Journal of pharmaceutical and biomedical analysis最新文献_第8页

Investigation and elimination of noncovalent artificial aggregates during non-reduced capillary electrophoresis-sodium dodecyl sulfate analysis of a multi-specific antibody 非还原毛细管电泳过程中非共价人工聚集体的研究与消除——一种多特异性抗体的十二烷基硫酸钠分析。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-08 DOI: 10.1016/j.jpba.2025.116673

Jianhui Cheng , Qianchuan Lv , Yuanzhao Ji , Chunling Zhou , Jifen Guo , Xinxin Li , Jianzhong Hu

Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) is widely used in the biopharmaceutical industry for monitoring purity and analyzing impurities. The accuracy of the method may be compromised by artificial species resulting from sample preparation or electrophoresis separation due to suboptimal conditions. During non-reduced CE-SDS analysis of a multispecific antibody (msAb), named as multispecific antibody C (msAb-C), a cluster of unexpected peaks was observed after the main peak. The corrected peak area ratio of these peaks showed a strong dependence on loaded protein concentration, which affected the accurate assessment of the purity of msAb-C. After investigation, the unexpected peaks were identified as artifacts produced during electrophoresis separation. These artifacts can be mitigated by three different strategies: 1) adding a more hydrophobic surfactant, sodium hexadecyl sulfate (SHS), to the sample and/or sieving gel buffer; 2) reducing the sample loading amount; and 3) increasing the capillary separation temperature to above 40 ℃. We adopted strategy 1) and strategy 3), and successfully developed an optimal non-reduced CE-SDS method for the accurate and reliable purity assessment of msAb-C samples. These strategies of optimizing non-reduced CE-SDS can be used in developing quality control methods for other therapeutic bispecific/multispecific antibodies.

毛细管电泳-十二烷基硫酸钠（CE-SDS）广泛应用于生物制药行业的纯度监测和杂质分析。由于次优条件，该方法的准确性可能因样品制备或电泳分离产生的人工物种而受到损害。在对多特异性抗体C （msAb-C）进行非还原CE-SDS分析时，在主峰后观察到一簇意想不到的峰。校正后的峰面积比与载蛋白浓度有很大关系，影响了对msAb-C纯度的准确评价。经过调查，意外的峰被确定为在电泳分离过程中产生的伪影。这些伪影可以通过三种不同的策略来减轻：1)在样品和/或筛分凝胶缓冲液中添加更疏水的表面活性剂，十六烷基硫酸钠（SHS）；2)减少样品装填量；3)提高毛细管分离温度至40℃以上。我们采用策略1)和策略3)，成功开发了一种最佳的非还原CE-SDS方法，可以准确可靠地评估mabs - c样品的纯度。这些优化非还原CE-SDS的策略可用于开发其他治疗性双特异性/多特异性抗体的质量控制方法。

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引用次数: 0

A reliable LC-MS/MS method for the quantification of natural amino acids in human plasma and its application in clinic 可靠的LC-MS/MS定量人血浆中天然氨基酸的方法及其临床应用。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-08 DOI: 10.1016/j.jpba.2025.116672

Junhuan Lin , Yibo Song , Yangrui Zhang , Tao Ke , Fengting Ou , Kui Zeng , Debo He , Li Li , Lushan Yu

