in biochemical assays. During the recrystallization process by using PDRX, a new polymorphic form of Losartan potassium was discovered and named "Form M". In addition, a twelve-month stability study showed no degradation for either API at storage conditions. These results suggest that expired APIs are chemically and biologically viable for repurposing, as synthetic precursors in chemical research and in biological assays. The protocol is scalable, adaptable to pharmaceutical companies, and applicable in both industrial and research settings, offering a sustainable solution to pharmaceutical waste. By recovering APIs for educational and industrial use, this approach promotes environmental stewardship and resource efficiency in pharmaceutical science.

由于药物污染物在水生态系统中积累，过期药片的处置，特别是来自家庭的药片的处置日益成为一个环境问题。尽管FDA建议丢弃过期的药物，但研究表明，许多这些药物在过期后很长时间内仍能保持其原始效力的90% %。这为回收和再利用这些过期药片中含有的活性药物成分（api）提供了可能性。本研究提出了一种简单有效的从过期盐酸二甲双胍和氯沙坦钾片中提取原料药的方法。该方法对氯沙坦和二甲双胍的回收率分别为88 %和37 %。通过NMR， HPLC， IR， HRMS，粉末和单晶x射线衍射进行结构表征，与化学标准相比，证实了所回收原料药的完整性和高纯度。在L929细胞系中进行的生物实验表明，提取的原料药与化学标准的原料药具有相当的活性，这表明它们在生化分析中具有重复使用的潜力。在PDRX重结晶过程中，发现了氯沙坦钾的一种新的多晶形态，命名为“M型”。此外，一项为期12个月的稳定性研究表明，在储存条件下，这两种API都没有退化。这些结果表明，过期的原料药在化学和生物学上是可行的，可以作为化学研究和生物分析的合成前体。该议定书可扩展，适用于制药公司，并适用于工业和研究环境，为制药废物提供了可持续的解决方案。通过回收原料药用于教育和工业用途，这种方法促进了制药科学的环境管理和资源效率。

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引用次数: 0

Validation and clinical application of an LC-MS/MS method designed to simultaneously measure seven second-line TB drugs and two metabolites in human lung tissue. LC-MS/MS同时测定人肺组织中7种二线结核病药物和2种代谢物的方法验证及临床应用

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-12-15 Epub Date: 2025-08-06 DOI: 10.1016/j.jpba.2025.117093

Katie Kriegler Foster, Anil Pooran, Marthinus van der Merwe, Sandra Castel, Anton Joubert, Edda Zangenberg, Keertan Dheda, Lubbe Wiesner

We developed and validated a novel bioanalytical method for the simultaneous quantification of levofloxacin, linezolid, moxifloxacin, delamanid, bedaquiline, clofazimine, and pretomanid, along with the metabolites of delamanid (DM-6705) and bedaquiline (N-desmethyl-bedaquiline, M2), in human lung tissue samples. Following homogenization by bead beating and extraction by protein precipitation, the analytes were separated on an Agilent 1260 Infinity II HPLC system using a Poroshell 120 C18 EC (2.1 mm×50 mm, 2.7 µm) column with gradient elution, applying a mobile phase consisting of 0.1 % formic acid in water and 0.1 % formic acid in a mixture of acetonitrile and methanol. Detection and quantification of the analytes and their stable isotope labelled internal standards were performed on a Sciex API 5500 QTrap mass spectrometer using positive electrospray ionization and multiple reaction monitoring. Validation according to the guidelines of the FDA and EMA proved the method to be precise, accurate, and robust with no significant influence of matrix components. The application of the method to the analysis of clinical samples demonstrated the feasibility of quantifying the second-line anti-tuberculosis drugs in human lung tissue and the potential to provide insights into the drug distribution across the infection sites in the lung.

我们开发并验证了一种新的生物分析方法，用于同时定量人肺组织样品中左氧氟沙星、利奈唑胺、莫西沙星、德拉马尼、贝达喹啉、氯法齐明和普雷托马尼，以及德拉马尼（DM-6705）和贝达喹啉（n -去甲基贝达喹啉，M2）的代谢物。通过打珠匀浆和蛋白沉淀提取，在Agilent 1260 Infinity II高效液相色谱系统上使用Poroshell 120 C18 EC（2.1 mm×50 mm, 2.7 µm）柱进行梯度洗脱，流动相为0.1 %甲酸水溶液和0.1 %甲酸乙腈-甲醇混合物。在Sciex API 5500 QTrap质谱仪上使用正电喷雾电离和多重反应监测对分析物及其稳定同位素标记的内标进行检测和定量。根据FDA和EMA的指南进行验证，证明该方法精确、准确、稳健，没有基质成分的显著影响。将该方法应用于临床样本分析，证明了量化人肺组织中二线抗结核药物的可行性，并有可能深入了解肺部感染部位的药物分布。

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