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Fungi as a source of marker compounds for the control of illicit use of drugs: mesterolone as a case study

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Abstract

Introduction

Anabolic steroids are frequently misused for performance enhancement during sports competitions. One of the major bottlenecks in the confident analysis of steroids and their metabolites is the non-availability/cost of standard reference compounds.

Objective

The study objective was to identify the common metabolites of prohibited anabolic steroids that are produced in both fungi and human and thus can be synthesized in bulk using fungal cultures. Mesterolone is used as a case study.

Methods

The study was conducted in three steps; we first studied the fungal transformation of mesterolone. In the second step, these metabolites were used as references to detect in human urine after the oral use of mesterolone using LC-ESI-QqQ-MS/MS. In the third step, 12 fungal cultures were screened to evaluate their potential to produce reference markers.

Results

This led to the detection of two metabolites, 6α-hydroxymesterolone (M1) and 7α-hydroxymesterolone (M2) that were found to be common in both, fungal cultures and human urine samples. Moreover, Rhizopus stolonifer and Beauveria bassiana can be considered as good candidates to produce M1 and M2 metabolites, respectively.

Conclusion

This approach can be employed for the synthesis of marker compounds of other prohibited anabolic steroids thus can be detected efficiently during national and international sports competitions.

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Acknowledgements

The authors are thankful to Mr. Muhammad Noman Khan, Mr. Faraz Ul Haq, Ms. Hamna Shadab and Ms. Adeeba Abbas for their assistance in UHPLC-MS/MS analysis and sample preparation. The authors are also thankful to the volunteers for taking part in this study. The authors would like to acknowledge the financial support by the Pakistan Academy of Sciences for the Project No. 5-9/EAS 54.

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Authors and Affiliations

  1. H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan

    Syed Ghulam Musharraf, Qamar ul Arfeen, Arslan Ali, Fareeha Siddiqi, Malik Shoaib Ahmad, Ghazanfar Sultan, M. Iqbal Choudhary &  Atta-ur-Rahman

  2. Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan

    Syed Ghulam Musharraf & M. Iqbal Choudhary

  3. Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, 21412, Saudi Arabia

    M. Iqbal Choudhary

Authors
  1. Syed Ghulam Musharraf
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  2. Qamar ul Arfeen
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  3. Arslan Ali
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  4. Fareeha Siddiqi
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  5. Malik Shoaib Ahmad
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  6. Ghazanfar Sultan
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  7. M. Iqbal Choudhary
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  8. Atta-ur-Rahman
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Contributions

SGM conceived the idea, supervised the study and wrote the manuscript. MIC, AR, SA conducted the fungal transformation, purification and characterization of metabolites. GS participated in fugal extract preparations. SGM, QA, AA, and FS contributed in the quantitative method development and sample analysis. All authors contributed and approved the final manuscript.

Corresponding author

Correspondence to Syed Ghulam Musharraf.

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Conflict of interest

All the authors declare that they have no conflict of interests relevant to this study.

Ethical approval

The present study was approved by the Independent Ethic Committee of the principal investigating institute (ICCBS) (-027-HB-2017). All procedures performed in this study involving human participants were in accordance with the 1964 Helsinki declaration and its later amendments.

Informed consent

Informed consent was obtained from all individual participants of this study.

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Musharraf, S.G., Arfeen, Q.u., Ali, A. et al. Fungi as a source of marker compounds for the control of illicit use of drugs: mesterolone as a case study. Metabolomics 13, 150 (2017). https://doi.org/10.1007/s11306-017-1287-7

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  • DOI: https://doi.org/10.1007/s11306-017-1287-7

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