title: Detection and identification of 2-nitro-morphine and 2-nitro-morphine-6-glucuronide in nitrite adulterated urine specimens containing morphine and its glucuronides.
abstract: In vitro urine adulteration is a well-documented practice adopted by individuals aiming to evade detection of drug use, when required to undergo mandatory sports and workplace drug testing. Potassium nitrite is an effective urine adulterant due to its oxidizing potential, and has been shown to mask the presence of many drugs of abuse. However, limited research has been conducted to understand its mechanism of action, and to explore the possibility of the drugs undergoing direct oxidation to form stable reaction products. In this study, opiates including morphine, codeine, morphine-3-glucuronide and morphine-6-glucuronide were exposed to potassium nitrite in water and urine to mimic the process of nitrite adulteration. It was found that two stable reaction products were detected by liquid chromatography-mass spectrometry (LC-MS) when morphine and morphine-6-glucuronide were exposed to nitrite. Isolation and elucidation using spectrometric and spectroscopic techniques revealed that they were 2-nitro-morphine and 2-nitro-morphine-6-glucuronide, respectively. These reaction products were also formed when an authentic morphine-positive urine specimen was fortified with nitrite. 2-Nitro-morphine was found to be stable enough to undergo the enzymatic hydrolysis procedure and also detectable by gas chromatography-mass spectrometry (GC-MS) after forming a trimethylsilyl derivative. On the contrary, morphine-3-glucuronide did not appear to be chemically manipulated when exposed to potassium nitrite in urine. These reaction products are not endogenously produced, are relatively stable and can be monitored with both LC-MS and GC-MS confirmatory techniques. As a result, these findings have revealed the possibility for the use of 2-nitro-morphine and 2-nitro-morphine-6-glucuronide as markers for the indirect monitoring of morphine and morphine-6-glucuronide in urine specimens adulterated with nitrite.
authors.affiliation: Centre for Forensic Science, School of Chemistry and Forensic Science, University of Technology, Sydney (UTS), Ultimo, NSW 2007, Australia.
authors.forename: Susan
authors.initials: S
authors.lastname: Luong
authors.forename: Shanlin
authors.initials: S
authors.lastname: Fu
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009020:Morphine / Q000652:urine*; D009022:Morphine Derivatives / Q000652:urine*; D009294:Narcotics / Q000652:urine*; D009573:Nitrites / Q000652:urine*; D009574:Nitro Compounds / Q000652:urine; D015813:Substance Abuse Detection / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D009022:Morphine Derivatives; D009294:Narcotics; D009573:Nitrites; D009574:Nitro Compounds; C035349:morphine-6-glucuronide; D009020:Morphine; C041349:potassium nitrite
keywords: 2-nitro-morphine; 2-nitro-morphine-6-glucuronide; morphine; morphine-3-glucuronide; morphine-6-glucuronide; nitrite; urine adulteration