Application Note

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Opioids compose a chemical class of both naturally occurring compounds produced from the opium poppy plant (Papaver somniferum) and similar synthetic analogues. Substances within this class are commonly recognized as potential drugs of abuse. Biological matrices containing heroin (diacetyl morphine), codeine, morphine, hydrocodone, hydromorphone, oxycodone and oxymorphone and thebaine are regularly monitored by drugs of abuse analysis laboratories. Structural similarities of opiate metabolites introduce challenges to the accurate separation, confirmation and quantitation of these substances.

A forensic toxicology method for the confirmation and quantitation of codeine, morphine, hydrocodone, hydromorphone, oxycodone and oxymorphone in human urine was developed using the Thermo Scientific ISQ single quadrupole GC-MS system. Multiple analytical procedures can be replaced by this method, which combines a dual derivatization technique with the selection of the correct analytical column.

All samples were prepared as batches using a 2 mL sample size. Standard materials were obtained for calibration, and separate sources of the opiates were used as controls. Deuterated internal standards were employed. Batches included a matrix-matched single point calibrator (300 ng/mL), quality control samples set to contain each target compound at 40% and 125% of the calibrator (120 ng/mL and 375 ng/mL respectively), and a negative control, which was blank urine with internal standard only.

A hydrolysis step was performed to release the bound drug from the glucuronide. Samples underwent a first derivitization with hydroxylamine to convert the keto moieties to their oxime derivatives. Thermo Scientific HyperSep Verify-CX solid-phase extraction columns (200 mg, 10 mL) were used for sample extraction. Sample extracts were derivatized a second time with Thermo Scientific BSTFA and 1% TMCS.