Simultaneous determination of four PDE5 inhibitors and metabolites in rat plasma by UPLC-MS/MS

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-04-09 DOI:10.1016/j.jpba.2025.116883

Xiaonan Xia , Zhe Song , Weiwen He , Zhou Qiao , Fanglin Wang , Shaoyuan Li

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引用次数: 0

Abstract

Herein, we developed a sensitive UPLC-MS/MS method for the simultaneous determination of four PDE5 inhibitor and twelve metabolites in rat plasma. A total of 16 chemicals were separated on gradient elution with mobile phase A (0.1 % formate acetonitrile) and mobile phase B (0.1 % formate and 2 mmol/L ammonium formate aqueous solution) on an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm) after protein precipitation, followed by the detection on the positive ion mode of electrospray ion source (ESI) via dynamic multi-reaction monitoring mode (MRM). The method provided good linearity over the range of 0.25–20 ng/mL for all compounds with correlation coefficients R greater than 0.99. The recoveries were higher than 75.5 %, the matrix factors were 2.1 %-20.4 %. The precision, accuracy and stability were also within acceptable criteria. A rat model was established to investigate the metabolic profile of Sildenafil, Vardenafil, Acetildenafil and Tadalafil. Blood samples from the experimental group were analyzed using ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). Notably, desmethyltadalafil and methyl-desmethyltadalafil were not detected in the plasma. However, their glucuronide conjugates, desmethyltadalafil glucuronide and methyl-desmethyltadalafil glucuronide, were identified and characterized via tandem mass spectrometry (MS/MS). At present, cases of poisoning or even death due to overdose of phosphodiesterase type 5 (PDE5) inhibitors are often encountered in the practice of forensic science. Few studies on PDE5 inhibitors have been reported in forensic science toxicology identification, and the existing detection methods cannot meet the need for simultaneous and rapid analysis of multiple trace PDE5 inhibitors in vivo. Therefore, it is of great importance to establish and optimize the detection methods for PDE5 inhibitors drugs in biological samples. The significance of this method is to provide a reference for the determination of PDE5 inhibitors poisoning cases.

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利用 UPLC-MS/MS 同时测定大鼠血浆中的四种 PDE5 抑制剂和代谢物

为此，我们建立了一种灵敏的UPLC-MS/MS方法，用于同时测定大鼠血浆中4种PDE5抑制剂和12种代谢物。蛋白质沉淀后，在ACQUITY UPLC HSS T3柱（100 mm×2.1 mm, 1.8 μm）上，以流动相A（0.1 %甲酸乙腈）和流动相B（0.1 %甲酸乙酯和2 mmol/L甲酸铵水溶液）梯度洗脱，共分离出16种化学物质，并通过动态多反应监测模式（MRM）对电喷雾离子源（ESI）正离子模式进行检测。该方法在0.25 ~ 20 ng/mL范围内具有良好的线性关系，相关系数R大于0.99。加样回收率大于75.5 %，基质因子为2.1 % ~ 20.4 %。精密度、准确度和稳定性均在可接受的标准范围内。建立大鼠模型，研究西地那非、伐地那非、乙酰地那非和他达拉非的代谢谱。实验组血样采用超高效液相色谱-高分辨率质谱（UPLC-HRMS）分析。值得注意的是，血浆中未检测到去甲基他达拉非和甲基去甲基他达拉非。然而，它们的葡萄糖醛酸缀合物去甲基他达拉非葡萄糖醛酸和甲基-去甲基他达拉非葡萄糖醛酸通过串联质谱（MS/MS）进行了鉴定和表征。目前，在法医学实践中经常遇到因过量使用5型磷酸二酯酶（PDE5）抑制剂而中毒甚至死亡的案例。法医学毒理学鉴定中关于PDE5抑制剂的研究报道较少，现有的检测方法无法满足同时快速分析体内多种痕量PDE5抑制剂的需要。因此，建立和优化生物样品中PDE5抑制剂药物的检测方法具有重要意义。本方法的意义在于为PDE5抑制剂中毒病例的检测提供参考。

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来源期刊

Journal of pharmaceutical and biomedical analysis 医学-分析化学

CiteScore

6.70

自引率

5.90%

发文量

588

审稿时长

37 days

期刊介绍： This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.