A simple and fast LC-MS/MS method was developed and validated for simultaneous quantification of 20 L-amino acids (AAs) in human plasma. Chromatographic separation was achieved on an Agilent AdvanceBio Hilic column within 15 min via gradient elution with an aqueous solution containing 5 mM ammonium formate, 5 mM ammonium acetate and 0.1 % formic acid and an organic mobile phase containing 0.1 % formic acid, 5 mM ammonium formate and 5 mM ammonium acetate acetonitrile-water (90:10, v/v) at the flow rate of 0.25 mL/min. Individual AAs and internal standard were analyzed by multiple reaction monitoring (MRM) in positive ion mode under optimized conditions. Method validation consisted of linearity, sensitivity, accuracy and precision, recovery, matrix effect, and stability, and the results demonstrated this LC-MS/MS method as a specific, accurate, and reliable assay. The method was thus utilized to compare the dynamics of individual plasma AAs between healthy females and patients with ovarian tumors. Our results revealed that, in cancer group, plasma 3-Methyl-L-Histidine, L-Proline, L-Phenylalanine and L-Lysine concentrations were significantly increased in patients with malignant ovarian tumors while L-Leucine and L-Isoleucine levels were sharply decreased. These findings support the utilities of this LC-MS/MS method and the promise of specific AAs as possible biomarkers for ovarian cancer.

建立了一种简便、快速的LC-MS/MS同时定量人血浆中20种l -氨基酸（AAs）的方法。在Agilent AdvanceBio Hilic色谱柱上，以含有5 mM甲酸铵、5 mM乙酸铵和0.1 %甲酸的水溶液和含有0.1 %甲酸、5 mM甲酸铵和5 mM乙酸铵的有机流动相（90:10,v/v）梯度洗脱，在15 min内实现色谱分离，流速为0.25 mL/min。在优化条件下，采用正离子模式多反应监测（MRM）对单个原子吸收剂和内标进行分析。方法验证包括线性度、灵敏度、准确度和精密度、回收率、基质效应和稳定性，结果表明该LC-MS/MS方法具有特异性、准确性和可靠性。因此，该方法被用于比较健康女性和卵巢肿瘤患者个体血浆AAs的动态。结果显示，恶性卵巢肿瘤患者血浆3-甲基- l-组氨酸、l-脯氨酸、l-苯丙氨酸和l-赖氨酸浓度显著升高，l-亮氨酸和l-异亮氨酸水平显著降低。这些发现支持了LC-MS/MS方法的实用性，以及特异性AAs作为卵巢癌生物标志物的前景。

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引用次数: 0

Research on the metabolites and key metabolic enzymes of allocryptopine in chicken liver microsomes via stable isotope tracing technology 利用稳定同位素示踪技术研究异隐碱在鸡肝微粒体中的代谢产物及关键代谢酶。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-06 DOI: 10.1016/j.jpba.2025.116667

Zhiyong Wu , Xinhao Wang , Lin Wang , Na Sun , Zihui Yang , Jianguo Zeng

Allocryptopine (ALL), a principal active component of the novel veterinary medicine Bopu Powder®, has gained widespread application in the poultry farming sector for the effective management of Escherichia coli (E. coli) diarrhea. In order to explore the metabolites and the pivotal enzymes associated with ALL, this study was conducted employing an in vitro chicken liver microsomal incubation. The metabolites of ALL were analyzed and identified by combining isotope tracing technology with the application of mass spectrometry fragmentation patterns. The key metabolic enzymes involved in the biotransformation of ALL were explored using the CYP450 recombinant enzyme method, which facilitated the identification of the enzymes contributing to ALL's metabolic pathway. The liver microsomal metabolism investigation revealed a total of five metabolites, with the predominant being M2 (harmol or 3-hydroxy-4-methoxy-6-methyl-5,7,8,15-tetrahydro-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-g]benzo[c]azecin-14(6 H)-one). The recombinant enzyme analysis conclusively identified CYP2D6 as the pivotal CYP450 isoenzyme that plays a central role in the metabolic pathway of the principal ALL metabolite, M2. This research not only expands our comprehension of the biotransformation process of ALL but also provides significant scientific evidence for the clinical safety of ALL, which was of great importance for guiding the application of ALL in the field of veterinary medicine.

