RESUMEN
BACKGROUND: Benzodiazepines are used in hypnotics, sedation, and anti-anxiety. Recently liquid chromatography tandem mass spectrometry (LC-MS/MS) has been vastly developed for drug analysis in biological samples. METHODS: We developed and validated a LC-MS/MS method for simultaneous quantification of flunitrazepam (FM2), nimetazepam and nitrazepam levels in 87 benzodiazepine positive human urine specimens by enzyme immunoassay. Deuterated analogues were used as internal standard. RESULTS: The limits of quantification were found to be 0.25, 2.5, 5, 5 and 1ng/ml for FM2, 7-aminoFM2, nimetazepam, 7-amino-nimetazepam and nitrazepam, respectively. The intraday and inter-day CVs ranged from 0.6 to 4.6% and 1.2-9.4%, respectively. The within-day accuracy ranged from 80.8 to 108.7% and the between-day accuracy ranged from 80.5 to 118.0%. The recovery rate ranged from 70.5 to 96.7% for five different analytes. A group of 34 urine samples previously gas chromatography-mass spectrometry determined to contain 7-aminoFM2 was analyzed by this new LC-MS/MS approach. Quantitative data produced by both methods agreed well. CONCLUSIONS: The LC-MS/MS method has proved to be robust and specific for the quantification of FM2, nimetazepam and nitrazepam in urine samples. This study also confirmed that nitrazepam and 7-aminonimetazepam are the metabolic products of nimetazepam.
Asunto(s)
Benzodiazepinonas/orina , Cromatografía Liquida , Espectrometría de Masas en Tándem , Flunitrazepam/orina , Humanos , Límite de Detección , Nitrazepam/análogos & derivados , Nitrazepam/orinaRESUMEN
To facilitate the analysis of targeted drugs under high sample volume testing environment, an extraction, derivatization and gas chromatographic-mass spectrometric analysis method was developed for simultaneously determination of amphetamine (AMP), methamphetamine (MAMP), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), ketamine, and norketamine in urine. This method utilized solid-phase extraction in conjunction with derivatization using heptafluorobutyric anhydride (HFBA) as the derivatization reagent. Using a 1-mL sample, the limits of quantitation achieved for the analysis of AMP, MAMP, MDA, MDMA, MDEA, ketamine, and norketamine were 25, 15, 60, 60, 70, 25, and 30 ng/mL, respectively. Upper limits of quantitation were 8000 ng/mL for all amphetamines and 6000 ng/mL for ketamine and norketamine. Except for dehydronorketamine (DHNK), within-day and between-day precisions (as expressed in CV%) for quality control samples were ≤ 3.1% and ≤ 4.95%, respectively. Except DHNK, the within-day accuracy ranged between 96.0% and 110.7% and the between-day accuracy ranged between 96.9% and 108.7%. A group of 107 urine samples previously determined to contain the target analytes were analyzed by this new approach. Quantitative data produced by both methods agreed well. With this new approach, we were able to use a single analytical protocol to conduct the confirmation test for samples that preliminarily tested positive (by immunoassay) for amphetamines, ketamine, or both.