The Use of Bonded-Phase Extraction Columns for Rapid Sample Preparation of Benzodiazepines and Metabolites from Serum for HPLC Analysis

Abstract

A simple, rapid method for the determination of the benzodiazepines, diazepam and chlordiazepoxide, and their metabolites by high performance liquid chromatography (HPLC) has been developed. The procedure is applicable to the assay of other similar drugs in biological fluids. The method utilizes BondElut extraction columns to facilitate the extraction. BondElut columns selectively adsorb the benzodiazepines and metabolites from serum at a pH of 9.0. The compounds are eluted with 300 microliters of methanol which makes sample concentration rapid, if even necessary. Analysis is performed using isocratic reversed-phase chromatography, and quantitation is carried out by ultraviolet (UV) detection. Using this procedure, it is possible to determine drug and metabolite levels to as low as 25 ng/ml in 0.5 ml of serum.

Determination of several psychiatric drugs in whole blood using capillary gas–liquid chromatography with nitrogen phosphorus detection: comparison of two solid phase extraction procedures

“…There are numerous publications on SPE for relatively clean biological samples such as plasma, serum or urine [3][4][5][6][7][8][9][10][11][12]. Until now, only a few publications have described SPE methods for whole blood [13][14][15][16][17][18][19][20][21][22][23].…”

Rapid analysis of benzodiazepines in whole blood by high-performance liquid chromatography: use of a monolithic column

“…The choice of detectors offers several possibilities among which are the coupling of HPLC/UV or HPLC/DAD [13][14][15][16][17][18][19][20][21][22][23][24][25][26][27], and more recently the coupling of HPLC/MS which offers interesting selectivity and specificity [28].…”

A rapid solid-phase extraction and HPLC/DAD procedure for the simultaneous determination and quantification of different benzodiazepines in serum, blood and post-mortem blood

“…Screening and detection can be necessary under therapeutic aspects as well as in connection with forensic toxicology and traffic medicine [4]. Current methods for the determination of benzodiazepines available in clinical or forensic chemistry include thin-layer chromatography (TLC) [5,6], gas chromatography often coupled with mass spectrometry (GC/MS) [7][8][9][10][11], immunological methods [12][13][14][15][16] or high-performance liquid chromatography (HPLC) [11,[17][18][19][20][21][22]. The latter technique is particularly suitable for the quantitative determination of substances with intense UV absorption, such as the 1,4-benzodiazepines, particularly since some of these compounds exhibit thermal instability [23,24].…”