A novel analytical strategy based on gas chromatography-Orbitrap high-resolution mass spectrometry combined with solid-phase extraction for the monitoring of stanozolol misuse in human urine

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry Pub Date : 2024-10-30 DOI:10.1002/rcm.9935

Siying Zheng, Ziyi Ji, Yuqi Ge, Xian Fang, Mengpan Liu, Haoyi Sun, Xiaojun Deng, Lei Liao

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引用次数: 0

Abstract

Rationale

Stanozolol, an anabolic androgenic steroid listed in Part S1 of the World Anti-Doping Agency Prohibited List, exhibits a low response and significant matrix interference in urine samples when using liquid–liquid extraction–gas chromatography–mass spectrometry (GC–MS). Enhancing sample preparation techniques remains essential for the effective detection of stanozolol and its metabolites.

Methods

A method for determining stanozolol and its metabolites (3′-OH-stanozolol, 4β-OH-stanozolol, and 16β-OH-stanozolol) in human urine was developed and validated using GC-Orbitrap high-resolution MS combined with optimized mixed-mode solid-phase extraction (SPE). This method was applied to urine samples from two volunteers who orally administered a single dose of stanozolol, with samples collected over a 30-day period post-administration.

Results

The optimized mixed-mode SPE method reduced matrix interference and achieved satisfactory extraction efficiency and high sensitivity, enabling confident identification of all targets in human urine. Validation showed extraction recovery of 74% to 81% and limits of detection from 0.1 to 0.25 ng mL⁻¹. The method was successfully applied to detect urinary excretion profiles of stanozolol and its metabolites in positive volunteer samples.

Conclusion

This study presents a novel detection protocol for stanozolol and its metabolites, enhancing the monitoring of stanozolol abuse and contributing to the integrity of sports competitions. This protocol offers a robust tool for anti-doping laboratories, aiding in the accurate detection of stanozolol misuse and supporting the enforcement of fair play in athletics.

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一种基于气相色谱-轨道阱高分辨质谱法并结合固相萃取的新型分析策略，用于监测人体尿液中滥用司坦唑醇的情况。

理由：司坦唑醇是一种同化雄性类固醇，已被列入世界反兴奋剂机构禁用清单 S1 部分，在使用液液萃取-气相色谱-质谱联用技术（GC-MS）检测尿样时，司坦唑醇的响应度较低，且存在明显的基质干扰。加强样品制备技术对于有效检测司坦唑醇及其代谢物仍然至关重要：方法：采用GC-Orbitrap高分辨率质谱结合优化的混合模式固相萃取（SPE）技术，开发并验证了测定人体尿液中丹诺唑醇及其代谢物（3'-OH-丹诺唑醇、4β-OH-丹诺唑醇和16β-OH-丹诺唑醇）的方法。该方法适用于两名口服单剂量司坦唑醇的志愿者的尿样，并在用药后 30 天内采集尿样：结果：优化的混合模式固相萃取方法减少了基质干扰，萃取效率令人满意，灵敏度高，能够可靠地鉴定人体尿液中的所有目标物。验证表明萃取回收率为 74% 至 81%，检出限为 0.1 至 0.25 纳克 mL-1。该方法被成功应用于检测阳性志愿者样本中丹诺唑醇及其代谢物的尿液排泄曲线：本研究提出了一种新型的司坦唑醇及其代谢物检测方案，可加强对滥用司坦唑醇行为的监测，促进体育比赛的公正性。该方案为反兴奋剂实验室提供了一个强大的工具，有助于准确检测滥用司坦唑醇的情况，并支持田径运动中公平竞赛的实施。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Rapid Communications in Mass Spectrometry 化学-分析化学

CiteScore

4.10

自引率

5.00%

发文量

219

审稿时长

2.6 months

期刊介绍： Rapid Communications in Mass Spectrometry is a journal whose aim is the rapid publication of original research results and ideas on all aspects of the science of gas-phase ions; it covers all the associated scientific disciplines. There is no formal limit on paper length ("rapid" is not synonymous with "brief"), but papers should be of a length that is commensurate with the importance and complexity of the results being reported. Contributions may be theoretical or practical in nature; they may deal with methods, techniques and applications, or with the interpretation of results; they may cover any area in science that depends directly on measurements made upon gaseous ions or that is associated with such measurements.