Researchers affiliated with the University of South Carolina’s Departments of Chemistry and Biochemsitry investigated 6-monoacetlymorphine (6-MAM) conversion during enzyme hydrolysis. They used four different commercially available beta-glucuronidase solutions: IMCSzymeTM, Haliotis rufescens (Abalone), Snail (Helix pomatia), and Limpet (Patella vulgata). The researchers sought to find an enzyme that would be able to accurately quantitate 6-MAM because current methods involve processes that waste valuable time and resources in labs.
In department’s research, hydrolysis was performed at the recommended temperatures for each enzyme and measured at zero, one, and two hour hydrolysis time intervals. 6-MAM and morphine levels were compared at each time interval to determine the amount of 6-MAM loss and corresponding increase in morphine.
Data from the analysis of 6-MAM spiked urine samples before and after hydrolysis showed that the concentration of 6-MAM throughout the varying incubation times did not show significant changes for IMCSzyme. The data from Patella vulgata, Helix pomatia and the Abalone extract show a decreasing concentration of 6-MAM as incubation time increased, which indicates that these enzymes converted a significant amount of 6-MAM to morphine.
The investigation found that overall, significant conversion of 6-MAM to morphine occurs with Helix pomatia, Patella vulgata and Abalone enzymes during morphine glucuronide hydrolysis, which reduces the likelihood of detecting conclusive heroin use. The research also showed, however, that the unique properties of IMCSzyme allow for a more faster, more efficient hydrolysis of morphine glucuronide with negligible conversion of 6-MAM to morphine.
USC researchers found that IMCSzyme™ provides effective glucuronide hydrolysis without esterase activity. It is able to maintain the acetyl moiety of 6-MAM while rapidly cleaving the glucuronides in a time-effective manner. This allows for simultaneous LC-MS/MS detection of 6-MAM and morphine.
Information summarized from technical poster presented by Kaylee McDonald -University of South Carolina at SOFT 2014