Detection of 4-CPA and Its Related Metabolites Based on LC-QE-HF-MS Technology: A Study on the Impact of Doping Test on the Intake Pathways of Chlorphenesin Carbamate and Chlorphenesin

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry Pub Date : 2025-02-06 DOI:10.1002/rcm.10004

Wennuo Xu, Jiahui Cheng, Zhongquan Li, Bing Niu, Xiaojun Deng, Bing Liu

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引用次数: 0

Abstract

Objective

This study focused on meclofenoxate and its metabolite 4-CPA in the field of doping control and examined the urinary metabolism of chlorphenesin carbamate and chlorphenesin, two substances that may produce 4-CPA.

Methods

A liquid chromatography-Q Exactive-HF-Orbitrap-mass spectrometry (LC-QE-HF-MS)–based assay was developed and validated for the accurate detection of the metabolites of 4-CPA, chlorphenesin carbamate, and chlorphenesin and verified by human subject investigations. Human subjects were studied for three different modes of ingestion (chlorphenesin carbamate administration, sunscreen application containing chlorphenesin, and sunscreen spray application), and urine samples were collected before and after the administration to study the excretion profile of metabolites such as 4-CPA.

Results

The developed assay meets the routine testing requirements of the World Anti-Doping Agency. Following oral administration of chlorphenesin carbamate, 4-CPA levels in the urine ranged from 390 to 6929 ng·mL⁻¹, reaching their maximum concentrations in 12–24 h, with two volunteers having values over the reporting limit. The highest excretion happened 12–24 h after the treatment and continued until 168–264 h later. The C_max of 4-CPA was 1354–2063 and 340 ng·mL⁻¹ after application of sunscreen and spray sunscreen spray, respectively. Maximum excretion of other relevant metabolites was also concentrated at 12–24 h post-dose.

Conclusion

The results suggest that when measuring 4-CPA, the target analyte of meclofenoxate, in sports drug testing, special consideration needs to be given to whether volunteers have ingested normal therapeutic drugs, such as chlorphenesin carbamate, to avoid misreporting of meclofenoxate results. This finding offers crucial decision support for doping control.

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LC-QE-HF-MS技术检测4-CPA及其相关代谢物——掺杂检测对氨基甲酸氯苯菌素和氯苯菌素摄入途径影响的研究

目的研究甲氯芬诺酯及其代谢物4-CPA在兴奋剂控制领域的应用，并对可能产生4-CPA的氨基甲酸氯苯菌素和氯苯菌素两种物质的尿代谢进行检测。方法建立液相色谱- q精确- hf轨道谱-质谱（lc - q - hf - ms）分析方法，对4-CPA、氨基甲酸氯苯菌素和氯苯菌素的代谢物进行准确检测，并通过人体实验验证。研究了人体受试者三种不同的摄入方式（氨基甲酸氯苯菌素给药、含氯苯菌素的防晒霜涂抹和防晒霜喷雾涂抹），并在给药前后收集尿液样本，研究代谢物如4-CPA的排泄情况。结果该方法符合世界反兴奋剂机构的常规检测要求。口服氨基甲酸氯苯菌素后，尿液中的4-CPA水平在390至6929 ng·mL−1之间，在12-24小时内达到最高浓度，有两名志愿者的浓度超过了报告的限值。在给药后12 ~ 24 h达到最高，并持续到168 ~ 264 h。涂防晒霜和喷防晒霜后，4-CPA的Cmax分别为1354-2063和340 ng·mL−1。其他相关代谢物的最大排泄量也集中在给药后12-24 h。结论在运动药物检测中，测量甲氯芬酸靶物4-CPA时，应特别考虑志愿者是否摄入了正常的治疗药物，如氨基甲酸氯苯菌素，以避免误报甲氯芬酸结果。这一发现为兴奋剂控制提供了重要的决策支持。

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.