异隐碱（ALL）是新型兽药博普粉®的主要活性成分，已广泛应用于家禽养殖部门，用于有效管理大肠杆菌（E. coli）腹泻。为了探索与ALL相关的代谢产物和关键酶，本研究采用鸡肝微粒体体外孵育。采用同位素示踪技术和质谱破碎图谱相结合的方法对ALL的代谢物进行了分析鉴定。利用CYP450重组酶法对ALL生物转化过程中涉及的关键代谢酶进行了探索，从而促进了ALL代谢途径中相关酶的鉴定。肝微粒体代谢调查共发现5种代谢物，主要代谢物为M2 （harmol或3-羟基-4-甲氧基-6-甲基-5,7,8,15-四氢-[1,3]二唑罗[4',5':4,5]苯并[1,2-g]苯并[c]azecin-14(6 H)-one）。重组酶分析最终确定CYP2D6是关键的CYP450同工酶，在ALL主要代谢物M2的代谢途径中起核心作用。本研究不仅扩大了我们对ALL生物转化过程的认识，而且为ALL的临床安全性提供了重要的科学依据，对指导ALL在兽药领域的应用具有重要意义。

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引用次数: 0

Characterization of main degradation products from dendrobine under stress conditions by multistage cleavage of UPLC-ESI-IT-TOF UPLC-ESI-IT-TOF多级解理表征应力条件下石斛主要降解产物。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-05 DOI: 10.1016/j.jpba.2025.116663

Hengju Zhou , Meiling Zeng , Keyong Geng , Zaipeng Chen , Zhijia Tang , Jianwei Xu , Xiaoyan Zhang , Wei Zhou

Dendrobine is a sesquiterpene alkaloid primarily used in the treatment of inflammatory diseases, immune system disorders, and conditions related to oxidative stress. To understand the possible degradation pathways of dendrobine for its quality control, we conducted an in-depth investigation of its degradation products using forced degradation methods. The separation of dendrobine and its degradation products was achieved on a Shim-pack XR-ODS III (75 mm × 2 mm, 1.6 µm) column with a methanol-water mixture as the mobile phase under isocratic conditions, the isolated compounds were examined in positive ion mode with an ion trap-time of flight mass spectrometer (IT-TOF). In order to obtain in-depth structural information about the degradation products, mass spectrometry was performed using a five-stage fragmentation approach. This method allowed for thorough structural clarification via several rounds of selective fragmentation and high-resolution detection. System control and data acquisition were managed using LCMSsolution 3.81 software. The results showed that dendrobine undergoes significant degradation under oxidative, acidic, hydrolytic and thermal conditions, resulting in the formation of several degradation products with notable structural changes. Under oxidative conditions, dendrobine primarily generates two degradation products with mass increases of 16 Da and 32 Da, indicating mono-oxidation and di-oxidation reactions. Acidic degradation led to the identification of three degradation products, including a novel compound with an 18 Da mass increase, suggesting potential hydrolysis or dehydration reactions. Hydrolytic and thermal conditions resulted in the formation of two and three degradation products, respectively, with structural changes indicating possible molecular cleavage and reorganization mechanisms. In contrast, dendrobine exhibited strong stability under alkaline and photolytic conditions, with no significant degradation products detected. Detailed characterization of the degradation products via multi-stage mass spectrometry revealed key reaction pathways and mechanisms involved in dendrobine's degradation, providing critical insights for assessing its chemical stability and optimizing storage conditions.

