Solid-phase extraction and on-disc derivatization of the major benzodiazepines in urine using enzyme hydrolysis and Toxi-Lab VC MP3 column.
Because of the increase in use of the newer benzodiazepines, we explored the opportunity to develop a gas chromatographic-mass spectrometric (GC-MS) method that encompasses most of the widely prescribed benzodiazepines in use today. The benzodiazepines included in our study are nordiazepam, oxazepam, temazepam, lorazepam, alpha-hydroxyalprazolam, alpha-hydroxytriazolam, desalkylflurazepam, and 2-hydroxyethylflurazepam. Using 1.0 mL of urine as the matrix, we added the enzyme Glusulase and incubated the specimens for 2 h to obtain the free drugs. The hydrolyzed samples were then loaded onto a Toxi-Lab Spec VC MP3 column containing a 15-mg disc. On-disc derivatization was accomplished by adding N-methyl-N-(t-butyldimethylsilyl) trifluroacetamide (MTBSTFA) with 1% TBDMSCI to the disc. The derivatives were then placed in a GC vial and analyzed by GC-MS in the selected ion monitoring mode. These results were then compared to confirmed positives by the traditional acid hydrolysis GC-MS method.
King
J W
JW
Roche Biomedical Laboratories, Houston, Texas 77074, USA.
King
L J
LJ
D001569:Benzodiazepines / Q000302:isolation & purification* / Q000652:urine*; D008401:Gas Chromatography-Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D005966:Glucuronidase; D006801:Humans; D006868:Hydrolysis; D009097:Multienzyme Complexes; D013429:Sulfatases
D009097:Multienzyme Complexes; C023059:glusulase; D001569:Benzodiazepines; D013429:Sulfatases; D005966:Glucuronidase