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Clinical Chemistry 37: 1595-1601, 1991;
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Clinical Chemistry, Vol 37, 1595-1601, Copyright © 1991 by American Association for Clinical Chemistry

Enzymatic digestion, solid-phase extraction, and gas chromatography/mass spectrometry of derivatized intact oxazepam in urine

JG Langner, BK Gan, RH Liu, LD Baugh, P Chand, JL Weng, C Edwards and AS Walia
Environmental Health Research and Testing, Inc., Birmingham, AL 35205.

Enzymatic digestion with beta-glucuronidase (EC 3.2.1.31) was used to release intact oxazepam from urine samples containing the d5-analog internal standard. The resulting specimens were extracted with Du Pont PREP Type W cartridge (processed by a PREP Automated Sample Processor), Bond Elut Certify, and J.T. Baker "spe" columns for comparison of the columns' extraction recovery and overall effectiveness. Methyl iodide/tetrahexylammonium hydrogen sulfate and N,O- bis(trimethylsilyl)trifluoroacetamide/trimethylchlorosilane (10 g/L) were used for the methylation and trimethylsilylation studies. We used a Hewlett-Packard HP 5790 mass-selective detector equipped with a 13-m J & W DB-5 column (5% phenyl polysiloxane phase) for gas chromatography/mass spectroscopy (GC/MS) analysis and the Thru-Put Target software package for data processing. After several exploratory experiments, we adopted the Du Pont PREP system methylation procedure because of its effective recovery, the superior stability of the derivatization product, the possibility of incorporating a clean-up step, and the potential for high throughput. The extraction recovery from a set of control samples was 87%. Coefficients of variation obtained for six replicates of GC/MS analysis and for the overall procedure were 1% and 3%, respectively. Excellent linearity was established in the 50-8000 micrograms/L concentration range studied. With the use of 3-mL samples, a 20-microL final reconstitution volume, oxazepam at 50 micrograms/L was easily detected under the adopted operation conditions.


The following articles in journals at HighWire Press have cited this article:


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 Hum Exp ToxicolHome page
R. Griffin, L. Burka, and K. Demby
Oxidative biotransformation of oxazepam to reactive and nonreactive products in rat, mouse and human microsomes
Human and Experimental Toxicology, October 1, 1995; 14(10): 779 - 786.
[Abstract] [PDF]


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Copyright © 1991 by the American Association for Clinical Chemistry.