石斛碱是一种倍半萜生物碱，主要用于治疗炎症性疾病、免疫系统紊乱和与氧化应激相关的疾病。为了了解石斛石可能的降解途径并进行质量控制，我们采用强制降解方法对石斛石的降解产物进行了深入研究。采用Shim-pack XR-ODS III（75 mm × 2 mm, 1.6 µm）色谱柱，甲醇-水混合物为流动相，等压条件下对石斛碱及其降解产物进行分离，用离子捕获飞行时间质谱仪（IT-TOF）在正离子模式下对分离产物进行检测。为了获得降解产物的深入结构信息，质谱分析采用五段破碎法进行。这种方法允许通过几轮选择性破碎和高分辨率检测来彻底澄清结构。系统控制和数据采集采用LCMSsolution 3.81软件进行管理。结果表明，石斛石在氧化、酸性、水解和热条件下都发生了明显的降解，形成了几种结构变化明显的降解产物。在氧化条件下，石斛石主要生成两种降解产物，质量分别增加16 Da和32 Da，分别为单氧化和双氧化反应。酸性降解鉴定出三种降解产物，包括一种Da质量增加18 的新化合物，表明可能发生水解或脱水反应。水解和热条件下分别形成2种和3种降解产物，其结构变化表明可能的分子裂解和重组机制。相比之下，石斛石在碱性和光解条件下表现出很强的稳定性，没有检测到明显的降解产物。通过多级质谱分析对降解产物进行了详细表征，揭示了石斛降解的关键反应途径和机制，为评估石斛的化学稳定性和优化储存条件提供了重要见解。

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引用次数: 0

Implementation of a novel method for the determination of plasma protein binding of highly bound compounds using the equilibrium dialysis method coupled with extraction to the organic phase and its comparison to known methods 一种利用平衡透析法耦合萃取到有机相来测定血浆蛋白与高结合化合物结合的新方法的实施及其与已知方法的比较。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-05 DOI: 10.1016/j.jpba.2025.116665

Dawid Gogola , Sanja Novak Ratajczak , Ewelina Gabor-Worwa , Anna Kowal-Chwast , Nilesh Gaud , Gniewomir Latacz , Krzysztof Brzózka , Kamil Kuś

Accurate determination of plasma protein binding (PPB) is crucial in understanding the pharmacokinetics and pharmacodynamics of drugs, particularly for highly bound compounds where traditional methods may fall short. In this study, we present a pioneering approach for the precise determination of PPB that takes advantage of the lipophilicity of highly bound compounds. Twenty four highly bound compounds (with a fraction unbound (f_u) from 10⁻¹ to 10⁻⁶) were tested with the most commonly used method, i.e., the rapid equilibrium dialysis (RED) method, along with its modifications adapted especially for strongly bound compounds, including dilution, presaturation, competition, and flux-dialysis methods. The results of these methods were compared to data obtained with the modification of RED coupled with extraction to organic phase, and the correlations between them were presented. Comparison studies demonstrate the accuracy of our approach across a set of highly bound compounds within a twofold range. Moreover, PPB values were determined for venetoclax, amiodarone, montelukast, and fulvestrant, for which, to the best of our knowledge and after extensive review of the literature, it has not been possible with the standard RED method until now. In conclusion, our new method can be a big step forward in finding the PPB of strongly plasma protein-bound compounds. It gives us a better understanding of how drugs and proteins interact, which helps us make safer and more effective medicines.

血浆蛋白结合（PPB）的准确测定对于理解药物的药代动力学和药效学至关重要，特别是对于传统方法可能达不到的高结合化合物。在这项研究中，我们提出了一种开创性的方法来精确测定PPB，利用高结合化合物的亲脂性。用最常用的方法，即快速平衡透析（RED）方法，对24种高结合化合物（未结合分数（fu）从10-1到10-6）进行了测试，并对其进行了专门针对强结合化合物的修改，包括稀释、预饱和、竞争和通量透析方法。将这些方法得到的结果与RED改性和萃取有机相得到的结果进行了比较，并给出了它们之间的相关性。比较研究证明了我们的方法在两倍范围内跨越一组高度结合的化合物的准确性。此外，还测定了venetoclax、胺碘酮、孟鲁司特和氟维司汀的PPB值，据我们所知，经过广泛的文献回顾，迄今为止，标准的RED方法还无法测定这些药物的PPB值。总之，我们的新方法在寻找强血浆蛋白结合化合物的PPB方面迈出了一大步。它让我们更好地了解药物和蛋白质是如何相互作用的，这有助于我们制造更安全、更有效的药物。

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引用次数: 0

Equipmentless point-of-care testing of dengue antibodies using ELISA and smartphones 使用ELISA和智能手机对登革热抗体进行无设备即时检测。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-05 DOI: 10.1016/j.jpba.2025.116666

