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Analysis for higenamine in urine by means of UHPLC/MS/MS: Interpretation of results.

  • Grucza, Krzysztof1, 2
  • Kwiatkowska, Dorota2
  • Kowalczyk, Katarzyna2, 3
  • Wicka, Mariola2
  • Szutowski, Mirosław1
  • Chołbiński, Piotr2
  • 1 Department of Toxicology, Medical University of Warsaw, Warsaw, Poland. , (Poland)
  • 2 Department of Anti-Doping Research, Institute of Sport - National Research Institute, Warsaw, Poland. , (Poland)
  • 3 Biological and Chemical Research Centre, University of Warsaw, Warsaw, Poland. , (Poland)
Published Article
Drug Testing and Analysis
Wiley (John Wiley & Sons)
Publication Date
Oct 30, 2017
DOI: 10.1002/dta.2331
PMID: 29084416


Higenamine (Norcoclaurine) is a very popular substance in Chinese medicine and it is present in many plants. Moreover, the substance may be found in supplements or nutrients consumption of which may result in violation of anti-doping rules. Higenamine is prohibited in sport at all times and included in class S3 (β-2-agonists) of the World Anti-Doping Agency (WADA) "2017 Prohibited List". Presence of higenamine in urine samples at concentration greater than or equal to 10 ng/mL constitutes an adverse analytical finding (AAF). This work presents a new metabolite of higenamine in urine sample which was identified by means of UPLCMS/ MS. Samples were prepared according to two protocols - a Dilute and Shoot ("DaS") approach and a method involving acid hydrolysis and double liquid-liquid extraction. In order to meet the requirements typical for a confirmatory analysis, the screening procedure was further developed. In samples prepared by "DaS" method two peaks were observed the earlier one- specific for higenamine and the later one- "unknown". MS scan analysis showed mass about 80Da higher than that of higenamine. In turn, in samples prepared in accordance with the protocol involving hydrolysis an increase in the area under peak for higenamine was observed, while the second peak was absent. It seems that described strategy of detection of higenamine in urine allows to avoid false negative results.

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