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A high throughput approach for determination of dermorphin in human urine using liquid chromatography–mass spectrometry for doping control purposes
Author(s) -
Castro Juliana de L.,
Martucci Maria Elvira P.,
Pereira Henrique M.G.,
Sousa Valéria P.
Publication year - 2020
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.4593
Subject(s) - dermorphin , chemistry , chromatography , mass spectrometry , tandem mass spectrometry , selected reaction monitoring , detection limit , liquid chromatography–mass spectrometry , opioid peptide , opioid , receptor , biochemistry
Dermorphin is a peptide with analgesic actions similar to morphine, but with greater effect and less potential to cause tolerance. The use of dermorphin has been documented in race horses, and its use in humans has already been reported. Considering the potential advantages from the use of dermorphin over morphine, a method to monitor it, and its main metabolite dermorphin (1‐4) in humans becomes necessary for doping control. Here, we present two orthogonal methods for this purpose: a high‐throughput liquid chromatography coupled to high‐resolution mass spectrometry (HRMS) as an initial testing procedure and liquid chromatography–tandem mass spectrometry (MS/MS) in the selected reaction monitoring (SRM) acquisition mode for a confirmation procedure. For urine samples, pretreatment through a mixed‐mode weak cation‐exchange solid‐phase extraction emerged as an effective approach to extract peptides from the biological sample. For the HRMS analysis, a full‐MS scan acquisition mode was selected to detect the exact masses of dermorphin and dermorphin (1‐4) at m/z 803.37226 and 457.20816, respectively. The SRM method used in the MS/MS confirmation protocol presented high specificity and sensitivity. The selected product ions for dermorphin were 602.2, 202.1 and 574.3 and for dermorphin (1‐4) were 207.1, 223.1, and 235.1. Both methods were evaluated for specificity, repeatability, carryover, matrix effects, and recovery. No carryover and matrix effects were detected. The limit of detection for initial testing procedure and the limit of identification for confirmation procedure was 2.5 ng/ml. Also, specificity and robustness were acceptable for the application. Together, the developed methods proved to be efficient for the analysis of dermorphin and metabolite for human doping control purpose.