Diego Mendicino , Christian Avalos , Romina Chiaraviglio , Ludmila Bazán Domínguez , Federico Schaumburg

Infections with the dengue virus affect more than 100 million people every year. The infected can present a mild form of the disease or a severe form, which can, eventually, lead to death. Dengue prevails in tropical and subtropical regions, although increased incidence has been observed in the last years in tempered climates. Vaccines are available but testing for previous infection is often required prior to application. Commercially available ELISA and rapid tests for the diagnosis of dengue IgG do not fulfill individually the performance required by control agencies. In this context, rapid, simple and decentralized point-of-care testing (POCT) is highly desirable. However, POCT approaches available usually offer expensive solutions, often due to the complex complementary hardware required. In this article, an equipmentless system based on a commercial ELISA kit and a smartphone is developed for POCT of dengue antibodies. A customized app provides guiding, optical reading, result reporting and connectivity. The reading method employes an algorithm which requires no external information, other than the available on the digital images from the smartphone camera, to classify samples into positives, negatives or indeterminates. The full system operation, from sample extraction to result reporting, was tested in a low resource medical facility with real patients (n = 26). After comparison with an ELISA reader, a Cohen’s κ coefficient of 0.92 was obtained, showing very good agreement between both methods. These results show that it is possible to perform ELISA with no specific equipment, bringing massive testing at low resource facilities one step closer.

每年有1亿多人感染登革热病毒。感染者可以表现出轻微的疾病形式，也可以表现出严重的疾病形式，最终导致死亡。登革热流行于热带和亚热带地区，尽管近年来在温和气候中发现发病率有所增加。疫苗是可用的，但在接种前通常需要对以前的感染进行检测。市售的诊断登革热IgG的酶联免疫吸附试验和快速检测不能单独满足控制机构所要求的性能。在这种情况下，快速、简单和分散的护理点检测（POCT）是非常可取的。然而，可用的POCT方法通常提供昂贵的解决方案，这通常是由于需要复杂的补充硬件。在本文中，基于商用ELISA试剂盒和智能手机开发了一种用于登革热抗体POCT的无设备系统。一个定制的应用程序提供指导，光学读数，结果报告和连接。该读取方法采用一种算法，该算法不需要任何外部信息，除了智能手机相机上的数字图像上可用的信息，将样本分为阳性，阴性或不确定。从样本提取到结果报告的整个系统操作都在一个资源匮乏的医疗设施中进行了测试，有真正的病人（n = 26）。与ELISA阅读器比较，Cohen's κ系数为0.92，两种方法的一致性很好。这些结果表明，在没有特定设备的情况下进行酶联免疫吸附试验是可能的，这使在资源匮乏的设施中进行大规模检测更近了一步。

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引用次数: 0

Differentiation of the chronic lymphocytic leukemia response to ibrutinib and acalabrutinib treatment by single-cell MALDI-TOF MS imaging 单细胞MALDI-TOF MS显像鉴别慢性淋巴细胞白血病对依鲁替尼和阿卡拉布替尼治疗的反应。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-05 DOI: 10.1016/j.jpba.2025.116664

Ivana Marković , Iva Lukić , Maja Lukić , Ema Pavičić , Stefan Mrđenović , Ana Kotris , Branko Dmitrović , Željko Debeljak

