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Miniaturized Analysis of Methylhexanamine in Urine by Gas Chromatography Applying In Situ Derivatization

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Abstract

The use of stimulants by professional and amateur athletes, as well as in the general population, to enhance their athletic performance represents a human health hazard. Methylhexaneamine (MHA) is a stimulant found in numerous dietary supplements and detected frequently in urine samples. The analytical process accepted by most Drug Testing Programs that employ urine use two methods, the first is an initial screening test, followed by a more specific method for the confirmation of presumptive positive urine specimens. For this reason, two specific analytical methods were developed and validated to determine MHA in urine based on its in situ derivatization with isobutyl chloroformate; the first was salting out assisted liquid–liquid extraction (SALLE) with gas chromatography coupled to mass spectrometry (GC–MS) and the second headspace solid-phase microextraction (HS-SPME) followed by gas chromatography with flame ionization detector (GC–FID). Both methods were evaluated for precision (relative standard deviation < 10%), linearity (r2 > 0.98), accuracy at two concentration levels (recovery > 92%) and limit of detection (5–7 ng mL−1). The methods were successfully applied to analyze athletes’ urine samples from Comisión Nacional de Cultura Física y Deporte (CONADE, Mexico). The proposed methodologies are sensitive, precise, accurate, fast, and miniaturized, and they are suitable specific methods to analyze MHA in urine for anti-doping or forensic purposes even with an affordable GC–FID system.

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Acknowledgements

This work was supported by Dirección General de Asuntos del Personal Académico from the Universidad Nacional Autónoma de México (DGAPA-UNAM) [Grant PAPIIT: IN216114-2] and Faculty of Chemistry [PAIP: 5000-9026]. The authors thank Consejo Nacional de Ciencia y Tecnología (CONACYT) for the scholarship awarded to Félix Araujo-González (scholar number 273472), to Comisión Nacional de Cultura Física y Deporte (CONADE, México), and Dra. Evangelina Camacho for their contribution in providing the samples. The authors also thank Silvia Ocaña Rios for her support with the artwork and illustrations.

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Authors and Affiliations

  1. Facultad de Química, Departamento de Química Analítica, Universidad Nacional Autónoma de México, Mexico City, Mexico

    Iran Ocaña-Rios, Félix Araujo-González & Araceli Peña-Alvarez

  2. Sección de Química Analítica, Facultad de Estudios Superiores Cuautitlán, Universidad Nacional Autónoma de México, Cuautitlán Izcalli, Mexico

    José J. Olmos-Espejel

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  1. Iran Ocaña-Rios
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Correspondence to Araceli Peña-Alvarez.

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Ocaña-Rios, I., Araujo-González, F., Olmos-Espejel, J.J. et al. Miniaturized Analysis of Methylhexanamine in Urine by Gas Chromatography Applying In Situ Derivatization. Chromatographia 85, 95–104 (2022). https://doi.org/10.1007/s10337-021-04116-w

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