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Forensic
Applications
Benzodiazepines in Urine
Alternative Derivatization
Using 200mg 10mL HyperSep Verify-CX Extraction Column
(Part Number: 60108-742)
Derivatize
To 2mL of urine add internal standard(s)* and 1mL of
β
-Glucuronidase
β
-Glucuronidase: 5,000 F units/mL Patella vulgate in
100mM acetate buffer (pH 5.0)
Mix/vortex
Hydrolyze for 3 hours at 65°C
Centrifuge for 10 minutes at 2,000rpm and discard pellet
Analysis
Inject 1 to 2µL onto GC/MS
For mass spectrometry monitor the following ions:
Generic Name
Trade Name
Primary Ion**
Secondary
Tertiary
Nordiazepam -D5-TBDMS
332
334
333
Nordiazepam-TBDMS
327
328
329
Oxazepam-D5-TBDMS
462
519
462
Oxazepam-TBDMS
Serax
457
513
459
Temazepam-D5-TBDMS
362
390
288
Temazepam-TBDMS
Restoril
®
357
359
385
Lorazepam-TBDMS
Ativan
®
491
513
493
Clonazepam
Klonopin
®
372
374
326
7-Aminoclonazepam -TBMS
456
458
513
Diazepam
Valium
®
256
283
221
Desalkylflurazepam-TBDMS
345
347
402
Prazepam*
269
241
324
α
-Hydroxymidazolam-TBDMS
Versed
®
398
400
440
Desmethylflunitrazepam-TBDMS
357
310
356
7-Aminoflunitrazepam-TBDMS
397
324
398
Alprazolam
Xanax
®
308
279
204
α
-Hydroxyalprazolam-D5-TBDMS
386
388
387
α
-Hydroxyalprazolam-TBDMS
383
384
381
Triazolam
Halcion
®
313
314
342
α
-Hydroxytriazolam-TBDMS
415
417
190
* Suggested internal standard
** Quantitation ion
Recommended GC Columns
Part Number
TRACE TR-DoA5 Column, 15m x 0.25mm x 0.25µm
26AF130P
TRACE TR-DoA35 Column, 15m x 0.20mm x 0.33µm
26AC497P