7-Aminoclonazepam is superior to clonazepam for detection of clonazepam use in oral fluid by LC–MS/MS

Analytical Methods used for the detection and quantification of benzodiazepines

“…Some are qualitative and some others are quantitative methods. Indeed, developed methods are classified as chromatography (HPLC, TLC, and GC) [11][12][13][14][15]18], immunoassays [38,[43][44][45][46][47][48], photometric (nuclear magnetic resonance and ultraviolet-visible) [20,42], and electroanalytical methods (potentiometric, polarographic, and voltammetry) [49][50][51] have been increasingly used for drugs of abuse in forensic science.…”

“…Using any immunological technique, the risk of false positive by cross reaction with another drug is important, and therefore, it requires confirmation of positive samples by a quantitative method such as chromatographic or spectroscopic allowing unambiguous identification and exact determination of the concentrations of the BZD molecule. e biological specimen dedicated for BZDs confirmation remains the blood, but recent work describes potential effective confirmation techniques using saliva or urine [11,12,14,[17][18][19].…”

“…e detection and quantification of BZDs can be done in different biological matrices including human breast milk [9], saliva [10][11][12], blood [13][14][15], blood serum [16], plasma [9], urine [14,[17][18][19][20], nails [21], hair [22], meconium [23], edible animal tissues and feed [24], and expired air [25]. However, it is important to keep in mind that whatever the biological matrix used for the screening, the BZDs concentrations depend on many factors, namely, the consumed dose, the quality of the product, the mode of consumption, the metabolism of the consumer, the body weight of the user, and his state of health.…”

“…e contamination of the saliva sample by other substances such as certain foods, condiments, drinks, or bacteria cannot be formally excluded [36]. Also, there is no standard method for treating and collecting samples before analysis, and the choice of sampling device and the treatment of the saliva have been approached very differently by different authors [11,12,37].…”

“…e detection window in the blood is narrower than urine, and the concentrations are lower. erefore, a sensitive and a very specific confirmation technique is mandatory for the detection of BZDs and their metabolites in the blood like HPLC or LC-MS/MS [11,[17][18][19], gas chromatography, or GC-Mass spectrometry [10,[13][14][15]18]. ese matrices contain a multitude of substances endogenous (proteins in the blood or fatty acids in urine) in amounts well above those compounds and their metabolites to be quantified.…”