Ibrutinib and acalabrutinib, Bruton's tyrosine kinase inhibitors (BTKi) used for chronic lymphocytic leukemia (CLL) treatment, aim the same target but their off-target effects are different. The aim of this study was to use single-cell MALDI TOF mass spectrometry imaging to compare the CD19+ lymphocytes’ mass spectra in untreated and ibrutinib- or acalabrutinib-treated subjects in order to better understand the therapeutic effect of BTKi. 180 cells from 9 male subjects divided in 3 groups (untreated, ibrutinib-treated and acalabrutinib-treated) were analyzed using MALDI-TOF mass spectrometry analyzer. Mass spectra were acquired in the 300–600 Da mass range. Partial least squares discriminant analysis (PLS-DA) was used for evaluation of mass spectra, while Volcano plots and Venn diagram were used for a review of significantly altered m/z values. Cross-validated PLS-DA classification accuracy of cells was 85 %. Ibrutinib-treated cells overlap with the untreated cell population, while acalabrutinib-treated cells form a separate class. 13 m/z signals were specific for each BTKi group. In conclusion, single-cell MALDI-TOF mass spectrometry imaging detects differences in the mass spectra of CD19+ lymphocytes treated with two different BTKi. Drug-specific m/z signal clusters can be identified as a chemical fingerprint of the BTKi effect and represent candidates for the therapeutic response biomarkers.

用于治疗慢性淋巴细胞白血病（CLL）的布鲁顿酪氨酸激酶抑制剂（BTKi） Ibrutinib和acalabrutinib的靶点相同，但脱靶效果不同。本研究的目的是使用单细胞MALDI TOF质谱成像比较未治疗和伊鲁替尼或阿卡拉布替尼治疗的受试者的CD19+淋巴细胞的质谱，以便更好地了解BTKi的治疗效果。采用MALDI-TOF质谱分析仪分析9例男性受试者180个细胞，分为3组（未治疗组、依鲁替尼组和阿卡拉布替尼组）。获得了300-600 Da质量范围内的质谱。质谱评价采用偏最小二乘判别分析（PLS-DA），而m/z值显著变化的评价采用火山图和维恩图。交叉验证的PLS-DA细胞分类准确率为85 %。伊鲁替尼处理的细胞与未处理的细胞群重叠，而阿卡拉布替尼处理的细胞形成单独的一类。每个BTKi组有13个 m/z信号。总之，单细胞MALDI-TOF质谱成像检测了两种不同BTKi处理的CD19+淋巴细胞的质谱差异。药物特异性m/z信号簇可以被识别为BTKi效应的化学指纹，并代表治疗反应生物标志物的候选物。

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引用次数: 0

Development and validation of an LC-MS/MS method for quantification of Osimertinib and its two metabolites AZ7550 and AZ5104 in human plasma including long-time storage LC-MS/MS定量人血浆中奥西替尼及其两种代谢物AZ7550和AZ5104的方法的建立和验证

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-03 DOI: 10.1016/j.jpba.2025.116662

Eva Greibe , Boe Sorensen , Peter Meldgaard , Elke Hoffmann-Lücke

Osimertinib (AZD9291) is a widely used tyrosine kinase inhibitor for the treatment of non-small cell lung cancer patients with activating EGFR mutations. However, the correlation between dose and efficacy has been debated for several years. For this reason, there is a need for standardized methods for routine analysis, clinical studies on pharmacokinetics and dose-response relationships, and greater understanding of preanalytical conditions, such as sample storage stability. The objective of this study was to develop and validate a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of osimertinib and its two metabolites, AZ7550 and AZ5104, in human plasma and to investigate long-term storage stability of the analytes. Samples were prepared by protein precipitation and separated on a Kinetex EVO C18 column (2.1 × 150 mm, 2.6 µm). Electrospray ionization in positive mode and multiple reaction monitoring were used to monitor the ion transitions. The validated concentration ranges were from 1.25 to 3000 ng/mL. Interassay precisions and accuracies were all ≤ 15 %. Linearity, dilution integrity, and carry-over were also examined and satisfied the validation criteria. Stability was examined under different conditions, and the analytes were found to be stable for more than 3 years at −80°C (< 15 % decline). Finally, the analytical method was successfully applied in a clinical setting on plasma samples from 30 patients with non-small cell lung cancer in treatment with osimertinib, demonstrating its suitability for use in clinical studies and its potential for therapeutic drug monitoring.

