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基于衍生化和注射液相色谱-质谱/(高分辨质谱)方法对诺龙II期氧代代谢物进行完整定量和鉴定。

Simultaneous quantitation and identification of intact Nandrolone phase II oxo-metabolites based on derivatization and inject LC-MS/(HRMS) methodology.

作者信息

Sakellariou Panagiotis, Kiousi Polyxeni, Petrou Michael, Angelis Yiannis S

机构信息

Doping Control Laboratory of Athens, Institute of Biosciences and Applications, National Centre for Scientific Research "Demokritos", Athens, Greece.

Cyprus Anti-Doping Authority (CyADA), Nicosia, Cyprus.

出版信息

Drug Test Anal. 2025 Jan;17(1):118-130. doi: 10.1002/dta.3689. Epub 2024 Apr 6.

Abstract

Α sensitive and selective derivatization and inject method for the quantification of intact nandrolone phase II oxo-metabolites was developed and validated using liquid chromatography - (tandem high resolution) mass spectrometry (LC-MS/(HRMS)). For the derivatization, Girard's reagent T (GRT) was used directly in natural urine samples and the analysis of the metabolites of interest was performed by direct injection into LC-MS/(HRMS) system operating in positive ionization mode. Derivatization enabled the efficient detection of nandrolone oxo-metabolites, while at the same time producing intense product ions under collision-induced dissociation (CID) conditions that are related to metabolites of the steroid backbone and not to the conjugated moieties. Glucuronide and sulfate metabolites of nandrolone were chromatographically resolved and quantified in the same run in the range of 1-100 ng mL, while at the same time structure identification could be performed for each metabolite. Full validation of the method was performed according to the World Anti-Doping Agency (WADA) International Standard for Laboratories (ISL). Nandrolone oxo-metabolites were quantified in two sets of urine samples, the first set consisted of real urine samples previously detected as negative and the second set consisted of urine samples collected from two excretion studies after nandrolone decanoate administration. The results for 19-norandrosterone glucuronide (19-NAG) and 19-noretiocholanolone glucuronide (19-NEG) were compared with those obtained by traditional gas chromatography - (tandem) mass spectrometry (GC-MS/[MS]) method.

摘要

开发并验证了一种灵敏且具有选择性的衍生化和进样方法,用于定量完整的诺龙II期氧代代谢物,该方法采用液相色谱 - (串联高分辨率)质谱法(LC-MS/(HRMS))。对于衍生化,直接在天然尿液样本中使用吉拉德试剂T(GRT),并通过直接进样到以正离子模式运行的LC-MS/(HRMS)系统中对感兴趣的代谢物进行分析。衍生化能够有效检测诺龙氧代代谢物,同时在碰撞诱导解离(CID)条件下产生与甾体骨架代谢物相关而非与共轭部分相关的强产物离子。诺龙的葡萄糖醛酸苷和硫酸酯代谢物在1 - 100 ng/mL范围内的同一次运行中通过色谱法分离并定量,同时可以对每种代谢物进行结构鉴定。该方法根据世界反兴奋剂机构(WADA)实验室国际标准(ISL)进行了全面验证。在两组尿液样本中对诺龙氧代代谢物进行了定量,第一组由先前检测为阴性的真实尿液样本组成,第二组由癸酸诺龙给药后两项排泄研究收集的尿液样本组成。将19 - 去甲雄酮葡萄糖醛酸苷(19 - NAG)和19 - 去甲本胆烷醇酮葡萄糖醛酸苷(19 - NEG)的结果与通过传统气相色谱 - (串联)质谱法(GC-MS/[MS])获得的结果进行了比较。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f0a/11730825/45ec86b59efa/DTA-17-118-g005.jpg

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