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Simultaneous analysis of codeine, morphine, and heroin after B-glucuronidase hydrolysis
M Zezulak1, J J Snyder, S B Needleman
1Navy Drug Screening Laboratory, Great Lakes, IL.
Journal of Forensic Sciences
|November 1, 1993
Summary
This study introduces an enzyme-based method for simultaneously analyzing morphine, codeine, 6-monoacetylmorphine (6-MAM), and heroin in urine. This approach avoids issues associated with acid hydrolysis, simplifying drug testing procedures.
Area of Science:
- Forensic Toxicology
- Analytical Chemistry
- Pharmacology
Background:
- Opiate analysis, including morphine and codeine, typically involves acid hydrolysis to release morphine from its glucuronide metabolite.
- Heroin is rapidly metabolized to 6-monoacetylmorphine (6-MAM), with limited further conversion to morphine glucuronide, necessitating separate analytical procedures for heroin and morphine/codeine.
- Simultaneous analysis of these compounds is challenging due to potential loss of heroin identity during acid hydrolysis and significant concentration differences between parent drugs and metabolites.
Purpose of the Study:
- To develop a novel analytical method for the simultaneous detection of morphine, codeine, 6-MAM, and heroin in urine specimens.
- To overcome the limitations of traditional acid hydrolysis methods in opiate drug testing.
- To establish an efficient and cost-effective procedure for comprehensive opiate analysis.
Main Methods:
- Utilized beta-glucuronidase for hydrolysis of morphine glucuronide, followed by derivatization with propionic anhydride to form dipropionylmorphine.
- Heroin remained unhydrolyzed and was derivatized as diacetylmorphine.
- Employed a solid-phase column system for purification of the enzyme reaction mixture.
- Characterized optimal conditions for enzyme concentration, hydrolysis temperature, and pH, and identified ions for propionyl derivatives.
Main Results:
- The proposed enzyme-based method successfully enables simultaneous analysis of morphine, codeine, 6-MAM, and heroin.
- Eliminated complex steps associated with acid hydrolysis, making the enzyme procedure cost-comparable.
- Established optimal conditions for enzyme hydrolysis and derivative identification.
Conclusions:
- The developed method offers a simplified and effective approach for simultaneous opiate analysis in toxicology.
- Beta-glucuronidase hydrolysis provides an alternative to acid hydrolysis, preserving drug identities and simplifying the analytical workflow.
- This method enhances the efficiency and potentially reduces the cost of drug testing for a panel of common opiates.