奥西替尼（AZD9291）是一种广泛使用的酪氨酸激酶抑制剂，用于治疗具有激活EGFR突变的非小细胞肺癌患者。然而，剂量和疗效之间的关系已经争论了好几年。因此，需要标准化的常规分析方法、药代动力学和剂量-反应关系的临床研究，以及对分析前条件（如样品储存稳定性）的更多了解。本研究的目的是建立并验证一种灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量人血浆中奥西替尼及其两种代谢物AZ7550和AZ5104，并研究分析物的长期储存稳定性。样品采用蛋白沉淀法制备，在Kinetex EVO C18色谱柱（2.1 × 150 mm, 2.6 µm）上分离。采用正模式电喷雾电离和多反应监测来监测离子跃迁。验证浓度范围为1.25 ~ 3000 ng/mL。测定间精密度和准确度均≤ 15 %。还检查了线性、稀释完整性和携带性，并满足验证标准。在不同条件下测试了稳定性，发现分析物在-80°C下稳定超过3年（< 15 %下降）。最后，将该分析方法成功应用于临床，对30例接受奥西替尼治疗的非小细胞肺癌患者的血浆样本进行了分析，证明了该方法在临床研究中的适用性和治疗药物监测的潜力。

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引用次数: 0

Impact of sample heterogeneity on the evaluation of uncertainty from sampling and analytical steps in pharmaceutical analysis 样品异质性对药物分析中取样和分析步骤不确定度评定的影响。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-03 DOI: 10.1016/j.jpba.2025.116660

Victória Aparecida Vit Crivilari, Jean Bispo dos Santos, Felipe Rebello Lourenço

Measurement uncertainty is a critical factor in the reliability of pharmaceutical analyses, since it directly affects batch acceptance and regulatory compliance. While analytical uncertainty has been extensively studied, uncertainty arising from sampling remains less explored. This study aims to address this gap by evaluating the contributions of sampling and analytical uncertainties to the overall uncertainty for acetaminophen tablets and oral solution. The duplicate method and analysis of variance (ANOVA) were used to assess the uncertainty from sampling and analytical steps. Ten different batches of acetaminophen tablets and oral solution were analyzed, and the combined uncertainties were calculated. For tablets, uncertainty from sampling accounted for 89 % of the overall uncertainty, highlighting the critical role of sample heterogeneity. In contrast, for oral solutions, analytical uncertainty dominated (90 % of the total), reflecting the homogeneous nature of the solution. Considering the uncertainty from sampling can be important to ensure a reduced risk of false acceptance or false rejection, particularly for heterogeneous dosage forms. In conclusion, incorporating sampling uncertainty into uncertainty budgets can significantly improve decision-making in pharmaceutical quality control.

测量不确定度是影响药物分析可靠性的关键因素，因为它直接影响到批次的可接受性和法规符合性。虽然分析不确定度已经得到了广泛的研究，但采样产生的不确定度仍然很少被探索。本研究旨在通过评估采样和分析不确定度对扑热息痛片剂和口服液的总体不确定度的贡献来解决这一差距。采用重复法和方差分析（ANOVA）来评估抽样和分析步骤的不确定性。对10个不同批次的对乙酰氨基酚片剂和口服液进行分析，计算其综合不确定度。对于片剂，抽样不确定度占总体不确定度的89% %，突出了样品异质性的关键作用。相比之下，对于口服溶液，分析不确定度占主导地位（占总数的90 %），反映了溶液的均匀性。考虑取样的不确定性对于确保降低错误接受或错误拒绝的风险是很重要的，特别是对于异质剂型。综上所述，将抽样不确定性纳入不确定性预算可以显著改善药品质量控制决策。

