Abstract
The aim of this study was to validate a new method for determining benzodiazepines in urine via their benzophenone derivatives, based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). Selected benzodiazepines were analysed after acid hydrolysis of urine and extraction by ethyl acetate in the presence of an internal standard. Samples were analysed using electrospray ionization LC-MS/MS in a multiple reaction monitoring mode. The chromatographic run time on a reversed phase C18 analytical column was set for 9 min. This method was validated in 21 patients receiving methadone. Benzodiazepines intake was established in two out of three patients. LC-MS/MS results were also compared with the rapid immunoassay and the methods showed good agreement. However, in three cases benzodiazepines were detected by LC-MS/MS, but not by the immunoassay. The sensitivity of the developed LC-MS/MS method is comparable to or even higher than of previously reported methods, which makes it suitable as a confirmatory method.
Highlights
The aim of this study was to validate a new method for determining benzodiazepines in urine via their benzophenone derivatives, based on liquid chromatography-tandem mass spectrometry (LC-MS/MS)
The most convenient screening methods are based on the immunoassay and only positive results need to be confirmed, usually using gas chromatography or liquid chromatography coupled with mass spectrometry [5,6,7]
We have developed a liquid chromatographytandem mass spectrometry (LC-MS/MS) method that determines low levels of various benzodiazepines in urine at the same time via their products of hydrolysis, the corresponding benzophenones
Summary
The aim of this study was to validate a new method for determining benzodiazepines in urine via their benzophenone derivatives, based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). Benzodiazepines as part of a managed methadone program alleviate some of the withdrawal symptoms of treated heroin addicts such as insomnia, nausea, anxiety, and depression Their abuse is associated mostly with the extension of mitigation of withdrawal symptoms. Analysis of benzodiazepines and their metabolites is very important for establishing abuse or poisoning They are mainly determined in urine and less often in serum [6]. The most convenient screening methods are based on the immunoassay and only positive results need to be confirmed, usually using gas chromatography or liquid chromatography coupled with mass spectrometry [5,6,7] These methods are not suitable for quantification in some biological samples due to the presence of their different metabolites and/or other substances in the matrix with similar properties. There have been only a few reports so far on benzodiazepine analysis via their benzophenone derivatives, and these mainly refer to GC-MS [6] or HPLC with UV detection [9]
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