Although hydrolysis of conjugated species is sometimes required prior to analysis, desomorphine-glucuronide has not been fully investigated, so this study optimized and evaluated six hydrolysis procedures.
Desomorphine, the principal opioid in Krokodil, has an analgesic potency approximately ten-times that of morphine. Similar to other opioids, during phase II metabolism it undergoes conjugation with glucuronic acid to form desomorphine-glucuronide. Deconjugation efficiencies using chemical and enzymatic hydrolysis were evaluated and stability in aqueous solution was assessed. Acid hydrolysis was compared with five -glucuronidase sources (BGTurbo, IMCSzyme, Escherichia coli, Helix pomatia and Patella vulgata). At optimal conditions, each hydrolysis method produced complete hydrolysis (96 percent); however, under simulated challenging conditions, P. vulgata was the most efficient -glucuronidase for the hydrolysis of desomorphine-glucuronide. Both BGTurbo and IMCSzyme offered fast hydrolysis with no need for sample cleanup prior to liquid chromatography-quadrupole/time of flight-mass spectrometry (LC-Q/TOF-MS) analysis. Hydrolysates using E. coli, H. pomatia, and P. vulgata underwent additional sample treatment using Gone cartridges. In addition, the stability of free and conjugated drug was evaluated at elevated temperature (60 degrees C) in aqueous solutions between pH 4 and 10. No degradation was observed for either desomorphine or desomorphine-glucuronide under any of the conditions tested. (publisher abstract modified)