{"title":"Impact of sample heterogeneity on the evaluation of uncertainty from sampling and analytical steps in pharmaceutical analysis","authors":"Victória Aparecida Vit Crivilari, Jean Bispo dos Santos, Felipe Rebello Lourenço","doi":"10.1016/j.jpba.2025.116660","DOIUrl":"10.1016/j.jpba.2025.116660","url":null,"abstract":"<div><div>Measurement uncertainty is a critical factor in the reliability of pharmaceutical analyses, since it directly affects batch acceptance and regulatory compliance. While analytical uncertainty has been extensively studied, uncertainty arising from sampling remains less explored. This study aims to address this gap by evaluating the contributions of sampling and analytical uncertainties to the overall uncertainty for acetaminophen tablets and oral solution. The duplicate method and analysis of variance (ANOVA) were used to assess the uncertainty from sampling and analytical steps. Ten different batches of acetaminophen tablets and oral solution were analyzed, and the combined uncertainties were calculated. For tablets, uncertainty from sampling accounted for 89 % of the overall uncertainty, highlighting the critical role of sample heterogeneity. In contrast, for oral solutions, analytical uncertainty dominated (90 % of the total), reflecting the homogeneous nature of the solution. Considering the uncertainty from sampling can be important to ensure a reduced risk of false acceptance or false rejection, particularly for heterogeneous dosage forms. In conclusion, incorporating sampling uncertainty into uncertainty budgets can significantly improve decision-making in pharmaceutical quality control.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"255 ","pages":"Article 116660"},"PeriodicalIF":3.1,"publicationDate":"2025-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142965572","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}

引用次数: 0

Whole-cell aptamer-based techniques for rapid bacterial detection: Alternatives to traditional methods 基于全细胞适体的快速细菌检测技术：替代传统方法。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-03 DOI: 10.1016/j.jpba.2025.116661

Juliette Nourry , Pauline Chevalier , Emmanuelle Laurenceau , Xavier Cattoen , Xavier Bertrand , Basile Peres , Farid Oukacine , Eric Peyrin , Luc Choisnard

Controlling the spread of bacterial infectious diseases is a major public health issue, particularly in view of the pandemic of bacterial resistance to antibiotics. In this context, the detection and identification of pathogenic bacteria is a prerequisite for the implementation of control measures. Current reference methods are mainly based on culture methods, which generate a delay in obtaining a result and requires equipment. Consequently, focusing on the detection of the whole bacterium represents a very attractive alternative, since no culture is required. Several techniques have already been deployed to identify whole-cell bacteria. In recent decades, growing interest in nucleic acid aptamers has emerged as a viable alternative to antibodies as recognition elements, offering preferable stability, cost-efficiency, good specificity and affinity. This review explores current alternative methods for the detection of whole-cell bacteria, with particular emphasis on aptamer-based assays. These assays have shown promising results in various transduction mechanisms, including optical, electrochemical, and mechanical approaches, enhancing their versatility in different diagnostic platforms. The integration of aptamers in these detection methods offers rapid, sensitive, versatile and portable solutions for pathogen identification, positioning them as valuable tools in the fight against bacterial infections.

控制细菌传染病的传播是一项重大的公共卫生问题，特别是考虑到细菌对抗生素的耐药性普遍存在。在这种情况下，病原细菌的检测和鉴定是实施控制措施的先决条件。目前的参考方法主要是基于培养法，这种方法获得结果有一定的延迟，并且需要设备。因此，专注于整个细菌的检测是一个非常有吸引力的选择，因为不需要培养。已有几种技术被用于鉴定全细胞细菌。近几十年来，人们对核酸适体的兴趣日益浓厚，核酸适体作为一种可行的替代抗体的识别元件，具有更好的稳定性、成本效益、良好的特异性和亲和力。这篇综述探讨了目前检测全细胞细菌的替代方法，特别强调了基于适配体的测定。这些检测在各种转导机制中显示出有希望的结果，包括光学、电化学和机械方法，增强了它们在不同诊断平台中的通用性。这些检测方法中适配体的整合为病原体鉴定提供了快速、敏感、通用和便携的解决方案，使其成为对抗细菌感染的宝贵工具。

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引用次数: 0