Simultaneous determination of morphine, codeine and 6-acetyl morphine in human urine and blood samples using direct aqueous derivatisation: Validation and application to real cases

Opiates play a relevant role in forensic toxicology and their assay in urine or blood is usually performed for example in workplace drug-testing or toxicological investigation of drug impaired driving. The present work describes two new methods for detecting morphine, codeine and 6-monoacethyl morphine in human urine or blood using a single step derivatisation in aqueous phase. Propyl chloroformate is used as the dramatizing agent followed by liquid-liquid extraction and gas-chromatography-mass spectroscopy to detect the derivatives. The methods have been validated both for hydrolysed and unhydrolysed urine.For hydrolysed urine, the LOD and LOQ were 2.5. ng/ml and 8.5. ng/ml for codeine, and 5.2. ng/ml and 15.1. ng/ml for morphine, respectively. For unhydrolysed urine, the LOD and LOQ were 3.0. ng/ml and 10.1. ng/ml for codeine, 2.7. ng/ml and 8.1. ng/ml for morphine, 0.8. ng/ml and 1.5. ng/ml for 6-monoacetyl morphine, respectively. In blood, the LOD and LOQ were 0.44. ng/ml and 1.46. ng/ml for codeine, 0.29. ng/ml and 0.98. ng/ml for morphine, 0.15. ng/ml and 0.51. ng/ml for 6-monoacetyl morphine, respectively.The validated methods have been applied to 50 urine samples and 40 blood samples (both positive and negative) and they can be used in routine analyses. © 2013 Elsevier B.V.