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Extraction and Analysis of Clonazepam and 7-aminoclonazepam in Whole Blood Using a Dual Internal Standard Methodology

Extraction and Analysis of Clonazepam and 7-aminoclonazepam in Whole Blood Using a Dual Internal Standard Methodology

Jeffery Hackett, Albert A Elian

Forensic Sci Int. 2007 Mar 2;166(2-3):209-17.

PMID: 16809012

Abstract:

In this paper, a simple and robust method for the determination of clonazepam and its primary metabolite (7-aminoclonazepam) in whole blood is described. Clonazepam (klonopin) is a popular prescription drug that has been implicated in the field of drug facilitated sexual assaults (DFSA). Clonazepam, 7-aminoclonazepam and the internal standards (deuterated analogues for GC-MS analysis and nitrazepam for analysis by LC-PDA/GC-MS) were spiked into blood samples. The samples were buffered with a pH 6-phosphate solution (5 mL) and extracted from phenyl spe columns. The columns were washed with 5% acetonitrile in pH 6-phosphate buffer (3 mL) and eluted with ethyl acetate (2 x 3 mL). The eluents were evaporated for further chromatographic analysis. For GC-MS, the samples were derivatized prior to analysis. When performed with LC-PDA the samples were reconstituted in distilled water. From this method LOQs of 5 ng/mL of sample is easily achievable by either chromatographic system. By using GC-MS in EI mode, 1 ng/mL of sample can be detected. Data is presented here to show the simplicity and efficiency of the extraction scheme. By employing the properties of GC-MS and LC-PDA, this extraction and analysis procedure further extends the number of tools open to the forensic toxicologist for the analysis of this drug.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP4959175	7-Aminoclonazepam solution		4959-17-5	Price

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